TY - JOUR A1 - Schartl, Manfred A1 - Shen, Yingjia A1 - Maurus, Katja A1 - Walter, Ron A1 - Tomlinson, Chad A1 - Wilson, Richard K. A1 - Postlethwait, John A1 - Warren, Wesley C. T1 - Whole body melanoma transcriptome response in medaka JF - PLoS ONE N2 - The incidence of malignant melanoma continues to increase each year with poor prognosis for survival in many relapse cases. To reverse this trend, whole body response measures are needed to discover collaborative paths to primary and secondary malignancy. Several species of fish provide excellent melanoma models because fish and human melanocytes both appear in the epidermis, and fish and human pigment cell tumors share conserved gene expression signatures. For the first time, we have examined the whole body transcriptome response to invasive melanoma as a prelude to using transcriptome profiling to screen for drugs in a medaka (Oryzias latipes) model. We generated RNA-seq data from whole body RNA isolates for controls and melanoma fish. After testing for differential expression, 396 genes had significantly different expression (adjusted p-value <0.02) in the whole body transcriptome between melanoma and control fish; 379 of these genes were matched to human orthologs with 233 having annotated human gene symbols and 14 matched genes that contain putative deleterious variants in human melanoma at varying levels of recurrence. A detailed canonical pathway evaluation for significant enrichment showed the top scoring pathway to be antigen presentation but also included the expected melanocyte development and pigmentation signaling pathway. Results revealed a profound down-regulation of genes involved in the immune response, especially the innate immune system. We hypothesize that the developing melanoma actively suppresses the immune system responses of the body in reacting to the invasive malignancy, and that this mal-adaptive response contributes to disease progression, a result that suggests our whole-body transcriptomic approach merits further use. In these findings, we also observed novel genes not yet identified in human melanoma expression studies and uncovered known and new candidate drug targets for further testing in this malignant melanoma medaka model. KW - metastatic melanoma KW - expression KW - fish KW - cancer KW - stage III KW - melanogenesis KW - genome cells KW - gene KW - contributes Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144714 VL - 10 IS - 12 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 - Patil, Sandeep S. A1 - Gentschev, Ivaylo A1 - Adelfinger, Marion A1 - Donat, Ulrike A1 - Hess, Michael A1 - Weibel, Stephanie A1 - Nolte, Ingo A1 - Frentzen, Alexa A1 - Szalay, Aladar A. T1 - Virotherapy of Canine Tumors with Oncolytic Vaccinia Virus GLV-1h109 Expressing an Anti-VEGF Single-Chain Antibody JF - PLoS One N2 - Virotherapy using oncolytic vaccinia virus (VACV) strains is one promising new strategy for cancer therapy. We have previously reported that oncolytic vaccinia virus strains expressing an anti-VEGF (Vascular Endothelial Growth Factor) single-chain antibody (scAb) GLAF-1 exhibited significant therapeutic efficacy for treatment of human tumor xenografts. Here, we describe the use of oncolytic vaccinia virus GLV-1h109 encoding GLAF-1 for canine cancer therapy. In this study we analyzed the virus-mediated delivery and production of scAb GLAF-1 and the oncolytic and immunological effects of the GLV-1h109 vaccinia virus strain against canine soft tissue sarcoma and canine prostate carcinoma in xenograft models. Cell culture data demonstrated that the GLV-1h109 virus efficiently infect, replicate in and destroy both tested canine cancer cell lines. In addition, successful expression of GLAF-1 was demonstrated in virus-infected canine cancer cells and the antibody specifically recognized canine VEGF. In two different xenograft models, the systemic administration of the GLV-1h109 virus was found to be safe and led to anti-tumor and immunological effects resulting in the significant reduction of tumor growth in comparison to untreated control mice. Furthermore, tumor-specific virus infection led to a continued production of functional scAb GLAF-1, resulting in inhibition of angiogenesis. Overall, the GLV-1h109-mediated cancer therapy and production of immunotherapeutic anti-VEGF scAb may open the way for combination therapy concept i.e. vaccinia virus mediated oncolysis and intratumoral production of therapeutic drugs in canine cancer patients. KW - angiogenesis KW - microenvironment KW - model KW - cancer KW - therapy KW - pet dogs KW - nude-mice KW - breast-tumors KW - microvascular density KW - endothelial growth-factor Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130039 VL - 7 IS - 10 ER - TY - JOUR A1 - Schäfer, Simon A1 - Weibel, Stephanie A1 - Donat, Ulrike A1 - Zhang, Quian A1 - Aguilar, Richard J. A1 - Chen, Nanhai G. A1 - Szalay, Aladar A. T1 - Vaccinia virus-mediated intra-tumoral expression of matrix metalloproteinase 9 enhances oncolysis of PC-3 xenograft tumors JF - BMC Cancer N2 - Background Oncolytic viruses, including vaccinia virus (VACV), are a promising alternative to classical mono-cancer treatment methods such as surgery, chemo- or radiotherapy. However, combined therapeutic modalities may be more effective than mono-therapies. In this study, we enhanced the effectiveness of oncolytic virotherapy by matrix metalloproteinase (MMP-9)-mediated degradation of proteins of the tumoral extracellular matrix (ECM), leading to increased viral distribution within the tumors. Methods For this study, the oncolytic vaccinia virus GLV-1h255, containing the mmp-9 gene, was constructed and used to treat PC-3 tumor-bearing mice, achieving an intra-tumoral over-expression of MMP-9. The intra-tumoral MMP-9 content was quantified by immunohistochemistry in tumor sections. Therapeutic efficacy of GLV-1h255 was evaluated by monitoring tumor growth kinetics and intra-tumoral virus titers. Microenvironmental changes mediated by the intra-tumoral MMP-9 over-expression were investigated by microscopic quantification of the collagen IV content, the blood vessel density (BVD) and the analysis of lymph node metastasis formation. Results GLV-1h255-treatment of PC-3 tumors led to a significant over-expression of intra-tumoral MMP-9, accompanied by a marked decrease in collagen IV content in infected tumor areas, when compared to GLV-1h68-infected tumor areas. This led to considerably elevated virus titers in GLV-1h255 infected tumors, and to enhanced tumor regression. The analysis of the BVD, as well as the lumbar and renal lymph node volumes, revealed lower BVD and significantly smaller lymph nodes in both GLV-1h68- and GLV-1h255- injected mice compared to those injected with PBS, indicating that MMP-9 over-expression does not alter the metastasis-reducing effect of oncolytic VACV. Conclusions Taken together, these results indicate that a GLV-1h255-mediated intra-tumoral over-expression of MMP-9 leads to a degradation of collagen IV, facilitating intra-tumoral viral dissemination, and resulting in accelerated tumor regression. We propose that approaches which enhance the oncolytic effect by increasing the intra-tumoral viral load, may be an effective way to improve therapeutic outcome. KW - microenvironment KW - angiogenesis KW - therapy KW - cancer KW - breast-tumors KW - matrix metalloproteinases KW - adenovirus KW - carcinoma KW - prostate KW - mice Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-140800 VL - 12 IS - 366 ER - TY - THES A1 - Heß, Michael T1 - Vaccinia virus-encoded bacterial beta-glucuronidase as a diagnostic biomarker for oncolytic virotherapy T1 - Vaccinia Virus-codierte bakterielle Beta-Glucuronidase als diagnostischer Biomarker in der onkolytischen Virotherapie N2 - Oncolytic virotherapy represents a promising approach to revolutionize cancer therapy. Several preclinical and clinical trials display the safety of oncolytic viruses as wells as their efficiency against solid tumors. The development of complementary diagnosis and monitoring concepts as well as the optimization of anti-tumor activity are key points of current virotherapy research. Within the framework of this thesis, the diagnostic and therapeutic prospects of beta-glucuronidase expressed by the oncolytic vaccinia virus strain GLV-1h68 were evaluated. In this regard, a beta-glucuronidase-based, therapy-accompanying biomarker test was established which is currently under clinical validation. By using fluorescent substrates, the activity of virally expressed beta-glucuronidase could be detected and quantified. Thereby conclusions about the replication kinetics of oncolytic viruses in animal models and virus-induced cancer cell lysis could be drawn. These findings finally led to the elaboration and establishment of a versatile biomarker assay which allows statements regarding the replication of oncolytic viruses in mice based on serum samples. Besides the analysis of retrospective conditions, this test is able to serve as therapy-accompanying monitoring tool for virotherapy approaches with beta-glucuronidase-expressing viruses. The newly developed assay also served as complement to routinely used plaque assays as well as reference for virally expressed anti-angiogenic antibodies in additional preclinical studies. Further validation of this biomarker test is currently taking place in the context of clinical trials with GL-ONC1 (clinical grade GLV-1h68) and has already shown promising preliminary results. It was furthermore demonstrated that fluorogenic substrates in combination with beta-glucuronidase expressed by oncolytic viruses facilitated the optical detection of solid tumors in preclinical models. In addition to diagnostic purposes, virus-encoded enzymes could also be combined with prodrugs resulting in an improved therapeutic outcome of oncolytic virotherapy. In further studies, the visualization of virus-induced immune reactions as well as the establishment of innovative concepts to improve the therapeutic outcome of oncolytic virotherapy could be accomplished. In conclusion, the results of this thesis provide crucial findings about the influence of virally expressed beta-glucuronidase on various diagnostic concepts in the context of oncolytic virotherapy. In addition, innovative monitoring and therapeutic strategies could be established. Our preclinical findings have important clinical influence, particularly by the development of a therapy-associated biomarker assay which is currently used in different clinical trials. N2 - Onkolytische Viren stellen einen vielversprechenden Therapieansatz dar, der die Behandlung von Krebserkrankungen revolutionieren könnte. Intensive präklinische und klinische Studien zeigen sowohl die körperliche Verträglichkeit von onkolytischen Viren, als auch deren Wirksamkeit gegenüber soliden Tumoren. Die Entwicklung von therapiebegleitenden Diagnose- und Monitoringkonzepten sowie eine Optimierung der Antitumorwirkung onkolytischer Viren stellen Eckpunkte der aktuellen Forschung auf dem Gebiet der Virotherapie dar. Im Rahmen dieser Arbeit wurde untersucht, welche diagnostischen und therapeutischen Möglichkeiten die virale Expression von beta-Glucuronidase