TY  - JOUR
A1  - Gröbner, Susanne N.
A1  - Worst, Barbara C.
A1  - Weischenfeldt, Joachim
A1  - Buchhalter, Ivo
A1  - Kleinheinz, Kortine
A1  - Rudneva, Vasilisa A.
A1  - Johann, Pascal D.
A1  - Balasubramanian, Gnana Prakash
A1  - Segura-Wang, Maia
A1  - Brabetz, Sebastian
A1  - Bender, Sebastian
A1  - Hutter, Barbara
A1  - Sturm, Dominik
A1  - Pfaff, Elke
A1  - Hübschmann, Daniel
A1  - Zipprich, Gideon
A1  - Heinold, Michael
A1  - Eils, Jürgen
A1  - Lawerenz, Christian
A1  - Erkek, Serap
A1  - Lambo, Sander
A1  - Waszak, Sebastian
A1  - Blattmann, Claudia
A1  - Borkhardt, Arndt
A1  - Kuhlen, Michaela
A1  - Eggert, Angelika
A1  - Fulda, Simone
A1  - Gessler, Manfred
A1  - Wegert, Jenny
A1  - Kappler, Roland
A1  - Baumhoer, Daniel
A1  - Stefan, Burdach
A1  - Kirschner-Schwabe, Renate
A1  - Kontny, Udo
A1  - Kulozik, Andreas E.
A1  - Lohmann, Dietmar
A1  - Hettmer, Simone
A1  - Eckert, Cornelia
A1  - Bielack, Stefan
A1  - Nathrath, Michaela
A1  - Niemeyer, Charlotte
A1  - Richter, Günther H.
A1  - Schulte, Johannes
A1  - Siebert, Reiner
A1  - Westermann, Frank
A1  - Molenaar, Jan J.
A1  - Vassal, Gilles
A1  - Witt, Hendrik
A1  - Burkhardt, Birgit
A1  - Kratz, Christian P.
A1  - Witt, Olaf
A1  - van Tilburg, Cornelis M.
A1  - Kramm, Christof M.
A1  - Fleischhack, Gudrun
A1  - Dirksen, Uta
A1  - Rutkowski, Stefan
A1  - Frühwald, Michael
A1  - Hoff, Katja von
A1  - Wolf, Stephan
A1  - Klingebeil, Thomas
A1  - Koscielniak, Ewa
A1  - Landgraf, Pablo
A1  - Koster, Jan
A1  - Resnick, Adam C.
A1  - Zhang, Jinghui
A1  - Liu, Yanling
A1  - Zhou, Xin
A1  - Waanders, Angela J.
A1  - Zwijnenburg, Danny A.
A1  - Raman, Pichai
A1  - Brors, Benedikt
A1  - Weber, Ursula D.
A1  - Northcott, Paul A.
A1  - Pajtler, Kristian W.
A1  - Kool, Marcel
A1  - Piro, Rosario M.
A1  - Korbel, Jan O.
A1  - Schlesner, Matthias
A1  - Eils, Roland
A1  - Jones, David T. W.
A1  - Lichter, Peter
A1  - Chavez, Lukas
A1  - Zapatka, Marc
A1  - Pfister, Stefan M.
T1  - The landscape of genomic alterations across childhood cancers
JF  - Nature
N2  - Pan-cancer analyses that examine commonalities and differences among various cancer types have emerged as a powerful way to obtain novel insights into cancer biology. Here we present a comprehensive analysis of genetic alterations in a pan-cancer cohort including 961 tumours from children, adolescents, and young adults, comprising 24 distinct molecular types of cancer. Using a standardized workflow, we identified marked differences in terms of mutation frequency and significantly mutated genes in comparison to previously analysed adult cancers. Genetic alterations in 149 putative cancer driver genes separate the tumours into two classes: small mutation and structural/copy-number variant (correlating with germline variants). Structural variants, hyperdiploidy, and chromothripsis are linked to TP53 mutation status and mutational signatures. Our data suggest that 7–8% of the children in this cohort carry an unambiguous predisposing germline variant and that nearly 50% of paediatric neoplasms harbour a potentially druggable event, which is highly relevant for the design of future clinical trials.
KW  - cancer genomics
KW  - oncogenesis
KW  - paediatric cancer
KW  - predictive markers
KW  - translational research
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229579
VL  - 555
ER  - 
TY  - JOUR
A1  - Giampaolo, Sabrina
A1  - Wójcik, Gabriela
A1  - Klein-Hessling, Stefan
A1  - Serfling, Edgar
A1  - Patra, Amiya K.
T1  - B cell development is critically dependent on NFATc1 activity
JF  - Cellular & Molecular Immunology
N2  - B cell development in bone marrow is a precisely regulated complex process. Through successive stages of differentiation, which are regulated by a multitude of signaling pathways and an array of lineage-specific transcription factors, the common lymphoid progenitors ultimately give rise to mature B cells. Similar to early thymocyte development in the thymus, early B cell development in bone marrow is critically dependent on IL-7 signaling. During this IL-7-dependent stage of differentiation, several transcription factors, such as E2A, EBF1, and Pax5, among others, play indispensable roles in B lineage specification and maintenance. Although recent studies have implicated several other transcription factors in B cell development, the role of NFATc1 in early B cell developmental stages is not known. Here, using multiple gene-manipulated mouse models and applying various experimental methods, we show that NFATc1 activity is vital for early B cell differentiation. Lack of NFATc1 activity in pro-B cells suppresses EBF1 expression, impairs immunoglobulin gene rearrangement, and thereby preBCR formation, resulting in defective B cell development. Overall, deficiency in NFATc1 activity arrested the pro-B cell transition to the pre-B cell stage, leading to severe B cell lymphopenia. Our findings suggest that, along with other transcription factors, NFATc1 is a critical component of the signaling mechanism that facilitates early B cell differentiation.
KW  - differentiation
KW  - EBF1
KW  - NFATc1
KW  - pro-B
KW  - pre-B
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233006
VL  - 16
ER  - 
TY  - JOUR
A1  - El-Mesery, Mohamed
A1  - Rosenthal, Tina
A1  - Rauert-Wunderlich, Hilka
A1  - Schreder, Martin
A1  - Stühmer, Thorsten
A1  - Leich, Ellen
A1  - Schlosser, Andreas
A1  - Ehrenschwender, Martin
A1  - Wajant, Harald
A1  - Siegmund, Daniela
T1  - The NEDD8-activating enzyme inhibitor MLN4924 sensitizes a TNFR1+ subgroup of multiple myeloma cells for TNF-induced cell death
JF  - Cell Death & Disease
N2  - The NEDD8-activating enzyme (NAE) inhibitor MLN4924 inhibits cullin-RING ubiquitin ligase complexes including the SKP1-cullin-F-box E3 ligase βTrCP. MLN4924 therefore inhibits also the βTrCP-dependent activation of the classical and the alternative NFĸB pathway. In this work, we found that a subgroup of multiple myeloma cell lines (e.g., RPMI-8226, MM.1S, KMS-12BM) and about half of the primary myeloma samples tested are sensitized to TNF-induced cell death by MLN4924. This correlated with MLN4924-mediated inhibition of TNF-induced activation of the classical NFκB pathway and reduced the efficacy of TNF-induced TNFR1 signaling complex formation. Interestingly, binding studies revealed a straightforward correlation between cell surface TNFR1 expression in multiple myeloma cell lines and their sensitivity for MLN4924/TNF-induced cell death. The cell surface expression levels of TNFR1 in the investigated MM cell lines largely correlated with TNFR1 mRNA expression. This suggests that the variable levels of cell surface expression of TNFR1 in myeloma cell lines are decisive for TNF/MLN4924 sensitivity. Indeed, introduction of TNFR1 into TNFR1-negative TNF/MLN4924-resistant KMS-11BM cells, was sufficient to sensitize this cell line for TNF/MLN4924-induced cell death. Thus, MLN4924 might be especially effective in myeloma patients with TNFR1+ myeloma cells and a TNFhigh tumor microenvironment.
KW  - cancer therapy
KW  - tumour-necrosis factors
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226666
VL  - 10
ER  - 
TY  - JOUR
A1  - Breun, Maria
A1  - Flock, Katharina
A1  - Feldheim, Jonas
A1  - Nattmann, Anja
A1  - Monoranu, Camelia M.
A1  - Herrmann, Pia
A1  - Ernestus, Ralf-Ingo
A1  - Löhr, Mario
A1  - Hagemann, Carsten
A1  - Stein, Ulrike
T1  - Metastasis associated in colorectal cancer 1 (MACC1) mRNA expression is enhanced in sporadic vestibular schwannoma and correlates to deafness
JF  - Cancers
N2  - Vestibular schwannoma (VS) are benign cranial nerve sheath tumors of the vestibulocochlear nerve. Their incidence is mostly sporadic, but they can also be associated with NF2-related schwannomatosis (NF2), a hereditary tumor syndrome. Metastasis associated in colon cancer 1 (MACC1) is known to contribute to angiogenesis, cell growth, invasiveness, cell motility and metastasis of solid malignant cancers. In addition, MACC1 may be associated with nonsyndromic hearing impairment. Therefore, we evaluated whether MACC1 may be involved in the pathogenesis of VS. Sporadic VS, recurrent sporadic VS, NF2-associated VS, recurrent NF2-associated VS and healthy vestibular nerves were analyzed for MACC1 mRNA and protein expression by quantitative polymerase chain reaction and immunohistochemistry. MACC1 expression levels were correlated with the patients’ clinical course and symptoms. MACC1 mRNA expression was significantly higher in sporadic VS compared to NF2-associated VS (p < 0.001). The latter expressed similar MACC1 concentrations as healthy vestibular nerves. Recurrent tumors resembled the MACC1 expression of the primary tumors. MACC1 mRNA expression was significantly correlated with deafness in sporadic VS patients (p = 0.034). Therefore, MACC1 might be a new molecular marker involved in VS pathogenesis.
KW  - vestibular schwannoma
KW  - metastasis associated in colorectal cancer 1 (MACC1)
KW  - pathogenesis
KW  - deafness
KW  - NF2-related schwannomatosis (NF2)
KW  - mRNA expression
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-362543
SN  - 2072-6694
VL  - 15
IS  - 16
ER  - 
TY  - JOUR
A1  - Murti, Krisna
A1  - Fender, Hendrik
A1  - Glatzle, Carolin
A1  - Wismer, Rhoda
A1  - Sampere-Birlanga, Salvador
A1  - Wild, Vanessa
A1  - Muhammad, Khalid
A1  - Rosenwald, Andreas
A1  - Serfling, Edgar
A1  - Avots, Andris
T1  - Calcineurin-independent NFATc1 signaling is essential for survival of Burkitt lymphoma cells
JF  - Frontiers in Oncology
N2  - In Burkitt lymphoma (BL), a tumor of germinal center B cells, the pro-apoptotic properties of MYC are controlled by tonic B cell receptor (BCR) signals. Since BL cells do not exhibit constitutive NF-κB activity, we hypothesized that anti-apoptotic NFATc1 proteins provide a major transcriptional survival signal in BL. Here we show that post-transcriptional mechanisms are responsible for the calcineurin (CN) independent constitutive nuclear over-expression of NFATc1 in BL and Eµ-MYC – induced B cell lymphomas (BCL). Conditional inactivation of the Nfatc1 gene in B cells of Eµ-MYC mice leads to apoptosis of BCL cells in vivo and ex vivo. Inhibition of BCR/SYK/BTK/PI3K signals in BL cells results in cytosolic re-location of NFATc1 and apoptosis. Therefore, NFATc1 activity is an integrated part of tonic BCR signaling and an alternative target for therapeutic intervention in BL.
KW  - apoptosis
KW  - Burkitt lymphoma
KW  - cyclosporin A
KW  - nuclear localization
KW  - NFATc1
KW  - activated B cell-like diffuse large B-cell lymphoma (ABC-DLBCL)
KW  - B cell receptor (BCR)
KW  - Burkitt lymphoma (BL)
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-323103
VL  - 13
ER  - 
TY  - JOUR
A1  - Seal, Rishav
A1  - Schwab, Lara S. U.
A1  - Chiarolla, Cristina M.
A1  - Hundhausen, Nadine
A1  - Klose, Georg Heinrich
A1  - Reu-Hofer, Simone
A1  - Rosenwald, Andreas
A1  - Wiest, Johannes
A1  - Berberich-Siebelt, Friederike
T1  - Delayed and limited administration of the JAKinib tofacitinib mitigates chronic DSS-induced colitis
JF  - Frontiers in Immunology
N2  - In inflammatory bowel disease, dysregulated T cells express pro-inflammatory cytokines. Using a chronic azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced colitis model resembling ulcerative colitis, we evaluated whether and when treatment with the Janus kinase (JAK) inhibitor tofacitinib could be curative. Comparing the treatment with two and three cycles of tofacitinib medication in drinking water – intermittently with DSS induction – revealed that two cycles were not only sufficient but also superior over the 3-x regimen. The two cycles of the 2-x protocol paralleled the second and third cycles of the longer protocol. T cells were less able to express interferon gamma (IFN-γ) and the serum levels of IFN-γ, interleukin (IL)-2, IL-6, IL-17, and tumor necrosis factor (TNF) were significantly reduced in sera, while those of IL-10 and IL-22 increased under the 2-x protocol. Likewise, the frequency and effector phenotype of regulatory T cells (Tregs) increased. This was accompanied by normal weight gain, controlled clinical scores, and restored stool consistency. The general and histologic appearance of the colons revealed healing and tissue intactness. Importantly, two phases of tofacitinib medication completely prevented AOM-incited pseudopolyps and the hyper-proliferation of epithelia, which was in contrast to the 3-x regimen. This implies that the initial IBD-induced cytokine expression is not necessarily harmful as long as inflammatory signaling can later be suppressed and that time-restricted treatment allows for anti-inflammatory and tissue-healing cytokine activities.
KW  - anti-inflammatory cytokines
KW  - AOM/DSS
KW  - pro-inflammatory cytokines
KW  - effector Treg (eTreg)
KW  - chronic IBD model
KW  - JAK inhibitor
KW  - tofacitinib
KW  - treatment regimens
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-317815
VL  - 14
ER  - 
TY  - JOUR
A1  - Hung, Sophia
A1  - Kasperkowitz, Amelie
A1  - Kurz, Florian
A1  - Dreher, Liane
A1  - Diessner, Joachim
A1  - Ibrahim, Eslam S.
A1  - Schwarz, Stefan
A1  - Ohlsen, Knut
A1  - Hertlein, Tobias
T1  - Next-generation humanized NSG-SGM3 mice are highly susceptible to Staphylococcus aureus infection
JF  - Frontiers in Immunology
N2  - Humanized hemato-lymphoid system mice, or humanized mice, emerged in recent years as a promising model to study the course of infection of human-adapted or human-specific pathogens. Though Staphylococcus aureus infects and colonizes a variety of species, it has nonetheless become one of the most successful human pathogens of our time with a wide armory of human-adapted virulence factors. Humanized mice showed increased vulnerability to S. aureus compared to wild type mice in a variety of clinically relevant disease models. Most of these studies employed humanized NSG (NOD-scid IL2Rgnull) mice which are widely used in the scientific community, but show poor human myeloid cell reconstitution. Since this immune cell compartment plays a decisive role in the defense of the human immune system against S. aureus, we asked whether next-generation humanized mice, like NSG-SGM3 (NOD-scid IL2Rgnull-3/GM/SF) with improved myeloid reconstitution, would prove to be more resistant to infection. To our surprise, we found the contrary when we infected humanized NSG-SGM3 (huSGM3) mice with S. aureus: although they had stronger human immune cell engraftment than humanized NSG mice, particularly in the myeloid compartment, they displayed even more pronounced vulnerability to S. aureus infection. HuSGM3 mice had overall higher numbers of human T cells, B cells, neutrophils and monocytes in the blood and the spleen. This was accompanied by elevated levels of pro-inflammatory human cytokines in the blood of huSGM3 mice. We further identified that the impaired survival of huSGM3 mice was not linked to higher bacterial burden nor to differences in the murine immune cell repertoire. Conversely, we could demonstrate a correlation of the rate of humanization and the severity of infection. Collectively, this study suggests a detrimental effect of the human immune system in humanized mice upon encounter with S. aureus which might help to guide future therapy approaches and analysis of virulence mechanisms.
KW  - humanized mice
KW  - Staphylococcus aureus
KW  - MRSA
KW  - NSG
KW  - NSG-SGM3
KW  - staphylococcal abscess
KW  - Staphylococcus aureus immune response
KW  - humanized hemato-lymphoid mice
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-306966
VL  - 14
ER  - 
TY  - JOUR
A1  - Madrahimov, Nodir
A1  - Mutsenko, Vitalii
A1  - Natanov, Ruslan
A1  - Radaković, Dejan
A1  - Klapproth, André
A1  - Hassan, Mohamed
A1  - Rosenfeldt, Mathias
A1  - Kleefeldt, Florian
A1  - Aleksic, Ivan
A1  - Ergün, Süleyman
A1  - Otto, Christoph
A1  - Leyh, Rainer G.
A1  - Bening, Constanze
T1  - Multiorgan recovery in a cadaver body using mild hypothermic ECMO treatment in a murine model
JF  - Intensive Care Medicine Experimental
N2  - Background
Transplant candidates on the waiting list are increasingly challenged by the lack of organs. Most of the organs can only be kept viable within very limited timeframes (e.g., mere 4–6 h for heart and lungs exposed to refrigeration temperatures ex vivo). Donation after circulatory death (DCD) using extracorporeal membrane oxygenation (ECMO) can significantly enlarge the donor pool, organ yield per donor, and shelf life. Nevertheless, clinical attempts to recover organs for transplantation after uncontrolled DCD are extremely complex and hardly reproducible. Therefore, as a preliminary strategy to fulfill this task, experimental protocols using feasible animal models are highly warranted. The primary aim of the study was to develop a model of ECMO-based cadaver organ recovery in mice. Our model mimics uncontrolled organ donation after an “out-of-hospital” sudden unexpected death with subsequent “in-hospital” cadaver management post-mortem. The secondary aim was to assess blood gas parameters, cardiac activity as well as overall organ state. The study protocol included post-mortem heparin–streptokinase administration 10 min after confirmed death induced by cervical dislocation under full anesthesia. After cannulation, veno-arterial ECMO (V–A ECMO) was started 1 h after death and continued for 2 h under mild hypothermic conditions followed by organ harvest. Pressure- and flow-controlled oxygenated blood-based reperfusion of a cadaver body was accompanied by blood gas analysis (BGA), electrocardiography, and histological evaluation of ischemia–reperfusion injury. For the first time, we designed and implemented, a not yet reported, miniaturized murine hemodialysis circuit for the treatment of severe hyperkalemia and metabolic acidosis post-mortem.

Results
BGA parameters confirmed profound ischemia typical for cadavers and incompatible with normal physiology, including extremely low blood pH, profound negative base excess, and enormously high levels of lactate. Two hours after ECMO implantation, blood pH values of a cadaver body restored from < 6.5 to 7.3 ± 0.05, pCO2 was lowered from > 130 to 41.7 ± 10.5 mmHg, sO2, base excess, and HCO3 were all elevated from below detection thresholds to 99.5 ± 0.6%, − 4 ± 6.2 and 22.0 ± 6.0 mmol/L, respectively (Student T test, p < 0.05). A substantial decrease in hyperlactatemia (from > 20 to 10.5 ± 1.7 mmol/L) and hyperkalemia (from > 9 to 6.9 ± 1.0 mmol/L) was observed when hemodialysis was implemented. On balance, the first signs of regained heart activity appeared on average 10 min after ECMO initiation without cardioplegia or any inotropic and vasopressor support. This was followed by restoration of myocardial contractility with a heart rate of up to 200 beats per minute (bpm) as detected by an electrocardiogram (ECG). Histological examinations revealed no evidence of heart injury 3 h post-mortem, whereas shock-specific morphological changes relevant to acute death and consequent cardiac/circulatory arrest were observed in the lungs, liver, and kidney of both control and ECMO-treated cadaver mice.

Conclusions
Thus, our model represents a promising approach to facilitate studying perspectives of cadaveric multiorgan recovery for transplantation. Moreover, it opens new possibilities for cadaver organ treatment to extend and potentiate donation and, hence, contribute to solving the organ shortage dilemma.
KW  - extracorporeal membrane oxygenation
KW  - cadaver multiorgan preservation
KW  - mild hypothermia
KW  - post-mortem heart recovery
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-357381
VL  - 11
ER  - 
TY  - JOUR
A1  - Lisowski, Dominik
A1  - Hartrampf, Philipp E.
A1  - Hasenauer, Natalie
A1  - Nickl, Vera
A1  - Monoranu, Camelia-Maria
A1  - Tamihardja, Jörg
T1  - Complete loss of E-cadherin expression in a rare case of metastatic malignant meningioma: a case report
JF  - BMC Neurology
N2  - Background
Hematogenous tumor spread of malignant meningiomas occurs very rarely but is associated with very poor prognosis.

Case presentation
We report an unusual case of a patient with a malignant meningioma who developed multiple metastases in bones, lungs and liver after initial complete resection of the primary tumor. After partial hepatic resection, specimens were histologically analyzed, and a complete loss of E-cadherin adhesion molecules was found. No oncogenic target mutations were found. The patient received a combination of conventional radiotherapy and peptide receptor radionuclide therapy (PRRT). Due to aggressive tumor behavior and rapid spread of metastases, the patient deceased after initiation of treatment.

Conclusions
E-cadherin downregulation is associated with a higher probability of tumor invasion and distant metastasis formation in malignant meningioma. Up to now, the efficacy of systemic therapy, including PRRT, is very limited in malignant meningioma patients.
KW  - beta-catenin
KW  - E-cadherin
KW  - meningioma
KW  - peptide receptor radionuclide therapy (PRRT)
KW  - radiotherapy
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-357996
VL  - 23
ER  - 
TY  - JOUR
A1  - Meinert, Madlen
A1  - Jessen, Christina
A1  - Hufnagel, Anita
A1  - Kreß, Julia Katharina Charlotte
A1  - Burnworth, Mychal
A1  - Däubler, Theo
A1  - Gallasch, Till
A1  - Da Xavier Silva, Thamara Nishida
A1  - Dos Santos, Ancély Ferreira
A1  - Ade, Carsten Patrick
A1  - Schmitz, Werner
A1  - Kneitz, Susanne
A1  - Friedmann Angeli, José Pedro
A1  - Meierjohann, Svenja
T1  - Thiol starvation triggers melanoma state switching in an ATF4 and NRF2-dependent manner
JF  - Redox Biology
N2  - The cystine/glutamate antiporter xCT is an important source of cysteine for cancer cells. Once taken up, cystine is reduced to cysteine and serves as a building block for the synthesis of glutathione, which efficiently protects cells from oxidative damage and prevents ferroptosis. As melanomas are particularly exposed to several sources of oxidative stress, we investigated the biological role of cysteine and glutathione supply by xCT in melanoma. xCT activity was abolished by genetic depletion in the Tyr::CreER; Braf\(^{CA}\); Pten\(^{lox/+}\) melanoma model and by acute cystine withdrawal in melanoma cell lines. Both interventions profoundly impacted melanoma glutathione levels, but they were surprisingly well tolerated by murine melanomas in vivo and by most human melanoma cell lines in vitro. RNA sequencing of human melanoma cells revealed a strong adaptive upregulation of NRF2 and ATF4 pathways, which orchestrated the compensatory upregulation of genes involved in antioxidant defence and de novo cysteine biosynthesis. In addition, the joint activation of ATF4 and NRF2 triggered a phenotypic switch characterized by a reduction of differentiation genes and induction of pro-invasive features, which was also observed after erastin treatment or the inhibition of glutathione synthesis. NRF2 alone was capable of inducing the phenotypic switch in a transient manner. Together, our data show that cystine or glutathione levels regulate the phenotypic plasticity of melanoma cells by elevating ATF4 and NRF2.
KW  - thiol starvation
KW  - ATF4
KW  - NRF2
KW  - melanoma
Y1  - 2024
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-350328
VL  - 70
ER  - 
TY  - JOUR
A1  - Bassler, Miriam C.
A1  - Knoblich, Mona
A1  - Gerhard-Hartmann, Elena
A1  - Mukherjee, Ashutosh
A1  - Youssef, Almoatazbellah
A1  - Hagen, Rudolf
A1  - Haug, Lukas
A1  - Goncalves, Miguel
A1  - Scherzad, Agmal
A1  - Stöth, Manuel
A1  - Ostertag, Edwin
A1  - Steinke, Maria
A1  - Brecht, Marc
A1  - Hackenberg, Stephan
A1  - Meyer, Till Jasper
T1  - Differentiation of salivary gland and salivary gland tumor tissue via Raman imaging combined with multivariate data analysis
JF  - Diagnostics
N2  - Salivary gland tumors (SGTs) are a relevant, highly diverse subgroup of head and neck tumors whose entity determination can be difficult. Confocal Raman imaging in combination with multivariate data analysis may possibly support their correct classification. For the analysis of the translational potential of Raman imaging in SGT determination, a multi-stage evaluation process is necessary. By measuring a sample set of Warthin tumor, pleomorphic adenoma and non-tumor salivary gland tissue, Raman data were obtained and a thorough Raman band analysis was performed. This evaluation revealed highly overlapping Raman patterns with only minor spectral differences. Consequently, a principal component analysis (PCA) was calculated and further combined with a discriminant analysis (DA) to enable the best possible distinction. The PCA-DA model was characterized by accuracy, sensitivity, selectivity and precision values above 90% and validated by predicting model-unknown Raman spectra, of which 93% were classified correctly. Thus, we state our PCA-DA to be suitable for parotid tumor and non-salivary salivary gland tissue discrimination and prediction. For evaluation of the translational potential, further validation steps are necessary.
KW  - salivary gland tumor
KW  - confocal Raman imaging
KW  - principal component analysis
KW  - discriminant analysis
KW  - multivariate data analysis
KW  - molecular diagnostics
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-355558
SN  - 2075-4418
VL  - 14
IS  - 1
ER  - 
TY  - JOUR
A1  - Kreß, Julia Katharina Charlotte
A1  - Jessen, Christina
A1  - Hufnagel, Anita
A1  - Schmitz, Werner
A1  - Da Xavier Silva, Thamara Nishida
A1  - Ferreira Dos Santos, Ancély
A1  - Mosteo, Laura
A1  - Goding, Colin R.
A1  - Friedmann Angeli, José Pedro
A1  - Meierjohann, Svenja
T1  - The integrated stress response effector ATF4 is an obligatory metabolic activator of NRF2
JF  - Cell Reports
N2  - Highlights
• The integrated stress response leads to a general ATF4-dependent activation of NRF2
• ATF4 causes a CHAC1-dependent GSH depletion, resulting in NRF2 stabilization
• An elevation of NRF2 transcript levels fosters this effect
• NRF2 supports the ISR/ATF4 pathway by improving cystine and antioxidant supply

Summary
The redox regulator NRF2 becomes activated upon oxidative and electrophilic stress and orchestrates a response program associated with redox regulation, metabolism, tumor therapy resistance, and immune suppression. Here, we describe an unrecognized link between the integrated stress response (ISR) and NRF2 mediated by the ISR effector ATF4. The ISR is commonly activated after starvation or ER stress and plays a central role in tissue homeostasis and cancer plasticity. ATF4 increases NRF2 transcription and induces the glutathione-degrading enzyme CHAC1, which we now show to be critically important for maintaining NRF2 activation. In-depth analyses reveal that NRF2 supports ATF4-induced cells by increasing cystine uptake via the glutamate-cystine antiporter xCT. In addition, NRF2 upregulates genes mediating thioredoxin usage and regeneration, thus balancing the glutathione decrease. In conclusion, we demonstrate that the NRF2 response serves as second layer of the ISR, an observation highly relevant for the understanding of cellular resilience in health and disease.
KW  - NRF2
KW  - ATF4
KW  - integrated stress response
KW  - CHAC1
KW  - melanoma
KW  - SLC7A11
KW  - GSH
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-350312
VL  - 42
IS  - 7
ER  - 
TY  - JOUR
A1  - Böck, Julia
A1  - Maurus, Katja
A1  - Gerhard-Hartmann, Elena
A1  - Brändlein, Stephanie
A1  - Kurz, Katrin S.
A1  - Ott, German
A1  - Anagnostopoulos, Ioannis
A1  - Rosenwald, Andreas
A1  - Zamò, Alberto
T1  - Targeted panel sequencing in the routine diagnosis of mature T- and NK-cell lymphomas
BT  - report of 128 cases from two German reference centers
JF  - Frontiers in Oncology
N2  - Diagnosing any of the more than 30 types of T-cell lymphomas is considered a challenging task for many pathologists and currently requires morphological expertise as well as the integration of clinical data, immunophenotype, flow cytometry and clonality analyses. Even considering all available information, some margin of doubt might remain using the current diagnostic procedures. In recent times, the genetic landscape of most T-cell lymphomas has been elucidated, showing a number of diagnostically relevant mutations. In addition, recent data indicate that some of these genetic alterations might bear prognostic and predictive value. Extensive genetic analyses, such as whole exome or large panel sequencing are still expensive and time consuming, therefore limiting their application in routine diagnostic. We therefore devoted our effort to develop a lean approach for genetic analysis of T-cell lymphomas, focusing on maximum efficiency rather than exhaustively covering all possible targets. Here we report the results generated with our small amplicon-based panel that could be used routinely on paraffin-embedded and even decalcified samples, on a single sample basis in parallel with other NGS-panels used in our routine diagnostic lab, in a relatively short time and with limited costs. We tested 128 available samples from two German reference centers as part of our routine work up (among which 116 T-cell lymphomas), which is the largest routine diagnostic series reported to date. Our results showed that this assay had a very high rate of technical success (97%) and could detect mutations in the majority (79%) of tested T-cell lymphoma samples.
KW  - T-cell lymphoma
KW  - panel-sequencing
KW  - NGS
KW  - diagnostics
KW  - mutation
KW  - FFPE
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-326478
SN  - 2234-943X
VL  - 13
ER  - 
TY  - JOUR
A1  - Gschmack, Eva
A1  - Monoranu, Camelia-Maria
A1  - Marouf, Hecham
A1  - Meyer, Sarah
A1  - Lessel, Lena
A1  - Idris, Raja
A1  - Berg, Daniela
A1  - Maetzler, Walter
A1  - Steigerwald, Frank
A1  - Volkmann, Jens
A1  - Gerlach, Manfred
A1  - Riederer, Peter
A1  - Koutsilieri, Eleni
A1  - Scheller, Carsten
T1  - Plasma autoantibodies to glial fibrillary acidic protein (GFAP) react with brain areas according to Braak staging of Parkinson’s disease
JF  - Journal of Neural Transmission
N2  - Idiopathic Parkinson’s disease (PD) is characterized by a progredient degeneration of the brain, starting at deep subcortical areas such as the dorsal motor nucleus of the glossopharyngeal and vagal nerves (DM) (stage 1), followed by the coeruleus–subcoeruleus complex; (stage 2), the substantia nigra (SN) (stage 3), the anteromedial temporal mesocortex (MC) (stage 4), high-order sensory association areas and prefrontal fields (HC) (stage 5) and finally first-order sensory association areas, premotor areas, as well as primary sensory and motor field (FC) (stage 6). Autoimmunity might play a role in PD pathogenesis. Here we analyzed whether anti-brain autoantibodies differentially recognize different human brain areas and identified autoantigens that correlate with the above-described dissemination of PD pathology in the brain. Brain tissue was obtained from deceased individuals with no history of neurological or psychiatric disease and no neuropathological abnormalities. Tissue homogenates from different brain regions (DM, SN, MC, HC, FC) were subjected to SDS-PAGE and Western blot. Blots were incubated with plasma samples from 30 PD patients and 30 control subjects and stained with anti-IgG antibodies to detect anti-brain autoantibodies. Signals were quantified. Prominent autoantigens were identified by 2D-gel-coupled mass spectrometry sequencing. Anti-brain autoantibodies are frequent and occur both in healthy controls and individuals with PD. Glial fibrillary acidic protein (GFAP) was identified as a prominent autoantigen recognized in all plasma samples. GFAP immunoreactivity was highest in DM areas and lowest in FC areas with no significant differences in anti-GFAP autoantibody titers between healthy controls and individuals with PD. The anti-GFAP autoimmunoreactivity of different brain areas correlates with the dissemination of histopathological neurodegeneration in PD. We hypothesize that GFAP autoantibodies are physiological but might be involved as a cofactor in PD pathogenesis secondary to a leakage of the blood–brain barrier.
KW  - Parkinson
KW  - GFAP
KW  - autoantibodies
KW  - Braak
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325161
VL  - 129
IS  - 5-6
ER  - 
TY  - JOUR
A1  - Linz, Christian
A1  - Brands, Roman C.
A1  - Kertels, Olivia
A1  - Dierks, Alexander
A1  - Brumberg, Joachim
A1  - Gerhard-Hartmann, Elena
A1  - Hartmann, Stefan
A1  - Schirbel, Andreas
A1  - Serfling, Sebastian
A1  - Zhi, Yingjun
A1  - Buck, Andreas K.
A1  - Kübler, Alexander
A1  - Hohm, Julian
A1  - Lapa, Constantin
A1  - Kircher, Malte
T1  - Targeting fibroblast activation protein in newly diagnosed squamous cell carcinoma of the oral cavity – initial experience and comparison to [\(^{18}\)F]FDG PET/CT and MRI
JF  - European Journal of Nuclear Medicine and Molecular Imaging
N2  - Purpose
While [\(^{18}\)F]-fluorodeoxyglucose ([\(^{18}\)F]FDG) is the standard for positron emission tomography/computed tomography (PET/CT) imaging of oral squamous cell carcinoma (OSCC), diagnostic specificity is hampered by uptake in inflammatory cells such as neutrophils or macrophages. Recently, molecular imaging probes targeting fibroblast activation protein α (FAP), which is overexpressed in a variety of cancer-associated fibroblasts, have become available and might constitute a feasible alternative to FDG PET/CT.

Methods
Ten consecutive, treatment-naïve patients (8 males, 2 females; mean age, 62 ± 9 years) with biopsy-proven OSCC underwent both whole-body [\(^{18}\)F]FDG and [\(^{68}\)Ga]FAPI-04 (FAP-directed) PET/CT for primary staging prior to tumor resection and cervical lymph node dissection. Detection of the primary tumor, as well as the presence and number of lymph node and distant metastases was analysed. Intensity of tracer accumulation was assessed by means of maximum (SUV\(_{max}\)) and peak (SUV\(_{peak}\) standardized uptake values. Histological work-up including immunohistochemical staining for FAP served as standard of reference.

Results
[\(^{18}\)F]FDG and FAP-directed PET/CT detected all primary tumors with a SUVmax of 25.5 ± 13.2 (FDG) and 20.5 ± 6.4 (FAP-directed) and a SUVpeak of 16.1 ± 10.3 ([\(^{18}\)F]FDG) and 13.8 ± 3.9 (FAP-directed), respectively. Regarding cervical lymph node metastases, FAP-directed PET/CT demonstrated comparable sensitivity (81.3% vs. 87.5%; P = 0.32) and specificity (93.3% vs. 81.3%; P = 0.16) to [\(^{18}\)F]FDG PET/CT. FAP expression on the cell surface of cancer-associated fibroblasts in both primary lesions as well as lymph nodes metastases was confirmed in all samples.

Conclusion
FAP-directed PET/CT in OSCC seems feasible. Future research to investigate its potential to improve patient staging is highly warranted.
KW  - molecular imaging
KW  - fibroblast activation protein
KW  - head and neck cancer
KW  - PET
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-307246
SN  - 1619-7070
SN  - 1619-7089
VL  - 48
IS  - 12
ER  - 
TY  - JOUR
A1  - Zamò, Alberto
A1  - Gerhard-Hartmann, Elena
A1  - Ott, German
A1  - Anagnostopoulos, Ioannis
A1  - Scott, David W.
A1  - Rosenwald, Andreas
A1  - Rauert-Wunderlich, Hilka
T1  - Routine application of the Lymph2Cx assay for the subclassification of aggressive B-cell lymphoma: report of a prospective real-world series
JF  - Virchows Archiv
N2  - The subclassification of diffuse large B-cell lymphoma (DLBCL) into germinal center B-cell-like (GCB) and activated B-cell-like (ABC) subtypes has become mandatory in the 2017 update of the WHO classification of lymphoid neoplasms and will continue to be used in the WHO 5\(^{th}\) edition. The RNA-based Lymph2Cx assay has been validated as a reliable surrogate of high-throughput gene expression profiling assays for distinguishing between GCB and ABC DLBCL and provides reliable results from formalin-fixed, paraffin-embedded (FFPE) material. This test has been previously used in clinical trials, but experience from real-world routine application is rare. We routinely applied the Lymph2Cx assay to day-to-day diagnostics on a series of 147 aggressive B-cell lymphoma cases and correlated our results with the immunohistochemical subclassification using the Hans algorithm and fluorescence in situ hybridization findings using break-apart probes for MYC, BCL2, and BCL6. The routine use of the Lymph2Cx assay had a high technical success rate (94.6%) with a low rate of failure due to poor material and/or RNA quality. The Lymph2Cx assay was discordant with the Hans algorithm in 18% (23 of 128 cases). Discordant cases were mainly classified as GCB by the Hans algorithm and as ABC by Lymph2Cx (n = 11, 8.6%). Only 5 cases (3.9%) were classified as non-GCB by the Hans algorithm and as GCB by Lymph2Cx. Additionally, 5.5% of cases (n = 7) were left unclassified by Lymph2Cx, whereas they were defined as GCB (n = 4) or non-GCB (n = 3) by the Hans algorithm. Our data support the routine applicability of the Lymph2Cx assay.
KW  - diffuse large B-cell lymphoma
KW  - Hans algorithm
KW  - Lymph2Cx assay
KW  - cell of origin
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-324686
VL  - 481
IS  - 6
ER  - 
TY  - JOUR
A1  - Hrudka, Jan
A1  - Eis, Václav
A1  - Heřman, Josef
A1  - Prouzová, Zuzana
A1  - Rosenwald, Andreas
A1  - František, Duška
T1  - Panniculitis-like T-cell-lymphoma in the mesentery associated with hemophagocytic syndrome: autopsy case report
JF  - Diagnostic Pathology
N2  - Background
Panniculitis-like T-cell lymphoma is an uncommon type of non-Hodgkin lymphoma, occurring usually in the form of nodules within the subcutaneous fat tissue of the extremities or trunk. In the literature, subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is described as a distinct type of T-cell lymphoma with a variable clinical behavior, depending on molecular phenotype of T-cell receptor (TCR) and on the presence or absence of hemophagocytic syndrome.

Case presentation
We present a bioptic and autoptic case of a 65-years old Caucasian man with panniculitic T-cell lymphoma with morphological and immunohistochemical features of SPTCL, limited to the retroperitoneal and mesenteric mass, i.e. without any cutaneous involvement, and associated with severe hemophagocytic lymphohistiocytosis.

Conclusion
A panniculitic T-cell lymphoma with morphological and molecular features of SPTCL, which is limited to mesentery, i.e. does not involve subcutaneous fat, seems to be exceedingly rare.
KW  - panniculitis
KW  - T-cell lymphoma
KW  - mesentery
KW  - hemophagocytosis
KW  - lymphohistiocytosis
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-322665
VL  - 14
ER  - 
TY  - JOUR
A1  - Bohnert, Simone
A1  - Trella, Stefanie
A1  - Preiß, Ulrich
A1  - Heinsen, Helmut
A1  - Bohnert, Michael
A1  - Zwirner, Johann
A1  - Tremblay, Marie-Ève
A1  - Monoranu, Camelia-Maria
A1  - Ondruschka, Benjamin
T1  - Density of TMEM119-positive microglial cells in postmortem cerebrospinal fluid as a surrogate marker for assessing complex neuropathological processes in the CNS
JF  - International Journal of Legal Medicine
N2  - Routine coronal paraffin-sections through the dorsal frontal and parieto-occipital cortex of a total of sixty cases with divergent causes of death were immunohistochemically (IHC) stained with an antibody against TMEM119. Samples of cerebrospinal fluid (CSF) of the same cases were collected by suboccipital needle-puncture, subjected to centrifugation and processed as cytospin preparations stained with TMEM119. Both, cytospin preparations and sections were subjected to computer-assisted density measurements. The density of microglial TMEM119-positive cortical profiles correlated with that of cytospin results and with the density of TMEM119-positive microglial profiles in the medullary layer. There was no statistically significant correlation between the density of medullary TMEM119-positive profiles and the cytospin data. Cortical microglial cells were primarily encountered in supragranular layers I, II, and IIIa and in infragranular layers V and VI, the region of U-fibers and in circumscribed foci or spread in a diffuse manner and high density over the white matter. We have evidence that cortical microglia directly migrate into CSF without using the glympathic pathway. Microglia in the medullary layer shows a strong affinity to the adventitia of deep vessels in the myelin layer. Selected rapidly fatal cases including myocardial infarcts and drowning let us conclude that microglia in cortex and myelin layer can react rapidly and its reaction and migration is subject to pre-existing external and internal factors. Cytospin preparations proved to be a simple tool to analyze and assess complex changes in the CNS after rapid fatal damage. There is no statistically significant correlation between cytospin and postmortem interval. Therefore, the quantitative analyses of postmortem cytospins obviously reflect the neuropathology of the complete central nervous system. Cytospins provide forensic pathologists a rather simple and easy to perform method for the global assessment of CNS affliction.
KW  - cerebrospinal fluid
KW  - forensic neuropathology
KW  - forensic neurotraumatology
KW  - immunohistochemistry
KW  - immunocytochemistry
KW  - biomarker
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-325009
VL  - 136
IS  - 6
ER  - 
TY  - JOUR
A1  - Dirks, Johannes
A1  - Haase, Gabriele
A1  - Cantaert, Tineke
A1  - Frey, Lea
A1  - Klaas, Moritz
A1  - Rickert, Christian H.
A1  - Girschick, Hermann
A1  - Meffre, Eric
A1  - Morbach, Henner
T1  - A novel AICDA splice-site mutation in two siblings with HIGM2 permits somatic hypermutation but abrogates mutational targeting
JF  - Journal of Clinical Immunology
N2  - Hyper-IgM syndrome type 2 (HIGM2) is a B cell intrinsic primary immunodeficiency caused by mutations in AICDA encoding activation-induced cytidine deaminase (AID) which impair immunoglobulin class switch recombination (CSR) and somatic hypermutation (SHM). Whereas autosomal-recessive AID-deficiency (AR-AID) affects both CSR and SHM, the autosomal-dominant form (AD-AID) due to C-terminal heterozygous variants completely abolishes CSR but only partially affects SHM. AR-AID patients display enhanced germinal center (GC) reactions and autoimmune manifestations, which are not present in AD-AID, suggesting that SHM but not CSR regulates GC reactions and peripheral B cell tolerance. Herein, we describe two siblings with HIGM2 due to a novel homozygous AICDA mutation (c.428-1G > T) which disrupts the splice acceptor site of exon 4 and results in the sole expression of a truncated AID variant that lacks 10 highly conserved amino acids encoded by exon 4 (AID-ΔE4a). AID-ΔE4a patients suffered from defective CSR and enhanced GC reactions and were therefore indistinguishable from other AR-AID patients. However, the AID-ΔE4a variant only partially affected SHM as observed in AD-AID patients. In addition, AID-ΔE4a but not AD-AID patients revealed impaired targeting of mutational hotspot motives and distorted mutational patterns. Hence, qualitative defects in AID function and altered SHM rather than global decreased SHM activity may account for the disease phenotype in these patients.
KW  - hyper-IgM syndrome type 2 (HIGM2)
KW  - AICDA
KW  - AID-ΔE4a
KW  - AD-AID
KW  - mutational targeting
KW  - somatic hypermutation
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-324253
VL  - 42
IS  - 4
ER  - 
TY  - JOUR
A1  - Schulmeyer, Carla E.
A1  - Fasching, Peter A.
A1  - Häberle, Lothar
A1  - Meyer, Julia
A1  - Schneider, Michael
A1  - Wachter, David
A1  - Ruebner, Matthias
A1  - Pöschke, Patrik
A1  - Beckmann, Matthias W.
A1  - Hartmann, Arndt
A1  - Erber, Ramona
A1  - Gass, Paul
T1  - Expression of the immunohistochemical markers CK5, CD117, and EGFR in molecular subtypes of breast cancer correlated with prognosis
JF  - Diagnostics
N2  - Molecular-based subclassifications of breast cancer are important for identifying treatment options and stratifying the prognosis in breast cancer. This study aimed to assess the prognosis relative to disease-free survival (DFS) and overall survival (OS) in patients with triple-negative breast cancer (TNBC) and other subtypes, using a biomarker panel including cytokeratin 5 (CK5), cluster of differentiation 117 (CD117), and epidermal growth factor receptor (EGFR). This cohort–case study included histologically confirmed breast carcinomas as cohort arm. From a total of 894 patients, 572 patients with early breast cancer, sufficient clinical data, and archived tumor tissue were included. Using the immunohistochemical markers CK5, CD117, and EGFR, two subgroups were formed: one with all three biomarkers negative (TBN) and one with at least one of those three biomarkers positive (non-TBN). There were significant differences between the two biomarker subgroups (TBN versus non-TBN) in TNBC for DFS (p = 0.04) and OS (p = 0.02), with higher survival rates (DFS and OS) in the non-TBN subgroup. In this study, we found the non-TBN subgroup of TNBC lesions with at least one positive biomarker of CK5, CD117, and/or EGFR, to be associated with longer DFS and OS.
KW  - early breast cancer
KW  - therapy
KW  - prognosis
KW  - CK5
KW  - CD117
KW  - EGFR
KW  - triple-negative breast cancer
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-304987
SN  - 2075-4418
VL  - 13
IS  - 3
ER  - 
TY  - JOUR
A1  - Stefanakis, Mona
A1  - Bassler, Miriam C.
A1  - Walczuch, Tobias R.
A1  - Gerhard-Hartmann, Elena
A1  - Youssef, Almoatazbellah
A1  - Scherzad, Agmal
A1  - Stöth, Manuel Bernd
A1  - Ostertag, Edwin
A1  - Hagen, Rudolf
A1  - Steinke, Maria R.
A1  - Hackenberg, Stephan
A1  - Brecht, Marc
A1  - Meyer, Till Jasper
T1  - The impact of tissue preparation on salivary gland tumors investigated by Fourier-transform infrared microspectroscopy
JF  - Journal of Clinical Medicine
N2  - Due to the wide variety of benign and malignant salivary gland tumors, classification and malignant behavior determination based on histomorphological criteria can be difficult and sometimes impossible. Spectroscopical procedures can acquire molecular biological information without destroying the tissue within the measurement processes. Since several tissue preparation procedures exist, our study investigated the impact of these preparations on the chemical composition of healthy and tumorous salivary gland tissue by Fourier-transform infrared (FTIR) microspectroscopy. Sequential tissue cross-sections were prepared from native, formalin-fixed and formalin-fixed paraffin-embedded (FFPE) tissue and analyzed. The FFPE cross-sections were dewaxed and remeasured. By using principal component analysis (PCA) combined with a discriminant analysis (DA), robust models for the distinction of sample preparations were built individually for each parotid tissue type. As a result, the PCA-DA model evaluation showed a high similarity between native and formalin-fixed tissues based on their chemical composition. Thus, formalin-fixed tissues are highly representative of the native samples and facilitate a transfer from scientific laboratory analysis into the clinical routine due to their robust nature. Furthermore, the dewaxing of the cross-sections entails the loss of molecular information. Our study successfully demonstrated how FTIR microspectroscopy can be used as a powerful tool within existing clinical workflows.
KW  - formalin
KW  - fixation
KW  - tissue preparation
KW  - salivary gland neoplasia
KW  - FTIR spectroscopy
KW  - principal component analysis
KW  - discriminant analysis
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-304887
SN  - 2077-0383
VL  - 12
IS  - 2
ER  - 
TY  - JOUR
A1  - Feldheim, Jonas
A1  - Kessler, Almuth F.
A1  - Feldheim, Julia J.
A1  - Schmitt, Dominik
A1  - Oster, Christoph
A1  - Lazaridis, Lazaros
A1  - Glas, Martin
A1  - Ernestus, Ralf-Ingo
A1  - Monoranu, Camelia M.
A1  - Löhr, Mario
A1  - Hagemann, Carsten
T1  - BRMS1 in gliomas — an expression analysis
JF  - Cancers
N2  - The metastatic suppressor BRMS1 interacts with critical steps of the metastatic cascade in many cancer entities. As gliomas rarely metastasize, BRMS1 has mainly been neglected in glioma research. However, its interaction partners, such as NFκB, VEGF, or MMPs, are old acquaintances in neurooncology. The steps regulated by BRMS1, such as invasion, migration, and apoptosis, are commonly dysregulated in gliomas. Therefore, BRMS1 shows potential as a regulator of glioma behavior. By bioinformatic analysis, in addition to our cohort of 118 specimens, we determined BRMS1 mRNA and protein expression as well as its correlation with the clinical course in astrocytomas IDH mutant, CNS WHO grade 2/3, and glioblastoma IDH wild-type, CNS WHO grade 4. Interestingly, we found BRMS1 protein expression to be significantly decreased in the aforementioned gliomas, while BRMS1 mRNA appeared to be overexpressed throughout. This dysregulation was independent of patients’ characteristics or survival. The protein and mRNA expression differences cannot be finally explained at this stage. However, they suggest a post-transcriptional dysregulation that has been previously described in other cancer entities. Our analyses present the first data on BRMS1 expression in gliomas that can provide a starting point for further investigations.
KW  - glioblastoma
KW  - metastasis
KW  - suppressor
KW  - behavior
KW  - mRNA
KW  - protein
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-319225
SN  - 2072-6694
VL  - 15
IS  - 11
ER  - 
TY  - JOUR
A1  - Nickl, Vera
A1  - Eck, Juliana
A1  - Goedert, Nicolas
A1  - Hübner, Julian
A1  - Nerreter, Thomas
A1  - Hagemann, Carsten
A1  - Ernestus, Ralf-Ingo
A1  - Schulz, Tim
A1  - Nickl, Robert Carl
A1  - Keßler, Almuth Friederike
A1  - Löhr, Mario
A1  - Rosenwald, Andreas
A1  - Breun, Maria
A1  - Monoranu, Camelia Maria
T1  - Characterization and optimization of the tumor microenvironment in patient-derived organotypic slices and organoid models of glioblastoma
JF  - Cancers
N2  - While glioblastoma (GBM) is still challenging to treat, novel immunotherapeutic approaches have shown promising effects in preclinical settings. However, their clinical breakthrough is hampered by complex interactions of GBM with the tumor microenvironment (TME). Here, we present an analysis of TME composition in a patient-derived organoid model (PDO) as well as in organotypic slice cultures (OSC). To obtain a more realistic model for immunotherapeutic testing, we introduce an enhanced PDO model. We manufactured PDOs and OSCs from fresh tissue of GBM patients and analyzed the TME. Enhanced PDOs (ePDOs) were obtained via co-culture with PBMCs (peripheral blood mononuclear cells) and compared to normal PDOs (nPDOs) and PT (primary tissue). At first, we showed that TME was not sustained in PDOs after a short time of culture. In contrast, TME was largely maintained in OSCs. Unfortunately, OSCs can only be cultured for up to 9 days. Thus, we enhanced the TME in PDOs by co-culturing PDOs and PBMCs from healthy donors. These cellular TME patterns could be preserved until day 21. The ePDO approach could mirror the interaction of GBM, TME and immunotherapeutic agents and may consequently represent a realistic model for individual immunotherapeutic drug testing in the future.
KW  - glioblastoma
KW  - organoids
KW  - slice culture
KW  - tumormicroenvironment
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-319249
SN  - 2072-6694
VL  - 15
IS  - 10
ER  - 
TY  - JOUR
A1  - Löb, Sanja
A1  - Linsmeier, Eva
A1  - Herbert, Saskia-Laureen
A1  - Schlaiß, Tanja
A1  - Kiesel, Matthias
A1  - Wischhusen, Jörg
A1  - Salmen, Jessica
A1  - Kranke, Peter
A1  - Quenzer, Anne
A1  - Kurz, Florian
A1  - Weiss, Claire
A1  - Gerhard-Hartmann, Elena
A1  - Wöckel, Achim
A1  - Diessner, Joachim
T1  - Prognostic effect of HER2 evolution from primary breast cancer to breast cancer metastases
JF  - Journal of Cancer Research and Clinical Oncology
N2  - Purpose
Therapeutic options for breast cancer (BC) treatment are constantly evolving. The Human Epidermal Growth Factor 2 (HER2)-low BC entity is a new subgroup, representing about 55% of all BC patients. New antibody–drug conjugates demonstrated promising results for this BC subgroup. Currently, there is limited information about the conversion of HER2 subtypes between primary tumor and recurrent disease.

Methods
This retrospective study included women with BC at the University Medical Centre Wuerzburg from 1998 to 2021. Data were retrieved from patients' records. HER2 evolution from primary diagnosis to the first relapse and the development of secondary metastases was investigated.

Results
In the HR-positive subgroup without HER2 overexpression, HER2-low expression in primary BC was 56.7 vs. 14.6% in the triple-negative subgroup (p < 0.000). In the cohort of the first relapse, HER2-low represented 64.1% of HR-positive vs. 48.2% of the triple-negative cohort (p = 0.03). In patients with secondary metastases, HER2-low was 75.6% vs. 50% in the triple negative subgroup (p = 0.10). The subgroup of HER2-positive breast cancer patients numerically increased in the course of disease; the HER2-negative overall cohort decreased. A loss of HER2 expression from primary BC to the first relapse correlated with a better OS (p = 0.018). No clinicopathological or therapeutic features could be identified as potential risk factors for HER2 conversion.