durch den onkolytischen Vaccinia-Virus-Stamm GLV-1h68 eröffnet. In diesem Zusammenhang wurde ein, auf beta-Glucuronidase basierender, therapiebegleitender Biomarkertest entwickelt, dessen klinische Validierung derzeit stattfindet. Mit Hilfe von fluorogenen Substraten konnte die Aktivität viral exprimierter beta-Glucuronidase detektiert und quantifiziert werden. Dies lies direkte Rückschlüsse auf das Replikationsverhalten von onkolytischen Viren im Tiermodell zu und ermöglichte zudem Aussagen über die Zelllyse Virus-infizierter Krebszellen. Diese Erkenntnisse führten letztendlich zur Ausarbeitung und Etablierung eines vielseitig anwendbaren Biomarker-Assays, der es ermöglicht anhand von Blutproben Aussagen über das Replikationsverhalten onkolytischer Viren in Mäusen zu machen. Neben retrospektiven Analysen erlaubt dieser Test auch ein therapiebegleitendes Monitoring der onkolytischen Virotherapie mit beta-Glucuronidase-exprimierenden Viren. In weiteren präklinischen Untersuchungen diente der entwickelte Assay zudem als Ergänzung zum viralen Plaque Assays sowie als Referenz für Virus-exprimierte anti-angiogene Antikörper. Eine fortführende Validierung dieses neuartigen Biomarkertests findet derzeit im Rahmen humaner Studien mit der klinischen Formulierung von GLV-1h68, GL-ONC1, statt und zeigte bereits erste positive Resultate. Weiterhin konnte im Rahmen dieser Arbeit gezeigt werden, dass die Expression von beta-Glucuronidase durch onkolytische Viren in Verbindung mit fluoreszierenden Substraten eine optische Detektion von Karzinomen im präklinischen Tiermodell ermöglicht. Neben diagnostischen Zwecken, konnten Virus-kodierte Enzyme in Kombination mit Prodrugs genutzt werden, um den Therapieerfolg der onkolytischen Virotherapie zu verbessern. In zusätzlichen Studien konnten zudem Methoden zur Visualisierung der Virus-induzierten Immunantwort sowie neuartige Konzepte zur Therapieverbesserung etabliert werden. Zusammenfassend liefern die Ergebnisse der vorliegenden Arbeit wichtige Erkenntnisse über den Einfluss Virus-exprimierter beta-Glucuronidase auf unterschiedliche Diagnosekonzepte im Rahmen der onkolytischen Virotherapie. Daneben konnten entscheidende Erkenntnisse über den möglichen Einsatz neuer Monitoring- und Therapieansätze erzielt werden. Insbesondere durch die Entwicklung eines therapiebegleitenden Biomarkertests haben diese Resultate erheblichen Einfluss auf die weitere klinische Anwendung von onkolytischen Vaccinia-Viren. KW - Vaccinia-Virus KW - Glucuronidase KW - Krebs KW - cancer KW - oncolytic virus KW - biomarker KW - beta-glucuronidase Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-86789 ER - TY - JOUR A1 - Prieto-Garcia, Cristian A1 - Tomašković, Ines A1 - Shah, Varun Jayeshkumar A1 - Dikic, Ivan A1 - Diefenbacher, Markus T1 - USP28: oncogene or tumor suppressor? a unifying paradigm for squamous cell carcinoma JF - Cells N2 - Squamous cell carcinomas are therapeutically challenging tumor entities. Low response rates to radiotherapy and chemotherapy are commonly observed in squamous patients and, accordingly, the mortality rate is relatively high compared to other tumor entities. Recently, targeting USP28 has been emerged as a potential alternative to improve the therapeutic response and clinical outcomes of squamous patients. USP28 is a catalytically active deubiquitinase that governs a plethora of biological processes, including cellular proliferation, DNA damage repair, apoptosis and oncogenesis. In squamous cell carcinoma, USP28 is strongly expressed and stabilizes the essential squamous transcription factor ΔNp63, together with important oncogenic factors, such as NOTCH1, c-MYC and c-JUN. It is presumed that USP28 is an oncoprotein; however, recent data suggest that the deubiquitinase also has an antineoplastic effect regulating important tumor suppressor proteins, such as p53 and CHK2. In this review, we discuss: (1) The emerging role of USP28 in cancer. (2) The complexity and mutational landscape of squamous tumors. (3) The genetic alterations and cellular pathways that determine the function of USP28 in squamous cancer. (4) The development and current state of novel USP28 inhibitors. KW - USP28 KW - SCC KW - USP25 KW - FBXW7 KW - Tp63 KW - c-MYC KW - ΔNp63 KW - p53 KW - cancer KW - DUB inhibitor KW - squamous Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-248409 SN - 2073-4409 VL - 10 IS - 10 ER - TY - THES A1 - Gnamlin, Prisca T1 - Use of Tumor Vasculature for Successful Treatment of Carcinomas by Oncolytic Vaccinia Virus T1 - Die Tumorvasulatur in der erfolgreichen Therapie von Carcinomen durch onkolytische Vaccinia Viren N2 - Tumor-induced angiogenesis is of major interest for oncology research. Vascular endothelial growth factor (VEGF) is the most potent angiogenic factor characterized so far. VEGF blockade was shown to be sufficient for angiogenesis inhibition and subsequent tumor regression in several preclinical tumor models. Bevacizumab was the first treatment targeting specifically tumor-induced angiogenesis through VEGF blockade to be approved by the Food and Drugs Administration (FDA) for cancer treatment. However, after very promising results in preclinical evaluations, VEGF blockade did not show the expected success in patients. Some tumors became resistant to VEGF blockade. Several factors have been accounted responsible, the over-expression of other angiogenic factors, the noxious influence of VEFG blockade on normal tissues, the selection of hypoxia resistant neoplastic cells, the recruitment of hematopoietic progenitor cells and finally the transient nature of angiogenesis inhibition by VEGF blockade. The development of blocking agents against other angiogenic factors like placental growth factor (PlGF) and Angiopoietin-2 (Ang-2) allows the development of an anti-angiogenesis strategy adapted to the profile of the tumor. Oncolytic virotherapy uses the natural propensity of viruses to colonize tumors to treat cancer. The recombinant vaccinia virus GLV-1h68 was shown to infect, colonize and lyse several tumor types. Its descendant GLV-1h108, expressing an anti-VEGF antibody, was proved in previous studies to inhibit efficiently tumor induced angiogenesis. Additional VACVs expressing single chain antibodies (scAb) antibodies against PlGF and Ang-2 alone or in combination with anti VEGF scAb were designed. In this study, VACV-mediated anti-angiogenesis treatments have been evaluated in several preclinical tumor models. The efficiency of PlGF blockade, alone or in combination with VEGF, mediated by VACV has been established and confirmed. PlGF inhibition alone or with VEGF reduced tumor burden 5- and 2-folds more efficiently than the control virus, respectively. Ang-2 blockade efficiency for cancer treatment gave controversial results when tested in different laboratories. Here we demonstrated that unlike VEGF, the success of Ang-2 blockade is not only correlated to the strength of the blockade. A particular balance between Ang-2, VEGF and Ang-1 needs to be induced by the treatment to see a regression of the tumor and an improved survival. We saw that Ang-2 inhibition delayed tumor growth up to 3-folds compared to the control virus. These same viruses induced statistically significant tumor growth delays. This study unveiled the need to establish an angiogenic profile of the tumor to be treated as well as the necessity to better understand the synergic effects of VEGF and Ang-2. In addition angiogenesis inhibition by VACV-mediated PlGF and Ang-2 blockade was able to reduce the number of metastases and migrating tumor cells (even more efficiently than VEGF blockade). VACV colonization of tumor cells, in vitro, was limited by VEGF, when the use of the anti-VEGF VACV GLV-1h108 drastically improved the colonization efficiency up to 2-fold, 72 hours post-infection. These in vitro data were confirmed by in vivo analysis of tumors. Fourteen days post-treatment, the anti-VEGF virus GLV-1h108 was colonizing 78.8% of the tumors when GLV-1h68 colonization rate was 49.6%. These data confirmed the synergistic effect of VEGF blockade and VACV replication for tumor regression. Three of the tumor cell lines used to assess VACV-mediated angiogenesis inhibition were found, in certain conditions, to mimic either endothelial cell or pericyte functions, and participate directly to the vascular structure. The expression by these tumor cells of e-selectin, p-selectin, ICAM-1 and VCAM-1, normally expressed on activated endothelial cells, corroborates our findings. These proteins play an important role in immune cell recruitment, and there amount vary in presence of VEGF, PlGF and Ang-2, confirming the involvement of angiogenic factors in the immuno-modulatory abilities of tumors. In this study VACV-mediated angiogenesis blockade proved its potential as a therapeutic agent able to treat different tumor types and prevent resistance observed during bevacizumab treatment by acting on different factors. First, the expression of several antibodies by VACV would prevent another angiogenic factor to take over VEGF and stimulate angiogenesis. Then, the ability of VACV to infect tumor cells would prevent them to form blood vessel-like structures to sustain tumor growth, and the localized delivery of the antibody would decrease the risk of adverse effects. Next, the blockade of angiogenic factors would improve VACV replication and decrease the immune-modulatory effect of tumors. Finally the fact that angiogenesis blockade lasts until total regression of the tumor would prevent the recovery of the tumor-associated vasculature and the relapse of patients. N2 - Ein Hauptinteresse der onkologischen Forschung liegt auf dem Verständnis der Tumor-induzierten Angiogenese. Es wurde bereits festgestellt, dass die meisten Tumortypen eine abnorme Expression angiogener Faktoren zeigen. Der vascular endothelial growth factor (VEGF) wurde als der effektivste angiogene Faktor beschrieben. Es wurde gezeigt, dass die Hemmung des VEGF zur Inhibition der Angiogenese führt, das wiederum zu Tumorregression in vorklinischen Tumormodellen führte. Bevacizumab ist das erste FDA zugelassene Krebs-Therapeutikum, welches spezifisch auf die Tumor-induzierte Angiogenese durch VEGF-Inhibition abzielt. Der erwartete Erfolg durch VEGF-Hemmung konnte im Patienten allerdings nicht erzielt werden. Die Entwicklung von neuen Angiogenese hemmenden Stoffen wie gegen den placental growth factor (PIGF) oder Angiopoietin-2 (Ang-2), ermöglichen eine an das Tumor-Profil angepasste anti-angiogene Strategie. Die onkolytische Virustherapie die natürliche Eigenschaft der Viren Tumore zu kolonisieren. Das Vaccinia-Virus (VACV) gehört zur Familie der Poxviridae und wurde bereits lange Zeit als Vakzin zur Immunisierung gegen Pocken eingesetzt. Es konnte gezeigt werden, dass das rekombinante VACV GLV-1h68 effizient verschiedene Tumortypen infiziert, kolonisiert und lysiert. Das VACV GLV-1h108, welches auf der Basis des GLV-1h68 generiert wurde, kodiert einen anti-VEGF Antikörper. Dieses Virus ist in der Lage ist die Tumor-induzierte Angiogenese effizient zu inhibieren. Zusätzlich zu diesem VACV wurden weitere Konstrukte kloniert, welche für Antikörper gegen PIGF und Ang-2 kodieren. Zusätzlich