Conclusion
HER2 expression is rising during the progression of BC disease. In view of upcoming therapeutical options, the re-analysis of newly developed metastasis will become increasingly important.
KW  - breast cancer
KW  - HER2 conversion
KW  - HER2-low
KW  - trastuzumab deruxtecan
KW  - HER2 targeted therapy
KW  - trastuzumab
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-324068
VL  - 149
IS  - 8
ER  - 
TY  - JOUR
A1  - Marquardt, André
A1  - Hartrampf, Philipp
A1  - Kollmannsberger, Philip
A1  - Solimando, Antonio G.
A1  - Meierjohann, Svenja
A1  - Kübler, Hubert
A1  - Bargou, Ralf
A1  - Schilling, Bastian
A1  - Serfling, Sebastian E.
A1  - Buck, Andreas
A1  - Werner, Rudolf A.
A1  - Lapa, Constantin
A1  - Krebs, Markus
T1  - Predicting microenvironment in CXCR4- and FAP-positive solid tumors — a pan-cancer machine learning workflow for theranostic target structures
JF  - Cancers
N2  - (1) Background: C-X-C Motif Chemokine Receptor 4 (CXCR4) and Fibroblast Activation Protein Alpha (FAP) are promising theranostic targets. However, it is unclear whether CXCR4 and FAP positivity mark distinct microenvironments, especially in solid tumors. (2) Methods: Using Random Forest (RF) analysis, we searched for entity-independent mRNA and microRNA signatures related to CXCR4 and FAP overexpression in our pan-cancer cohort from The Cancer Genome Atlas (TCGA) database —  representing n = 9242 specimens from 29 tumor entities. CXCR4- and FAP-positive samples were assessed via StringDB cluster analysis, EnrichR, Metascape, and Gene Set Enrichment Analysis (GSEA). Findings were validated via correlation analyses in n = 1541 tumor samples. TIMER2.0 analyzed the association of CXCR4 / FAP expression and infiltration levels of immune-related cells. (3) Results: We identified entity-independent CXCR4 and FAP gene signatures representative for the majority of solid cancers. While CXCR4 positivity marked an immune-related microenvironment, FAP overexpression highlighted an angiogenesis-associated niche. TIMER2.0 analysis confirmed characteristic infiltration levels of CD8+ cells for CXCR4-positive tumors and endothelial cells for FAP-positive tumors. (4) Conclusions: CXCR4- and FAP-directed PET imaging could provide a non-invasive decision aid for entity-agnostic treatment of microenvironment in solid malignancies. Moreover, this machine learning workflow can easily be transferred towards other theranostic targets.
KW  - machine learning
KW  - tumor microenvironment
KW  - immune infiltration
KW  - angiogenesis
KW  - mRNA
KW  - miRNA
KW  - transcriptome
Y1  - 2023
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-305036
SN  - 2072-6694
VL  - 15
IS  - 2
ER  - 
TY  - JOUR
A1  - Seiler, Jonas
A1  - Ebert, Regina
A1  - Rudert, Maximilian
A1  - Herrmann, Marietta
A1  - Leich, Ellen
A1  - Weißenberger, Manuela
A1  - Horas, Konstantin
T1  - Bone metastases of diverse primary origin frequently express the VDR (vitamin D receptor) and CYP24A1
JF  - Journal of Clinical Medicine
N2  - Active vitamin D (1,25(OH)2D3) is known to exert direct anti-cancer actions on various malignant tissues through binding to the vitamin D receptor (VDR). These effects have been demonstrated in breast, prostate, renal and thyroid cancers, which all have a high propensity to metastasise to bone. In addition, there is evidence that vitamin D catabolism via 24-hydroxylase (CYP24A1) is altered in tumour cells, thus, reducing local active vitamin D levels in cancer cells. The aim of this study was to assess VDR and CYP24A1 expression in various types of bone metastases by using immunohistochemistry. Overall, a high total VDR protein expression was detected in 59% of cases (39/66). There was a non-significant trend of high-grade tumours towards the low nuclear VDR expression (p = 0.07). Notably, patients with further distant metastases had a reduced nuclear VDR expression (p = 0.03). Furthermore, a high CYP24A1 expression was detected in 59% (39/66) of bone metastases. There was a significant positive correlation between nuclear VDR and CYP24A1 expression (p = 0.001). Collectively, the VDR and CYP24A1 were widely expressed in a multitude of bone metastases, pointing to a potential role of vitamin D signalling in cancer progression. This is of high clinical relevance, as vitamin D deficiency is frequent in patients with bone metastases.
KW  - vitamin D receptor
KW  - VDR
KW  - CYP24A1
KW  - bone metastasis
KW  - vitamin D
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-297377
SN  - 2077-0383
VL  - 11
IS  - 21
ER  - 
TY  - JOUR
A1  - Wobser, Marion
A1  - Schummer, Patrick
A1  - Appenzeller, Silke
A1  - Kneitz, Hermann
A1  - Roth, Sabine
A1  - Goebeler, Matthias
A1  - Geissinger, Eva
A1  - Rosenwald, Andreas
A1  - Maurus, Katja
T1  - Panel sequencing of primary cutaneous B-cell lymphoma
JF  - Cancers
N2  - Background: Primary cutaneous follicular B-cell lymphoma (PCFBCL) represents an indolent subtype of Non-Hodgkin’s lymphomas, being clinically characterized by slowly growing tumors of the skin and common cutaneous relapses, while only exhibiting a low propensity for systemic dissemination or fatal outcome. Up to now, only few studies have investigated underlying molecular alterations of PCFBCL with respect to somatic mutations. Objectives: Our aim was to gain deeper insight into the pathogenesis of PCFBCL and to delineate discriminatory molecular features of this lymphoma subtype. Methods: We performed hybridization-based panel sequencing of 40 lymphoma-associated genes of 10 cases of well-characterized PCFBCL. In addition, we included two further ambiguous cases of atypical B-cell-rich lymphoid infiltrate/B-cell lymphoma of the skin for which definite subtype attribution had not been possible by routine investigations. Results: In 10 out of 12 analyzed cases, we identified genetic alterations within 15 of the selected 40 target genes. The most frequently detected alterations in PCFBCL affected the TNFRSF14, CREBBP, STAT6 and TP53 genes. Our analysis unrevealed novel mutations of the BCL2 gene in PCFBCL. All patients exhibited an indolent clinical course. Both the included arbitrary cases of atypical B-cell-rich cutaneous infiltrates showed somatic mutations within the FAS gene. As these mutations have previously been designated as subtype-specific recurrent alterations in primary cutaneous marginal zone lymphoma (PCMZL), we finally favored the diagnosis of PCMZL in these two cases based on these molecular findings. Conclusions: To conclude, our molecular data support that PCFBCL shows distinct somatic mutations which may aid to differentiate PCFBCL from pseudo-lymphoma as well as from other indolent and aggressive cutaneous B-cell lymphomas. While the detected genetic alterations of PCFBCL did not turn out to harbor any prognostic value in our cohort, our molecular data may add adjunctive discriminatory features for diagnostic purposes on a molecular level.
KW  - B-cell lymphoma
KW  - primary cutaneous follicular B-cell lymphoma
KW  - targeted sequencing
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-290330
SN  - 2072-6694
VL  - 14
IS  - 21
ER  - 
TY  - JOUR
A1  - Lüke, Florian
A1  - Haller, Florian
A1  - Utpatel, Kirsten
A1  - Krebs, Markus
A1  - Meidenbauer, Norbert
A1  - Scheiter, Alexander
A1  - Spoerl, Silvia
A1  - Heudobler, Daniel
A1  - Sparrer, Daniela
A1  - Kaiser, Ulrich
A1  - Keil, Felix
A1  - Schubart, Christoph
A1  - Tögel, Lars
A1  - Einhell, Sabine
A1  - Dietmaier, Wolfgang
A1  - Huss, Ralf
A1  - Dintner, Sebastian
A1  - Sommer, Sebastian
A1  - Jordan, Frank
A1  - Goebeler, Maria-Elisabeth
A1  - Metz, Michaela
A1  - Haake, Diana
A1  - Scheytt, Mithun
A1  - Gerhard-Hartmann, Elena
A1  - Maurus, Katja
A1  - Brändlein, Stephanie
A1  - Rosenwald, Andreas
A1  - Hartmann, Arndt
A1  - Märkl, Bruno
A1  - Einsele, Hermann
A1  - Mackensen, Andreas
A1  - Herr, Wolfgang
A1  - Kunzmann, Volker
A1  - Bargou, Ralf
A1  - Beckmann, Matthias W.
A1  - Pukrop, Tobias
A1  - Trepel, Martin
A1  - Evert, Matthias
A1  - Claus, Rainer
A1  - Kerscher, Alexander
T1  - Identification of disparities in personalized cancer care — a joint approach of the German WERA consortium
JF  - Cancers
N2  - (1) Background: molecular tumor boards (MTBs) are crucial instruments for discussing and allocating targeted therapies to suitable cancer patients based on genetic findings. Currently, limited evidence is available regarding the regional impact and the outreach component of MTBs; (2) Methods: we analyzed MTB patient data from four neighboring Bavarian tertiary care oncology centers in Würzburg, Erlangen, Regensburg, and Augsburg, together constituting the WERA Alliance. Absolute patient numbers and regional distribution across the WERA-wide catchment area were weighted with local population densities; (3) Results: the highest MTB patient numbers were found close to the four cancer centers. However, peaks in absolute patient numbers were also detected in more distant and rural areas. Moreover, weighting absolute numbers with local population density allowed for identifying so-called white spots—regions within our catchment that were relatively underrepresented in WERA MTBs; (4) Conclusions: investigating patient data from four neighboring cancer centers, we comprehensively assessed the regional impact of our MTBs. The results confirmed the success of existing collaborative structures with our regional partners. Additionally, our results help identifying potential white spots in providing precision oncology and help establishing a joint WERA-wide outreach strategy.
KW  - precision oncology
KW  - MTB
KW  - patient access
KW  - cancer care
KW  - outreach
KW  - real world data
KW  - outcomes research
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-290311
SN  - 2072-6694
VL  - 14
IS  - 20
ER  - 
TY  - JOUR
A1  - Feldheim, Jonas
A1  - Kessler, Almuth F.
A1  - Feldheim, Julia J.
A1  - Schulz, Ellina
A1  - Wend, David
A1  - Lazaridis, Lazaros
A1  - Kleinschnitz, Christoph
A1  - Glas, Martin
A1  - Ernestus, Ralf-Ingo
A1  - Brandner, Sebastian
A1  - Monoranu, Camelia M.
A1  - Löhr, Mario
A1  - Hagemann, Carsten
T1  - Effects of long-term temozolomide treatment on glioblastoma and astrocytoma WHO grade 4 stem-like cells
JF  - International Journal of Molecular Sciences
N2  - Glioblastoma leads to a fatal course within two years in more than two thirds of patients. An essential cornerstone of therapy is chemotherapy with temozolomide (TMZ). The effect of TMZ is counteracted by the cellular repair enzyme O\(^6\)-methylguanine-DNA methyltransferase (MGMT). The MGMT promoter methylation, the main regulator of MGMT expression, can change from primary tumor to recurrence, and TMZ may play a significant role in this process. To identify the potential mechanisms involved, three primary stem-like cell lines (one astrocytoma with the mutation of the isocitrate dehydrogenase (IDH), CNS WHO grade 4 (HGA)), and two glioblastoma (IDH-wildtype, CNS WHO grade 4) were treated with TMZ. The MGMT promoter methylation, migration, proliferation, and TMZ-response of the tumor cells were examined at different time points. The strong effects of TMZ treatment on the MGMT methylated cells were observed. Furthermore, TMZ led to a loss of the MGMT promoter hypermethylation and induced migratory rather than proliferative behavior. Cells with the unmethylated MGMT promoter showed more aggressive behavior after treatment, while HGA cells reacted heterogenously. Our study provides further evidence to consider the potential adverse effects of TMZ chemotherapy and a rationale for investigating potential relationships between TMZ treatment and change in the MGMT promoter methylation during relapse.
KW  - glioblastoma
KW  - astrocytoma
KW  - IDH
KW  - MGMT
KW  - therapy
KW  - temozolomide
KW  - cancer stem cells
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284417
SN  - 1422-0067
VL  - 23
IS  - 9
ER  - 
TY  - JOUR
A1  - An, Ran
A1  - Strissel, Pamela L.
A1  - Al-Abboodi, Majida
A1  - Robering, Jan W.
A1  - Supachai, Reakasame
A1  - Eckstein, Markus
A1  - Peddi, Ajay
A1  - Hauck, Theresa
A1  - Bäuerle, Tobias
A1  - Boccaccini, Aldo R.
A1  - Youssef, Almoatazbellah
A1  - Sun, Jiaming
A1  - Strick, Reiner
A1  - Horch, Raymund E.
A1  - Boos, Anja M.
A1  - Kengelbach-Weigand, Annika
T1  - An innovative arteriovenous (AV) loop breast cancer model tailored for cancer research
JF  - Bioengineering
N2  - Animal models are important tools to investigate the pathogenesis and develop treatment strategies for breast cancer in humans. In this study, we developed a new three-dimensional in vivo arteriovenous loop model of human breast cancer with the aid of biodegradable materials, including fibrin, alginate, and polycaprolactone. We examined the in vivo effects of various matrices on the growth of breast cancer cells by imaging and immunohistochemistry evaluation. Our findings clearly demonstrate that vascularized breast cancer microtissues could be engineered and recapitulate the in vivo situation and tumor-stromal interaction within an isolated environment in an in vivo organism. Alginate–fibrin hybrid matrices were considered as a highly powerful material for breast tumor engineering based on its stability and biocompatibility. We propose that the novel tumor model may not only serve as an invaluable platform for analyzing and understanding the molecular mechanisms and pattern of oncologic diseases, but also be tailored for individual therapy via transplantation of breast cancer patient-derived tumors.
KW  - arteriovenous loop
KW  - breast cancer
KW  - animal model
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-278919
SN  - 2306-5354
VL  - 9
IS  - 7
ER  - 
TY  - JOUR
A1  - Barahona de Brito, Carlotta
A1  - Klein-Hessling, Stefan
A1  - Serfling, Edgar
A1  - Patra, Amiya Kumar
T1  - Hematopoietic stem and progenitor cell maintenance and multiple lineage differentiation is an integral function of NFATc1
JF  - Cells
N2  - Hematopoietic stem and progenitor cell (HSPC) maintenance and the differentiation of various lineages is a highly complex but precisely regulated process. Multiple signaling pathways and an array of transcription factors influence HSPC maintenance and the differentiation of individual lineages to constitute a functional hematopoietic system. Nuclear factor of activated T cell (NFAT) family transcription factors have been studied in the context of development and function of multiple mature hematopoietic lineage cells. However, until now their contribution in HSPC physiology and HSPC differentiation to multiple hematopoietic lineages has remained poorly understood. Here, we show that NFAT proteins, specifically NFATc1, play an indispensable role in the maintenance of HSPCs. In the absence of NFATc1, very few HSPCs develop in the bone marrow, which are functionally defective. In addition to HSPC maintenance, NFATc1 also critically regulates differentiation of lymphoid, myeloid, and erythroid lineage cells from HSPCs. Deficiency of NFATc1 strongly impaired, while enhanced NFATc1 activity augmented, the differentiation of these lineages, which further attested to the vital involvement of NFATc1 in regulating hematopoiesis. Hematopoietic defects due to lack of NFATc1 activity can lead to severe pathologies such as lymphopenia, myelopenia, and a drastically reduced lifespan underlining the critical role NFATc1 plays in HSPC maintenance and in the differentaion of various lineages. Our findings suggest that NFATc1 is a critical component of the myriad signaling and transcriptional regulators that are essential to maintain normal hematopoiesis.
KW  - hematopoiesis
KW  - HSC
KW  - lineage differentiation
KW  - NFATc1
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-278809
SN  - 2073-4409
VL  - 11
IS  - 13
ER  - 
TY  - JOUR
A1  - Barahona de Brito, Carlotta
A1  - Patra, Amiya Kumar
T1  - NFAT factors are dispensable for the development but are critical for the maintenance of Foxp3\(^+\) regulatory T cells
JF  - Cells
N2  - The transcription factors of the nuclear factor of activated T cell (NFAT) family play a crucial role in multiple aspects of T cell function. It has recently been reported that NFATs play an important role in the suppressive function of CD4\(^+\)CD25\(^+\)Foxp3\(^+\) regulatory T (T\(_{reg}\)) cells. In this study, we have investigated the role of NFATs in the thymic development of T\(_{reg}\) cells in mice. We show that NFAT factors are dispensable for the development of Foxp3\(^+\) T\(_{reg}\) cells in the thymus but are critical for the maintenance of both the phenotype and survival of T\(_{reg}\) cells in the thymus as well as in peripheral lymphoid organs. Specifically, the homeostasis of CD4\(^+\)CD25\(^+\)Foxp3\(^+\) but not the CD4\(^+\)CD25\(^-\)Foxp3\(^+\) fraction is severely perturbed when NFAT signaling is blocked, leading to a strongly reduced T\(_{reg}\) population. We underscored this intriguing effect of NFAT on CD4\(^+\)CD25\(^+\)Foxp3\(^+\) T\(_{reg}\) cells to the disruption of survival signals provided by interleukin 2 (IL-2). Accordingly, blocking T\(_{reg}\) cell death by abolishing the activity of pro-apoptotic Bcl-2 family member Bim, compensated for the survival defects induced due to a lack of NFAT-IL-2-IL-2R signaling. Inhibition of NFAT activity led to a strong reduction in the number of Foxp3\(^+\) T\(_{reg}\) cells; however, it did not influence the level of Foxp3 expression on an individual cell basis. In addition, we show a differential effect of IL-2 and IL-7 signaling on Foxp3\(^+\) T\(_{reg}\) versus CD4\(^+\)CD25\(^-\) T cell development, again underlining the dispensability of NFAT signaling in the development, but not in the maintenance of Foxp3\(^+\) T\(_{reg}\) cells.
KW  - thymocytes
KW  - NFAT
KW  - cyclosporine A
KW  - T\(_{reg}\) and Foxp3
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-270668
SN  - 2073-4409
VL  - 11
IS  - 9
ER  - 
TY  - JOUR
A1  - Schümann, Franziska Lea
A1  - Groß, Elisabeth
A1  - Bauer, Marcus
A1  - Rohde, Christian
A1  - Sandmann, Sarah
A1  - Terziev, Denis
A1  - Müller, Lutz P.
A1  - Posern, Guido
A1  - Wienke, Andreas
A1  - Fend, Falko
A1  - Hansmann, Martin-Leo
A1  - Klapper, Wolfram
A1  - Rosenwald, Andreas
A1  - Stein, Harald
A1  - Dugas, Martin
A1  - Müller-Tidow, Carsten
A1  - Wickenhauser, Claudia
A1  - Binder, Mascha
A1  - Weber, Thomas
T1  - Divergent effects of EZH1 and EZH2 protein expression on the prognosis of patients with T-cell lymphomas
JF  - Biomedicines
N2  - T-cell lymphomas are highly heterogeneous and their prognosis is poor under the currently available therapies. Enhancers of zeste homologue 1 and 2 (EZH1/2) are histone H3 lysine-27 trimethyltransferases (H3K27me3). Despite the rapid development of new drugs inhibiting EZH2 and/or EZH1, the molecular interplay of these proteins and the impact on disease progression and prognosis of patients with T-cell lymphomas remains insufficiently understood. In this study, EZH1/2 mutation status was evaluated in 33 monomorphic epitheliotropic intestinal T-cell lymphomas by next generation sequencing and EZH1/2 and H3K27me3 protein expression levels were detected by immunohistochemistry in 46 T-cell lymphomas. Correlations with clinicopathologic features were analyzed and survival curves generated. No EZH1 mutations and one (3%) EZH2 missense mutation were identified. In univariable analysis, high EZH1 expression was associated with an improved overall survival (OS) and progression-free survival (PFS) whereas high EZH2 and H3K27me3 expression were associated with poorer OS and PFS. Multivariable analysis revealed EZH1 (hazard ratio (HR) = 0.183; 95% confidence interval (CI): 0.044–0.767; p = 0.020;) and EZH2 (HR = 8.245; 95% CI: 1.898–35.826; p = 0.005) to be independent, divergent prognostic markers for OS. In conclusion, EZH1/2 protein expression had opposing effects on the prognosis of T-cell lymphoma patients.
KW  - T-cell non-Hodgkin's lymphomas
KW  - PTCL
KW  - epigenetics
KW  - EZH1
KW  - EZH2
KW  - H3K27me3
KW  - immunohistochemistry
KW  - next generation sequencing
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-252155
SN  - 2227-9059
VL  - 9
IS  - 12
ER  - 
TY  - JOUR
A1  - Wobser, Marion
A1  - Roth, Sabine
A1  - Appenzeller, Silke
A1  - Houben, Roland
A1  - Schrama, David
A1  - Goebeler, Matthias
A1  - Geissinger, Eva
A1  - Rosenwald, Andreas
A1  - Maurus, Katja
T1  - Targeted deep sequencing of mycosis fungoides reveals intracellular signaling pathways associated with aggressiveness and large cell transformation
JF  - Cancers
N2  - Introduction: Large-cell transformation (LCT) of mycosis fungoides (MF) has been associated with a higher risk of relapse and progression and, consequently, restricted prognosis. Its molecular pathogenesis has not been elucidated yet. Materials and Methods: In order to address molecular mechanisms of LCT, we performed hybrid capture panel-based sequencing of skin biopsies from 10 patients suffering from MF with LCT versus 17 patients without LCT including follow-up biopsies during clinical course, respectively (51 samples in total). The analyzed patients were attributed to three different groups based on the presence of LCT and clinical behavior. Results: While indolent MF cases without LCT did not show pathogenic driver mutations, a high rate of oncogenic alterations was detected in patients with LCT and aggressive clinical courses. Various genes of different oncogenic signaling pathways, including the MAPK and JAK-STAT signaling pathways, as well as epigenetic modifiers were affected. A high inter-individual and distinctive intra-individual mutation diversity was observed. Oncogenic RAS mutations were exclusively detected in patients with LCT. Conclusion: Our data demonstrate that LCT transition of MF is associated with increased frequency of somatic mutations in cancer-associated genes. In particular, the activation of RAS signaling — together with epigenetic dysregulation — may crucially contribute to the molecular pathogenesis of the LCT phenotype, thus conveying its adverse clinical behavior.
KW  - mycosis fungoides
KW  - cutaneous T-cell-lymphoma
KW  - panel sequencing
KW  - large cell transformation
KW  - CD30
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250094
SN  - 2072-6694
VL  - 13
IS  - 21
ER  - 
TY  - JOUR
A1  - Fuhr, Viktoria
A1  - Heidenreich, Shanice
A1  - Srivastava, Mugdha
A1  - Riedel, Angela
A1  - Düll, Johannes
A1  - Gerhard-Hartmann, Elena
A1  - Rosenwald, Andreas
A1  - Rauert-Wunderlich, Hilka
T1  - CD52 and OXPHOS-potential targets in ibrutinib-treated mantle cell lymphoma
JF  - Cell Death Discovery
N2  - Altered features of tumor cells acquired across therapy can result in the survival of treatment-resistant clones that may cause minimal residual disease (MRD). Despite the efficacy of ibrutinib in treating relapsed/refractory mantle cell lymphoma, the obstacle of residual cells contributes to relapses of this mature B-cell neoplasm, and the disease remains incurable. RNA-seq analysis of an ibrutinib-sensitive mantle cell lymphoma cell line following ibrutinib incubation of up to 4 d, corroborated our previously postulated resistance mechanism of a metabolic switch to reliance on oxidative phosphorylation (OXPHOS) in surviving cells. Besides, we had shown that treatment-persisting cells were characterized by increased CD52 expression. Therefore, we hypothesized that combining ibrutinib with another agent targeting these potential escape mechanisms could minimize the risk of survival of ibrutinib-resistant cells. Concomitant use of ibrutinib with OXPHOS-inhibitor IACS-010759 increased toxicity compared to ibrutinib alone. Targeting CD52 was even more efficient, as addition of CD52 mAb in combination with human serum following ibrutinib pretreatment led to rapid complement-dependent-cytotoxicity in an ibrutinib-sensitive cell line. In primary mantle cell lymphoma cells, a higher toxic effect with CD52 mAb was obtained, when cells were pretreated with ibrutinib, but only in an ibrutinib-sensitive cohort. Given the challenge of treating multi-resistant mantle cell lymphoma patients, this work highlights the potential use of anti-CD52 therapy as consolidation after ibrutinib treatment in patients who responded to the BTK inhibitor to achieve MRD negativity and prolong progression-free survival.
KW  - cancer metabolism
KW  - cell death
KW  - target validation
KW  - targeted therapies
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-300817
SN  - 2058-7716
VL  - 8
ER  - 
TY  - JOUR
A1  - Fischer, Thomas
A1  - Hartmann, Oliver
A1  - Reissland, Michaela
A1  - Prieto-Garcia, Cristian
A1  - Klann, Kevin
A1  - Pahor, Nikolett
A1  - Schülein-Völk, Christina
A1  - Baluapuri, Apoorva
A1  - Polat, Bülent
A1  - Abazari, Arya
A1  - Gerhard-Hartmann, Elena
A1  - Kopp, Hans-Georg
A1  - Essmann, Frank
A1  - Rosenfeldt, Mathias
A1  - Münch, Christian
A1  - Flentje, Michael
A1  - Diefenbacher, Markus E.
T1  - PTEN mutant non-small cell lung cancer require ATM to suppress pro-apoptotic signalling and evade radiotherapy
JF  - Cell & Bioscience
N2  - Background
Despite advances in treatment of patients with non-small cell lung cancer, carriers of certain genetic alterations are prone to failure. One such factor frequently mutated, is the tumor suppressor PTEN. These tumors are supposed to be more resistant to radiation, chemo- and immunotherapy.

Results
We demonstrate that loss of PTEN led to altered expression of transcriptional programs which directly regulate therapy resistance, resulting in establishment of radiation resistance. While PTEN-deficient tumor cells were not dependent on DNA-PK for IR resistance nor activated ATR during IR, they showed a significant dependence for the DNA damage kinase ATM. Pharmacologic inhibition of ATM, via KU-60019 and AZD1390 at non-toxic doses, restored and even synergized with IR in PTEN-deficient human and murine NSCLC cells as well in a multicellular organotypic ex vivo tumor model.

Conclusion
PTEN tumors are addicted to ATM to detect and repair radiation induced DNA damage. This creates an exploitable bottleneck. At least in cellulo and ex vivo we show that low concentration of ATM inhibitor is able to synergise with IR to treat PTEN-deficient tumors in genetically well-defined IR resistant lung cancer models.
KW  - PTEN
KW  - ATM
KW  - IR
KW  - NSCLC
KW  - radiotherapy
KW  - cancer
KW  - DNA-PK
KW  - PI3K
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-299865
SN  - 2045-3701
VL  - 12
ER  - 
TY  - JOUR
A1  - Polat, Bülent
A1  - Wohlleben, Gisela
A1  - Kosmala, Rebekka
A1  - Lisowski, Dominik
A1  - Mantel, Frederick
A1  - Lewitzki, Victor
A1  - Löhr, Mario
A1  - Blum, Robert
A1  - Herud, Petra
A1  - Flentje, Michael
A1  - Monoranu, Camelia-Maria
T1  - Differences in stem cell marker and osteopontin expression in primary and recurrent glioblastoma
JF  - Cancer Cell International
N2  - Background
Despite of a multimodal approach, recurrences can hardly be prevented in glioblastoma. This may be in part due to so called glioma stem cells. However, there is no established marker to identify these stem cells.

Methods
Paired samples from glioma patients were analyzed by immunohistochemistry for expression of the following stem cell markers: CD133, Musashi, Nanog, Nestin, octamer-binding transcription factor 4 (Oct4), and sex determining region Y-box 2 (Sox2). In addition, the expression of osteopontin (OPN) was investigated. The relative number of positively stained cells was determined. By means of Kaplan–Meier analysis, a possible association with overall survival by marker expression was investigated.

Results
Sixty tissue samples from 30 patients (17 male, 13 female) were available for analysis. For Nestin, Musashi and OPN a significant increase was seen. There was also an increase (not significant) for CD133 and Oct4. Patients with mutated Isocitrate Dehydrogenase-1/2 (IDH-1/2) status had a reduced expression for CD133 and Nestin in their recurrent tumors. Significant correlations were seen for CD133 and Nanog between OPN in the primary and recurrent tumor and between CD133 and Nestin in recurrent tumors. By confocal imaging we could demonstrate a co-expression of CD133 and Nestin within recurrent glioma cells. Patients with high CD133 expression had a worse prognosis (22.6 vs 41.1 months, p = 0.013). A similar trend was seen for elevated Nestin levels (24.9 vs 41.1 months, p = 0.08).

Conclusions
Most of the evaluated markers showed an increased expression in their recurrent tumor. CD133 and Nestin were associated with survival and are candidate markers for further clinical investigation.
KW  - Glioblastoma
KW  - Glioma stem cells
KW  - Osteopontin
KW  - CD133
KW  - Nestin
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-301240
SN  - 1475-2867
VL  - 22
ER  - 
TY  - JOUR
A1  - Loeffler-Wirth, Henry
A1  - Kreuz, Markus
A1  - Hopp, Lydia
A1  - Arakelyan, Arsen
A1  - Haake, Andrea
A1  - Cogliatti, Sergio B.
A1  - Feller, Alfred C.
A1  - Hansmann, Martin-Leo
A1  - Lenze, Dido
A1  - Möller, Peter
A1  - Müller-Hermelink, Hans Konrad
A1  - Fortenbacher, Erik
A1  - Willscher, Edith
A1  - Ott, German
A1  - Rosenwald, Andreas
A1  - Pott, Christiane
A1  - Schwaenen, Carsten
A1  - Trautmann, Heiko
A1  - Wessendorf, Swen
A1  - Stein, Harald
A1  - Szczepanowski, Monika
A1  - Trümper, Lorenz
A1  - Hummel, Michael
A1  - Klapper, Wolfram
A1  - Siebert, Reiner
A1  - Loeffler, Markus
A1  - Binder, Hans
T1  - A modular transcriptome map of mature B cell lymphomas
JF  - Genome Medicine
N2  - Background
Germinal center-derived B cell lymphomas are tumors of the lymphoid tissues representing one of the most heterogeneous malignancies. Here we characterize the variety of transcriptomic phenotypes of this disease based on 873 biopsy specimens collected in the German Cancer Aid MMML (Molecular Mechanisms in Malignant Lymphoma) consortium. They include diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), Burkitt’s lymphoma, mixed FL/DLBCL lymphomas, primary mediastinal large B cell lymphoma, multiple myeloma, IRF4-rearranged large cell lymphoma, MYC-negative Burkitt-like lymphoma with chr. 11q aberration and mantle cell lymphoma.