wurden Virusstämme konstruiert, die gleichzeitig zwei Angiogenesefaktoren anzielen. Es wurde verschiedene VACV-vermittelte anti-Angiogenese Therapien in vorklinischen Tumormodellen wie Lungenadenokarzinome, KolonKarzinom, Melanom und Lungenadenokarzinome evaluiert. Die Effizienz der VACV-vermittelten Hemmung von PIGF und Ang-2, singulär oder in Kombination mit VEGF, wurde mit Tumor-Xenotransplantaten ermittelt. Die Inhibition von PlGF alleine oder in Kombination mit VEGF reduzierten die Tumorbelastung bis zu fünf, beziehungsweise zwei mal effizienter als GLV-1h68. Weiterhin wurde gezeigt, dass anders als VEGF, der Erfolg der Ang-2 Hemmung nicht nur mit der Stärke der Hemmung korreliert. Um Tumorregression sowie eine verbesserte Überlebensrate zu verursachen muss eine Balance zwischen Ang-2, VEGF und Ang-1 induziert werden. GLV-1h68 behandelte Tumore waren drei mal gröβer als Tumore, die mit den anti-Ang2 exprimierenden Viren behandelt wurden. Dieselben Virusstämme verursachten eine erhebliche Verspätung des Wachstums der Tumoren. Ausserdem hat diese Arbeit die Notwendigkeit enthüllt, ein angiogenes Profil des zu behandelnden Tumors zu etablieren sowie den Bedarf die synergistischen Effekte von VEGF und Ang-2 besser zu verstehen. Durch die Inhibition der Angiogenese durch VACV-verursachte PIGF und Ang-2 Hemmung wurde die Anzahl der Metastasen und der migrierenden Tumorzellen reduziert. Es wurde gezeigt, dass VEGF die VACV-Kolonisierung von Tumorzellen limitiert, da der Einsatz eines anti-VEGF VACV zu einer Verbesserung der Kolonisierung führt. In vivo Analysen bestätigten diese in vitro Daten. Nach vierzehn Tagen kolonisierte das anti-VEGF Virus 78,85% der Tumoren während die Kolonizationsquote des Kontrollviruses 49,64 % war. Dies resultierte in Tumorregression. Drei der getesteten Tumorzelllinien, in welchen die VACV-vermittelte Angiogenese-Inhibition untersucht wurde, waren in der Lage als Teil der Vaskulatur zu fungieren. Die Expression von Adhäsionsproteinen in diesen Tumorzellen untermauert die Ergebnisse. Weiterhin konnte ein unterschiedliches Expressionsmuster in Anwesenheit von VEGF, PIGF und Ang-2 festgestellt werden, wodurch die Beteiligung angiogener Faktoren bei den immunmodulatorischen Eigenschaften von Tumoren gezeigt werden konnte. In dieser Arbeit konnte gezeigt werden, dass eine VACV-vermittelte anti-angiogene Behandlung für verschiedene Tumorvarianten erfolgsversprechend ist. Die Möglichkeit verschiedene Antikörper gegen unterschiedliche angiogene Faktoren zu exprimieren würde verhindern, dass diese die Angiogenese stimulierende Wirkung des VEGF übernehmen. Die Eigenschaft von VACV Tumorzellen zu infizieren verhindert, dass diese Blutgefäß-ähnliche Strukturen bilden, welche das Tumorwachstum gewährleisten würde. Weiterhin würde die lokal begrenzte Antikörper-Freisetzung das Risiko von Nebenwirkungen senken. Die Inhibition angiogener Faktoren würde die VACV Replikationsrate steigern und den immunmodulatorischen Effekt der Tumore abschwächen. Letztlich würde die Hemmung der Angiogenese bis zur völligen Regression des Tumors aufrechterhalten, die Neubildung Tumor-assoziierter Vaskulatur verhindern und somit den Rückfall des Patienten. KW - Vaccinia-Virus KW - cancer KW - vaccinia virus KW - virotherapy KW - tumor vascularization KW - oncolytic virotherapy KW - Onkolyse KW - Angiogenese Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119019 ER - TY - THES A1 - Klingelhöffer, Christoph T1 - Untersuchungen zur Ascorbinsäure-vermittelten anti-Tumor-Wirkung: Oxidativer Stress als Auslöser selektiver Zytotoxizität T1 - Ascorbic acid-induced anticancer effect based on the inability of tumour cells to detoxify ROS N2 - Die Bedeutung von Ascorbinsäure als „Krebsschutzfaktor“ wird auch weiterhin kontrovers diskutiert. Seit einiger Zeit wird vermutet, dass Ascorbinsäure oxidativen Stress auslöst. In der vorliegenden Untersuchung wurde die Wirkung von Ascorbinsäure auf 12 maligne und 3 benigne Zelllinien in vitro untersucht. Die Zellen wurden für 2 bzw. 14 Stunden mit unterschiedlichen Konzentrationen von Ascorbinsäure (5 bis 100 mmol/L) inkubiert und 24, 48 und 72 Stunden nach Versuchsbeginn der Anteil vitaler Zellen bestimmt. Die hierfür verwendeten Assays, WST-8 und Kristallviolett-Assay, ließen zudem Aussagen über die Stoffwechselaktivität (WST-8) und Zellvitalität (Kristallviolett) zu. Die schädigende Wirkung von Ascorbinsäure wurde als EC50-Wert angegeben, bei dieser Ascorbinsäure-Konzentration sind 50 % der Zellen zerstört. Ascorbinsäure wirkte nach 2 Stunden Inkubation kaum zelltoxisch, während nach 14 Stunden Inkubation eindeutige zelltoxische Effekte bei 6 der 12 malignen Zelllinien zu beobachten waren. So waren die drei getesteten Glioblastomzelllinien allesamt bereits bei einer Ascorbinsäure-Konzentrationen von 5 mmol/L nahezu vollkommen zerstört (EC50: 2,6-5,5 mmol/L). Die Mammakarzinomzelllinie BT-20 hingegen war am widerstandsfähigsten gegenüber dem zelltoxischen Effekt der Ascorbinsäure (EC50: 95 mmol/L). Als wesentliches Effektormolekül der zelltoxischen Wirkung der Ascorbinsäure wurde Wasserstoffperoxid identifiziert. Die Zugabe von Katalase schützt Ascorbinsäure- sensitive Zellen, in dem es Wasserstoffperoxid abbaut. Ein weiteres Indiz hierfür ist, dass Zelllinien, die gegenüber dem Ascorbinsäure-vermittelten Effekt unempfindlich waren, dies auch gegenüber Wasserstoffperoxid waren. Umgekehrt waren Zelllinien, die empfindlich gegenüber dem Ascorbinsäurevermittelten zelltoxischen Effekt reagierten, auch empfindlich gegenüber Wasserstoffperoxid. 45 Eine wesentliche sich aus den Daten dieser Arbeit ergebende Frage ist die, worin sich Ascorbinsäure-resistente Tumorzellen von Ascorbinsäure-empfindlichen Tumorzellen unterscheiden. Da Ascorbinsäure-empfindliche Zellen durch Zugabe von Katalase vor der zelltoxischen Wirkung der Ascorbinsäure geschützt werden, liegt die Vermutung nahe, dass eine wesentliche Ursache hierfür in der zelleigenen Katalase begründet liegt. Somit sollten Ascorbinsäureresistente Zellen mehr bzw. aktivere Katalase aufweisen, als Ascorbinsäureempfindliche Zellen. Diese Vermutung ist in weiteren Experimenten zu überprüfen. N2 - Ascorbic acid (Asc) has a controversial history in supportive cancer treatment. The Asc-mediated cytotoxic effect is based on the local production of hydrogen peroxide (H2O2). Tumour cells lines were screened for their susceptibility or resistance to Asc-mediated cytotoxicity. The purpose of this study was to analyse the impact of ascorbic acid modifying tumour cell vitality. A panel of twelve tumour cell lines, carcinomas and glioblastoma, were exposed to serial dilutions of Asc (0-100 mmol/L) up to 2 and 14hrs in vitro. The Asc concentration that effectively decreased survival by 50% (EC50) was determined by wst-8 and crystal violet assay. The tested cell lines demonstrated obvious differences in their resistance to Asc. Fifty percent of the cancer cell lines had an EC50 >20mM and were resistant to Asc, whereas fifty percent had an EC50 <10mM; they were susceptible to Asc. In addition, Asc resistant cell lines were also cross-resistant to H2O2. Glioblastoma cell lines were found to be the most susceptible cell lines (EC50: 2.5-6mM). External catalase (250U) neutralizes the cytotoxic effect of Asc and protects against cell death. KW - Vitamin C KW - Krebs KW - Oxidativer Stress KW - ascorbic acid KW - cancer KW - oxidative stress Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-66402 ER - TY - THES A1 - Schneider, Tim Frederik T1 - Untersuchung des EGF-Rezeptor-Signalwegs an Karzinomen der Kopfspeicheldrüsen T1 - Examination of EGF-Receptor-pathway of salivary-gland carcinomas N2 - Bei vielen Karzinomen spielt EGFR und das KRAS-Onkogen eine wichtige Rolle in der Tumorentstehung. Da bei den seltenen Karzinomen an Kopfspeicheldrüsen sehr wenig über molekulare Mechanismen der Tumorgenese bekannt ist, war es das Ziel der Arbeit den EGFR-Signalweg zu untersuchen. Es wurden Paraffinschnitte von 43 Speicheldrüsenkarzinomen von den Typen ACC, MEC und Adeno-Ca NOS mit dem phosphorylierten EGFR-Antikörper gefärbt und mit klinisch-pathologischen Daten korreliert. Weiterhin wurde eine Mutationsanalyse der kras-Gensequenz durchgeführt. In allen Fällen war das kras-Gen vom Wildtyp. Bei der Expressionsanalyse von EGFR stellte sich heraus, dass 79% der Proben einen aktivierten EGF-Rezeptor besitzen. Statistisch signifikante Korrelationen gab es zwischen der EGFR-Expression und dem Patientenalter, dem zervikalen Lymphknotenbefall und der Tumorgröße. Der EGF-Signaltransduktionsweg ist bei den untersuchten Karzinomen der Kopfspeicheldrüsen im überwiegenden Masse aktiviert, ohne dass eine autonome Aktivierung beim KRAS-Onkogen vorliegt. N2 - EGFR and the KRAS-onkogen plays an important role in tumor-genesis of many cancers. Little is known about the tumor-genesis of the rare entity of salivary gland cancers. The intention of this examination was to explore the EGFR-Pathway of adenoid-cystic carcinomas, mucoepidermoid carcinomas and the adeno-Ca not otherwise specified. 43 cases of paraffin-embedded tissue were analyzed, if there were EGFR-Expressions and mutations in the kras-gene. This data was correlated with clinical and pathological data. In none of the cases mutations were found. The EGF-receptor was activated in 79% of the samples and there were significant correlations between the receptor-activation and age as well as cervical lymphnode status. Result: The EGFR-Pathway in the examined malignancies of salivary gland cancers is predominant activated withour autonomous activation of this pathway by kras-mutation. KW - Epidermaler Wachstumsfaktor-Rezeptor KW - Speicheldrüse KW - Krebs KW - Speicheldrüsenkrebs KW - kras-Mutation KW - cancer KW - salivary gland KW - egfr KW - kras Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-65278 ER - TY - THES A1 - Wassermann, Veronika T1 - Ueber die Beteiligung der Hitzeschockproteine HSP27 und HSP70 an einer Resistenz, Resistenzentwicklung unter CMF-Therapie T1 - Is heat shock protein expression an important faktor in drug resistance? N2 - Ein Problem der Therapie maligner Tumore ist die Resistez gegenüber Chemotherapeutika. Diskutiert wird ein Zusammenhang zur Expression von Hitzeschockproteinen (HSP). Insbesondere das in Mamma-Karzinomen stark exprimierte HSP27 und HSP70 scheinen hier beteiligt. Hitzeschockproteine sind Teil eines durch Noxen induzierten Mechanismus, welcher Schutz vor weiterer Noxenexposition verleiht, also die entsprechenen Zellen im Unterschied zu nicht exponierten Kontrollzellen zu überleben befähigt. Es kommt nach einem Streßereignis zu einer Veränderung von Zelltod unter nachfolgenden Bedingungen, z.B. Verhütung von Apoptose (physiologischer Zelltod). Tumortherapie mittels Zytostatika stellt eine „kontrollierte Apoptose“ dar. Ziel dieser Therapie muß es also folglich sein, eine HSP-Induktion zu vermeiden, da eine solche den Therapieerfolg und Benefit für den Patienten schmälert. Die Exposition einer Tumorzelle mit einem chemotherapeutisch wirksamen Medikament bedeutet für diese Zelle toxischen/oxidativen Streß, den sie nur durch Induktion entsprechender Abwehrmechanismen überleben kann. Diese Mechanismen haben letztlich einen wesentlichen Einfluß auf die Wirksamkeit des Medikamentes und Profit des Patienten durch die ihm angebotene Therapie. Eine frühe adaptive Zellantwort von Säugerzellen auf toxische Einflüsse stellt die Expression von Hitzeschockproteinen dar. Den Fokus der vorliegenden Untersuchungen stellen einerseits diejenigen Substanzen dar, welche heutzutage in der Therapie des Mamma-Karzinoms eingesetzt werden, den drei Einzelsubstanzen des CMF-Protokolls Methotrexat, 5-Fluorouracil und Cyclophosphamid, andererseits die Hitzeschockproteine mit der höchsten bekannten Bedeutung für Tumorwachstum. In einem ersten Schritt wurde in vitro mit einem Zellsystem, welches durch Transfektion mit dem humanen hsp27-Gen bei bekannter HSP70-Induzierbarkeit als einfaches isoliertes Zellsystem ein gutes Werkzeug zur spezifischen Untersuchung dieser beiden Proteine darstellt, untersucht werden, inwieweit sich diese speziellen Proteine durch die unterschiedlichen Substanzgruppen des CMF-Protokolls induzieren lassen. Es wurde also nach einer adaptiven Zellantwort in Form einer Induktion von HSP27 und HSP70 nachfolgend einer Noxenexposition gesucht werden. In einem zweiten Schritt wurde untersucht, ob die für diese beiden HSPs postulierte zytoprotektive Eigenschaften auch für Zytostatika gelten. Es wurde überprüft , inwieweit Chemotherapeutika unter dem Einfluß von HSP70 und HSP27 stehen, also inwieweit die Expression von HSP27 und HSP70 einen Zellschutz gegen toxischen Streß (durch CMF) in den verwendeten L929-Mausfibroblasten darstellen kann. N2 - Resistance to cytotoxic drugs is a major problem in tumor therapy. Paradoxycally both aquired and inhärent resistance to chemotherapeutica seem to be more common in tumors than in normal cells. A correllation to the expression of heat schock proteins (HSPs) is discussed. In particular, HSP27 and HSP70 seem to controbute to this phenomen. Our study focussed on the role of both HSPs in cellular response to methotrexate, 5-FU an cyclophosphamide, which are actually used in breast cancer therapy. KW - Hietzeschockprotein KW - Resistenzentwicklung KW - CMF-Therapie KW - Mammakarzinom KW - heat shock protein KW - resistance KW - cancer Y1 - 2005 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-12292 ER - TY - JOUR A1 - Montelius, Mikael A1 - Ljungberg, Maria A1 - Horn, Michael A1 - Forssell-Aronsson, Eva T1 - Tumour size measurement in a mouse model using high resolution MRI JF - BMC Medical Imaging N2 - Background Animal models are frequently used to assess new treatment methods in cancer research. MRI offers a non-invasive in vivo monitoring of tumour tissue and thus allows longitudinal measurements of treatment effects, without the need for large cohorts of animals. Tumour size is an important biomarker of the disease development, but to our knowledge, MRI based size measurements have not yet been verified for small tumours (10−2–10−1 g). The aim of this study was to assess the accuracy of MRI based tumour size measurements of small tumours on mice. Methods 2D and 3D T2-weighted RARE images of tumour bearing mice were acquired in vivo using a 7 T dedicated animal MR system. For the 3D images the acquired image resolution was varied. The images were exported to a PC workstation where the tumour mass was determined assuming a density of 1 g/cm3, using an in-house developed tool for segmentation and delineation. The resulting data were compared to the weight of the resected tumours after sacrifice of the animal using regression analysis. Results Strong correlations were demonstrated between MRI- and necropsy determined masses. In general, 3D acquisition was not a prerequisite for high accuracy. However, it was slightly more accurate than 2D when small (<0.2 g) tumours were assessed for inter- and intraobserver variation. In 3D images, the voxel sizes could be increased from 1603 μm3 to 2403 μm3 without affecting the results significantly, thus reducing acquisition time substantially. Conclusions 2D MRI was sufficient for accurate tumour size measurement, except for small tumours (<0.2 g) where 3D acquisition was necessary to reduce interobserver variation. Acquisition times between 15 and 50 minutes, depending on tumour size, were sufficient for accurate tumour volume measurement. Hence, it is possible to include further MR investigations of the tumour, such as tissue perfusion, diffusion or metabolic composition in the same MR session. KW - cancer KW - magnetic resonance KW - animal model KW - volume determination Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-124049 VL - 12 IS - 12 ER - TY - JOUR A1 - Muturi, Harrison T. A1 - Dreesen, Janine D. A1 - Nilewski, Elena A1 - Jastrow, Holger A1 - Giebel, Bernd A1 - Ergun, Suleyman A1 - Singer, Berhard B. T1 - Tumor and Endothelial Cell-Derived Microvesicles Carry Distinct CEACAMs and Influence T-Cell Behavior JF - PLOS ONE N2 - Normal and malignant cells release a variety of different vesicles into their extracellular environment. The most prominent vesicles are the microvesicles (MVs, 100-1 000 nm in diameter), which are shed of the plasma membrane, and the exosomes (70-120 nm in diameter), derivates of the endosomal system. MVs have been associated with intercellular communication processes and transport numerous proteins, lipids and RNAs. As essential component of immune-escape mechanisms tumor-derived MVs suppress immune responses. Additionally, tumor-derived MVs have been found to promote metastasis, tumor-stroma interactions and angiogenesis. Since members of the carcinoembryonic antigen related cell adhesion molecule (CEACAM)-family have been associated with similar processes, we studied the distribution and function of CEACAMs in MV fractions of different human epithelial tumor cells and of human and murine endothelial cells. Here we demonstrate that in association to their cell surface phenotype, MVs released from different human epithelial tumor cells contain CEACAM1, CEACAM5 and CEACAM6, while human and murine endothelial cells were positive for CEACAM1 only. Furthermore, MVs derived from CEACAM1 transfected CHO cells carried CEACAM1. In terms of their secretion kinetics, we show that MVs are permanently released in low doses, which are extensively increased upon cellular starvation stress. Although CEACAM1 did not transmit signals into MVs it served as ligand for CEACAM expressing cell types. We gained evidence that CEACAM1-positive MVs significantly increase the CD3 and CD3/CD28-induced T-cell proliferation. All together, our data demonstrate that MV-bound forms of CEACAMs play important roles in intercellular communication processes, which can modulate immune response, tumor progression, metastasis and angiogenesis. KW - carcinoembryonic anitgen family KW - biliary glycoprotein CD66A KW - adhesion molecule-1 KW - epithelial cells KW - membrane vesicles KW - growth-factor KW - cancer KW - expression KW - proliferation Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-128373 SN - 1932-6203 VL - 8 IS - 9 ER - TY - JOUR A1 - Mousset, Sabine A1 - Buchheidt, Dieter A1 - Heinz, Werner A1 - Ruhnke, Markus A1 - Cornely, Oliver A. A1 - Egerer, Gerlinde A1 - Krüger, William A1 - Link, Hartmut A1 - Neumann, Silke A1 - Ostermann, Helmut A1 - Panse, Jens A1 - Penack, Olaf A1 - Rieger, Christina A1 - Schmidt-Hieber, Martin A1 - Silling, Gerda A1 - Südhoff, Thomas A1 - Ullmann, Andrew J. A1 - Wolf, Hans-Heinrich A1 - Maschmeyer, Georg A1 - Böhme, Angelika T1 - Treatment of invasive fungal infections in cancer patients—updated recommendations of the Infectious Diseases Working Party (AGIHO) of the German Society of Hematology and Oncology (DGHO) JF - Annals of Hematology N2 - Invasive fungal infections are a main cause of morbidity and mortality in cancer patients undergoing intensive chemotherapy regimens. Early antifungal treatment is mandatory to improve survival. Today, a number of effective and better-tolerated but more expensive antifungal agents compared to the former gold standard amphotericin B deoxycholate are available. Clinical decision-making must consider results from numerous studies and published guidelines, as well as licensing status and cost pressure. New developments in antifungal prophylaxis improving survival rates result in a continuous need for actualization. The treatment options for invasive Candida infections include fluconazole, voriconazole, and amphotericin B and its lipid formulations, as well as echinocandins. Voriconazole, amphotericin B, amphotericin B lipid formulations, caspofungin, itraconazole, and posaconazole are available for the treatment of invasive aspergillosis. Additional procedures, such as surgical interventions, immunoregulatory therapy, and granulocyte transfusions, have to be considered. The Infectious Diseases Working Party of the German Society of Hematology and Oncology here presents its 2008 recommendations discussing the dos and do-nots, as well as the problems and possible solutions, of evidence criteria selection. KW - cancer KW - invasive fungal infections KW - antifungals KW - mycoses KW - hematologic malignancies KW - aspergillosis KW - antifungal agents KW - invasive candidiasis Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-121340 VL - 96 ER - TY - JOUR A1 - Paschke, Ralf A1 - Lincke, Thomas A1 - Müller, Stefan P. A1 - Kreissl, Michael C. A1 - Dralle, Henning A1 - Fassnacht, Martin T1 - The Treatment of Well-Differentiated Thyroid Carcinoma JF - Deutsches Ärzteblatt International N2 - Background: Recent decades have seen a rise in the incidence of well-differentiated (mainly papillary) thyroid carcinoma around the world. In Germany, the age-adjusted incidence of well-differentiated thyroid carcinoma in 2010 was 3.5 per 100 000 men and 8.7 per 100 000 women per year. Method: This review is based on randomized, controlled trials and multicenter trials on the treatment of well-differentiated thyroid carcinoma that were retrieved by a selective literature search, as well as on three updated guidelines issued in the past two years. Results: The recommended extent of surgical resection depends on whether the tumor is classified as low-risk or high-risk, so that papillary microcar cinomas, which carry a highly favorable prognosis, will not be overtreated. More than 90% of localized, well-differentiated thyroid carcinomas can be cured with a combination of surgery and radioactive iodine therapy. Radio active iodine therapy is also effective in the treatment of well-differentiated thyroid carcinomas with distant metastases, yielding a 10-year survival rate of 90%, as long as there is good iodine uptake and the tumor goes into remission after treatment; otherwise, the 10-year survival rate is only 10%. In the past two years, better treatment options have become available for radioactive-iodine-resistant thyroid carcinoma. Phase 3 studies of two different tyrosine kinase inhibitors have shown that either one can markedly prolong progression-free survival, but not overall survival. Their more common clinically significant side effects are hand-foot syndrome, hypertension, diarrhea, proteinuria, and weight loss. Conclusion: Slow tumor growth, good resectability, and susceptibility to radioactive iodine therapy lend a favorable prognosis to most