Methods
We apply self-organizing map (SOM) machine learning to microarray-derived expression data to generate a holistic view on the transcriptome landscape of lymphomas, to describe the multidimensional nature of gene regulation and to pursue a modular view on co-expression. Expression data were complemented by pathological, genetic and clinical characteristics.

Results
We present a transcriptome map of B cell lymphomas that allows visual comparison between the SOM portraits of different lymphoma strata and individual cases. It decomposes into one dozen modules of co-expressed genes related to different functional categories, to genetic defects and to the pathogenesis of lymphomas. On a molecular level, this disease rather forms a continuum of expression states than clearly separated phenotypes. We introduced the concept of combinatorial pattern types (PATs) that stratifies the lymphomas into nine PAT groups and, on a coarser level, into five prominent cancer hallmark types with proliferation, inflammation and stroma signatures. Inflammation signatures in combination with healthy B cell and tonsil characteristics associate with better overall survival rates, while proliferation in combination with inflammation and plasma cell characteristics worsens it. A phenotypic similarity tree is presented that reveals possible progression paths along the transcriptional dimensions. Our analysis provided a novel look on the transition range between FL and DLBCL, on DLBCL with poor prognosis showing expression patterns resembling that of Burkitt’s lymphoma and particularly on ‘double-hit’ MYC and BCL2 transformed lymphomas.

Conclusions
The transcriptome map provides a tool that aggregates, refines and visualizes the data collected in the MMML study and interprets them in the light of previous knowledge to provide orientation and support in current and future studies on lymphomas and on other cancer entities.
KW  - tumor heterogeneity
KW  - B cell malignancies
KW  - gene regulation
KW  - molecular subtypes
KW  - machine learning
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-237262
VL  - 11
ER  - 
TY  - JOUR
A1  - Otto, Christoph
A1  - Kastner, Carolin
A1  - Schmidt, Stefanie
A1  - Uttinger, Konstantin
A1  - Baluapuri, Apoorva
A1  - Denk, Sarah
A1  - Rosenfeldt, Mathias T.
A1  - Rosenwald, Andreas
A1  - Roehrig, Florian
A1  - Ade, Carsten P.
A1  - Schuelein-Voelk, Christina
A1  - Diefenbacher, Markus E.
A1  - Germer, Christoph-Thomas
A1  - Wolf, Elmar
A1  - Eilers, Martin
A1  - Wiegering, Armin
T1  - RNA polymerase I inhibition induces terminal differentiation, growth arrest, and vulnerability to senolytics in colorectal cancer cells
JF  - Molecular Oncology
N2  - Ribosomal biogenesis and protein synthesis are deregulated in most cancers, suggesting that interfering with translation machinery may hold significant therapeutic potential. Here, we show that loss of the tumor suppressor adenomatous polyposis coli (APC), which constitutes the initiating event in the adenoma carcinoma sequence for colorectal cancer (CRC), induces the expression of RNA polymerase I (RNAPOL1) transcription machinery, and subsequently upregulates ribosomal DNA (rDNA) transcription. Targeting RNAPOL1 with a specific inhibitor, CX5461, disrupts nucleolar integrity, and induces a disbalance of ribosomal proteins. Surprisingly, CX5461-induced growth arrest is irreversible and exhibits features of senescence and terminal differentiation. Mechanistically, CX5461 promotes differentiation in an MYC-interacting zinc-finger protein 1 (MIZ1)- and retinoblastoma protein (Rb)-dependent manner. In addition, the inhibition of RNAPOL1 renders CRC cells vulnerable towards senolytic agents. We validated this therapeutic effect of CX5461 in murine- and patient-derived organoids, and in a xenograft mouse model. These results show that targeting ribosomal biogenesis together with targeting the consecutive, senescent phenotype using approved drugs is a new therapeutic approach, which can rapidly be transferred from bench to bedside.
KW  - CRC
KW  - CX5461
KW  - MIZ1
KW  - MYC
KW  - ribosome
KW  - RNAPOL1
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-312806
VL  - 16
IS  - 15
ER  - 
TY  - JOUR
A1  - Prieto-Garcia, Cristian
A1  - Hartmann, Oliver
A1  - Reissland, Michaela
A1  - Braun, Fabian
A1  - Bozkurt, Süleyman
A1  - Pahor, Nikolett
A1  - Fuss, Carmina
A1  - Schirbel, Andreas
A1  - Schülein-Völk, Christina
A1  - Buchberger, Alexander
A1  - Calzado Canale, Marco A.
A1  - Rosenfeldt, Mathias
A1  - Dikic, Ivan
A1  - Münch, Christian
A1  - Diefenbacher, Markus E.
T1  - USP28 enables oncogenic transformation of respiratory cells, and its inhibition potentiates molecular therapy targeting mutant EGFR, BRAF and PI3K
JF  - Molecular Oncology
N2  - Oncogenic transformation of lung epithelial cells is a multistep process, frequently starting with the inactivation of tumour suppressors and subsequent development of activating mutations in proto-oncogenes, such as members of the PI3K or MAPK families. Cells undergoing transformation have to adjust to changes, including altered metabolic requirements. This is achieved, in part, by modulating the protein abundance of transcription factors. Here, we report that the ubiquitin carboxyl-terminal hydrolase 28 (USP28) enables oncogenic reprogramming by regulating the protein abundance of proto-oncogenes such as c-JUN, c-MYC, NOTCH and ∆NP63 at early stages of malignant transformation. USP28 levels are increased in cancer compared with in normal cells due to a feed-forward loop, driven by increased amounts of oncogenic transcription factors such as c-MYC and c-JUN. Irrespective of oncogenic driver, interference with USP28 abundance or activity suppresses growth and survival of transformed lung cells. Furthermore, inhibition of USP28 via a small-molecule inhibitor resets the proteome of transformed cells towards a ‘premalignant’ state, and its inhibition synergizes with clinically established compounds used to target EGFR\(^{L858R}\)-, BRAF\(^{V600E}\)- or PI3K\(^{H1047R}\)-driven tumour cells. Targeting USP28 protein abundance at an early stage via inhibition of its activity is therefore a feasible strategy for the treatment of early-stage lung tumours, and the observed synergism with current standard-of-care inhibitors holds the potential for improved targeting of established tumours.
KW  - buparlisib
KW  - c-MYC
KW  - gefitinib
KW  - lung cancer
KW  - USP28
KW  - vemurafenib
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-312777
VL  - 16
IS  - 17
ER  - 
TY  - JOUR
A1  - Nickl, Vera
A1  - Schulz, Ellina
A1  - Salvador, Ellaine
A1  - Trautmann, Laureen
A1  - Diener, Leopold
A1  - Kessler, Almuth F.
A1  - Monoranu, Camelia M.
A1  - Dehghani, Faramarz
A1  - Ernestus, Ralf-Ingo
A1  - Löhr, Mario
A1  - Hagemann, Carsten
T1  - Glioblastoma-derived three-dimensional ex vivo models to evaluate effects and efficacy of Tumor Treating Fields (TTFields)
JF  - Cancers
N2  - Simple Summary
In glioblastoma, tumor recurrence is inevitable and the prognosis of patients is poor, despite multidisciplinary treatment approaches involving surgical resection, radiotherapy and chemotherapy. Recently, Tumor Treating Fields (TTFields) have been added to the therapeutic set-up. These alternating electric fields are applied to glioblastoma at 200 kHz frequency via arrays placed on the shaved scalp of patients. Patients show varying response to this therapy. Molecular effects of TTFields have been investigated largely in cell cultures and animal models, but not in patient tissue samples. Acquisition of matched treatment-naïve and recurrent patient tissues is a challenge. Therefore, we suggest three reliable patient-derived three-dimensional ex vivo models (primary cells grown as microtumors on murine organotypic hippocampal slices, organoids and tumor slice cultures) which may facilitate prediction of patients’ treatment responses and provide important insights into clinically relevant cellular and molecular alterations under TTFields.
        
Abstract
Glioblastoma (GBM) displays a wide range of inter- and intra-tumoral heterogeneity contributing to therapeutic resistance and relapse. Although Tumor Treating Fields (TTFields) are effective for the treatment of GBM, there is a lack of ex vivo models to evaluate effects on patients’ tumor biology or to screen patients for treatment efficacy. Thus, we adapted patient-derived three-dimensional tissue culture models to be compatible with TTFields application to tissue culture. Patient-derived primary cells (PDPC) were seeded onto murine organotypic hippocampal slice cultures (OHSC), and microtumor development with and without TTFields at 200 kHz was observed. In addition, organoids were generated from acute material cultured on OHSC and treated with TTFields. Lastly, the effect of TTFields on expression of the Ki67 proliferation marker was evaluated on cultured GBM slices. Microtumors exhibited increased sensitivity towards TTFields compared to monolayer cell cultures. TTFields affected tumor growth and viability, as the size of microtumors and the percentage of Ki67-positive cells decreased after treatment. Nevertheless, variability in the extent of the response was preserved between different patient samples. Therefore, these pre-clinical GBM models could provide snapshots of the tumor to simulate patient treatment response and to investigate molecular mechanisms of response and resistance.
KW  - glioblastoma
KW  - Tumor Treating Fields (TTFields)
KW  - organotypic hippocampal slice cultures (OHSC)
KW  - organoids
KW  - tumor slice cultures
KW  - 3D ex vivo models
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-290340
SN  - 2072-6694
VL  - 14
IS  - 21
ER  - 
TY  - JOUR
A1  - Marquardt, André
A1  - Kollmannsberger, Philip
A1  - Krebs, Markus
A1  - Argentiero, Antonella
A1  - Knott, Markus
A1  - Solimando, Antonio Giovanni
A1  - Kerscher, Alexander Georg
T1  - Visual clustering of transcriptomic data from primary and metastatic tumors — dependencies and novel pitfalls
JF  - Genes
N2  - Personalized oncology is a rapidly evolving area and offers cancer patients therapy options that are more specific than ever. However, there is still a lack of understanding regarding transcriptomic similarities or differences of metastases and corresponding primary sites. Applying two unsupervised dimension reduction methods (t-Distributed Stochastic Neighbor Embedding (t-SNE) and Uniform Manifold Approximation and Projection (UMAP)) on three datasets of metastases (n = 682 samples) with three different data transformations (unprocessed, log10 as well as log10 + 1 transformed values), we visualized potential underlying clusters. Additionally, we analyzed two datasets (n = 616 samples) containing metastases and primary tumors of one entity, to point out potential familiarities. Using these methods, no tight link between the site of resection and cluster formation outcome could be demonstrated, or for datasets consisting of solely metastasis or mixed datasets. Instead, dimension reduction methods and data transformation significantly impacted visual clustering results. Our findings strongly suggest data transformation to be considered as another key element in the interpretation of visual clustering approaches along with initialization and different parameters. Furthermore, the results highlight the need for a more thorough examination of parameters used in the analysis of clusters.
KW  - visual clustering
KW  - t-SNE
KW  - UMAP
KW  - transcriptomic analysis
KW  - cancer
KW  - metastasis
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-281872
SN  - 2073-4425
VL  - 13
IS  - 8
ER  - 
TY  - JOUR
A1  - Feldheim, Jonas
A1  - Wend, David
A1  - Lauer, Mara J.
A1  - Monoranu, Camelia M.
A1  - Glas, Martin
A1  - Kleinschnitz, Christoph
A1  - Ernestus, Ralf-Ingo
A1  - Braunger, Barbara M.
A1  - Meybohm, Patrick
A1  - Hagemann, Carsten
A1  - Burek, Malgorzata
T1  - Protocadherin Gamma C3 (PCDHGC3) is strongly expressed in glioblastoma and its high expression is associated with longer progression-free survival of patients
JF  - International Journal of Molecular Sciences
N2  - Protocadherins (PCDHs) belong to the cadherin superfamily and represent the largest subgroup of calcium-dependent adhesion molecules. In the genome, most PCDHs are arranged in three clusters, α, β, and γ on chromosome 5q31. PCDHs are highly expressed in the central nervous system (CNS). Several PCDHs have tumor suppressor functions, but their individual role in primary brain tumors has not yet been elucidated. Here, we examined the mRNA expression of PCDHGC3, a member of the PCDHγ cluster, in non-cancerous brain tissue and in gliomas of different World Health Organization (WHO) grades and correlated it with the clinical data of the patients. We generated a PCDHGC3 knockout U343 cell line and examined its growth rate and migration in a wound healing assay. We showed that PCDHGC3 mRNA and protein were significantly overexpressed in glioma tissue compared to a non-cancerous brain specimen. This could be confirmed in glioma cell lines. High PCDHGC3 mRNA expression correlated with longer progression-free survival (PFS) in glioma patients. PCDHGC3 knockout in U343 resulted in a slower growth rate but a significantly faster migration rate in the wound healing assay and decreased the expression of several genes involved in WNT signaling. PCDHGC3 expression should therefore be further investigated as a PFS-marker in gliomas. However, more studies are needed to elucidate the molecular mechanisms underlying the PCDHGC3 effects.
KW  - glioblastoma multiforme
KW  - glioma
KW  - astrocytoma
KW  - recurrence
KW  - relapse
KW  - mRNA
KW  - protein
KW  - brain
KW  - expression
KW  - PCDHGC3
KW  - WNT signaling
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284433
SN  - 1422-0067
VL  - 23
IS  - 15
ER  - 
TY  - JOUR
A1  - Löhr, Mario
A1  - Härtig, Wolfgang
A1  - Schulze, Almut
A1  - Kroiß, Matthias
A1  - Sbiera, Silviu
A1  - Lapa, Constantin
A1  - Mages, Bianca
A1  - Strobel, Sabrina
A1  - Hundt, Jennifer Elisabeth
A1  - Bohnert, Simone
A1  - Kircher, Stefan
A1  - Janaki-Raman, Sudha
A1  - Monoranu, Camelia-Maria
T1  - SOAT1: A suitable target for therapy in high-grade astrocytic glioma?
JF  - International Journal of Molecular Sciences
N2  - Targeting molecular alterations as an effective treatment for isocitrate dehydrogenase-wildtype glioblastoma (GBM) patients has not yet been established. Sterol-O-Acyl Transferase 1 (SOAT1), a key enzyme in the conversion of endoplasmic reticulum cholesterol to esters for storage in lipid droplets (LD), serves as a target for the orphan drug mitotane to treat adrenocortical carcinoma. Inhibition of SOAT1 also suppresses GBM growth. Here, we refined SOAT1-expression in GBM and IDH-mutant astrocytoma, CNS WHO grade 4 (HGA), and assessed the distribution of LD in these tumors. Twenty-seven GBM and three HGA specimens were evaluated by multiple GFAP, Iba1, IDH1 R132H, and SOAT1 immunofluorescence labeling as well as Oil Red O staining. To a small extent SOAT1 was expressed by tumor cells in both tumor entities. In contrast, strong expression was observed in glioma-associated macrophages. Triple immunofluorescence labeling revealed, for the first time, evidence for SOAT1 colocalization with Iba1 and IDH1 R132H, respectively. Furthermore, a notable difference in the amount of LD between GBM and HGA was observed. Therefore, SOAT1 suppression might be a therapeutic option to target GBM and HGA growth and invasiveness. In addition, the high expression in cells related to neuroinflammation could be beneficial for a concomitant suppression of protumoral microglia/macrophages.
KW  - SOAT1
KW  - glioblastoma
KW  - astrocytoma
KW  - IDH1/2
KW  - lipid droplets
KW  - mitotane
KW  - targeted therapy
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-284178
SN  - 1422-0067
VL  - 23
IS  - 7
ER  - 
TY  - JOUR
A1  - Kuai, Yue
A1  - Gong, Xin
A1  - Ding, Liya
A1  - Li, Fang
A1  - Lei, Lizhen
A1  - Gong, Yuqi
A1  - Liu, Qingmeng
A1  - Tan, Huajiao
A1  - Zhang, Xinxia
A1  - Liu, Dongyu
A1  - Ren, Guoping
A1  - Pan, Hongyang
A1  - Shi, Yaoyao
A1  - Berberich-Siebelt, Friederike
A1  - Ma, Zhengrong
A1  - Zhou, Ren
T1  - Wilms’ tumor 1-associating protein plays an aggressive role in diffuse large B-cell lymphoma and forms a complex with BCL6 via Hsp90
JF  - Cell Communication and Signaling
N2  - Background
Wilms’ tumor 1-associating protein (WTAP) is a nuclear protein, which is ubiquitously expressed in many tissues. Furthermore, in various types of malignancies WTAP is overexpressed and plays a role as an oncogene. The function of WTAP in diffuse large B-cell lymphoma (DLBCL), however, remains unclear.

Methods
Immunohistochemistry was applied to evaluate the levels of WTAP expression in DLBCL tissues and normal lymphoid tissues. Overexpression and knock-down of WTAP in DLBCL cell lines, verified on mRNA and protein level served to analyze cell proliferation and apoptosis in DLBCL cell lines by flow cytometry. Finally, co-immunoprecipitation (Co-IP), IP, and GST-pull down assessed the interaction of WTAP with Heat shock protein 90 (Hsp90) and B-cell lymphoma 6 (BCL6) as well as determined the extend of its ubiquitinylation.

Results
WTAP protein levels were consistently upregulated in DLBCL tissues. WTAP promoted DLBCL cell proliferation and improved the ability to confront apoptosis, while knockdown of WTAP in DLBCL cell lines allowed a significant higher apoptosis rate after treatment with Etoposide, an anti-tumor drug. The stable expression of WTAP was depended on Hsp90. In line, we demonstrated that WTAP could form a complex with BCL6 via Hsp90 in vivo and in vitro.

Conclusion
WTAP is highly expressed in DLBCL, promoting growth and anti-apoptosis in DLBCL cell lines. WTAP is a client protein of Hsp90 and can appear in a complex with BCL6 and Hsp90 in DLBCL. Down-regulation of WTAP could improve the chemotherapeutic treatments in DLBCL.
KW  - WTAP
KW  - BCL6
KW  - Hsp90
KW  - complex
KW  - DLBCL
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230168
VL  - 16
ER  - 
TY  - JOUR
A1  - Karikari, Akua A.
A1  - McFleder, Rhonda L.
A1  - Ribechini, Eliana
A1  - Blum, Robert
A1  - Bruttel, Valentin
A1  - Knorr, Susanne
A1  - Gehmeyr, Mona
A1  - Volkmann, Jens
A1  - Brotchie, Jonathan M.
A1  - Ahsan, Fadhil
A1  - Haack, Beatrice
A1  - Monoranu, Camelia-Maria
A1  - Keber, Ursula
A1  - Yeghiazaryan, Rima
A1  - Pagenstecher, Axel
A1  - Heckel, Tobias
A1  - Bischler, Thorsten
A1  - Wischhusen, Jörg
A1  - Koprich, James B.
A1  - Lutz, Manfred B.
A1  - Ip, Chi Wang
T1  - Neurodegeneration by α-synuclein-specific T cells in AAV-A53T-α-synuclein Parkinson’s disease mice
JF  - Brain, Behavior, and Immunity
N2  - Background
Antigen-specific neuroinflammation and neurodegeneration are characteristic for neuroimmunological diseases. In Parkinson’s disease (PD) pathogenesis, α-synuclein is a known culprit. Evidence for α-synuclein-specific T cell responses was recently obtained in PD. Still, a causative link between these α-synuclein responses and dopaminergic neurodegeneration had been lacking. We thus addressed the functional relevance of α-synuclein-specific immune responses in PD in a mouse model.
Methods
We utilized a mouse model of PD in which an Adeno-associated Vector 1/2 serotype (AAV1/2) expressing human mutated A53T-α-Synuclein was stereotactically injected into the substantia nigra (SN) of either wildtype C57BL/6 or Recombination-activating gene 1 (RAG1)\(^{-/-}\) mice. Brain, spleen, and lymph node tissues from different time points following injection were then analyzed via FACS, cytokine bead assay, immunohistochemistry and RNA-sequencing to determine the role of T cells and inflammation in this model. Bone marrow transfer from either CD4\(^{+}\)/CD8\(^{-}\), CD4\(^{-}\)/CD8\(^{+}\), or CD4\(^{+}\)/CD8\(^{+}\) (JHD\(^{-/-}\)) mice into the RAG-1\(^{-/-}\) mice was also employed. In addition to the in vivo studies, a newly developed A53T-α-synuclein-expressing neuronal cell culture/immune cell assay was utilized.
Results
AAV-based overexpression of pathogenic human A53T-α-synuclein in dopaminergic neurons of the SN stimulated T cell infiltration. RNA-sequencing of immune cells from PD mouse brains confirmed a pro-inflammatory gene profile. T cell responses were directed against A53T-α-synuclein-peptides in the vicinity of position 53 (68–78) and surrounding the pathogenically relevant S129 (120–134). T cells were required for α-synuclein-induced neurodegeneration in vivo and in vitro, while B cell deficiency did not protect from dopaminergic neurodegeneration.
Conclusions
Using T cell and/or B cell deficient mice and a newly developed A53T-α-synuclein-expressing neuronal cell culture/immune cell assay, we confirmed in vivo and in vitro that pathogenic α-synuclein peptide-specific T cell responses can cause dopaminergic neurodegeneration and thereby contribute to PD-like pathology.
KW  - Parkinson’s disease
KW  - α-synuclein-specific T cells
KW  - neurodegeneration
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-300600
VL  - 101
SP  - 194
EP  - 210
ER  - 
TY  - JOUR
A1  - Schulze, Markus
A1  - Hutterer, Maria
A1  - Sabo, Anja
A1  - Hoja, Sabine
A1  - Lorenz, Julia
A1  - Rothhammer-Hampl, Tanja
A1  - Herold-Mende, Christel
A1  - Floßbach, Lucia
A1  - Monoranu, Camelia
A1  - Riemenschneider, Markus J.
T1  - Chronophin regulates active vitamin B6 levels and transcriptomic features of glioblastoma cell lines cultured under non-adherent, serum-free conditions
JF  - BMC Cancer
N2  - Background
The phosphatase chronophin (CIN/PDXP) has been shown to be an important regulator of glioma cell migration and invasion. It has two known substrates: p-Ser3-cofilin, the phosphorylated form of the actin binding protein cofilin, and pyridoxal 5′-phosphate, the active form of vitamin B6. Phosphoregulation of cofilin, among other functions, plays an important role in cell migration, whereas active vitamin B6 is a cofactor for more than one hundred enzymatic reactions. The role of CIN has yet only been examined in glioblastoma cell line models derived under serum culture conditions.

Results
We found that CIN is highly expressed in cells cultured under non-adherent, serum-free conditions that are thought to better mimic the in vivo situation. Furthermore, the substrates of CIN, p-Ser3-cofilin and active vitamin B6, were significantly reduced as compared to cell lines cultured in serum-containing medium. To further examine its molecular role we stably knocked down the CIN protein with two different shRNA hairpins in the glioblastoma cell lines NCH421k and NCH644. Both cell lines did not show any significant alterations in proliferation but expression of differentiation markers (such as GFAP or TUBB3) was increased in the knockdown cell lines. In addition, colony formation was significantly impaired in NCH644. Of note, in both cell lines CIN knockdown increased active vitamin B6 levels with vitamin B6 being known to be important for S-adenosylmethionine biosynthesis. Nevertheless, global histone and DNA methylation remained unaltered as was chemoresistance towards temozolomide. To further elucidate the role of phosphocofilin in glioblastoma cells we applied inhibitors for ROCK1/2 and LIMK1/2 to our model. LIMK- and ROCK-inhibitor treatment alone was not toxic for glioblastoma cells. However, it had profound, but antagonistic effects in NCH421k and NCH644 under chemotherapy.

Conclusion
In non-adherent glioblastoma cell lines cultured in serum-free medium, chronophin knockdown induces phenotypic changes, e.g. in colony formation and transcription, but these are highly dependent on the cellular background. The same is true for phenotypes observed after treatment with inhibitors for kinases regulating cofilin phosphorylation (ROCKs and LIMKs). Targeting the cofilin phosphorylation pathway might therefore not be a straightforward therapeutic option in glioblastoma.
KW  - chronophin
KW  - cofilin
KW  - vitamin metabolism
KW  - glioma
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-234645
VL  - 18
ER  - 
TY  - JOUR
A1  - Appeltshauser, Luise
A1  - Messinger, Julia
A1  - Starz, Katharina
A1  - Heinrich, David
A1  - Brunder, Anna-Michelle
A1  - Stengel, Helena
A1  - Fiebig, Bianca
A1  - Ayzenberg, Ilya
A1  - Birklein, Frank
A1  - Dresel, Christian
A1  - Dorst, Johannes
A1  - Dvorak, Florian
A1  - Grimm, Alexander
A1  - Joerk, Alexander
A1  - Leypoldt, Frank
A1  - Mäurer, Mathias
A1  - Merl, Patrick
A1  - Michels, Sebastian
A1  - Pitarokoili, Kalliopi
A1  - Rosenfeldt, Mathias
A1  - Sperfeld, Anne-Dorte
A1  - Weihrauch, Marc
A1  - Welte, Gabriel Simon
A1  - Sommer, Claudia
A1  - Doppler, Kathrin
T1  - Diabetes Mellitus Is a Possible Risk Factor for Nodo-paranodopathy With Antiparanodal Autoantibodies
JF  - Neurology: Neuroimmunology & Neuroinflammation
N2  - Background and Objectives 
Nodo-paranodopathies are peripheral neuropathies with dysfunction of the node of Ranvier. Affected patients who are seropositive for antibodies against adhesion molecules like contactin-1 and neurofascin show distinct clinical features and a disruption of the paranodal complex. An axoglial dysjunction is also a characteristic finding of diabetic neuropathy. Here, we aim to investigate a possible association of antibody-mediated nodo-paranodopathy and diabetes mellitus (DM).

Methods 
We retrospectively analyzed clinical data of 227 patients with chronic inflammatory demyelinating polyradiculoneuropathy and Guillain-Barré syndrome from multiple centers in Germany who had undergone diagnostic testing for antiparanodal antibodies targeting neurofascin-155, pan-neurofascin, contactin-1–associated protein 1, and contactin-1. To study possible direct pathogenic effects of antiparanodal antibodies, we performed immunofluorescence binding assays on human pancreatic tissue sections.

Results The frequency of DM was 33.3% in seropositive patients and thus higher compared with seronegative patients (14.1%, OR = 3.04, 95% CI = 1.31–6.80). The relative risk of DM in seropositive patients was 3.4-fold higher compared with the general German population. Seropositive patients with DM most frequently harbored anti–contactin-1 antibodies and had higher antibody titers than seropositive patients without DM. The diagnosis of DM preceded the onset of neuropathy in seropositive patients. No immunoreactivity of antiparanodal antibodies against pancreatic tissue was detected.