cases of well-differentiated thyroid carcinoma. The treatment should be risk-adjusted and interdisciplinary, in accordance with the current treatment guidelines. Even metastatic thyroid carcinoma has a favorable prognosis as long as there is good iodine uptake. The newly available medical treatment options for radioactive-iodine-resistant disease need to be further studied. KW - BRAF(V600E) mutation KW - distant metastases KW - papillary KW - guidelines KW - surgery KW - dissection KW - management KW - association KW - cancer KW - radioiodine therapy Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151636 VL - 112 SP - 452 EP - 458 ER - TY - JOUR A1 - Haider, Malik Salman A1 - Ahmad, Taufiq A1 - Groll, Jürgen A1 - Scherf-Clavel, Oliver A1 - Kroiss, Matthias A1 - Luxenhofer, Robert T1 - The Challenging Pharmacokinetics of Mitotane: An Old Drug in Need of New Packaging JF - European Journal of Drug Metabolism and Pharmacokinetics N2 - Adrenocortical carcinoma (ACC) is a malignant tumor originating from the adrenal gland cortex with a heterogeneous but overall dismal prognosis in advanced stages. For more than 50 years, mitotane has remained a cornerstone for the treatment of ACC as adjuvant and palliative therapy. It has a very poor aqueous solubility of 0.1 mg/l and high partition coefficient in octanol/water (log P) value of 6. The commercially available dosage form is 500 mg tablets (Lysodren®). Even at doses up to 6 g/day (12 tablets in divided doses) for several months, > 50% patients do not achieve therapeutic plasma concentration > 14 mg/l due to poor water solubility, large volume of distribution and inter/intra-individual variability in bioavailability. This article aims to give a concise update of the clinical challenges associated with the administration of high-dose mitotane oral therapy which encompass the issues of poor bioavailability, difficult-to-predict pharmacokinetics and associated adverse events. Moreover, we present recent efforts to improve mitotane formulations. Their success has been limited, and we therefore propose an injectable mitotane formulation instead of oral administration, which could bypass many of the main issues associated with high-dose oral mitotane therapy. A parenteral administration of mitotane could not only help to alleviate the adverse effects but also circumvent the variable oral absorption, give better control over therapeutic plasma mitotane concentration and potentially shorten the time to achieve therapeutic drug plasma concentrations considerably. Mitotane as tablet form is currently the standard treatment for adrenocortical carcinoma. It has been used for 5 decades but suffers from highly variable responses in patients, subsequent adverse effects and overall lower response rate. This can be fundamentally linked to the exceedingly poor water solubility of mitotane itself. In terms of enhancing water solubility, a few research groups have attempted to develop better formulations of mitotane to overcome the issues associated with tablet dosage form. However, the success rate was limited, and these formulations did not make it into the clinics. In this article, we have comprehensively reviewed the properties of these formulations and discuss the reasons for their limited utility. Furthermore, we discuss a recently developed mitotane nanoformulation that led us to propose a novel approach to mitotane therapy, where intravenous delivery supplements the standard oral administration. With this article, we combine the current state of knowledge as a single piece of information about the various problems associated with the use of mitotane tablets, and herein we postulate the development of a new injectable mitotane formulation, which can potentially circumvent the major problems associated to mitotane's poor water solubility. KW - Mitotane KW - cancer KW - adrenal gland Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-270476 SN - 2107-0180 VL - 46 IS - 5 ER - TY - JOUR A1 - Wajant, Harald A1 - Beilhack, Andreas T1 - Targeting regulatory T cells by addressing tumor necrosis factor and its receptors in allogeneic hematopoietic cell transplantation and cancer JF - Frontiers in Immunology N2 - An intricate network of molecular and cellular actors orchestrates the delicate balance between effector immune responses and immune tolerance. The pleiotropic cytokine tumor necrosis factor-alpha (TNF) proves as a pivotal protagonist promoting but also suppressing immune responses. These opposite actions are accomplished through specialist cell types responding to TNF via TNF receptors TNFR1 and TNFR2. Recent findings highlight the importance of TNFR2 as a key regulator of activated natural FoxP3+ regulatory T cells (Tregs) in inflammatory conditions, such as acute graft-vs.-host disease (GvHD) and the tumor microenvironment. Here we review recent advances in our understanding of TNFR2 signaling in T cells and discuss how these can reconcile seemingly conflicting observations when manipulating TNF and TNFRs. As TNFR2 emerges as a new and attractive target we furthermore pinpoint strategies and potential pitfalls for therapeutic targeting of TNFR2 for cancer treatment and immune tolerance after allogeneic hematopoietic cell transplantation. KW - GVHD KW - graft vs. host disease KW - cancer KW - Tregs (regulatory T cells) KW - TNFR family costimulatory receptors KW - TNFR2 agonists KW - TNFR2 antagonism Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201578 VL - 10 IS - 2040 ER - TY - THES A1 - Grebinyk, Anna T1 - Synergistic Chemo- and Photodynamic Treatment of Cancer Cells with C\(_{60}\) Fullerene Nanocomplexes T1 - Synergistische chemo- und photodynamische Behandlung von Krebszellen mit C\(_{60}\)-Fulleren-Nanokomplexen N2 - Recent progress in nanotechnology has attracted interest to a biomedical application of the carbon nanoparticle C60 fullerene (C60) due to its unique structure and versatile biological activity. In the current study the dual functionality of C60 as a photosensitizer and a drug nanocarrier was exploited to improve the efficiency of chemotherapeutic drugs towards human leukemic cells. Pristine C60 demonstrated time-dependent accumulation with predominant mitochondrial localization in leukemic cells. C60’s effects on leukemic cells irradiated with high power single chip LEDs of different wavelengths were assessed to find out the most effective photoexcitation conditions. A C60-based noncovalent nanosized system as a carrier for an optimized drug delivery to the cells was evaluated in accordance to its physicochemical properties and toxic effects. Finally, nanomolar amounts of C60-drug nanocomplexes in 1:1 and 2:1 molar ratios were explored to improve the efficiency of cell treatment, complementing it with photodynamic approach. A proposed treatment strategy was developed for C60 nanocomplexes with the common chemotherapeutic drug Doxorubicin, whose intracellular accumulation and localization, cytotoxicity and mechanism of action were investigated. The developed strategy was revealed to be transferable to an alternative potent anticancer drug – the herbal alkaloid Berberine. Hereafter, a strong synergy of treatments arising from the combination of C60-mediated drug delivery and C60 photoexcitation was revealed. Presented data indicate that a combination of chemo- and photodynamic treatments with C60-drug nanoformulations could provide a promising synergetic approach for cancer treatment. N2 - Kürzliche Fortschritte in der Nanotechnologie haben Interesse an einer biomedizinischen Anwendung des Kohlenstoffnanopartikels C60 Fulleren (C60) aufgrund seiner einzigartigen Struktur und breiten biologischen Aktivität geweckt. In der aktuellen Studie wurde die doppelte Funktionalität von C60 als Photosensibilisator und als Wirkstoff-Nanoträger genutzt, um die Wirkung von Chemotherapeutika auf menschliche Leukämiezellen zu verbessern. C60 alleine zeigte in den Zellen eine zeitabhängige Akkumulation mit vorherrschender mitochondrialer Lokalisation. Die Wirkung von C60 auf Leukämiezellen, die mit unterschiedlicher Wellenlänge bestrahlt wurden, wurde bewertet, um die effektivsten Photoanregungsbedingungen zu finden. Die physikochemischen Eigenschaften und toxischen Wirkungen von C60 auf die Leukämiezellen wurden nach nicht kovalenter Bindung von Arzneistoffen bewertet. Schließlich wurden nanomolare Mengen von C60-Wirkstoff-Nanokomplexen in Molverhältnissen von 1:1 und 2:1 untersucht, um die Effizienz der Behandlung von Zellen zu verbessern und sie durch photodynamischen Ansatz zu ergänzen. Mit dem gängigen Chemotherapeutikum Doxorubicin wurde eine Behandlungsstrategie entwickelt und dessen intrazelluläre Akkumulation und Lokalisation, Zytotoxizität und Wirkmechanismus untersucht wurden. Es wurde gezeigt, dass die entwickelte Strategie auch auf ein alternatives Krebsmedikament übertragbar ist – das pflanzliche Alkaloid Berberin. Die erhaltenen Daten deuten darauf hin, dass eine Kombination von chemo- und photodynamischen Behandlungen mit C60-Nanokomplexen einen vielversprechenden synergetischen Ansatz für die Krebsbehandlung bieten könnte. KW - cancer KW - drug delivery KW - photodynamic therapy KW - fullerene Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-222075 ER - TY - JOUR A1 - Wilhelm, Martin A1 - Smetak, Manfred A1 - Schaefer-Eckart, Kerstin A1 - Kimmel, Brigitte A1 - Birkmann, Josef A1 - Einsele, Hermann A1 - Kunzmann, Volker T1 - Successful adoptive transfer and in vivo expansion of haploidentical γδ T cells JF - Journal of Translational Medicine N2 - Background: The primary aim of this pilot study was to determine the feasibility and safety of an adoptive transfer and in vivo expansion of human haploidentical gamma delta T lymphocytes. Methods: Patients with advanced haematological malignancies who are not eligible for allogeneic transplantation received peripheral blood mononuclear cells from half-matched family donors. For that, a single unstimulated leukapheresis product was incubated with both the anti-CD4 and anti-CD8 antibodies conjugated to paramagnetic particles. The depletion procedure was performed on a fully automated CliniMACS (R) device according to the manufacturer's instructions. On average, patients received 2.17 x 10(6)/kg (range 0.9-3.48) γδ T cells with <1% CD4-or CD8-positive cells remaining in the product. All patients received prior lymphopenia-inducing chemotherapy (fludarabine 20-25 mg/m(2) day -6 until day -2 and cyclophosphamide 30-60 mg/kg day -6 and -5) and were treated with 4 mg zoledronate on day 0 and 1.0x10(6) IU/m(2) IL-2 on day +1 until day +6 for the induction of gamma delta T cell proliferation in vivo. Results: This resulted in a marked in vivo expansion of donor γδ T cells and, to a lower extent, natural killer cells and double-negative αβ T cells (mean 68-fold, eight-fold, and eight-fold, respectively). Proliferation peaked by around day +8 and donor cells persisted up to 28 days. Although refractory to all prior therapies, three out of four patients achieved a complete remission, which lasted for 8 months in a patient with plasma cell leukaemia. One patient died from an infection 6 weeks after treatment. Conclusion: This pilot study shows that adoptive transfer