Discussion 
We report an association of nodo-paranodopathy and DM. Our results suggest that DM may be a potential risk factor for predisposing to developing nodo-paranodopathy and argue against DM being induced by the autoantibodies. Our findings set the basis for further research investigating underlying immunopathogenetic connections.
KW  - Diabetes mellitus
KW  - Nodo-parandopathy
KW  - Antiparanodal Autoantibodies
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-300551
VL  - 9
IS  - 3
ER  - 
TY  - JOUR
A1  - Ritter, Julia
A1  - Zimmermann, Karin
A1  - Jöhrens, Korinna
A1  - Mende, Stefanie
A1  - Seegebarth, Anke
A1  - Siegmund, Britta
A1  - Hennig, Steffen
A1  - Todorova, Kremena
A1  - Rosenwald, Andreas
A1  - Daum, Severin
A1  - Hummel, Michael
A1  - Schumann, Michael
T1  - T-cell repertoires in refractory coeliac disease
JF  - Gut
N2  - Objective Refractory coeliac disease (RCD) is a potentially hazardous complication of coeliac disease (CD). In contrast to RCD type I, RCD type II is a precursor entity of enteropathy-associated T-cell lymphoma (EATL), which is associated with clonally expanding T-cells that are also found in the sequentially developing EATL. Using high-throughput sequencing (HTS), we aimed to establish the small-intestinal T-cell repertoire (TCR) in CD and RCD to unravel the role of distinct T-cell clonotypes in RCD pathogenesis. Design DNA extracted from duodenal mucosa specimens of controls (n=9), active coeliacs (n=10), coeliacs on a gluten-free diet (n=9), RCD type I (n= 8), RCD type II (n= 8) and unclassified Marsh I cases (n= 3) collected from 2002 to 2013 was examined by TCR beta-complementarity- determining regions 3 (CDR3) multiplex PCR followed by HTS of the amplicons. Results On average, 106 sequence reads per sample were generated consisting of up to 900 individual TCR beta rearrangements. In RCD type II, the most frequent clonotypes (ie, sequence reads with identical CDR3) represent in average 42.6% of all TCR beta rearrangements, which was significantly higher than in controls (6.8%; p<0.01) or RCD type I (6.7%; p<0.01). Repeat endoscopies in individual patients revealed stability of clonotypes for up to several years without clinical symptoms of EATL. Dominant clonotypes identified in individual patients with RCD type II were unique and not related between patients. CD-associated, gliad-independent CDR3 motifs were only detectable at low frequencies. Conclusions TCR beta-HTS analysis unravels the TCR in CD and allows detailed analysis of individual TCR beta rearrangements. Dominant TCR beta sequences identified in patients with RCD type II are unique and not homologous to known gliadin-specific TCR sequences, supporting the assumption that these clonal T-cells expand independent of gluten stimulation.
KW  - Intestinal Intraepithelial Lymphocy
KW  - Delta Repertoire
KW  - Lymphoma
KW  - Usage
KW  - Clonality
KW  - Sprue
KW  - Chains
KW  - Design
KW  - Epitope
KW  - Frequency
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226350
VL  - 67
IS  - 4
ER  - 
TY  - JOUR
A1  - Peindl, Matthias
A1  - Göttlich, Claudia
A1  - Crouch, Samantha
A1  - Hoff, Niklas
A1  - Lüttgens, Tamara
A1  - Schmitt, Franziska
A1  - Pereira, Jesús Guillermo Nieves
A1  - May, Celina
A1  - Schliermann, Anna
A1  - Kronenthaler, Corinna
A1  - Cheufou, Danjouma
A1  - Reu-Hofer, Simone
A1  - Rosenwald, Andreas
A1  - Weigl, Elena
A1  - Walles, Thorsten
A1  - Schüler, Julia
A1  - Dandekar, Thomas
A1  - Nietzer, Sarah
A1  - Dandekar, Gudrun
T1  - EMT, stemness, and drug resistance in biological context: a 3D tumor tissue/in silico platform for analysis of combinatorial treatment in NSCLC with aggressive KRAS-biomarker signatures
JF  - Cancers
N2  - Epithelial-to-mesenchymal transition (EMT) is discussed to be centrally involved in invasion, stemness, and drug resistance. Experimental models to evaluate this process in its biological complexity are limited. To shed light on EMT impact and test drug response more reliably, we use a lung tumor test system based on a decellularized intestinal matrix showing more in vivo-like proliferation levels and enhanced expression of clinical markers and carcinogenesis-related genes. In our models, we found evidence for a correlation of EMT with drug resistance in primary and secondary resistant cells harboring KRAS\(^{G12C}\) or EGFR mutations, which was simulated in silico based on an optimized signaling network topology. Notably, drug resistance did not correlate with EMT status in KRAS-mutated patient-derived xenograft (PDX) cell lines, and drug efficacy was not affected by EMT induction via TGF-β. To investigate further determinants of drug response, we tested several drugs in combination with a KRAS\(^{G12C}\) inhibitor in KRAS\(^{G12C}\) mutant HCC44 models, which, besides EMT, display mutations in P53, LKB1, KEAP1, and high c-MYC expression. We identified an aurora-kinase A (AURKA) inhibitor as the most promising candidate. In our network, AURKA is a centrally linked hub to EMT, proliferation, apoptosis, LKB1, and c-MYC. This exemplifies our systemic analysis approach for clinical translation of biomarker signatures.
KW  - EMT
KW  - drug resistance
KW  - invasion
KW  - stemness
KW  - 3D lung tumor tissue models
KW  - KRAS biomarker signatures
KW  - boolean in silico models
KW  - targeted combination therapy
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-270744
SN  - 2072-6694
VL  - 14
IS  - 9
ER  - 
TY  - JOUR
A1  - Gerhard‐Hartmann, Elena
A1  - Jöhrens, Korinna
A1  - Schinagl, Lisa‐Marie
A1  - Zamó, Alberto
A1  - Rosenwald, Andreas
A1  - Anagnostopoulos, Ioannis
A1  - Rosenfeldt, Mathias
T1  - Epstein–Barr virus infection patterns in nodular lymphocyte‐predominant Hodgkin lymphoma
JF  - Histopathology
N2  - Aims
To investigate Epstein‐Barr virus (EBV) latency types in 19 cases of EBV‐positive nodular lymphocyte‐predominant Hodgkin lymphoma (NLPHL), as such information is currently incomplete.
Methods and results
Immunohistochemistry (IHC) for CD20, CD79a, PAX5, OCT2, CD30, CD15, CD3 and programmed cell death protein 1 was performed. For EBV detection, in‐situ hybridisation (ISH) for EBV‐encoded RNA (EBER) was employed combined with IHC for EBV‐encoded latent membrane protein (LMP)‐1, EBV‐encoded nuclear antigen (EBNA)‐2, and EBV‐encoded BZLF1. In 95% of the cases, neoplastic cells with features of Hodgkin and Reed–Sternberg (HRS) cells were present, mostly showing expression of CD30. In all cases, the B‐cell phenotype was largely intact, and delineation from classic Hodgkin lymphoma (CHL) was further supported by myocyte enhancer factor 2B (MEF2B) detection. All tumour cells were EBER‐positive except in two cases. EBV latency type II was most frequent (89%) and type I was rare. Cases with latency type I were CD30‐negative. Five cases contained some BZLF1‐positive and/or EBNA‐2‐positive bystander lymphocytes.
Conclusions
As HRS morphology of neoplastic cells and CD30 expression are frequent features of EBV‐positive NLPHL, preservation of the B‐cell transcription programme, MEF2B expression combined with NLPHL‐typical architecture and background composition facilitate distinction from CHL. EBER ISH is the method of choice to identify these cases. The majority present with EBV latency type II, and only rare cases present with latency type I, which can be associated with missing CD30 expression. The presence of occasional bystander lymphocytes expressing BZLF1 and/or EBNA‐2 and the partial EBV infection of neoplastic cells in some cases could indicate that EBV is either not primarily involved or is only a transient driver in the pathogenesis of EBV‐positive NLPHL.
KW  - EBV
KW  - Hodgkin lymphoma
KW  - latency type
KW  - NLPHL
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-276327
VL  - 80
IS  - 7
SP  - 1071
EP  - 1080
ER  - 
TY  - JOUR
A1  - Mainz, Laura
A1  - Sarhan, Mohamed A. F. E.
A1  - Roth, Sabine
A1  - Sauer, Ursula
A1  - Maurus, Katja
A1  - Hartmann, Elena M.
A1  - Seibert, Helen-Desiree
A1  - Rosenwald, Andreas
A1  - Diefenbacher, Markus E.
A1  - Rosenfeldt, Mathias T.
T1  - Autophagy blockage reduces the incidence of pancreatic ductal adenocarcinoma in the context of mutant Trp53
JF  - Frontiers in Cell and Developmental Biology
N2  - Macroautophagy (hereafter referred to as autophagy) is a homeostatic process that preserves cellular integrity. In mice, autophagy regulates pancreatic ductal adenocarcinoma (PDAC) development in a manner dependent on the status of the tumor suppressor gene Trp53. Studies published so far have investigated the impact of autophagy blockage in tumors arising from Trp53-hemizygous or -homozygous tissue. In contrast, in human PDACs the tumor suppressor gene TP53 is mutated rather than allelically lost, and TP53 mutants retain pathobiological functions that differ from complete allelic loss. In order to better represent the patient situation, we have investigated PDAC development in a well-characterized genetically engineered mouse model (GEMM) of PDAC with mutant Trp53 (Trp53\(^{R172H}\)) and deletion of the essential autophagy gene Atg7. Autophagy blockage reduced PDAC incidence but had no impact on survival time in the subset of animals that formed a tumor. In the absence of Atg7, non-tumor-bearing mice reached a similar age as animals with malignant disease. However, the architecture of autophagy-deficient, tumor-free pancreata was effaced, normal acinar tissue was largely replaced with low-grade pancreatic intraepithelial neoplasias (PanINs) and insulin expressing islet β-cells were reduced. Our data add further complexity to the interplay between Atg7 inhibition and Trp53 status in tumorigenesis.
KW  - pancreatic cancer
KW  - autophagy
KW  - p53
KW  - metastasis
KW  - ATG7
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-266005
SN  - 2296-634X
VL  - 10
ER  - 
TY  - JOUR
A1  - Richter, Julia
A1  - Hüttmann, Andreas
A1  - Rekowski, Jan
A1  - Schmitz, Christine
A1  - Gärtner, Selina
A1  - Rosenwald, Andreas
A1  - Hansmann, Martin-Leo
A1  - Hartmann, Sylvia
A1  - Möller, Peter
A1  - Wacker, Hans-Heinrich
A1  - Feller, Alfred
A1  - Thorns, Christoph
A1  - Müller, Stefan
A1  - Dührsen, Ulrich
A1  - Klapper, Wolfram
T1  - Molecular characteristics of diffuse large B-cell lymphoma in the Positron Emission Tomography-Guided Therapy of Aggressive Non-Hodgkin lymphomas (PETAL) trial: correlation with interim PET and outcome
JF  - Blood Cancer Journal
N2  - No abstract available
KW  - Cancer genetics
KW  - Medical research
KW  - Translational research
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226185
VL  - 9
ER  - 
TY  - JOUR
A1  - Rapp, Christina K.
A1  - Van Dijck, Ine
A1  - Laugwitz, Lucia
A1  - Boon, Mieke
A1  - Briassoulis, George
A1  - Ilia, Stavroula
A1  - Kammer, Birgit
A1  - Reu, Simone
A1  - Hornung, Stefanie
A1  - Buchert, Rebecca
A1  - Sofan, Linda
A1  - Froukh, Tawfiq
A1  - Witters, Peter
A1  - Rymen, Daisy
A1  - Haack, Tobias B.
A1  - Proesmans, Marijke
A1  - Griese, Matthias
T1  - Expanding the phenotypic spectrum of FINCA (fibrosis, neurodegeneration, and cerebral angiomatosis) syndrome beyond infancy
JF  - Clinical Genetics
N2  - Fibrosis, neurodegeneration, and cerebral angiomatosis (FINCA, MIM#618278) is a rare clinical condition caused by bi‐allelic variants in NHL repeat containing protein 2 (NHLRC2, MIM*618277). Pulmonary disease may be the presenting sign and the few patients reported so far, all deceased in early infancy. Exome sequencing was performed on patients with childhood interstitial lung disease (chILD) and additional neurological features. The chILD‐EU register database and an in‐house database were searched for patients with NHLRC2 variants and clinical features overlapping FINCA syndrome. Six patients from three families were identified with bi‐allelic variants in NHLRC2. Two of these children died before the age of two while four others survived until childhood. Interstitial lung disease was pronounced in almost all patients during infancy and stabilized over the course of the disease with neurodevelopmental delay (NDD) evolving as the key clinical finding. We expand the phenotype of FINCA syndrome to a multisystem disorder with variable severity. FINCA syndrome should also be considered in patients beyond infancy with NDD and a history of distinct interstitial lung disease. Managing patients in registers for rare diseases helps identifying new diagnostic entities and advancing care for these patients.
KW  - cerebropulmonary disease
KW  - childhood interstitial lung disease
KW  - cholesterol pneumonia
KW  - FINCA
KW  - lung fibrosis
KW  - lipoid pneumonitis
KW  - multi‐organ disease
KW  - NHLRC2
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-262732
VL  - 100
IS  - 4
SP  - 453
EP  - 461
ER  - 
TY  - JOUR
A1  - Cadar, Dániel
A1  - Jellinger, Kurt A.
A1  - Riederer, Peter
A1  - Strobel, Sabrina
A1  - Monoranu, Camelia-Maria
A1  - Tappe, Dennis
T1  - No metagenomic evidence of causative viral pathogens in postencephalitic parkinsonism following encephalitis lethargica
JF  - Microorganisms
N2  - Postencephalitic parkinsonism (PEP) is a disease of unknown etiology and pathophysiology following encephalitis lethargica (EL), an acute-onset polioencephalitis of cryptic cause in the 1920s. PEP is a tauopathy with multisystem neuronal loss and gliosis, clinically characterized by bradykinesia, rigidity, rest tremor, and oculogyric crises. Though a viral cause of EL is likely, past polymerase chain reaction-based investigations in the etiology of both PEP and EL were negative. PEP might be caused directly by an unknown viral pathogen or the consequence of a post-infectious immunopathology. The development of metagenomic next-generation sequencing in conjunction with bioinformatic techniques has generated a broad-range tool for the detection of unknown pathogens in the recent past. Retrospective identification and characterization of pathogens responsible for past infectious diseases can be successfully performed with formalin-fixed paraffin-embedded (FFPE) tissue samples. In this study, we analyzed 24 FFPE brain samples from six patients with PEP by unbiased metagenomic next-generation sequencing. Our results show that no evidence for the presence of a specific or putative (novel) viral pathogen was found, suggesting a likely post-infectious immune-mediated etiology of PEP.
KW  - postencephalitic parkinsonism
KW  - encephalitis lethargica
KW  - von Economo
KW  - metagenomics
KW  - neuropathology
KW  - tauopathy
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-245074
SN  - 2076-2607
VL  - 9
IS  - 8
ER  - 
TY  - JOUR
A1  - Metzner, Valentin
A1  - Herzog, Gloria
A1  - Heckel, Tobias
A1  - Bischler, Thorsten
A1  - Hasinger, Julia
A1  - Otto, Christoph
A1  - Fassnacht, Martin
A1  - Geier, Andreas
A1  - Seyfried, Florian
A1  - Dischinger, Ulrich
T1  - Liraglutide + PYY\(_{3-36}\) combination therapy mimics effects of Roux-en-Y bypass on early NAFLD whilst lacking-behind in metabolic improvements
JF  - Journal of Clinical Medicine
N2  - Background: Treatment options for NAFLD are still limited. Bariatric surgery, such as Roux-en-Y gastric bypass (RYGB), has been shown to improve metabolic and histologic markers of NAFLD. Glucagon-like-peptide-1 (GLP-1) analogues lead to improvements in phase 2 clinical trials. We directly compared the effects of RYGB with a treatment using liraglutide and/or peptide tyrosine tyrosine 3-36 (PYY\(_{3-36}\)) in a rat model for early NAFLD. Methods: Obese male Wistar rats (high-fat diet (HFD)-induced) were randomized into the following treatment groups: RYGB, sham-operation (sham), liraglutide (0.4 mg/kg/day), PYY\(_{3-36}\) (0.1 mg/kg/day), liraglutide+PYY\(_{3-36}\), and saline. After an observation period of 4 weeks, liver samples were histologically evaluated, ELISAs and RNA sequencing + RT-qPCRs were performed. Results: RYGB and liraglutide+PYY\(_{3-36}\) induced a similar body weight loss and, compared to sham/saline, marked histological improvements with significantly less steatosis. However, only RYGB induced significant metabolic improvements (e.g., adiponectin/leptin ratio 18.8 ± 11.8 vs. 2.4 ± 1.2 in liraglutide+PYY\(_{3-36}\)- or 1.4 ± 0.9 in sham-treated rats). Furthermore, RNA sequencing revealed a high number of differentially regulated genes in RYGB treated animals only. Conclusions: The combination therapy of liraglutide+PYY\(_{3-36}\) partly mimics the positive effects of RYGB on weight reduction and on hepatic steatosis, while its effects on metabolic function lack behind RYGB.
KW  - liraglutide
KW  - GLP-1
KW  - peptide tyrosine tyrosine (PYY)
KW  - peptide tyrosine tyrosine 3-36 (PYY\(_{3-36}\))
KW  - RYGB
KW  - gastric bypass
KW  - obesity
KW  - NASH
KW  - NAFLD
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-255244
SN  - 2077-0383
VL  - 11
IS  - 3
ER  - 
TY  - JOUR
A1  - Pietro-Garcia, Christian
A1  - Hartmann, Oliver
A1  - Reissland, Michaela
A1  - Fischer, Thomas
A1  - Maier, Carina R.
A1  - Rosenfeldt, Mathias
A1  - Schülein-Völk, Christina
A1  - Klann, Kevin
A1  - Kalb, Reinhard
A1  - Dikic, Ivan
A1  - Münch, Christian
A1  - Diefenbacher, Markus E.
T1  - Inhibition of USP28 overcomes Cisplatin-resistance of squamous tumors by suppression of the Fanconi anemia pathway
JF  - Cell Death and Differentiation
N2  - Squamous cell carcinomas (SCC) frequently have an exceptionally high mutational burden. As consequence, they rapidly develop resistance to platinum-based chemotherapy and overall survival is limited. Novel therapeutic strategies are therefore urgently required. SCC express ∆Np63, which regulates the Fanconi Anemia (FA) DNA-damage response in cancer cells, thereby contributing to chemotherapy-resistance. Here we report that the deubiquitylase USP28 is recruited to sites of DNA damage in cisplatin-treated cells. ATR phosphorylates USP28 and increases its enzymatic activity. This phosphorylation event is required to positively regulate the DNA damage repair in SCC by stabilizing ∆Np63. Knock-down or inhibition of USP28 by a specific inhibitor weakens the ability of SCC to cope with DNA damage during platin-based chemotherapy. Hence, our study presents a novel mechanism by which ∆Np63 expressing SCC can be targeted to overcome chemotherapy resistance. Limited treatment options and low response rates to chemotherapy are particularly common in patients with squamous cancer. The SCC specific transcription factor ∆Np63 enhances the expression of Fanconi Anemia genes, thereby contributing to recombinational DNA repair and Cisplatin resistance. Targeting the USP28-∆Np63 axis in SCC tones down this DNA damage response pathways, thereby sensitizing SCC cells to cisplatin treatment.
KW  - USP28
KW  - Cisplatin
KW  - squamous tumors
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-273014
SN  - 1476-5403
VL  - 29
IS  - 3
ER  - 
TY  - JOUR
A1  - Steinhardt, Maximilian J.
A1  - Krummenast, Franziska C.
A1  - Rosenwald, Andreas
A1  - Gerhard-Hartmann, Elena
A1  - Heidemeier, Anke
A1  - Einsele, Hermann
A1  - Topp, Max S.
A1  - Duell, Johannes
T1  - R-CHOP intensification with mid-cycle methotrexate and consolidating AraC/TT with BCNU/aHSCT in primary aggressive lymphoma with CNS involvement
JF  - Journal of Cancer Research and Clinical Oncology
N2  - Purpose
Patients suffering from aggressive systemic peripheral lymphoma with primary central nervous system involvement (PCL) are a rare and sparsely investigated population. Recommended treatment regimens include a combination of intrathecal and systemic chemotherapy as well as whole brain radiotherapy while offering relatively poor survival.
Methods
We conducted a single-center retrospective study that analyzed safety and outcome of 4 + 4 cycles Rituximab (R)-CHOP and R-high-dose Methotrexate (HD-MTX) for newly diagnosed, transplant-eligible patients ("Ping-Pong"), followed by Cytarabine (AraC)/Thiotepa (TT), BCNU/TT, and autologous hematologic stem cell transplantation (aHSCT). We retrospectively analyzed a set of 16 patients with high-intermediate or high-risk IPI status.
Results
Overall response rate to Ping-Pong was 100% measured by CT/MRI, including 93.75% complete remissions after BCNU/TT followed by PBSCT. One patient failed to qualify for high-dose chemotherapy due to progression when receiving Cytarabine/TT. All patients experienced grade III adverse events, 3 of them a grade IV adverse event. Estimated progression-free survival is 93.75% after a 4.8-year follow-up currently.
Conclusion
Our study suggests high effectivity of R-CHOP with mid-cycle MTX with aHSCT consolidation towards acceptable OS results in this challenging patient population.
KW  - lymphoma
KW  - HD
KW  - MTX
KW  - R-CHOP
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-267731
SN  - 1432-1335
VL  - 148
IS  - 1
ER  - 
TY  - JOUR
A1  - Kader, Hidaya A.
A1  - Azeem, Muhammad
A1  - Jwayed, Suhib A.
A1  - Al-Shehhi, Aaesha
A1  - Tabassum, Attia
A1  - Ayoub, Mohammed Akli
A1  - Hetta, Helal F.
A1  - Waheed, Yasir
A1  - Iratni, Rabah
A1  - Al-Dhaheri, Ahmed
A1  - Muhammad, Khalid
T1  - Current insights into immunology and novel therapeutics of atopic dermatitis
JF  - Cells
N2  - Atopic dermatitis (AD) is one of the most prevalent inflammatory disease among non-fatal skin diseases, affecting up to one fifth of the population in developed countries. AD is characterized by recurrent pruritic and localized eczema with seasonal fluctuations. AD initializes the phenomenon of atopic march, during which infant AD patients are predisposed to progressive secondary allergies such as allergic rhinitis, asthma, and food allergies. The pathophysiology of AD is complex; onset of the disease is caused by several factors, including strong genetic predisposition, disrupted epidermal barrier, and immune dysregulation. AD was initially characterized by defects in the innate immune system and a vigorous skewed adaptive Th2 response to environmental agents; there are compelling evidences that the disorder involves multiple immune pathways. Symptomatic palliative treatment is the only strategy to manage the disease and restore skin integrity. Researchers are trying to more precisely define the contribution of different AD genotypes and elucidate the role of various immune axes. In this review, we have summarized the current knowledge about the roles of innate and adaptive immune responsive cells in AD. In addition, current and novel treatment strategies for the management of AD are comprehensively described, including some ongoing clinical trials and promising therapeutic agents. This information will provide an asset towards identifying personalized targets for better therapeutic outcomes.
KW  - atopic dermatitis
KW  - immune system
KW  - T cells
KW  - B cells
KW  - keratinocytes
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241008
SN  - 2073-4409
VL  - 10
IS  - 6
ER  - 
TY  - JOUR
A1  - Danhof, Sophia
A1  - Rasche, Leo
A1  - Mottok, Anja
A1  - Steinmüller, Tabea
A1  - Zhou, Xiang
A1  - Schreder, Martin
A1  - Kilian, Teresa
A1  - Strifler, Susanne
A1  - Rosenwald, Andreas
A1  - Hudecek, Michael
A1  - Einsele, Hermann
A1  - Gerhard-Hartmann, Elena
T1  - Elotuzumab for the treatment of extramedullary myeloma: a retrospective analysis of clinical efficacy and SLAMF7 expression patterns
JF  - Annals of Hematology
N2  - Extramedullary disease (EMD) represents a high-risk state of multiple myeloma (MM) associated with poor prognosis. While most anti-myeloma therapeutics demonstrate limited efficacy in this setting, some studies exploring the utility of chimeric antigen receptor (CAR)-modified T cells reported promising results. We have recently designed SLAMF7-directed CAR T cells for the treatment of MM. SLAMF7 is a transmembrane receptor expressed on myeloma cells that plays a role in myeloma cell homing to the bone marrow. Currently, the only approved anti-SLAMF7 therapeutic is the monoclonal antibody elotuzumab, but its efficacy in EMD has not been investigated thoroughly. Thus, we retrospectively analyzed the efficacy of elotuzumab-based combination therapy in a cohort of 15 patients with EMD. Moreover, since the presence of the target antigen is an indispensable prerequisite for effective targeted therapy, we investigated the SLAMF7 expression on extramedullary located tumor cells before and after treatment. We observed limited efficacy of elotuzumab-based combination therapies, with an overall response rate of 40% and a progression-free and overall survival of 3.8 and 12.9 months, respectively. Before treatment initiation, all available EMD tissue specimens (n = 3) demonstrated a strong and consistent SLAMF7 surface expression by immunohistochemistry. Furthermore, to investigate a potential antigen reduction under therapeutic selection pressure, we analyzed samples of de novo EMD (n = 3) outgrown during elotuzumab treatment. Again, immunohistochemistry documented strong and consistent SLAMF7 expression in all samples. In aggregate, our data point towards a retained expression of SLAMF7 in EMD and encourage the development of more potent SLAMF7-directed immunotherapies, such as CAR T cells.
KW  - plasma cells
KW  - extramedullary disease
KW  - monoclonal antibody
KW  - CD319
KW  - CS1
KW  - antigen loss
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-266468
SN  - 1432-0584
VL  - 100
IS  - 6
ER  - 
TY  - JOUR
A1  - Porubsky, Stefan
A1  - Popovic, Zoran V.
A1  - Badve, Sunil
A1  - Banz, Yara
A1  - Berezowska, Sabina
A1  - Borchert, Dietmar
A1  - Brüggemann, Monika
A1  - Gaiser, Timo
A1  - Graeter, Thomas
A1  - Hollaus, Peter
A1  - Huettl, Katrin S.
A1  - Kotrova, Michaela
A1  - Kreft, Andreas
A1  - Kugler, Christian
A1  - Lötscher, Fabian
A1  - Möller, Burkhard
A1  - Ott, German
A1  - Preissler, Gerhard
A1  - Roessner, Eric
A1  - Rosenwald, Andreas
A1  - Ströbel, Philipp
A1  - Marx, Alexander
T1  - Thymic hyperplasia with lymphoepithelial sialadenitis (LESA)-like features: strong association with lymphomas and non-myasthenic autoimmune diseases
JF  - Cancers
N2  - Thymic hyperplasia (TH) with lymphoepithelial sialadenitis (LESA)-like features (LESA-like TH) has been described as a tumor-like, benign proliferation of thymic epithelial cells and lymphoid follicles. We aimed to determine the frequency of lymphoma and autoimmunity in LESA-like TH and performed retrospective analysis of cases with LESA-like TH and/or thymic MALT-lymphoma. Among 36 patients (21 males) with LESA-like TH (age 52 years, 32–80; lesion diameter 7.0 cm, 1–14.5; median, range), five (14%) showed associated lymphomas, including four (11%) thymic MALT lymphomas and one (3%) diffuse large B-cell lymphoma. One additional case showed a clonal B-cell-receptor rearrangement without evidence of lymphoma. Twelve (33%) patients (7 women) suffered from partially overlapping autoimmune diseases: systemic lupus erythematosus (n = 4, 11%), rheumatoid arthritis (n = 3, 8%), myasthenia gravis (n = 2, 6%), asthma (n = 2, 6%), scleroderma, Sjögren syndrome, pure red cell aplasia, Grave’s disease and anti-IgLON5 syndrome (each n = 1, 3%). Among 11 primary thymic MALT lymphomas, remnants of LESA-like TH were found in two cases (18%). In summary, LESA-like TH shows a striking association with autoimmunity and predisposes to lymphomas. Thus, a hematologic and rheumatologic workup should become standard in patients diagnosed with LESA-like TH. Radiologists and clinicians should be aware of LESA-like TH as a differential diagnosis for mediastinal mass lesions in patients with autoimmune diseases.
KW  - autoimmune disease
KW  - imaging
KW  - LESA
KW  - lymphoma
KW  - myasthenia
KW  - pathology
KW  - surgery
KW  - thymus
KW  - thymic epithelial tumor
KW  - thymitis
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-223049
SN  - 2072-6694
VL  - 13
IS  - 2
ER  - 
TY  - JOUR
A1  - Riederer, P.
A1  - Monoranu, C.
A1  - Strobel, S.
A1  - Iordache, T.
A1  - Sian-Hülsmann, J.
T1  - Iron as the concert master in the pathogenic orchestra playing in sporadic Parkinson's disease
JF  - Journal of Neural Transmission
N2  - About 60 years ago, the discovery of a deficiency of dopamine in the nigro-striatal system led to a variety of symptomatic therapeutic strategies to supplement dopamine and to substantially improve the quality of life of patients with Parkinson's disease (PD). Since these seminal developments, neuropathological, neurochemical, molecular biological and genetic discoveries contributed to elucidate the pathology of PD. Oxidative stress, the consequences of reactive oxidative species, reduced antioxidative capacity including loss of glutathione, excitotoxicity, mitochondrial dysfunction, proteasomal dysfunction, apoptosis, lysosomal dysfunction, autophagy, suggested to be causal for ɑ-synuclein fibril formation and aggregation and contributing to neuroinflammation and neural cell death underlying this devastating disorder. However, there are no final conclusions about the triggered pathological mechanism(s) and the follow-up of pathological dysfunctions. Nevertheless, it is a fact, that iron, a major component of oxidative reactions, as well as neuromelanin, the major intraneuronal chelator of iron, undergo an age-dependent increase. And ageing is a major risk factor for PD. Iron is significantly increased in the substantia nigra pars compacta (SNpc) of PD. Reasons for this finding include disturbances in iron-related import and export mechanisms across the blood-brain barrier (BBB), localized opening of the BBB at the nigro-striatal tract including brain vessel pathology. Whether this pathology is of primary or secondary importance is not known. We assume that there is a better fit to the top-down hypotheses and pathogens entering the brain via the olfactory system, then to the bottom-up (gut-brain) hypothesis of PD pathology. Triggers for the bottom-up, the dual-hit and the top-down pathologies include chemicals, viruses and bacteria. If so, hepcidin, a regulator of iron absorption and its distribution into tissues, is suggested to play a major role in the pathogenesis of iron dyshomeostasis and risk for initiating and progressing ɑ-synuclein pathology. The role of glial components to the pathology of PD is still unknown. However, the dramatic loss of glutathione (GSH), which is mainly synthesized in glia, suggests dysfunction of this process, or GSH uptake into neurons. Loss of GSH and increase in SNpc iron concentration have been suggested to be early, may be even pre-symptomatic processes in the pathology of PD, despite the fact that they are progression factors. The role of glial ferritin isoforms has not been studied so far in detail in human post-mortem brain tissue and a close insight into their role in PD is called upon. In conclusion, "iron" is a major player in the pathology of PD. Selective chelation of excess iron at the site of the substantia nigra, where a dysfunction of the BBB is suggested, with peripherally acting iron chelators is suggested to contribute to the portfolio and therapeutic armamentarium of anti-Parkinson medications.
KW  - SARS-CoV-2
KW  - iron in parkinsonism
KW  - parkinson’s disease
KW  - iiron transporter
KW  - neuromelanin
KW  - iron pathology
KW  - neuroinflammation
KW  - iron model
KW  - ferroptosis
KW  - ɑ-Synuclein and iron
KW  - virus–iron interaction
KW  - COVID-19
KW  - hepcidin
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-268539
SN  - 1435-1463
VL  - 128
IS  - 10
ER  - 
TY  - JOUR
A1  - Bohnert, Simone
A1  - Georgiades, Kosmas
A1  - Monoranu, Camelia-Maria
A1  - Bohnert, Michael
A1  - Büttner, Andreas
A1  - Ondruschka, Benjamin
T1  - Quantitative evidence of suppressed TMEM119 microglial immunohistochemistry in fatal morphine intoxications
JF  - International Journal of Legal Medicine
N2  - The aim of this pilot study was to investigate the diagnostic potential of TMEM119 as a useful microglia-specific marker in combination with immunostainings for phagocytic function and infiltrating capacity of monocytes in cases of lethal monosubstance intoxications by morphine (MOR), methamphetamine (METH), and of ethanol-associated death (ETH) respectively. Human brain tissue samples were obtained from forensic autopsies of cases with single substance abuse (MOR, n = 8; ETH, n = 10; METH, n = 9) and then compared to a cohort of cardiovascular fatalities as controls (n = 9). Brain tissue samples of cortex, white matter, and hippocampus were collected and stained immunohistochemically with antibodies against TMEM119, CD68KiM1P, and CCR2. We could document the lowest density of TMEM119-positive cells in MOR deaths with highly significant differences to the control densities in all three regions investigated. In ETH and METH deaths, the expression of TMEM119 was comparable to cell densities in controls. The results indicate that the immunoreaction in brain tissue is different in these groups depending on the drug type used for abuse.
KW  - immunohistochemistry
KW  - drug abuse
KW  - forensic neuropathology
KW  - neuroinflammation
KW  - neurotoxicity
KW  - microglia
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-266934
SN  - 1437-1596
VL  - 135
IS  - 6
ER  - 
TY  - JOUR
A1  - Bohnert, Simone
A1  - Wirth, Christoph
A1  - Schmitz, Werner
A1  - Trella, Stefanie
A1  - Monoranu, Camelia-Maria
A1  - Ondruschka, Benjamin
A1  - Bohnert, Michael
T1  - Myelin basic protein and neurofilament H in postmortem cerebrospinal fluid as surrogate markers of fatal traumatic brain injury
JF  - International Journal of Legal Medicine
N2  - The aim of this study was to investigate if the biomarkers myelin basic protein (MBP) and neurofilament-H (NF-H) yielded informative value in forensic diagnostics when examining cadaveric cerebrospinal fluid (CSF) biochemically via an enzyme-linked immunosorbent assay (ELISA) and comparing the corresponding brain tissue in fatal traumatic brain injury (TBI) autopsy cases by immunocytochemistry versus immunohistochemistry. In 21 trauma and 19 control cases, CSF was collected semi-sterile after suboccipital puncture and brain specimens after preparation. The CSF MBP (p = 0.006) and NF-H (p = 0.0002) levels after TBI were significantly higher than those in cardiovascular controls. Immunohistochemical staining against MBP and against NF-H was performed on cortical and subcortical samples from also biochemically investigated cases (5 TBI cases/5 controls). Compared to the controls, the TBI cases showed a visually reduced staining reaction against MBP or repeatedly ruptured neurofilaments against NF-H. Immunocytochemical tests showed MBP-positive phagocytizing macrophages in CSF with a survival time of > 24 h. In addition, numerous TMEM119-positive microglia could be detected with different degrees of staining intensity in the CSF of trauma cases. As a result, we were able to document that elevated levels of MBP and NF-H in the CSF should be considered as useful neuroinjury biomarkers of traumatic brain injury.
KW  - biofluid
KW  - CSF
KW  - cerebrospinal fluid
KW  - forensic neuropathology
KW  - forensic neurotraumatology
KW  - biomarker
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-266929
SN  - 1437-1596
VL  - 135
IS  - 4
ER  - 
TY  - JOUR
A1  - Meyer, Till Jasper
A1  - Gerhard-Hartmann, Elena
A1  - Lodes, Nina
A1  - Scherzad, Agmal
A1  - Hagen, Rudolf
A1  - Steinke, Maria
A1  - Hackenberg, Stephan
T1  - Pilot study on the value of Raman spectroscopy in the entity assignment of salivary gland tumors
JF  - PLoS One
N2  - Background
The entity assignment of salivary gland tumors (SGT) based on histomorphology can be challenging. Raman spectroscopy has been applied to analyze differences in the molecular composition of tissues. The aim of this study was to evaluate the suitability of RS for entity assignment in SGT.

Methods
Raman data were collected in deparaffinized sections of pleomorphic adenomas (PA) and adenoid cystic carcinomas (ACC). Multivariate data and chemometric analysis were completed using the Unscrambler software.

Results
The Raman spectra detected in ACC samples were mostly assigned to nucleic acids, lipids, and amides. In a principal component-based linear discriminant analysis (LDA) 18 of 20 tumor samples were classified correctly.

Conclusion
In this proof of concept study, we show that a reliable SGT diagnosis based on LDA algorithm appears possible, despite variations in the entity-specific mean spectra. However, a standardized workflow for tissue sample preparation, measurement setup, and chemometric algorithms is essential to get reliable results.
KW  - Head and neck cancers
KW  - salivary gland tumors
KW  - salivary glands
KW  - cancers and neoplasms
KW  - malignant tumors
KW  - lipids
KW  - raman spectroscopy
KW  - surgical oncology
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-264736
VL  - 16
IS  - 9
ER  - 
TY  - JOUR
A1  - Frings, Verena G.
A1  - Roth, Sabine
A1  - Rosenwald, Andreas
A1  - Goebeler, Matthias
A1  - Geissinger, Eva
A1  - Wobser, Marion
T1  - EBER in situ hybridization in subcutaneous aluminum granulomas/lymphoid hyperplasia: A diagnostic clue to differentiate injection-associated lymphoid hyperplasia from other forms of pseudolymphomas and cutaneous lymphomas
JF  - Journal of Cutaneous Pathology
N2  - Background
Subcutaneous vaccination or desensitization may induce persistent nodules at the injection sites. Without the knowledge of prior injection, histopathological work-up may be challenging.

Objective
Aim of this study was to contribute to the histopathological work-up of unclear subcutaneous nodules, especially their differentiation from cutaneous lymphoma.

Methods
We retrospectively reviewed clinical data and histopathological slides of four patients with subcutaneous nodules, which were suspected to suffer from cutaneous T- or B-cell lymphoma. Sections of these cases and 12 negative controls were stained with hematoxylin and eosin and a standardized immunohistochemical panel of B- and T-cell markers including EBER in situ hybridization as well as electron microscopy.

Results
In all cases, large histiocytes with granular cytoplasm compatible with intracellular aluminum hydroxide were present. EBER in situ hybridization revealed positive staining of these granular histiocytes while staining was absent in negative controls.

Limitations
Post hoc completion of medical history revealed that vaccination or specific immunotherapy had been applied before at the biopsy site in only three out of four patients; one patient was lost to follow-up.

Conclusion
EBER in situ hybridization is an adjunctive tool to differentiate aluminum-induced granuloma/lymphoid hyperplasia from other forms of pseudolymphoma and cutaneous B- or T-cell lymphomas.
KW  - RNA probe
KW  - aluminum granuloma
KW  - EBER in situ hybridization
KW  - lymphoid hyperplasia
KW  - pseudolymphoma
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258405
VL  - 48
IS  - 5
ER  - 
TY  - JOUR
A1  - Heuser, Christoph
A1  - Gotot, Janine
A1  - Piotrowski, Eveline Christina
A1  - Philipp, Marie-Sophie
A1  - Courrèges, Christina Johanna Felicia
A1  - Otte, Martin Sylvester
A1  - Guo, Linlin
A1  - Schmid-Burgk, Jonathan Leo
A1  - Hornung, Veit
A1  - Heine, Annkristin
A1  - Knolle, Percy Alexander
A1  - Garbi, Natalio
A1  - Serfling, Edgar
A1  - Evaristo, César
A1  - Thaiss, Friedrich
A1  - Kurts, Christian
T1  - Prolonged IKK\(\beta\) Inhibition Improves Ongoing CTL Antitumor Responses by Incapacitating Regulatory T Cells
JF  - Cell Reports
N2  - Regulatory T cells (Tregs) prevent autoimmunity but limit antitumor immunity. The canonical NF-\(\kappa\)B signaling pathway both activates immunity and promotes thymic Treg development. Here, we report that mature Tregs continue to require NF-\(\kappa\)B signaling through I\(\kappa\)B-kinase \(\beta\) (IKK\(\beta\)) after thymic egress. Mice lacking IKK\(\beta\) in mature Tregs developed scurfy-like immunopathology due to death of peripheral FoxP3\(^+\) Tregs. Also, pharmacological IKK\(\beta\) inhibition reduced Treg numbers in the circulation by ~50% and downregulated FoxP3 and CD25 expression and STAT5 phosphorylation. In contrast, activated cytotoxic T lymphocytes (CTLs) were resistant to IKK\(\beta\) inhibition because other pathways, in particular nuclear factor of activated T cells (NFATc1) signaling, sustained their survival and expansion. In a melanoma mouse model, IKK\(\beta\) inhibition after CTL cross-priming improved the antitumor response and delayed tumor growth. In conclusion, prolonged IKK\(\beta\) inhibition decimates circulating Tregs and improves CTL responses when commenced after tumor vaccination, indicating that IKK\(\beta\) represents a druggable checkpoint.
KW  - medicine
KW  - regulatory T cells
KW  - NF-\(\kappa\)B pathway
KW  - tumor vaccination
KW  - checkpoint inhibition
KW  - cytotoxic T cells
KW  - cross-priming
KW  - apoptosis
KW  - tumor immunology
KW  - melanoma
Y1  - 2017
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-173643
VL  - 21
IS  - 3
ER  - 
TY  - JOUR
A1  - Garitano-Trojaola, Andoni
A1  - Sancho, Ana
A1  - Götz, Ralph
A1  - Eiring, Patrick
A1  - Walz, Susanne
A1  - Jetani, Hardikkumar
A1  - Gil-Pulido, Jesus
A1  - Da Via, Matteo Claudio
A1  - Teufel, Eva
A1  - Rhodes, Nadine
A1  - Haertle, Larissa
A1  - Arellano-Viera, Estibaliz
A1  - Tibes, Raoul
A1  - Rosenwald, Andreas
A1  - Rasche, Leo
A1  - Hudecek, Michael
A1  - Sauer, Markus
A1  - Groll, Jürgen
A1  - Einsele, Hermann
A1  - Kraus, Sabrina
A1  - Kortüm, Martin K.
T1  - Actin cytoskeleton deregulation confers midostaurin resistance in FLT3-mutant acute myeloid leukemia
JF  - Communications Biology
N2  - The presence of FMS-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) is one of the most frequent mutations in acute myeloid leukemia (AML) and is associated with an unfavorable prognosis. FLT3 inhibitors, such as midostaurin, are used clinically but fail to entirely eradicate FLT3-ITD+AML. This study introduces a new perspective and highlights the impact of RAC1-dependent actin cytoskeleton remodeling on resistance to midostaurin in AML. RAC1 hyperactivation leads resistance via hyperphosphorylation of the positive regulator of actin polymerization N-WASP and antiapoptotic BCL-2. RAC1/N-WASP, through ARP2/3 complex activation, increases the number of actin filaments, cell stiffness and adhesion forces to mesenchymal stromal cells (MSCs) being identified as a biomarker of resistance. Midostaurin resistance can be overcome by a combination of midostaruin, the BCL-2 inhibitor venetoclax and the RAC1 inhibitor Eht1864 in midostaurin-resistant AML cell lines and primary samples, providing the first evidence of a potential new treatment approach to eradicate FLT3-ITD+AML. Garitano-Trojaola et al. used a combination of human acute myeloid leukemia (AML) cell lines and primary samples to show that RAC1-dependent actin cytoskeleton remodeling through BCL2 family plays a key role in resistance to the FLT3 inhibitor, Midostaurin in AML. They showed that by targeting RAC1 and BCL2, Midostaurin resistance was diminished, which potentially paves the way for an innovate treatment approach for FLT3 mutant AML.
KW  - actin
KW  - acute myeloid leukaemia
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260709
VL  - 4
IS  - 1
ER  - 
TY  - JOUR
A1  - Gerhard-Hartmann, Elena
A1  - Goergen, Helen
A1  - Bröckelmann, Paul J.
A1  - Mottok, Anja
A1  - Steinmüller, Tabea
A1  - Grund, Johanna
A1  - Zamò, Alberto
A1  - Ben-Neriah, Susana
A1  - Sasse, Stephanie
A1  - Borchmann, Sven
A1  - Fuchs, Michael
A1  - Borchmann, Peter
A1  - Reinke, Sarah
A1  - Engert, Andreas
A1  - Veldman, Johanna
A1  - Diepstra, Arjan
A1  - Klapper, Wolfram
A1  - Rosenwald, Andreas
T1  - 9p24.1 alterations and programmed cell death 1 ligand 1 expression in early stage unfavourable classical Hodgkin lymphoma: an analysis from the German Hodgkin Study Group NIVAHL trial
JF  - British Journal of Haematology
N2  - High programmed cell death 1 ligand 1 (PD-L1) protein expression and copy number alterations (CNAs) of the corresponding genomic locus 9p24.1 in Hodgkin- and Reed–Sternberg cells (HRSC) have been shown to be associated with favourable response to anti-PD-1 checkpoint inhibition in relapsed/refractory (r/r) classical Hodgkin lymphoma (cHL). In the present study, we investigated baseline 9p24.1 status as well as PD-L1 and major histocompatibility complex (MHC) class I and II protein expression in 82 biopsies from patients with early stage unfavourable cHL treated with anti-PD-1-based first-line treatment in the German Hodgkin Study Group (GHSG) NIVAHL trial (ClinicalTrials.gov Identifier: NCT03004833). All evaluated specimens showed 9p24.1 CNA in HRSC to some extent, but with high intratumoral heterogeneity and an overall smaller range of alterations than reported in advanced-stage or r/r cHL. All but two cases (97%) showed PD-L1 expression by the tumour cells in variable amounts. While MHC-I was rarely expressed in >50% of HRSC, MHC-II expression in >50% of HRSC was found more frequently. No obvious impact of 9p24.1 CNA or PD-L1 and MHC-I/II expression on early response to the highly effective anti-PD-1-based NIVAHL first-line treatment was observed. Further studies evaluating an expanded panel of potential biomarkers are needed to optimally stratify anti-PD-1 first-line cHL treatment.
KW  - fluorescence in situ hybridisation
KW  - major histocompatibility complex
KW  - immune checkpoint blockade
KW  - classical Hodgkin lymphoma
KW  - CD274
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-258358
VL  - 196
IS  - 1
ER  - 
TY  - JOUR
A1  - Meir, Michael
A1  - Maurus, Katja
A1  - Kuper, Jochen
A1  - Hankir, Mohammed
A1  - Wardelmann, Eva
A1  - Rosenwald, Andreas
A1  - Germer, Christoph-Thomas
A1  - Wiegering, Armin
T1  - The novel KIT exon 11 germline mutation K558N is associated with gastrointestinal stromal tumor, mastocytosis, and seminoma development
JF  - Genes, Chromosomes & Cancer
N2  - Familial gastrointestinal stromal tumors (GIST) are dominant genetic disorders that are caused by germline mutations of the type III receptor tyrosine kinase KIT. While sporadic mutations are frequently found in mastocytosis and GISTs, germline mutations of KIT have only been described in 39 families until now. We detected a novel germline mutation of KIT in exon 11 (p.Lys-558-Asn; K558N) in a patient from a kindred with several GISTs harboring different secondary somatic KIT mutations. Structural analysis suggests that the primary germline mutation alone is not sufficient to release the autoinhibitory region of KIT located in the transmembrane domain. Instead, the KIT kinase module becomes constitutively activated when K558N combines with different secondary somatic mutations. The identical germline mutation in combination with an additional somatic KIT mutation was detected in a second patient of the kindred with seminoma while a third patient within the family had a cutaneous mastocytosis. These findings suggest that the K558N mutation interferes with the juxtamembranous part of KIT, since seminoma and mastocystosis are usually not associated with exon 11 mutations.
KW  - germline mutation
KW  - GIST
KW  - KIT
KW  - mastocytosis
KW  - seminoma
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-257476
VL  - 60
IS  - 12
ER  - 
TY  - JOUR
A1  - Kreß, Julia Katharina Charlotte
A1  - Jessen, Christina
A1  - Marquardt, André
A1  - Hufnagel, Anita
A1  - Meierjohann, Svenja
T1  - NRF2 enables EGFR signaling in melanoma cells
JF  - International Journal of Molecular Sciences
N2  - Receptor tyrosine kinases (RTK) are rarely mutated in cutaneous melanoma, but the expression and activation of several RTK family members are associated with a proinvasive phenotype and therapy resistance. Epidermal growth factor receptor (EGFR) is a member of the RTK family and is only expressed in a subgroup of melanomas with poor prognosis. The insight into regulators of EGFR expression and activation is important for the understanding of the development of this malignant melanoma phenotype. Here, we describe that the transcription factor NRF2, the master regulator of the oxidative and electrophilic stress response, mediates the expression and activation of EGFR in melanoma by elevating the levels of EGFR as well as its ligands EGF and TGFα. ChIP sequencing data show that NRF2 directly binds to the promoter of EGF, which contains a canonical antioxidant response element. Accordingly, EGF is induced by oxidative stress and is also increased in lung adenocarcinoma and head and neck carcinoma with mutationally activated NRF2. In contrast, regulation of EGFR and TGFA occurs by an indirect mechanism, which is enabled by the ability of NRF2 to block the activity of the melanocytic lineage factor MITF in melanoma. MITF effectively suppresses EGFR and TGFA expression and therefore serves as link between NRF2 and EGFR. As EGFR was previously described to stimulate NRF2 activity, the mutual activation of NRF2 and EGFR pathways was investigated. The presence of NRF2 was necessary for full EGFR pathway activation, as NRF2-knockout cells showed reduced AKT activation in response to EGF stimulation compared to controls. Conversely, EGF led to the nuclear localization and activation of NRF2, thereby demonstrating that NRF2 and EGFR are connected in a positive feedback loop in melanoma. In summary, our data show that the EGFR-positive melanoma phenotype is strongly supported by NRF2, thus revealing a novel maintenance mechanism for this clinically challenging melanoma subpopulation.
KW  - EGFR
KW  - NRF2
KW  - NFE2L2
KW  - KEAP1
KW  - MITF-low
KW  - TGF-alpha
KW  - EGF
KW  - NSCLC
KW  - HNSC
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260222
SN  - 1422-0067
VL  - 22
IS  - 8
ER  - 
TY  - JOUR
A1  - Gerhard-Hartmann, Elena
A1  - Wiegering, Verena
A1  - Benoit, Clemens
A1  - Meyer, Thomas
A1  - Rosenwald, Andreas
A1  - Maurus, Katja
A1  - Ernestus, Karen
T1  - A large retroperitoneal lipoblastoma as an incidental finding: a case report
JF  - BMC Pediatrics
N2  - Background
Lipoblastoma is a rare benign mesenchymal neoplasm of infancy that most commonly occurs on the extremities and trunk but can arise at variable sites of the body. Retroperitoneal lipoblastomas are particularly rare but can grow to enormous size, and preoperative diagnosis is difficult with diverse, mostly malignant differential diagnoses that would lead to aggressive therapy. Since lipoblastoma is a benign tumor that has an excellent prognosis after resection, correct diagnosis is crucial.

Case presentation
A case of a large retroperitoneal tumor of a 24-month old infant that was clinically suspicious of a malignant tumor is presented. Due to proximity to the right kidney, clinically most probably a nephroblastoma or clear cell sarcoma of the kidney was suspected. Radiological findings were ambiguous. Therefore, the mass was biopsied, and histology revealed an adipocytic lesion. Although mostly composed of mature adipocytes, in view of the age of the patient, the differential diagnosis of a (maturing) lipoblastoma was raised, which was supported by molecular analysis demonstrating a HAS2-PLAG1 fusion. The tumor was completely resected, and further histopathological workup led to the final diagnosis of a 13 cm large retroperitoneal maturing lipoblastoma. The child recovered promptly from surgery and showed no evidence of recurrence so far.