and in vivo expansion of haploidentical γδ T lymphocytes is feasible and suggests a potential role of these cells in the treatment of haematological diseases. KW - NK cells KW - in vivo cell expansion KW - haploidentical γδ T lymphocytes KW - adoptive transfer KW - CD4(+) KW - innate immunity KW - stimulation KW - acute myeloid-leukemia KW - immunotherapy KW - cancer KW - infusion KW - Interleukin-2 KW - biophosphonate Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117290 VL - 12 IS - 45 ER - TY - JOUR A1 - Cardani, Diego A1 - Sardi, Claudia A1 - La Ferla, Barbara A1 - D'Orazio, Guiseppe A1 - Sommariva, Michele A1 - Marcucci, Fabrizio A1 - Olivero, Daniela A1 - Tagliabue, Elda A1 - Koepsell, Hermann A1 - Nicotra, Francesco A1 - Balsari, Andrea A1 - Rumio, Christiano T1 - Sodium glucose cotransporter 1 ligand BLF501 as a novel tool for management of gastrointestinal mucositis JF - Molecular Cancer N2 - Background: Recent studies demonstrated that engagement of sodium glucose transporter 1 (SGLT-1) by orally administered D-glucose protects the intestinal mucosa from lipopolysaccharide (LPS)-induced injury. We tested whether SGLT-1 engagement might protect the intestinal mucosa from doxorubicin (DXR)- and 5-fluorouracil (5-FU)-induced injury in animal models mimicking acute or chronic mucositis. Methods: Mice were treated intraperitoneally with DXR, alone or in combination with 5-FU, and orally with BLF501, a glucose-derived synthetic compound with high affinity for SGLT-1. Intestinal mucosal epithelium integrity was assessed by histological analysis, cellular proliferation assays, real-time PCR gene expression assays and Western blot assays. Student's t-test (paired two-tailed) and X-2 analyses were used for comparisons between groups. Differences were considered significant at p < 0.05. Results: BLF501 administration in mice treated with DXR and/or 5-FU decreased the injuries to the mucosa in terms of epithelial integrity and cellular proliferative ability. Co-treatment with BLF501 led to a normal expression and distribution of both zonula occludens-1 (ZO-1) and beta-catenin, which were underexpressed after treatment with either chemotherapeutic agent alone. BLF501 administration also restored normal expression of caspase-3 and ezrin/radixin/moesin (ERM), which were overexpressed after treatment with DXR and 5-FU. In SGLT1-/- mice, BLF501 had no detectable effects. BLF501 administration in wild-type mice with growing A431 tumors did not modify antitumor activity of DXR. Conclusions: BLF501-induced protection of the intestinal mucosa is a promising novel therapeutic approach to reducing the severity of chemotherapy-induced mucositis. KW - apoptosis KW - prevention KW - doxorubicin KW - cancer KW - gastrointestinal mucositis KW - SGLT-1 KW - synthetic D-glucose analogy KW - chemotherapy KW - inflammation KW - clinical practice guidelines KW - intestinal mucositis KW - epithelial cells KW - oral mucositis KW - gene-expression Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117352 SN - 1476-4598 VL - 13 IS - 23 ER - TY - JOUR A1 - Esser, Peter A1 - Mehnert‐Theuerkauf, Anja A1 - Friedrich, Michael A1 - Johansen, Christoffer A1 - Brähler, Elmar A1 - Faller, Hermann A1 - Härter, Martin A1 - Koch, Uwe A1 - Schulz, Holger A1 - Wegscheider, Karl A1 - Weis, Joachim A1 - Kuba, Katharina A1 - Hinz, Andreas A1 - Hartung, Tim T1 - Risk and associated factors of depression and anxiety in men with prostate cancer: Results from a German multicenter study JF - Psycho‐Oncology N2 - Objective In order to optimize psycho‐oncological care, studies that quantify the extent of distress and identify certain risk groups are needed. Among patients with prostate cancer (PCa), findings on depression and anxiety are limited. Methods We analyzed data of PCa patients selected from a German multi‐center study. Depression and anxiety were assessed with the PHQ‐9 and the GAD‐7 (cut‐off ≥7). We provided physical symptom burden, calculated absolute and relative risk (AR and RR) of depression and anxiety across patient subsets and between patients and the general population (GP) and tested age as a moderator within the relationship of disease‐specific symptoms with depression and anxiety. Results Among 636 participants, the majority reported disease‐specific problems (sexuality: 60%; urination: 52%). AR for depression and anxiety was 23% and 22%, respectively. Significant RR were small, with higher risks of distress in patients who are younger (eg, RR\(_{depression}\) = 1.15; 95%‐CI: 1.06‐1.26), treated with chemotherapy (RR\(_{depression}\)n = 1.46; 95%‐CI: 1.09‐1.96) or having metastases (RR\(_{depression}\) = 1.30; 95%‐CI: 1.02‐1.65). Risk of distress was slightly elevated compared to GP (eg, RR\(_{depression}\) = 1.13; 95%‐CI: 1.07‐1.19). Age moderated the relationship between symptoms and anxiety (B\(_{urination}\) = −0.10, P = .02; B\(_{sexuality}\) = −0.11, P = .01). Conclusions Younger patients, those with metastases or treatment with chemotherapy seem to be at elevated risk for distress and should be closely monitored. Many patients suffer from disease‐specific symptom burden, by which younger patients seem to be particularly distressed. Support of coping mechanisms associated with disease‐specific symptom burden seems warranted. KW - anxiety KW - cancer KW - depression KW - oncology KW - prostatic neoplasms Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-218277 VL - 29 IS - 10 SP - 1604 EP - 1612 ER - TY - THES A1 - Robubi, Armin T1 - RAF Kinases: Pathway, Modulation and Modeling T1 - RAF Kinase: Signalweg, Modulation und Modellierung N2 - The Ras/RAF/MEK/ERK cascade is a central cellular signal transduction pathway involved in cell proliferation, differentiation, and survival where RAF kinases are pivotal kinases implicated in cancer. The development of specific irreversible kinase inhibitors is a rewarding but difficult aim. CI-1033 was developed to irreversibly inhibit erbB receptor tyrosine kinases by reacting to the Cys113 residue (p38alpha MAP kinase numbering) of the kinase domain. In this study we tried a similar approach to target the RAF oncoproteins which posses a similar cysteine at position 108 in the hinge region between the small n-lobe and the large c-lobe of the kinase domain. A novel synthetic approach including a lyophilization step allowed us the synthesis of a diphenyl urea compound with an epoxide moiety (compound 1). Compound 1 possessed inhibitory activity in vitro. However our time kinetics experiments and mass spectroscopic studies clearly indicate that compound 1 does not react covalently with the cysteine residue in the hinge region. Moreover, in cell culture experiments, a strong activation of the RAF signaling pathway was observed, an effect which is known from several other RAF kinase inhibitors and is here reported for the first time for a diphenyl urea compound, to which the clinically used unspecific kinase inhibitor BAY 43-9006 (Sorafinib, Nexavar) belongs. Although activation was apparently independent on B- and C-RAF hetero-oligomerization in vitro, in vivo experiments support such a mechanism as the activation did not occur in starved knockout cells lacking either B-RAF or C-RAF. Furthermore, we developed a mathematical model of the Ras/RAF/MEK/ERK cascade demonstrating how stimuli induce different signal patterns and thereby different cellular responses, depending on cell type and the ratio between B-RAF and C-RAF. Based on biochemical data for activation and dephosphorylation, we set up differential equations for a dynamical model of the Ras/RAF/MEK/ERK cascade. We find a different signaling pattern and response result for B-RAF (strong activation, sustained signal) and C-RAF (steep activation, transient signal). We further support the significance of such differential modulatory signaling by showing different RAF isoform expression in various cell lines and experimental testing of the predicted kinase activities in B-RAF, C-RAF as well as mutated versions. Additionally the effect of the tumor suppressor DiRas3 (also known as Noey2 or ARHI) on RAF signaling was studied. I could show that DiRas3 down-regulates the mitogenic pathway by inhibition of MEK, a basis for a refined model of the Ras/RAF/MEK/ERK cascade. N2 - Die Ras/RAF/MEK/ERK Kaskade ist ein zentraler zellulärer Signalweg, der bei der Regulierung der Proliferation, Differenzierung und Überleben der Zelle eine entscheide Rolle spielt. Dabei kommt den RAF Kinasen eine Schlüsselrolle bei der Tumorgenese zu. Die Entwicklung von spezifischen irreversiblen Kinasehemmern stellt einen attraktiven, jedoch schwierigen Ansatz zur Tumorsupression dar. CI-1033 wurde erfolgreich mit dem Ziel entwickelt, ErbB-Rezeptor-Tyrosinkinasen irreversibel zu inhibieren, indem es kovalent mit dem Cys113 (p38alpha MAP Kinase Nummerierung) in der Kinase-Domäne reagiert. In dieser Arbeit wird ein vergleichbarer Ansatz gegen die RAF-Onkoproteine verfolgt, die einen analogen Cystein-Rest in der Position 108 aufweisen. Dieser ist in der Hinge-Region zwischen dem kleinen n-lobe und dem großen c-lobe der Kinase-Domäne lokalisiert. Ein neuer synthetischer Ansatz, der einen Lyophilisierungsschritt mit einschloss, erlaubte hierfür die Synthese einer Diphenylharnstoff-Verbindung mit einer Epoxidgruppe (Verbindung 1). Verbindung 1 zeigt in vitro tatsächlich eine inhibitorische Aktivität gegen RAF-Kinasen. Jedoch zeigen unsere zeitkinetischen Experimente, sowie unsere massenspektrometrischen Analysen, dass Verbindung 1 keine kovalente Bindung mit dem Cystein-Rest in der Hinge-Region bildet. Außerdem stellten wir in Zellkulturexperimenten eine starke Aktivierung des RAF-induzierten Signalweges fest; ein Effekt, der bereits für andere RAF-Kinase-Inhibitoren beschrieben wurde, jedoch hier erstmalig auch für eine Diphenylharnstoff-Verbindung, zu der auch BAY 43-9006 (Sarafinib, Nexavar) gehört. BAY 43-9006 ist ein unspezifischer, für die Behandlung von Krebs zugelassener, Kinase Inhibitor. Obwohl die Aktivierung in vitro scheinbar unabhängig von einer Heterooligomerisierung von B-RAF und C-RAF war, unterstützen in vivo Experimente einen solchen Mechanismus, da in gehungerten knockout Zellen, in denen B-RAF oder C-RAF fehlte, keine Aktivierung beobachtet werden konnte. Des Weiteren zeigten wir in einem mathematischen Modell, wie abhängig vom B-RAF/C-RAF-Verhältnis verschiedene Zellantworten durch unterschiedliche Stimuli induzierbar werden. Basierend auf biochemischen Daten über Aktivierung und Dephosphorylierung sowie auf den Differentialgleichungen unseres Rechenmodells fanden wir eine unterschiedliche Signalkinetik für B-RAF (starke Aktivierung, anhaltendes Signal) und C-RAF (schwache Aktivierung, transientes Signal). Die Bedeutung dieser differenzierten Signalmodifikation wurde auch durch unterschiedliche Expression der RAF Isoformen in verschiedenen Zelllinien und durch die experimentelle Messung der Kinaseaktivität von B- und C-RAF sowie mutierte Formen überprüft. Zusätzlich wurde der Effekt des Tumorsupressorproteins DiRas3 (auch bekannt als Noey2 oder ARHI) auf den RAF-Signalweg untersucht. Wir konnten zeigen, dass DiRas3 den mitogenen Signalweges durch Inhibierung der mitogen-aktivierten Proteinkinase