Conclusion
Although rare, lipoblastoma should be included in the differential diagnoses of retroperitoneal tumors in infants and children, and molecular diagnostic approaches could be a helpful diagnostic adjunct in challenging cases.
KW  - retroperitoneal tumor
KW  - pediatric
KW  - lipoblastoma
KW  - PLAG1 rearrangement
KW  - case report
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-260173
VL  - 21
ER  - 
TY  - JOUR
A1  - Wallaschek, Nina
A1  - Reuter, Saskia
A1  - Silkenat, Sabrina
A1  - Wolf, Katharina
A1  - Niklas, Carolin
A1  - Özge, Kayisoglu
A1  - Aguilar, Carmen
A1  - Wiegering, Armin
A1  - Germer, Christoph-Thomas
A1  - Kircher, Stefan
A1  - Rosenwald, Andreas
A1  - Shannon-Lowe, Claire
A1  - Bartfeld, Sina
T1  - Ephrin receptor A2, the epithelial receptor for Epstein-Barr virus entry, is not available for efficient infection in human gastric organoids
JF  - PLoS Pathogens
N2  - Epstein-Barr virus (EBV) is best known for infection of B cells, in which it usually establishes an asymptomatic lifelong infection, but is also associated with the development of multiple B cell lymphomas. EBV also infects epithelial cells and is associated with all cases of undifferentiated nasopharyngeal carcinoma (NPC). EBV is etiologically linked with at least 8% of gastric cancer (EBVaGC) that comprises a genetically and epigenetically distinct subset of GC. Although we have a very good understanding of B cell entry and lymphomagenesis, the sequence of events leading to EBVaGC remains poorly understood. Recently, ephrin receptor A2 (EPHA2) was proposed as the epithelial cell receptor on human cancer cell lines. Although we confirm some of these results, we demonstrate that EBV does not infect healthy adult stem cell-derived gastric organoids. In matched pairs of normal and cancer-derived organoids from the same patient, EBV only reproducibly infected the cancer organoids. While there was no clear pattern of differential expression between normal and cancer organoids for EPHA2 at the RNA and protein level, the subcellular location of the protein differed markedly. Confocal microscopy showed EPHA2 localization at the cell-cell junctions in primary cells, but not in cancer cell lines. Furthermore, histologic analysis of patient tissue revealed the absence of EBV in healthy epithelium and presence of EBV in epithelial cells from inflamed tissue. These data suggest that the EPHA2 receptor is not accessible to EBV on healthy gastric epithelial cells with intact cell-cell contacts, but either this or another, yet to be identified receptor may become accessible following cellular changes induced by inflammation or transformation, rendering changes in the cellular architecture an essential prerequisite to EBV infection.
KW  - Organoids
KW  - ephitelial cells
KW  - gastrointestinal infections
KW  - cancers and neoplasms
KW  - Epstein-Barr virus
KW  - flow cytometry
KW  - epithelium
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259206
VL  - 17
IS  - 2
ER  - 
TY  - JOUR
A1  - Vaiopoulos, Aristeidis G.
A1  - Kanakis, Meletios A.
A1  - Kapsimali, Violetta
A1  - Vaiopoulos, Georgios
A1  - Kaklamanis, Phedon G.
A1  - Zouboulis, Christos C.
T1  - Juvenile Adamantiades-Behçet disease
JF  - Dermatology
N2  - Adamantiades-Behçet disease (ABD) is a chronic, multisystemic, recurrent, inflammatory vascular disorder of unknown etiology. Patients with symptoms initially appearing at the age of 16 or less are considered as cases of juvenile-onset ABD (JABD). JABD is relatively rare compared to ABD of adults, and only case reports and case studies have been published regarding this subtype of the disease. Epidemiology, clinical features, diagnosis and treatment of JABD are discussed in this review.
KW  - Aphthae
KW  - Childhood
KW  - Epidemiological study
KW  - Genitoanal region
KW  - Adamantiades-Behçet disease
KW  - Behçet’s disease
KW  - Uveitis
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-196616
SN  - 1018-8665
SN  - 1421-9832
N1  - This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.
VL  - 232
IS  - 2
SP  - 129
EP  - 136
ER  - 
TY  - JOUR
A1  - Sbiera, Iuliu
A1  - Kircher, Stefan
A1  - Altieri, Barbara
A1  - Lenz, Kerstin
A1  - Hantel, Constanze
A1  - Fassnacht, Martin
A1  - Sbiera, Silviu
A1  - Kroiss, Matthias
T1  - Role of FGF Receptors and Their Pathways in Adrenocortical Tumors and Possible Therapeutic Implications
JF  - Frontiers in Endocrinology
N2  - Adrenocortical carcinoma (ACC) is a rare endocrine malignancy and treatment of advanced disease is challenging. Clinical trials with multi-tyrosine kinase inhibitors in the past have yielded disappointing results. Here, we investigated fibroblast growth factor (FGF) receptors and their pathways in adrenocortical tumors as potential treatment targets. We performed real-time RT-PCR of 93 FGF pathway related genes in a cohort of 39 fresh frozen benign and malignant adrenocortical, 9 non-adrenal tissues and 4 cell lines. The expression of FGF receptors was validated in 166 formalin-fixed paraffin embedded (FFPE) tissues using RNA in situ hybridization (RNAscope) and correlated with clinical data. In malignant compared to benign adrenal tumors, we found significant differences in the expression of 16/94 FGF receptor pathway related genes. Genes involved in tissue differentiation and metastatic spread through epithelial to mesechymal transition were most strongly altered. The therapeutically targetable FGF receptors 1 and 4 were upregulated 4.6- and 6-fold, respectively, in malignant compared to benign adrenocortical tumors, which was confirmed by RNAscope in FFPE samples. High expression of FGFR1 and 4 was significantly associated with worse patient prognosis in univariate analysis. After multivariate adjustment for the known prognostic factors Ki-67 and ENSAT tumor stage, FGFR1 remained significantly associated with recurrence-free survival (HR=6.10, 95%CI: 1.78 – 20.86, p=0.004) and FGFR4 with overall survival (HR=3.23, 95%CI: 1.52 – 6.88, p=0.002). Collectively, our study supports a role of FGF pathways in malignant adrenocortical tumors. Quantification of FGF receptors may enable a stratification of ACC for the use of FGFR inhibitors in future clinical trials.
KW  - normal adrenal glands
KW  - adrenocortical tumors
KW  - FGF-pathway
KW  - FGFR
KW  - RNA Expression
KW  - RNAScope
KW  - unsupervised clustering
KW  - patient survival
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-251953
SN  - 1664-2392
VL  - 12
ER  - 
TY  - JOUR
A1  - Shaikh, Haroon
A1  - Vargas, Juan Gamboa
A1  - Mokhtari, Zeinab
A1  - Jarick, Katja J.
A1  - Ulbrich, Maria
A1  - Mosca, Josefina Peña
A1  - Viera, Estibaliz Arellano
A1  - Graf, Caroline
A1  - Le, Duc-Dung
A1  - Heinze, Katrin G.
A1  - Büttner-Herold, Maike
A1  - Rosenwald, Andreas
A1  - Pezoldt, Joern
A1  - Huehn, Jochen
A1  - Beilhack, Andreas
T1  - Mesenteric Lymph Node Transplantation in Mice to Study Immune Responses of the Gastrointestinal Tract
JF  - Frontiers in Immunology
N2  - Mesenteric lymph nodes (mLNs) are sentinel sites of enteral immunosurveillance and immune homeostasis. Immune cells from the gastrointestinal tract (GIT) are constantly recruited to the mLNs in steady-state and under inflammatory conditions resulting in the induction of tolerance and immune cells activation, respectively. Surgical dissection and transplantation of lymph nodes (LN) is a technique that has supported seminal work to study LN function and is useful to investigate resident stromal and endothelial cell biology and their cellular interactions in experimental disease models. Here, we provide a detailed protocol of syngeneic mLN transplantation and report assays to analyze effective mLN engraftment in congenic recipients. Transplanted mLNs allow to study T cell activation and proliferation in preclinical mouse models. Donor mLNs proved viable and functional after surgical transplantation and regenerated blood and lymphatic vessels. Immune cells from the host completely colonized the transplanted mLNs within 7-8 weeks after the surgical intervention. After allogeneic hematopoietic cell transplantation (allo-HCT), adoptively transferred allogeneic CD4+ T cells from FVB/N (H-2q) mice homed to the transplanted mLNs in C57BL/6 (H-2b) recipients during the initiation phase of acute graft-versus-host disease (aGvHD). These CD4+ T cells retained full proliferative capacity and upregulated effector and gut homing molecules comparable to those in mLNs from unmanipulated wild-type recipients. Wild type mLNs transplanted into MHCII deficient syngeneic hosts sufficed to activate alloreactive T cells upon allogeneic hematopoietic cell transplantation, even in the absence of MHCII+ CD11c+ myeloid cells. These data support that orthotopically transplanted mLNs maintain physiological functions after transplantation. The technique of LN transplantation can be applied to study migratory and resident cell compartment interactions in mLNs as well as immune reactions from and to the gut under inflammatory and non-inflammatory conditions.
KW  - acute graft-versus host disease
KW  - alloreactive T cells
KW  - mesenteric lymph node
KW  - lymph node transplantation
KW  - mouse models
KW  - lymph node stromal cells
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-244869
SN  - 1664-3224
VL  - 12
ER  - 
TY  - JOUR
A1  - Majumder, Snigdha
A1  - Jugovic, Isabelle
A1  - Saul, Domenica
A1  - Bell, Luisa
A1  - Hundhausen, Nadine
A1  - Seal, Rishav
A1  - Beilhack, Andreas
A1  - Rosenwald, Andreas
A1  - Mougiakakos, Dimitrios
A1  - Berberich-Siebelt, Friederike
T1  - Rapid and Efficient Gene Editing for Direct Transplantation of Naive Murine Cas9\(^+\) T Cells
JF  - Frontiers in Immunology
N2  - Gene editing of primary T cells is a difficult task. However, it is important for research and especially for clinical T-cell transfers. CRISPR/Cas9 is the most powerful gene-editing technique. It has to be applied to cells by either retroviral transduction or electroporation of ribonucleoprotein complexes. Only the latter is possible with resting T cells. Here, we make use of Cas9 transgenic mice and demonstrate nucleofection of pre-stimulated and, importantly, of naive CD3\(^+\) T cells with guideRNA only. This proved to be rapid and efficient with no need of further selection. In the mixture of Cas9\(^+\)CD3\(^+\) T cells, CD4\(^+\) and CD8\(^+\) conventional as well as regulatory T cells were targeted concurrently. IL-7 supported survival and naivety in vitro, but T cells were also transplantable immediately after nucleofection and elicited their function like unprocessed T cells. Accordingly, metabolic reprogramming reached normal levels within days. In a major mismatch model of GvHD, not only ablation of NFATc1 and/or NFATc2, but also of the NFAT-target gene IRF4 in naïve primary murine Cas9\(^+\)CD3\(^+\) T cells by gRNA-only nucleofection ameliorated GvHD. However, pre-activated murine T cells could not achieve long-term protection from GvHD upon single NFATc1 or NFATc2 knockout. This emphasizes the necessity of gene-editing and transferring unstimulated human T cells during allogenic hematopoietic stem cell transplantation.
KW  - CRISPR/Cas9
KW  - gRNA-only
KW  - GvHD
KW  - metabolism
KW  - NFAT
KW  - naive T-cell gene editing
KW  - T-cell transfer
KW  - IRF4
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242896
SN  - 1664-3224
VL  - 12
ER  - 
TY  - JOUR
A1  - Hartmann, Oliver
A1  - Reissland, Michaela
A1  - Maier, Carina R.
A1  - Fischer, Thomas
A1  - Prieto-Garcia, Cristian
A1  - Baluapuri, Apoorva
A1  - Schwarz, Jessica
A1  - Schmitz, Werner
A1  - Garrido-Rodriguez, Martin
A1  - Pahor, Nikolett
A1  - Davies, Clare C.
A1  - Bassermann, Florian
A1  - Orian, Amir
A1  - Wolf, Elmar
A1  - Schulze, Almut
A1  - Calzado, Marco A.
A1  - Rosenfeldt, Mathias T.
A1  - Diefenbacher, Markus E.
T1  - Implementation of CRISPR/Cas9 Genome Editing to Generate Murine Lung Cancer Models That Depict the Mutational Landscape of Human Disease
JF  - Frontiers in Cell and Developmental Biology
N2  - Lung cancer is the most common cancer worldwide and the leading cause of cancer-related deaths in both men and women. Despite the development of novel therapeutic interventions, the 5-year survival rate for non-small cell lung cancer (NSCLC) patients remains low, demonstrating the necessity for novel treatments. One strategy to improve translational research is the development of surrogate models reflecting somatic mutations identified in lung cancer patients as these impact treatment responses. With the advent of CRISPR-mediated genome editing, gene deletion as well as site-directed integration of point mutations enabled us to model human malignancies in more detail than ever before. Here, we report that by using CRISPR/Cas9-mediated targeting of Trp53 and KRas, we recapitulated the classic murine NSCLC model Trp53fl/fl:lsl-KRasG12D/wt. Developing tumors were indistinguishable from Trp53fl/fl:lsl-KRasG12D/wt-derived tumors with regard to morphology, marker expression, and transcriptional profiles. We demonstrate the applicability of CRISPR for tumor modeling in vivo and ameliorating the need to use conventional genetically engineered mouse models. Furthermore, tumor onset was not only achieved in constitutive Cas9 expression but also in wild-type animals via infection of lung epithelial cells with two discrete AAVs encoding different parts of the CRISPR machinery. While conventional mouse models require extensive husbandry to integrate new genetic features allowing for gene targeting, basic molecular methods suffice to inflict the desired genetic alterations in vivo. Utilizing the CRISPR toolbox, in vivo cancer research and modeling is rapidly evolving and enables researchers to swiftly develop new, clinically relevant surrogate models for translational research.
KW  - non-small cell lung cancer
KW  - CRISPR-Cas9
KW  - mouse model
KW  - lung cancer
KW  - MYC
KW  - JUN
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-230949
SN  - 2296-634X
VL  - 9
ER  - 
TY  - JOUR
A1  - Lenschow, Christina
A1  - Fuss, Carmina Teresa
A1  - Kircher, Stefan
A1  - Buck, Andreas
A1  - Kickuth, Ralph
A1  - Reibetanz, Joachim
A1  - Wiegering, Armin
A1  - Stenzinger, Albrecht
A1  - Hübschmann, Daniel
A1  - Germer, Christoph Thomas
A1  - Fassnacht, Martin
A1  - Fröhling, Stefan
A1  - Schlegel, Nicolas
A1  - Kroiss, Matthias
T1  - Case Report: Abdominal Lymph Node Metastases of Parathyroid Carcinoma: Diagnostic Workup, Molecular Diagnosis, and Clinical Management
JF  - Frontiers in Endocrinology
N2  - Parathyroid carcinoma (PC) is an orphan malignancy accounting for only ~1% of all cases with primary hyperparathyroidism. The localization of recurrent PC is of critical importance and can be exceedingly difficult to diagnose and sometimes futile when common sites of recurrence in the neck and chest cannot be confirmed. Here, we present the diagnostic workup, molecular analysis and multimodal therapy of a 46-year old woman with the extraordinary manifestation of abdominal lymph node metastases 12 years after primary diagnosis of PC. The patient was referred to our endocrine tumor center in 2016 with the aim to localize the tumor causative of symptomatic biochemical recurrence. In view of the extensive previous workup we decided to perform [18F]FDG-PET-CT. A pathological lymph node in the liver hilus showed slightly increased FDG-uptake and hence was suspected as site of recurrence. Selective venous sampling confirmed increased parathyroid hormone concentration in liver veins. Abdominal lymph node metastasis was resected and histopathological examination confirmed PC. Within four months, the patient experienced biochemical recurrence and based on high tumor mutational burden detected in the surgical specimen by whole exome sequencing the patient received immunotherapy with pembrolizumab that led to a biochemical response. Subsequent to disease progression repeated abdominal lymph node resection was performed in 10/2018, 01/2019 and in 01/2020. Up to now (12/2020) the patient is biochemically free of disease. In conclusion, a multimodal diagnostic approach and therapy in an interdisciplinary setting is needed for patients with rare endocrine tumors. Molecular analyses may inform additional treatment options including checkpoint inhibitors such as pembrolizumab.
KW  - parathyroid carcinoma
KW  - abdominal lymph node metastases
KW  - molecular diagnostics
KW  - repeated surgery
KW  - [18F]FDG-PET-CT
KW  - immune check inhibitor
KW  - pembrolizumab
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233362
SN  - 1664-2392
VL  - 12
ER  - 
TY  - JOUR
A1  - Marquardt, André
A1  - Solimando, Antonio Giovanni
A1  - Kerscher, Alexander
A1  - Bittrich, Max
A1  - Kalogirou, Charis
A1  - Kübler, Hubert
A1  - Rosenwald, Andreas
A1  - Bargou, Ralf
A1  - Kollmannsberger, Philip
A1  - Schilling, Bastian
A1  - Meierjohann, Svenja
A1  - Krebs, Markus
T1  - Subgroup-Independent Mapping of Renal Cell Carcinoma — Machine Learning Reveals Prognostic Mitochondrial Gene Signature Beyond Histopathologic Boundaries
JF  - Frontiers in Oncology
N2  - Background: Renal cell carcinoma (RCC) is divided into three major histopathologic groups—clear cell (ccRCC), papillary (pRCC) and chromophobe RCC (chRCC). We performed a comprehensive re-analysis of publicly available RCC datasets from the TCGA (The Cancer Genome Atlas) database, thereby combining samples from all three subgroups, for an exploratory transcriptome profiling of RCC subgroups.
Materials and Methods: We used FPKM (fragments per kilobase per million) files derived from the ccRCC, pRCC and chRCC cohorts of the TCGA database, representing transcriptomic data of 891 patients. Using principal component analysis, we visualized datasets as t-SNE plot for cluster detection. Clusters were characterized by machine learning, resulting gene signatures were validated by correlation analyses in the TCGA dataset and three external datasets (ICGC RECA-EU, CPTAC-3-Kidney, and GSE157256).
Results: Many RCC samples co-clustered according to histopathology. However, a substantial number of samples clustered independently from histopathologic origin (mixed subgroup)—demonstrating divergence between histopathology and transcriptomic data. Further analyses of mixed subgroup via machine learning revealed a predominant mitochondrial gene signature—a trait previously known for chRCC—across all histopathologic subgroups. Additionally, ccRCC samples from mixed subgroup presented an inverse correlation of mitochondrial and angiogenesis-related genes in the TCGA and in three external validation cohorts. Moreover, mixed subgroup affiliation was associated with a highly significant shorter overall survival for patients with ccRCC—and a highly significant longer overall survival for chRCC patients.
Conclusions: Pan-RCC clustering according to RNA-sequencing data revealed a distinct histology-independent subgroup characterized by strengthened mitochondrial and weakened angiogenesis-related gene signatures. Moreover, affiliation to mixed subgroup went along with a significantly shorter overall survival for ccRCC and a longer overall survival for chRCC patients. Further research could offer a therapy stratification by specifically addressing the mitochondrial metabolism of such tumors and its microenvironment.
KW  - kidney cancer
KW  - pan-RCC
KW  - machine learning
KW  - mitochondrial DNA
KW  - mtDNA
KW  - mTOR
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232107
SN  - 2234-943X
VL  - 11
ER  - 
TY  - JOUR
A1  - Friedmann Angeli, José Pedro
A1  - Meierjohann, Svenja
T1  - NRF2‐dependent stress defense in tumor antioxidant control and immune evasion
JF  - Pigment Cell & Melanoma Research
N2  - The transcription factor NRF2 is known as the master regulator of the oxidative stress response. Tumor entities presenting oncogenic activation of NRF2, such as lung adenocarcinoma, are associated with drug resistance, and accumulating evidence demonstrates its involvement in immune evasion. In other cancer types, the KEAP1/NRF2 pathway is not commonly mutated, but NRF2 is activated by other means such as radiation, oncogenic activity, cytokines, or other pro‐oxidant triggers characteristic of the tumor niche. The obvious effect of stress‐activated NRF2 is the protection from oxidative or electrophilic damage and the adaptation of the tumor metabolism to changing conditions. However, data from melanoma also reveal a role of NRF2 in modulating differentiation and suppressing anti‐tumor immunity. This review summarizes the function of NRF2 in this tumor entity and discusses the implications for current tumor therapies.
KW  - immune evasion
KW  - KEAP1
KW  - Nrf2
KW  - oxidative stress
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224536
VL  - 34
IS  - 2
SP  - 268
EP  - 279
ER  - 
TY  - JOUR
A1  - Hartmann, Sylvia
A1  - Plütschow, Annette
A1  - Mottok, Anja
A1  - Bernd, Heinz‐Wolfram
A1  - Feller, Alfred C.
A1  - Ott, German
A1  - Cogliatti, Sergio
A1  - Fend, Falko
A1  - Quintanilla‐Martinez, Leticia
A1  - Stein, Harald
A1  - Klapper, Wolfram
A1  - Möller, Peter
A1  - Rosenwald, Andreas
A1  - Engert, Andreas
A1  - Hansmann, Martin‐Leo
A1  - Eichenauer, Dennis A.
T1  - The time to relapse correlates with the histopathological growth pattern in nodular lymphocyte predominant Hodgkin lymphoma
JF  - American Journal of Hematology
N2  - Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) can present with different histopathological growth patterns. The impact of these histopathological growth patterns on relapse characteristics is unknown. We therefore analyzed paired biopsies obtained at initial diagnosis and relapse from 33 NLPHL patients who had received first‐line treatment within German Hodgkin Study Group (GHSG) trial protocols, and from a second cohort of 41 relapsed NLPHL patients who had been treated outside GHSG studies. Among the 33 GHSG patients, 21 patients presented with a typical growth pattern at initial diagnosis, whereas 12 patients had a variant histology. The histopathological growth patterns at initial diagnosis and at relapse were consistent in 67% of cases. A variant histology at initial diagnosis was associated with a shorter median time to lymphoma recurrence (2.8 vs 5.2 years; P = .0219). A similar tendency towards a shorter median time to lymphoma recurrence was observed for patients presenting with a variant histology at relapse, irrespective of the growth pattern at initial diagnosis. Results obtained from the 41 NLPHL patients who had been treated outside GHSG studies were comparable (median time to lymphoma recurrence for variant histology vs typical growth pattern at initial diagnosis: 1.5 vs 7.0 years). In conclusion, the histopathological growth pattern remains consistent at relapse in the majority of NLPHL cases, and has major impact on the time of relapse.
KW  - Hodgkin lymphoma
KW  - relapse
KW  - growth patterns
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212594
VL  - 94
IS  - 11
SP  - 1208
EP  - 1213
ER  - 
TY  - JOUR
A1  - Marquardt, André
A1  - Landwehr, Laura-Sophie
A1  - Ronchi, Cristina L.
A1  - di Dalmazi, Guido
A1  - Riester, Anna
A1  - Kollmannsberger, Philip
A1  - Altieri, Barbara
A1  - Fassnacht, Martin
A1  - Sbiera, Silviu
T1  - Identifying New Potential Biomarkers in Adrenocortical Tumors Based on mRNA Expression Data Using Machine Learning
JF  - Cancers
N2  - Simple Summary
Using a visual-based clustering method on the TCGA RNA sequencing data of a large adrenocortical carcinoma (ACC) cohort, we were able to classify these tumors in two distinct clusters largely overlapping with previously identified ones. As previously shown, the identified clusters also correlated with patient survival. Applying the visual clustering method to a second dataset also including benign adrenocortical samples additionally revealed that one of the ACC clusters is more closely located to the benign samples, providing a possible explanation for the better survival of this ACC cluster. Furthermore, the subsequent use of machine learning identified new possible biomarker genes with prognostic potential for this rare disease, that are significantly differentially expressed in the different survival clusters and should be further evaluated.

Abstract
Adrenocortical carcinoma (ACC) is a rare disease, associated with poor survival. Several “multiple-omics” studies characterizing ACC on a molecular level identified two different clusters correlating with patient survival (C1A and C1B). We here used the publicly available transcriptome data from the TCGA-ACC dataset (n = 79), applying machine learning (ML) methods to classify the ACC based on expression pattern in an unbiased manner. UMAP (uniform manifold approximation and projection)-based clustering resulted in two distinct groups, ACC-UMAP1 and ACC-UMAP2, that largely overlap with clusters C1B and C1A, respectively. However, subsequent use of random-forest-based learning revealed a set of new possible marker genes showing significant differential expression in the described clusters (e.g., SOAT1, EIF2A1). For validation purposes, we used a secondary dataset based on a previous study from our group, consisting of 4 normal adrenal glands and 52 benign and 7 malignant tumor samples. The results largely confirmed those obtained for the TCGA-ACC cohort. In addition, the ENSAT dataset showed a correlation between benign adrenocortical tumors and the good prognosis ACC cluster ACC-UMAP1/C1B. In conclusion, the use of ML approaches re-identified and redefined known prognostic ACC subgroups. On the other hand, the subsequent use of random-forest-based learning identified new possible prognostic marker genes for ACC.
KW  - adrenocortical carcinoma
KW  - in silico analysis
KW  - machine learning
KW  - bioinformatic clustering
KW  - biomarker prediction
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-246245
SN  - 2072-6694
VL  - 13
IS  - 18
ER  - 
TY  - JOUR
A1  - Sbiera, Iuliu
A1  - Kircher, Stefan
A1  - Altieri, Barbara
A1  - Fassnacht, Martin
A1  - Kroiss, Matthias
A1  - Sbiera, Silviu
T1  - Epithelial and Mesenchymal Markers in Adrenocortical Tissues: How Mesenchymal Are Adrenocortical Tissues?
JF  - Cancers
N2  - A clinically relevant proportion of adrenocortical carcinoma (ACC) cases shows a tendency to metastatic spread. The objective was to determine whether the epithelial to mesenchymal transition (EMT), a mechanism associated with metastasizing in several epithelial cancers, might play a crucial role in ACC. 138 ACC, 29 adrenocortical adenomas (ACA), three normal adrenal glands (NAG), and control tissue samples were assessed for the expression of epithelial (E-cadherin and EpCAM) and mesenchymal (N-cadherin, SLUG and SNAIL) markers by immunohistochemistry. Using real-time RT-PCR we quantified the alternative isoform splicing of FGFR 2 and 3, another known indicator of EMT. We also assessed the impact of these markers on clinical outcome. Results show that both normal and neoplastic adrenocortical tissues lacked expression of epithelial markers but strongly expressed mesenchymal markers N-cadherin and SLUG. FGFR isoform splicing confirmed higher similarity of adrenocortical tissues to mesenchymal compared to epithelial tissues. In ACC, higher SLUG expression was associated with clinical markers indicating aggressiveness, while N-cadherin expression inversely associated with these markers. In conclusion, we could not find any indication of EMT as all adrenocortical tissues lacked expression of epithelial markers and exhibited closer similarity to mesenchymal tissues. However, while N-cadherin might play a positive role in tissue structure upkeep, SLUG seems to be associated with a more aggressive phenotype.
KW  - adrenocortical tissues
KW  - EMT
KW  - epithelial markers
KW  - mesenchymal markers
KW  - recurrence-free survival
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-236486
SN  - 2072-6694
VL  - 13
IS  - 7
ER  - 
TY  - JOUR
A1  - Detomas, Mario
A1  - Altieri, Barbara
A1  - Schlötelburg, Wiebke
A1  - Appenzeller, Silke
A1  - Schlaffer, Sven
A1  - Coras, Roland
A1  - Schirbel, Andreas
A1  - Wild, Vanessa
A1  - Kroiss, Matthias
A1  - Sbiera, Silviu
A1  - Fassnacht, Martin
A1  - Deutschbein, Timo
T1  - Case Report: Consecutive Adrenal Cushing’s Syndrome and Cushing’s Disease in a Patient With Somatic CTNNB1, USP8, and NR3C1 Mutations
JF  - Frontiers in Endocrinology
N2  - The occurrence of different subtypes of endogenous Cushing’s syndrome (CS) in single individuals is extremely rare. We here present the case of a female patient who was successfully cured from adrenal CS 4 years before being diagnosed with Cushing’s disease (CD). The patient was diagnosed at the age of 50 with ACTH-independent CS and a left-sided adrenal adenoma, in January 2015. After adrenalectomy and histopathological confirmation of a cortisol-producing adrenocortical adenoma, biochemical hypercortisolism and clinical symptoms significantly improved. However, starting from 2018, the patient again developed signs and symptoms of recurrent CS. Subsequent biochemical and radiological workup suggested the presence of ACTH-dependent CS along with a pituitary microadenoma. The patient underwent successful transsphenoidal adenomectomy, and both postoperative adrenal insufficiency and histopathological workup confirmed the diagnosis of CD. Exome sequencing excluded a causative germline mutation but showed somatic mutations of the β-catenin protein gene (CTNNB1) in the adrenal adenoma, and of both the ubiquitin specific peptidase 8 (USP8) and the glucocorticoid receptor (NR3C1) genes in the pituitary adenoma. In conclusion, our case illustrates that both ACTH-independent and ACTH-dependent CS may develop in a single individual even without evidence for a common genetic background.
KW  - Cushing’s syndrome
KW  - Cushing’s disease
KW  - hypercortisolism
KW  - glucocorticoid excess
KW  - USP8
KW  - CTNNB1
KW  - NR3C1
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-244596
SN  - 1664-2392
VL  - 12
ER  - 
TY  - JOUR
A1  - Adam, Pia
A1  - Kircher, Stefan
A1  - Sbiera, Iuliu
A1  - Koehler, Viktoria Florentine
A1  - Berg, Elke
A1  - Knösel, Thomas
A1  - Sandner, Benjamin
A1  - Fenske, Wiebke Kristin
A1  - Bläker, Hendrik
A1  - Smaxwil, Constantin
A1  - Zielke, Andreas
A1  - Sipos, Bence
A1  - Allelein, Stephanie
A1  - Schott, Matthias
A1  - Dierks, Christine
A1  - Spitzweg, Christine
A1  - Fassnacht, Martin
A1  - Kroiss, Matthias
T1  - FGF-Receptors and PD-L1 in Anaplastic and Poorly Differentiated Thyroid Cancer: Evaluation of the Preclinical Rationale
JF  - Frontiers in Endocrinology
N2  - Background
Treatment options for poorly differentiated (PDTC) and anaplastic (ATC) thyroid carcinoma are unsatisfactory and prognosis is generally poor. Lenvatinib (LEN), a multi-tyrosine kinase inhibitor targeting fibroblast growth factor receptors (FGFR) 1-4 is approved for advanced radioiodine refractory thyroid carcinoma, but response to single agent is poor in ATC. Recent reports of combining LEN with PD-1 inhibitor pembrolizumab (PEM) are promising.


Materials and Methods
Primary ATC (n=93) and PDTC (n=47) tissue samples diagnosed 1997-2019 at five German tertiary care centers were assessed for PD-L1 expression by immunohistochemistry using Tumor Proportion Score (TPS). FGFR 1-4 mRNA was quantified in 31 ATC and 14 PDTC with RNAscope in-situ hybridization. Normal thyroid tissue (NT) and papillary thyroid carcinoma (PTC) served as controls. Disease specific survival (DSS) was the primary outcome variable.


Results
PD-L1 TPS≥50% was observed in 42% of ATC and 26% of PDTC specimens. Mean PD-L1 expression was significantly higher in ATC (TPS 30%) than in PDTC (5%; p<0.01) and NT (0%, p<0.001). 53% of PDTC samples had PD-L1 expression ≤5%. FGFR mRNA expression was generally low in all samples but combined FGFR1-4 expression was significantly higher in PDTC and ATC compared to NT (each p<0.001). No impact of PD-L1 and FGFR 1-4 expression was observed on DSS.


Conclusion
High tumoral expression of PD-L1 in a large proportion of ATCs and a subgroup of PDTCs provides a rationale for immune checkpoint inhibition. FGFR expression is low thyroid tumor cells. The clinically observed synergism of PEM with LEN may be caused by immune modulation.
KW  - tyrosine kinase inhibitor (TKI)
KW  - immune checkpoint inhibitor (ICI)
KW  - immunohistochemistry
KW  - immunotherapy
KW  - PD-L1
KW  - FGFR
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-244653
SN  - 1664-2392
VL  - 12
ER  - 
TY  - JOUR
A1  - Schuch, Luise A.
A1  - Forstner, Maria
A1  - Rapp, Christina K.
A1  - Li, Yang
A1  - Smith, Desiree E. C.
A1  - Mendes, Marisa I.
A1  - Delhommel, Florent
A1  - Sattler, Michael
A1  - Emiralioğlu, Nagehan
A1  - Taskiran, Ekim Z.
A1  - Orhan, Diclehan
A1  - Kiper, Nural
A1  - Rohlfs, Meino
A1  - Jeske, Tim
A1  - Hastreiter, Maximilian
A1  - Gerstlauer, Michael
A1  - Torrent‐Vernetta, Alba
A1  - Moreno‐Galdó, Antonio
A1  - Kammer, Birgit
A1  - Brasch, Frank
A1  - Reu‐Hofer, Simone
A1  - Griese, Matthias
T1  - FARS1‐related disorders caused by bi‐allelic mutations in cytosolic phenylalanyl‐tRNA synthetase genes: Look beyond the lungs!
JF  - Clinical Genetics
N2  - Aminoacyl‐tRNA synthetases (ARSs) catalyze the first step of protein biosynthesis (canonical function) and have additional (non‐canonical) functions outside of translation. Bi‐allelic pathogenic variants in genes encoding ARSs are associated with various recessive mitochondrial and multisystem disorders. We describe here a multisystem clinical phenotype based on bi‐allelic mutations in the two genes (FARSA, FARSB) encoding distinct subunits for tetrameric cytosolic phenylalanyl‐tRNA synthetase (FARS1). Interstitial lung disease with cholesterol pneumonitis on histology emerged as an early characteristic feature and significantly determined disease burden. Additional clinical characteristics of the patients included neurological findings, liver dysfunction, and connective tissue, muscular and vascular abnormalities. Structural modeling of newly identified missense mutations in the alpha subunit of FARS1, FARSA, showed exclusive mapping to the enzyme's conserved catalytic domain. Patient‐derived mutant cells displayed compromised aminoacylation activity in two cases, while remaining unaffected in another. Collectively, these findings expand current knowledge about the human ARS disease spectrum and support a loss of canonical and non‐canonical function in FARS1‐associated recessive disease.
KW  - aminoacyl‐tRNA synthetases
KW  - cholesterol pneumonitis
KW  - FARS1
KW  - children´s interstitial lung disease (chILD) lipoid pneumonia
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-238827
VL  - 99
IS  - 6
SP  - 789
EP  - 801
ER  - 
TY  - JOUR
A1  - Romero‐Olmedo, Addi J.
A1  - Schulz, Axel R.
A1  - Huber, Magdalena
A1  - Brehm, Corinna U.
A1  - Chang, Hyun‐Dong
A1  - Chiarolla, Cristina M.
A1  - Bopp, Tobias
A1  - Skevaki, Chrysanthi
A1  - Berberich‐Siebelt, Friederike
A1  - Radbruch, Andreas
A1  - Mei, Henrik E.
A1  - Lohoff, Michael
T1  - Deep phenotypical characterization of human CD3\(^{+}\)CD56\(^{+}\) T cells by mass cytometry
JF  - European Journal of Immunology
N2  - CD56\(^{+}\) T cells are a group of pro‐inflammatory CD3\(^{+}\) lymphocytes with characteristics of natural killer cells, being involved in antimicrobial immune defense. Here, we performed deep phenotypic profiling of CD3\(^{+}\)CD56\(^{+}\) cells in peripheral blood of normal human donors and individuals sensitized to birch‐pollen or/and house dust mite by high‐dimensional mass cytometry combined with manual and computational data analysis. A co‐regulation between major conventional T‐cell subsets and their respective CD3\(^{+}\)CD56\(^{+}\) cell counterparts appeared restricted to CD8\(^{+}\), MAIT, and TCRγδ\(^{+}\) T‐cell compartments. Interestingly, we find a co‐regulation of several CD3\(^{+}\)CD56\(^{+}\) cell subsets in allergic but not in healthy individuals. Moreover, using FlowSOM, we distinguished a variety of CD56\(^{+}\) T‐cell phenotypes demonstrating a hitherto underestimated heterogeneity among these cells. The novel CD3\(^{+}\)CD56\(^{+}\) subset description comprises phenotypes superimposed with naive, memory, type 1, 2, and 17 differentiation stages, in part represented by a phenotypical continuum. Frequencies of two out of 19 CD3\(^{+}\)CD56\(^{+}\) FlowSOM clusters were significantly diminished in allergic individuals, demonstrating less frequent presence of cells with cytolytic, presumably protective, capacity in these donors consistent with defective expansion or their recruitment to the affected tissue. Our results contribute to defining specific cell populations to be targeted during therapy for allergic conditions.
KW  - allergy
KW  - CD56
KW  - human
KW  - mass cytometry
KW  - T cells
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-225699
VL  - 51
IS  - 3
SP  - 672
EP  - 681
ER  - 
TY  - JOUR
A1  - Weich, Alexander
A1  - Werner, Rudolf A.
A1  - Buck, Andreas K.
A1  - Hartrampf, Philipp E.
A1  - Serfling, Sebastian E.
A1  - Scheurlen, Michael
A1  - Wester, Hans-Jürgen
A1  - Meining, Alexander
A1  - Kircher, Stefan
A1  - Higuchi, Takahiro
A1  - Pomper, Martin G.
A1  - Rowe, Steven P.
A1  - Lapa, Constantin
A1  - Kircher, Malte
T1  - CXCR4-Directed PET/CT in Patients with Newly Diagnosed Neuroendocrine Carcinomas
JF  - Diagnostics
N2  - We aimed to elucidate the diagnostic potential of the C-X-C motif chemokine receptor 4 (CXCR4)-directed positron emission tomography (PET) tracer \(^{68}\)Ga-Pentixafor in patients with poorly differentiated neuroendocrine carcinomas (NEC), relative to the established reference standard \(^{18}\)F-FDG PET/computed tomography (CT). In our database, we retrospectively identified 11 treatment-naïve patients with histologically proven NEC, who underwent \(^{18}\)F-FDG and CXCR4-directed PET/CT for staging and therapy planning. The images were analyzed on a per-patient and per-lesion basis and compared to immunohistochemical staining (IHC) of CXCR4 from PET-guided biopsies. \(^{68}\)Ga-Pentixafor visualized tumor lesions in 10/11 subjects, while \(^{18}\)F-FDG revealed sites of disease in all 11 patients. Although weak to moderate CXCR4 expression could be corroborated by IHC in 10/11 cases, \(^{18}\)F-FDG PET/CT detected significantly more tumor lesions (102 vs. 42; total lesions, n = 107; p < 0.001). Semi-quantitative analysis revealed markedly higher 18F-FDG uptake as compared to \(^{68}\)Ga-Pentixafor (maximum and mean standardized uptake values (SUV) and tumor-to-background ratios (TBR) of cancerous lesions, SUVmax: 12.8 ± 9.8 vs. 5.2 ± 3.7; SUVmean: 7.4 ± 5.4 vs. 3.1 ± 3.2, p < 0.001; and, TBR 7.2 ± 7.9 vs. 3.4 ± 3.0, p < 0.001). Non-invasive imaging of CXCR4 expression in NEC is inferior to the reference standard \(^{18}\)F-FDG PET/CT.
KW  - CXCR4
KW  - NET
KW  - NEC
KW  - <sup>68</sup>Ga-Pentixafor
KW  - <sup>18</sup>F-FDG
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-234231
SN  - 2075-4418
VL  - 11
IS  - 4
ER  - 
TY  - JOUR
A1  - Rosenfeldt, Mathias T.
A1  - Hartmann, Elena M.
A1  - Leng, Corinna
A1  - Rosenwald, Andreas
A1  - Anagnostopoulos, Ioannis
T1  - A case of nodular lymphocyte predominant Hodgkin lymphoma with unexpected EBV-latency type
JF  - Annals of Hematology
N2  - No abstract available.
KW  - nodular lymphcyte
KW  - Hodgkin lymphoma
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232571
SN  - 0939-5555
VL  - 100
ER  - 
TY  - JOUR
A1  - Zamò, Alberto
A1  - Johnston, Peter
A1  - Attygalle, Ayoma D
A1  - Laurent, Camille
A1  - Arber, Daniel A
A1  - Fend, Falko
T1  - Aggressive B‐cell lymphomas with a primary bone marrow presentation
JF  - Histopathology
KW  - aggressive B‐cell lymphoma
KW  - bone marrow biopsy
KW  - bone marrow–spleen–liver large B‐cell lymphoma
KW  - diffuse large B‐cell lymphoma
KW  - EAHP/SH bone marrow workshop
KW  - high‐grade B‐cell lymphoma
KW  - intravascular large B‐cell lymphoma
KW  - primary bone marrow presentation
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-218327
VL  - 77
IS  - 3
SP  - 369
EP  - 379
ER  - 
TY  - JOUR
A1  - Rauert-Wunderlich, Hilka
A1  - Berberich, Ingolf
A1  - Rosenwald, Andreas
A1  - Rudelius, Martina
T1  - CD40L mediated alternative NF kappa B-signaling induces resistance to BCR-inhibitors in patients with mantle cell lymphoma
JF  - Cell Death & Disease
N2  - Drug resistance is a significant obstacle in cancer treatment and therefore a frequent subject of research. Developed or primary resistance limits the treatment success of inhibitors of the B cell receptor (BCR) pathway in mantle cell lymphoma (MCL) patients. Recent research has highlighted the role of the nuclear factor-kappa B (NF kappa B) pathway in the context of resistance to BCR inhibitors in MCL. In this study, we analyzed the dependency of MCL cell lines on NF kappa B signaling and illustrated the ability of CD40L to activate the alternative NF kappa B pathway in MCL. This activation leads to independency of classical NF kappa B signaling and results in resistance to BCR inhibitors. Therefore, ligands (such as CD40L) and their activation of the alternative NF kappa B pathway have a major impact on the drug response in MCL. Furthermore, this study indicates a protective role for cells expressing specific ligands as microenvironmental niches for MCL cells and underlines the significance of therapeutically targeting alternative NF kappa B signaling in MCL.
KW  - Bruton Tyrosine Kinase
KW  - Tumor Microenvironment
KW  - Targeted Therapies
KW  - Ibrutinib
KW  - Pathway
KW  - Malignancies
KW  - Activation
KW  - Ligand
KW  - Proliferation
KW  - PCI-32765
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-225027
IS  - 9
ER  - 
TY  - JOUR
A1  - Upcin, Berin
A1  - Henke, Erik
A1  - Kleefeldt, Florian
A1  - Hoffmann, Helene
A1  - Rosenwald, Andreas
A1  - Irmak-Sav, Ster
A1  - Aktas, Huseyin Bertal
A1  - Rückschloß, Uwe
A1  - Ergün, Süleyman
T1  - Contribution of adventitia-derived stem and progenitor cells to new vessel formation in tumors
JF  - Cells
N2  - Blocking tumor vascularization has not yet come to fruition to the extent it was hoped for, as angiogenesis inhibitors have shown only partial success in the clinic. We hypothesized that under- appreciated vascular wall-resident stem and progenitor cells (VW-SPCs) might be involved in tumor vascularization and influence effectiveness of anti-angiogenic therapy. Indeed, in patient samples, we observed that vascular adventitia-resident CD34\(^+\) VW-SPCs are recruited to tumors in situ from co-opted vessels. To elucidate this in detail, we established an ex vivo model using concomitant embedding of multi-cellular tumor spheroids (MCTS) and mouse aortic rings (ARs) into collagen
gels, similar to the so-called aortic ring assay (ARA). Moreover, ARA was modified by removing the ARs’ adventitia that harbors VW-SPCs. Thus, this model enabled distinguishing the contribution of VW-SPCs from that of mature endothelial cells (ECs) to new vessel formation. Our results show that the formation of capillary-like sprouts is considerably delayed, and their number and network formation were significantly reduced by removing the adventitia. Substituting iPSC-derived neural spheroids for MCTS resulted in distinct sprouting patterns that were also strongly influenced by the
presence or absence of VW-SPCs, also underlying the involvement of these cells in non-pathological vascularization. Our data suggest that more comprehensive approaches are needed in order to block all of the mechanisms contributing to tumor vascularization.
KW  - vascularization model
KW  - tumor spheroids
KW  - vascular wall stem and progenitor cells
KW  - aortic adventitia
KW  - vasculogenesis
KW  - tumor-vessel wall-interface model
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-242577
VL  - 10
IS  - 7
ER  - 
TY  - JOUR
A1  - Almanzar, Giovanni
A1  - Klein, Matthias
A1  - Schmalzing, Marc
A1  - Hilligardt, Deborah
A1  - El Hajj, Nady
A1  - Kneitz, Hermann
A1  - Wild, Vanessa
A1  - Rosenwald, Andreas
A1  - Benoit, Sandrine
A1  - Hamm, Henning
A1  - Tony, Hans-Peter
A1  - Haaf, Thomas
A1  - Goebeler, Matthias
A1  - Prelog, Martina
T1  - Disease Manifestation and Inflammatory Activity as Modulators of Th17/Treg Balance and RORC/FoxP3 Methylation in Systemic Sclerosis
JF  - International Archives of Allergy and Immunology
N2  - Background: There is much evidence that T cells are strongly involved in the pathogenesis of localized and systemic forms of scleroderma (SSc). A dysbalance between FoxP3+ regulatory CD4+ T cells (Tregs) and inflammatory T-helper (Th) 17 cells has been suggested. Methods: The study aimed (1) to investigate the phenotypical and functional characteristics of Th17 and Tregs in SSc patients depending on disease manifestation (limited vs. diffuse cutaneous SSc, dcSSc) and activity, and (2) the transcriptional level and methylation status of Th17- and Treg-specific transcription factors. Results: There was a concurrent accumulation of circulating peripheral IL-17-producing CCR6+ Th cells and FoxP3+ Tregs in patients with dcSSc. At the transcriptional level, Th17- and Treg-associated transcription factors were elevated in SSc. A strong association with high circulating Th17 and Tregs was seen with early, active, and severe disease presentation. However, a diminished suppressive function on autologous lymphocytes was found in SSc-derived Tregs. Significant relative hypermethylation was seen at the gene level for RORC1 and RORC2 in SSc, particularly in patients with high inflammatory activity. Conclusions: Besides the high transcriptional activity of T cells, attributed to Treg or Th17 phenotype, in active SSc disease, Tregs may be insufficient to produce high amounts of IL-10 or to control proliferative activity of effector T cells in SSc. Our results suggest a high plasticity of Tregs strongly associated with the Th17 phenotype. Future directions may focus on enhancing Treg functions and stabilization of the Treg phenotype.
KW  - methylation
KW  - systemic sclerosis
KW  - suppression
KW  - Tregs
KW  - Th17
Y1  - 2016
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-196577
SN  - 1018-2438
SN  - 1423-0097
N1  - This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.
VL  - 171
IS  - 2
ER  - 
TY  - JOUR
A1  - Zhou, Xiang
A1  - Steinhardt, Maximilian Johannes
A1  - Düll, Johannes
A1  - Krummenast, Franziska
A1  - Danhof, Sophia
A1  - Meckel, Katharina
A1  - Nickel, Katharina
A1  - Grathwohl, Denise
A1  - Leicht, Hans‐Benno
A1  - Rosenwald, Andreas
A1  - Einsele, Hermann
A1  - Rasche, Leo
A1  - Kortüm, Martin
T1  - Obinutuzumab and venetoclax induced complete remission in a patient with ibrutinib‐resistant non‐nodal leukemic mantle cell lymphoma
JF  - European Journal of Haematology
N2  - We herein report the case of a 73‐year‐old male patient who was diagnosed with leukemic non‐nodal MCL. This patient had received six cycles of bendamustine, which resulted in a transient remission, and a second‐line therapy with ibrutinib, which unfortunately failed to induce remission. We started a treatment with single‐agent obinutuzumab at a dose of 20 mg on day 1, 50 mg on day 2‐4, 330 mg on day 5, and 1000 mg on day 6. The laboratory analysis showed a rapid decrease of leukocyte count. Four weeks later, we repeated the treatment with obinutuzumab at a dose of 1000 mg q4w and started a therapy with venetoclax at a dose of 400 mg qd, which could be increased to 800 mg qd from the third cycle. This combination therapy was well tolerated. The patient achieved a complete remission (CR) after three cycles of obinutuzumab and venetoclax. To date, the patient has a progression‐free survival of 17 months under ongoing obinutuzumab maintenance q4w. This is the first report about obinutuzumab and venetoclax induced CR in rituximab‐intolerant patient with an ibrutinib‐resistant MCL. This case suggests that obinutuzumab‐ and venetoclax‐based combination therapy might be salvage therapy in patients with ibrutinib‐resistant MCL.
KW  - mantle cell lymphoma
KW  - obinutuzumab
KW  - venetoclax
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-215513
VL  - 104
IS  - 4
SP  - 352
EP  - 355
ER  - 
TY  - JOUR
A1  - Bohnert, Simone
A1  - Seiffert, Anja
A1  - Trella, Stefanie
A1  - Bohnert, Michael
A1  - Distel, Luitpold
A1  - Ondruschka, Benjamin
A1  - Monoranu, Camelia-Marie
T1  - TMEM119 as a specific marker of microglia reaction in traumatic brain injury in postmortem examination
JF  - International Journal of Legal Medicine
N2  - The aim of the present study was a refined analysis of neuroinflammation including TMEM119 as a useful microglia-specific marker in forensic assessments of traumatic causes of death, e.g., traumatic brain injury (TBI). Human brain tissue samples were obtained from autopsies and divided into cases with lethal TBI (n = 25) and subdivided into three groups according to their trauma survival time and compared with an age-, gender-, and postmortem interval-matched cohort of sudden cardiovascular fatalities as controls (n = 23). Brain tissue samples next to cortex contusions and surrounding white matter as well as samples of the ipsilateral uninjured brain stem and cerebellum were collected and stained immunohistochemically with antibodies against TMEM119, CD206, and CCR2. We could document the highest number of TMEM119-positive cells in acute TBI death with highly significant differences to the control numbers. CCR2-positive monocytes showed a significantly higher cell count in the cortex samples of TBI cases than in the controls with an increasing number of immunopositive cells over time. The number of CD206-positive M2 microglial cells increased survival time-dependent. After 3 days of survival, the cell number increased significantly in all four regions investigated compared with controls. In sum, we validate a specific and robustly expressed as well as fast reacting microglia marker, TMEM119, which distinguishes microglia from resident and infiltrating macrophages and thus offers a great potential for the estimation of the minimum survival time after TBI.
KW  - cerebrospinal fluid
KW  - forensic neuropathology
KW  - forensic neurotraumatology
KW  - immunohistochemistry
KW  - biomarker
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-235346
SN  - 0937-9827
VL  - 134
ER  - 
TY  - JOUR
A1  - Serfling, S.
A1  - Zhi, Y.
A1  - Schirbel, A.
A1  - Lindner, T.
A1  - Meyer, T.
A1  - Gerhard-Hartmann, E.
A1  - Lappa, C.
A1  - Hagen, R.
A1  - Hackenberg, S.
A1  - Buck, A. K.
A1  - Scherzad, A.
T1  - Improved cancer detection in Waldeyer’s tonsillar ring by \(^{68}\)Ga-FAPI PET/CT imaging
JF  - European Journal of Nuclear Medicine and Molecular Imaging
N2  - Purpose
In cancer of unknown primary (CUP), positron emission tomography/computed tomography (PET/CT) with the glucose analog [\(^{18}\)F]FDG represents the standard imaging approach for localization of the malignant primary. Frequently, however, [\(^{18}\)F]FDG PET/CT cannot precisely distinguish between small occult tumors and chronic inflammation, especially in Waldeyer’s tonsillar ring. To improve the accuracy for detecting primary tumors in the Waldeyer’s tonsillar ring, the novel PET tracer [\(^{68}\)Ga]Ga-FAPI-4 for specific imaging of fibroblast activation protein (FAP) expression was used as a more specific target for cancer imaging.

Methods
Eight patients with suspicion of a malignant tumor in Waldeyer’s tonsillar ring or a CUP syndrome were examined. PET/CT scans with [\(^{18}\)F]-FDG and [\(^{68}\)Ga]Ga-FAPI-4 were performed for pre-operative tumor localization. After surgical resection, histopathological and immunohistochemical results were compared to PET/CT findings.

Results
Histopathology revealed a palatine or lingual tonsil carcinoma in all patients. In case of lymph node metastases smaller than 7 mm in size, the [\(^{18}\)F]FDG PET/CT detection rate of cervical lymph node metastases was higher than that of [\(^{68}\)Ga]FAPI PET/CT, while both tracers identified the primary tumors in all eight cases. The size of the primary and the lymph node metastases was directly correlated to the respective FAP expression, as detected by immunohistochemistry. The mean SUVmax for the primary tumors was 21.29 ± 7.97 for \(^{18}\)F-FDG and 16.06 ± 6.29 for \(^{68}\)Ga-FAPI, respectively (p = 0.2). The mean SUVmax for the healthy contralateral tonsils was 8.38 ± 2.45 for [\(^{18}\)F]FDG and 3.55 ± 0.47 for [\(^{68}\)Ga]FAPI (p < 0.001). The SUVmax ratio of [68Ga]FAPI was significantly different from [\(^{18}\)F] FDG (p = 0.03). Mean TBRmax for the [\(^{68}\)Ga]Ga-FAPI-4 tracer was markedly higher in comparison to [\(^{18}\)F]FDG (10.90 vs. 4.11).

Conclusion
Non-invasive imaging of FAP expression by [\(^{68}\)Ga]FAPI PET/CT resulted in a better visual detection of the malignant primary in CUP, as compared to [\(^{18}\)F]FDG imaging. However, the detection rate of lymph node metastases was inferior, presumably due to low FAP expression in small metastases. Nevertheless, by offering a detection method for primary tumors with the potential of lower false positive rates and thus avoiding biopsies, patients with CUP syndrome may benefit from [\(^{68}\)Ga]FAPI PET/CT imaging.
KW  - Waldeyer’s tonsillar ring
KW  - cancer of unknown primary (CUP)
KW  - positron emission tomography/computed tomography
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-235271
SN  - 1619-7070
VL  - 48
ER  - 
TY  - JOUR
A1  - Kessler, Almuth F.
A1  - Feldheim, Jonas
A1  - Schmitt, Dominik
A1  - Feldheim, Julia J.
A1  - Monoranu, Camelia M.
A1  - Ernestus, Ralf-Ingo
A1  - Löhr, Mario
A1  - Hagemann, Carsten
T1  - Monopolar Spindle 1 Kinase (MPS1/TTK) mRNA Expression is Associated with Earlier Development of Clinical Symptoms, Tumor Aggressiveness and Survival of Glioma Patients
JF  - Biomedicines
N2  - Inhibition of the protein kinase MPS1, a mitotic spindle-checkpoint regulator, reinforces the effects of multiple therapies against glioblastoma multiforme (GBM) in experimental settings. We analyzed MPS1 mRNA-expression in gliomas WHO grade II, III and in clinical subgroups of GBM. Data were obtained by qPCR analysis of tumor and healthy brain specimens and correlated with the patients’ clinical data. MPS1 was overexpressed in all gliomas on an mRNA level (ANOVA, p < 0.01) and correlated with tumor aggressiveness. We explain previously published conflicting results on survival: high MPS1 was associated with poorer long term survival when all gliomas were analyzed combined in one group (Cox regression: t < 24 months, p = 0.009, Hazard ratio: 8.0, 95% CI: 1.7–38.4), with poorer survival solely in low-grade gliomas (LogRank: p = 0.02, Cox regression: p = 0.06, Hazard-Ratio: 8.0, 95% CI: 0.9–66.7), but not in GBM (LogRank: p > 0.05). This might be due to their lower tumor volume at the therapy start. GBM patients with high MPS1 mRNA-expression developed clinical symptoms at an earlier stage. This, however, did not benefit their overall survival, most likely due to the more aggressive tumor growth. Since MPS1 mRNA-expression in gliomas was enhanced with increasing tumor aggressiveness, patients with the worst outcome might benefit best from a treatment directed against MPS1.
KW  - glioblastoma multiforme
KW  - low-grade glioma
KW  - astrocytoma
KW  - recurrence
KW  - multifocal growth
KW  - mRNA expression
KW  - MPS1
KW  - TTK
KW  - therapy
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-236105
VL  - 8
IS  - 7
ER  - 
TY  - JOUR
A1  - Nattmann, Anja
A1  - Breun, Maria
A1  - Monoranu, Camelia M.
A1  - Matthies, Cordula
A1  - Ernestus, Ralf-Ingo
A1  - Löhr, Mario
A1  - Hagemann, Carsten
T1  - Analysis of ADAM9 regulation and function in vestibular schwannoma primary cells
JF  - BMC Research Notes
N2  - Objective
Recently, we described a disintegrin and metalloproteinase 9 (ADAM9) overexpression by Schwann cells of vestibular schwannoma (VS) and suggested that it might be a marker for VS tumor growth and invasiveness. This research note provides additional data utilizing a small cohort of VS primary cultures and tissue samples. We examined whether reconstitution of Merlin expression in VS cells regulates ADAM9 protein expression and performed lentiviral ADAM9 knock down to investigate possible effects on VS cells numbers. Moreover, the co-localization of ADAM9 and Integrins α6 and α2β1, respectively, was examined by immunofluorescence double staining.

Results
ADAM9 expression was not regulated by Merlin in VS. However, ADAM9 knock down led to 58% reduction in cell numbers in VS primary cell cultures (p < 0.0001). While ADAM9 and Integrin α2β1 were co-localized in only 22% (2 of 9) of VS, ADAM9 and Integrin α6 were co-localized in 91% (10 of 11) of VS. Therefore, we provide first observations on possible regulatory functions of ADAM9 expression in VS.
KW  - vestibular schwannoma
KW  - pathogenesis
KW  - ADAM9
KW  - knock down
KW  - integrin
KW  - immunofuorescence double staining
KW  - Merlin
KW  - primary cell culture
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-231213
VL  - 13
ER  - 
TY  - JOUR
A1  - Wallstabe, Julia
A1  - Bussemer, Lydia
A1  - Groeber-Becker, Florian
A1  - Freund, Lukas
A1  - Alb, Mirian
A1  - Dragan, Mariola
A1  - Waaga-Gasser, Ana Maria
A1  - Jakubietz, Rafael
A1  - Kneitz, Hermann
A1  - Rosenwald, Andreas
A1  - Rebhan, Silke
A1  - Walles, Heike
A1  - Mielke, Stephan
T1  - Inflammation-Induced Tissue Damage Mimicking GvHD in Human Skin Models as Test Platform for Immunotherapeutics
JF  - ALTEX
N2  - Due to the rapidly increasing development and use of cellular products, there is a rising demand for non-animal-based test platforms to predict, study and treat undesired immunity. Here, we generated human organotypic skin models from human biopsies by isolating and expanding keratinocytes, fibroblasts and microvascular endothelial cells and seeding these components on a collagen matrix or a biological vascularized scaffold matrix in a bioreactor. We then were able to induce inflammation-mediated tissue damage by adding pre-stimulated, mismatched allogeneic lymphocytes and/or inflammatory cytokine-containing supernatants histomorphologically mimicking severe graft versus host disease (GvHD) of the skin. This could be prevented by the addition of immunosuppressants to the models. Consequently, these models harbor a promising potential to serve as a test platform for the prediction, prevention and treatment of GvHD. They also allow functional studies of immune effectors and suppressors including but not limited to allodepleted lymphocytes, gamma-delta T cells, regulatory T cells and mesenchymal stromal cells, which would otherwise be limited to animal models. Thus, the current test platform, developed with the limitation that no professional antigen presenting cells are in place, could greatly reduce animal testing for investigation of novel immune therapies.
KW  - inflammation-induced tissue demage
KW  - immunotherapeutics
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229974
VL  - 37
IS  - 3
ER  -