Kinase (MEK) negativ reguliert, eine Basis für ein verfeinertes Modell der Ras/RAF/MEK/ERK Kaskade. KW - Systembiologie KW - RAf KW - BAY 43-9006 KW - Sorafinib KW - Nexavar KW - DiRas3 KW - Noey2 KW - ARHI KW - Diphenylharnstoff KW - Krebs KW - Melanom KW - Kinase Inhibitor KW - RAF KW - BAY 43-9006 KW - Sorafinib KW - Nexavar KW - DiRas3 KW - Noey2 KW - ARHI KW - diphenyl urea KW - cancer KW - melanoma KW - kinase inhibitor Y1 - 2007 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-26953 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 - THES A1 - Weber, Natalia T1 - Psychosoziale Aspekte bei hereditärer Mamma/Ovarial-Ca-Belastung T1 - Psychosocial aspects of hereditary breast/ovarian cancer exposure N2 - Ziel dieser Arbeit war die Untersuchung der psychischen Befindlichkeit und anderer gesundheitsbezogenen Konditionen der Frauen und Männer mit familiären Mamma- und Ovarialkarzinomrisiko sowie die Klärung hinsichtlich der Bewältigung und Auswirkung genetischer Risikoinformation. Es wurden Risikowahrnehmung, Informationsstand, Inanspruchnahme der Beratungsangebote sowie der Früherkennungsmaßnahmen, Einstellung gegenüber genetischer Brustkrebsdiagnostik und familiärer/sozialer Kommunikation untersucht. Die vollständig ausgefüllten Fragebögen von Ratsuchenden und Betroffenen, die an der Beratung und Befragung im Zentrum für „Familiären Brust-/Eierstockkrebs“ teilgenommen haben, wurden von uns ausgewertet. Für die beratenden Institutionen ist das Wissen der vielfältigen psychischen und sozialen Folgen bei den Testsuchenden und deren Familien sehr wichtig. Nur so kann das Betreuungskonzept und das Beratungsangebot verbessert werden. N2 - Goal of this dissertation was to examine the mental state and other health related conditions of women and men with familial breast and ovarian cancer risk and the clarification with regard to coping and impact of genetic risk information. Risk perception, level of information, usage of advisory services and early diagnosis measures, attitude towards genetic breast cancer diagnosis and familial/social communication have been investigated. The completely filled questionnaires of medical advice seeking and concerned persons having participated in the counseling and questioning in the center of familial breast and ovarian cancer have been evaluated by us. For the counseling institutions it is very important to have knowledge about the multifaceted mental and social implications of persons undergoing tests and their families. This is the only way to improve the care concept and the advisory services. KW - Brustkrebs KW - Eierstockkrebs KW - Psychosoziale Situation KW - Psychosoziale Beratung KW - Genetik KW - psychosoziale Aspekte KW - Krebserkrankungen KW - psychosocial aspects KW - cancer Y1 - 2007 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-28330 ER - TY - JOUR A1 - Heß, Verena A1 - Meng, Karin A1 - Schulte, Thomas A1 - Neuderth, Silke A1 - Bengel, Jürgen A1 - Faller, Hermann A1 - Schuler, Michael T1 - Prevalence and predictors of cancer patients' unexpressed needs in the admission interview of inpatient rehabilitation JF - Psycho‐Oncology N2 - Objective The admission interview in oncological inpatient rehabilitation might be a good opportunity to identify cancer patients' needs present after acute treatment. However, a relevant number of patients may not express their needs. In this study, we examined (a) the proportion of cancer patients with unexpressed needs, (b) topics of unexpressed needs and reasons for not expressing needs, (c) correlations of not expressing needs with several patient characteristics, and (d) predictors of not expressing needs. Methods We enrolled 449 patients with breast, prostate, and colon cancer at beginning and end of inpatient rehabilitation. We obtained self‐reports about unexpressed needs and health‐related variables (quality of life, depression, anxiety, adjustment disorder, and health literacy). We estimated frequencies and conducted correlation and ordinal logistic regression analyses. Results A quarter of patients stated they had “rather not” or “not at all” expressed all relevant needs. Patients mostly omitted fear of cancer recurrence. Most frequent reasons for not expressing needs were being focused on physical consequences of cancer, concerns emerging only later, and not knowing about the possibility of talking about distress. Not expressing needs was associated with several health‐related outcomes, for example, emotional functioning, adjustment disorder, fear of progression, and health literacy. Depression measured at the beginning of rehabilitation showed only small correlations and is therefore not sufficient to identify patients with unexpressed needs. Conclusions A relevant proportion of cancer patients reported unexpressed needs in the admission interview. This was associated with decreased mental health. Therefore, it seems necessary to support patients in expressing needs. KW - cancer KW - inpatient rehabilitation KW - oncology KW - prediction KW - prevalence KW - unexpressed needs Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228369 VL - 29 IS - 10 SP - 1549 EP - 1556 ER - TY - JOUR A1 - Adelfinger, Marion A1 - Bessler, Simon A1 - Cecil, Alexander A1 - Langbein-Laugwitz, Johanna A1 - Frentzen, Alexa A1 - Gentschev, Ivaylo A1 - Szalay, Aladar A. T1 - Preclinical Testing Oncolytic Vaccinia Virus Strain GLV-5b451 Expressing an Anti-VEGF Single-Chain Antibody for Canine Cancer Therapy JF - Viruses N2 - Virotherapy on the basis of oncolytic vaccinia virus (VACV) strains is a novel approach for canine cancer therapy. Here we describe, for the first time, the characterization and the use of VACV strain GLV-5b451 expressing the anti-vascular endothelial growth factor (VEGF) single-chain antibody (scAb) GLAF-2 as therapeutic agent against different canine cancers. Cell culture data demonstrated that GLV-5b451 efficiently infected and destroyed all four tested canine cancer cell lines including: mammary carcinoma (MTH52c), mammary adenoma (ZMTH3), prostate carcinoma (CT1258), and soft tissue sarcoma (STSA-1). The GLV-5b451 virus-mediated production of GLAF-2 antibody was observed in all four cancer cell lines. In addition, this antibody specifically recognized canine VEGF. Finally, in canine soft tissue sarcoma (CSTS) xenografted mice, a single systemic administration of GLV-5b451 was found to be safe and led to anti-tumor effects resulting in the significant reduction and substantial long-term inhibition of tumor growth. A CD31-based immuno-staining showed significantly decreased neo-angiogenesis in GLV-5b451-treated tumors compared to the controls. In summary, these findings indicate that GLV-5b451 has potential for use as a therapeutic agent in the treatment of CSTS. KW - canine cancer therapy KW - canine soft tissue sarcoma (CSTS) KW - oncolytic virus KW - cancer KW - canine cancer cell lines KW - antibody production KW - angiogenesis Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-125705 VL - 7 ER - TY - JOUR A1 - Gentschev, Ivaylo A1 - Adelfinger, Marion A1 - Josupeit, Rafael A1 - Rudolph, Stephan A1 - Ehrig, Klaas A1 - Donat, Ulrike A1 - Weibel, Stephanie A1 - Chen, Nanhai G. A1 - Yu, Yong A. A1 - Zhang, Qian A1 - Heisig, Martin A1 - Thamm, Douglas A1 - Stritzker, Jochen A1 - MacNeill, Amy A1 - Szalay, Aladar A. T1 - Preclinical Evaluation of Oncolytic Vaccinia Virus for Therapy of Canine Soft Tissue Sarcoma JF - PLoS One N2 - Virotherapy using oncolytic vaccinia virus (VACV) strains is one promising new strategy for canine cancer therapy. In this study we describe the establishment of an in vivo model of canine soft tissue sarcoma (CSTS) using the new isolated cell line STSA-1 and the analysis of the virus-mediated oncolytic and immunological effects of two different Lister VACV LIVP1.1.1 and GLV-1h68 strains against CSTS. Cell culture data demonstrated that both tested VACV strains efficiently infected and destroyed cells of the canine soft tissue sarcoma line STSA-1. In addition, in our new canine sarcoma tumor xenograft mouse model, systemic administration of LIVP1.1.1 or GLV-1h68 viruses led to significant inhibition of tumor growth compared to control mice. Furthermore, LIVP1.1.1 mediated therapy resulted in almost complete tumor regression and resulted in long-term survival of sarcoma-bearing mice. The replication of the tested VACV strains in tumor tissues led to strong oncolytic effects accompanied by an intense intratumoral infiltration of host immune cells, mainly neutrophils. These findings suggest that the direct viral oncolysis of tumor cells and the virus-dependent activation of tumor-associated host immune cells could be crucial parts of anti-tumor mechanism in STSA-1 xenografts. In summary, the data showed that both tested vaccinia virus strains and especially LIVP1.1.1 have great potential for effective treatment of CSTS. KW - breast-tumors KW - animal-model KW - nude-mice KW - cell-line KW - in-vitro KW - glv-1h68 KW - cancer KW - virotherapy KW - dogs KW - neutrophils Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-129998 VL - 7 IS - 5 ER - TY - JOUR A1 - Metzenmacher, Martin A1 - Váraljai, Renáta A1 - Hegedüs, Balazs A1 - Cima, Igor A1 - Forster, Jan A1 - Schramm, Alexander A1 - Scheffler, Björn A1 - Horn, Peter A. A1 - Klein, Christoph A. A1 - Szarvas, Tibor A1 - Reis, Hennig A1 - Bielefeld, Nicola A1 - Roesch, Alexander A1 - Aigner, Clemens A1 - Kunzmann, Volker A1 - Wiesweg, Marcel A1 - Siveke, Jens T. A1 - Schuler, Martin A1 - Lueong, Smiths S. T1 - Plasma Next Generation Sequencing and Droplet Digital-qPCR-Based Quantification of Circulating Cell-Free RNA for Noninvasive Early Detection of Cancer JF - Cancers N2 - Early detection of cancer holds high promise for reducing cancer-related mortality. Detection of circulating tumor-specific nucleic acids holds promise, but sensitivity and specificity issues remain with current technology. We studied cell-free RNA (cfRNA) in patients with non-small cell lung cancer (NSCLC; n = 56 stage IV, n = 39 stages I-III), pancreatic cancer (PDAC, n = 20 stage III), malignant melanoma (MM, n = 12 stage III-IV), urothelial bladder cancer (UBC, n = 22 stage II and IV), and 65 healthy controls by means of next generation sequencing (NGS) and real-time droplet digital PCR (RT-ddPCR). We identified 192 overlapping upregulated transcripts in NSCLC and PDAC by NGS, more than 90% of which were noncoding. Previously reported transcripts (e.g., HOTAIRM1) were identified. Plasma cfRNA transcript levels of POU6F2-AS2 discriminated NSCLC from healthy donors (AUC = 0.82 and 0.76 for stages IV and I–III, respectively) and significantly associated (p = 0.017) with the established tumor marker Cyfra 21-1. cfRNA yield and POU6F2-AS transcript abundance discriminated PDAC patients from healthy donors (AUC = 1.0). POU6F2-AS2 transcript was significantly higher in MM (p = 0.044). In summary, our findings support further validation of cfRNA detection by RT-ddPCR as a biomarker for early detection of solid cancers. KW - liquid biopsy KW - cfRNA KW - cancer KW - ddPCR KW - NGS KW - POU6F2-AS2 KW - early detection Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-200553 SN - 2072-6694 VL - 12 IS - 2 ER - TY - JOUR A1 - Vollmer, Andreas A1 - Nagler, Simon A1 - Hörner, Marius A1 - Hartmann, Stefan A1 - Brands, Roman C. A1 - Breitenbücher, Niko A1 - Straub, Anton A1 - Kübler, Alexander A1 - Vollmer, Michael A1 - Gubik, Sebastian A1 - Lang, Gernot A1 - Wollborn, Jakob A1 - Saravi, Babak T1 - Performance of artificial intelligence-based algorithms to predict prolonged length of stay after head and neck cancer surgery JF - Heliyon N2 - Background Medical resource management can be improved by assessing the likelihood of prolonged length of stay (LOS) for head and neck cancer surgery patients. The objective of this study was to develop predictive models that could be used to determine whether a patient's LOS after cancer surgery falls within the normal range of the cohort. Methods We conducted a retrospective analysis of a dataset consisting of 300 consecutive patients who underwent head and neck cancer surgery between 2017 and 2022 at a single university medical center. Prolonged LOS was defined as LOS exceeding the 75th percentile of the cohort. Feature importance analysis was performed to evaluate the most important predictors for prolonged LOS. We then constructed 7 machine learning and deep learning algorithms for the prediction modeling of prolonged LOS. Results The algorithms reached accuracy values of 75.40 (radial basis function neural network) to 97.92 (Random Trees) for the training set and 64.90 (multilayer perceptron neural network) to 84.14 (Random Trees) for the testing set. The leading parameters predicting prolonged LOS were operation time, ischemia time, the graft used, the ASA score, the intensive care stay, and the pathological stages. The results revealed that patients who had a higher number of harvested lymph nodes (LN) had a lower probability of recurrence but also a greater LOS. However, patients with prolonged LOS were also at greater risk of recurrence, particularly when fewer (LN) were extracted. Further, LOS was more strongly correlated with the overall number of extracted lymph nodes than with the number of positive lymph nodes or the ratio of positive to overall extracted lymph nodes, indicating that particularly unnecessary lymph node extraction might be associated with prolonged LOS. Conclusions The results emphasize the need for a closer follow-up of patients who experience prolonged LOS. Prospective trials are warranted to validate the present results. KW - prediction KW - head and neck cancer KW - machine learning KW - deep learning KW - artificial intelligence KW - length of stay KW - cancer Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-350416 SN - 2405-8440 VL - 9 IS - 11 ER - TY - JOUR A1 - Henriksson, Sofia A1 - Calderón-Montaño, José Manuel A1 - Solvie, Daniel A1 - Warpman Berglund, Ulrika A1 - Helleday, Thomas T1 - Overexpressed c-Myc sensitizes cells to TH1579, a mitotic arrest and oxidative DNA damage inducer JF - Biomolecules N2 - Previously, we reported that MTH1 inhibitors TH588 and TH1579 selectively induce oxidative damage and kill Ras-expressing or -transforming cancer cells, as compared to non-transforming immortalized or primary cells. While this explains the impressive anti-cancer properties of the compounds, the molecular mechanism remains elusive. Several oncogenes induce replication stress, resulting in under replicated DNA and replication continuing into mitosis, where TH588 and TH1579 treatment causes toxicity and incorporation of oxidative damage. Hence, we hypothesized that oncogene-induced replication stress explains the cancer selectivity. To test this, we overexpressed c-Myc in human epithelial kidney cells (HA1EB), resulting in increased proliferation, polyploidy and replication stress. TH588 and TH1579 selectively kill c-Myc overexpressing clones, enforcing the cancer cell selective killing of these compounds. Moreover, the toxicity of TH588 and TH1579 in c-Myc overexpressing cells is rescued by transcription, proteasome or CDK1 inhibitors, but not by nucleoside supplementation. We conclude that the molecular toxicological mechanisms of how TH588 and TH1579 kill c-Myc overexpressing cells have several components and involve MTH1-independent proteasomal degradation of c-Myc itself, c-Myc-driven transcription and CDK activation. KW - MTH1 KW - TH588 KW - TH1579 KW - c-Myc KW - replication stress KW - DNA damage KW - cell death KW - cancer Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-297547 SN - 2218-273X VL - 12 IS - 12 ER - TY - JOUR A1 - Kastner, Carolin A1 - Hendricks, Anne A1 - Deinlein, Hanna A1 - Hankir, Mohammed A1 - Germer, Christoph-Thomas A1 - Schmidt, Stefanie A1 - Wiegering, Armin T1 - Organoid Models for Cancer Research — From Bed to Bench Side and Back JF - Cancers N2 - Simple Summary Despite significant strides in multimodal therapy, cancers still rank within the first three causes of death especially in industrial nations. A lack of individualized approaches and accurate preclinical models are amongst the major barriers that limit the development of novel therapeutic options and drugs. Recently, the 3D culture system of organoids was developed which stably retains the genetic and phenotypic characteristics of the original tissue, healthy as well as diseased. In this review, we summarize current data and evidence on the relevance and reliability of such organoid culture systems in cancer research, focusing on their role in drug investigations (in a personalized manner). Abstract Organoids are a new 3D ex vivo culture system that have been applied in various fields of biomedical research. First isolated from the murine small intestine, they have since been established from a wide range of organs and tissues, both in healthy and diseased states. Organoids genetically, functionally and phenotypically retain the characteristics of their tissue of origin even after multiple passages, making them a valuable tool in studying various physiologic and pathophysiologic processes. The finding that organoids can also be established from tumor tissue or can be engineered to recapitulate tumor tissue has dramatically increased their use in cancer research. In this review, we discuss the potential of organoids to close the gap between preclinical in vitro and in vivo models as well as clinical trials in cancer research focusing on drug investigation and development. KW - cancer KW - tumor disease KW - organoid KW - patient-derived organoid (PDOs) KW - patient-derived tumor organoid (PDTO) Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-246307 SN - 2072-6694 VL - 13 IS - 19 ER - TY - JOUR A1 - Gentschev, Ivaylo A1 - Patil, Sadeep S. A1 - Petrov, Ivan A1 - Cappello, Joseph A1 - Adelfinger, Marion A1 - Szalay, Aladar A. T1 - Oncolytic Virotherapy of Canine and Feline Cancer JF - Viruses N2 - Cancer is the leading cause of disease-related death in companion animals such as dogs and cats. Despite recent progress in the diagnosis and treatment of advanced canine and feline cancer, overall patient treatment outcome has not been substantially improved. Virotherapy using oncolytic viruses is one promising new strategy for cancer therapy. Oncolytic viruses (OVs) preferentially infect and lyse cancer cells, without causing excessive damage to surrounding healthy tissue, and initiate tumor-specific immunity. The current review describes the use of different oncolytic viruses for cancer therapy and their application to canine and feline cancer. KW - oncolytic virus KW - oncolysis KW - cancer KW - canine and feline cancer therapy Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119753 VL - 6 IS - 5 ER - TY - JOUR A1 - Patil, Sandeep S. A1 - Gentschev, Ivaylo A1 - Nolte, Ingo A1 - Ogilvie, Gregory A1 - Szalay, Aladar A. T1 - Oncolytic virotherapy in veterinary medicine: current status and future prospects for canine patients N2 - Oncolytic viruses refer to those that are able to eliminate malignancies by direct targeting and lysis of cancer cells, leaving non-cancerous tissues unharmed. Several oncolytic viruses including adenovirus strains, canine distemper virus and vaccinia virus strains have been used for canine cancer therapy in preclinical studies. However, in contrast to human studies, clinical trials with oncolytic viruses for canine cancer patients have not been reported. An ‘ideal’ virus has yet to be identified. This review is focused on the prospective use of oncolytic viruses in the treatment of canine tumors - a knowledge that will undoubtedly contribute to the development of oncolytic viral agents for canine cancer therapy in the future. KW - Medizin KW - cancer KW - canine cancer therapy KW - oncolytic virus KW - oncolysis KW - target molecule KW - combination therapy Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-75128 ER - TY - JOUR A1 - Elkon, Ran A1 - Loayza-Puch, Fabricio A1 - Korkmaz, Gozde A1 - Lopes, Rui A1 - van Breugel, Pieter C A1 - Bleijerveld, Onno B A1 - Altelaar, AF Maarten A1 - Wolf, Elmar A1 - Lorenzin, Francesca A1 - Eilers, Martin A1 - Agami, Reuven T1 - Myc coordinates transcription and translation to enhance transformation and suppress invasiveness JF - EMBO reports N2 - c‐Myc is one of the major human proto‐oncogenes and is often associated with tumor aggression and poor clinical outcome. Paradoxically, Myc was also reported as a suppressor of cell motility, invasiveness, and metastasis. Among the direct targets of Myc are many components of the protein synthesis machinery whose induction results in an overall increase in protein synthesis that empowers tumor cell growth. At present, it is largely unknown whether beyond the global enhancement of protein synthesis, Myc activation results in translation modulation of specific genes. Here, we measured Myc‐induced global changes in gene expression at the transcription, translation, and protein levels and uncovered extensive transcript‐specific regulation of protein translation. Particularly, we detected a broad coordination between regulation of transcription and translation upon modulation of Myc activity and showed the connection of these responses to mTOR signaling to enhance oncogenic transformation and to the TGFβ pathway to modulate cell migration and invasiveness. Our results elucidate novel facets of Myc‐induced cellular responses and provide a more comprehensive view of the consequences of its activation in cancer cells. KW - c‐Myc KW - transcriptional responses KW - translational regulation KW - transcription KW - transformation KW - metastasis KW - cancer KW - protein biosynthesis & quality control Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-150373 VL - 16 IS - 12 ER - TY - JOUR A1 - Kienle-Garrido, Melina-Lorén A1 - Breitenbach, Tim A1 - Chudej, Kurt A1 - Borzì, Alfio T1 - Modeling and numerical solution of a cancer therapy optimal control problem JF - Applied Mathematics N2 - A mathematical optimal-control tumor therapy framework consisting of radio- and anti-angiogenesis control strategies that are included in a tumor growth model is investigated. The governing system, resulting from the combination of two well established models, represents the differential constraint of a non-smooth optimal control problem that aims at reducing the volume of the tumor while keeping the radio- and anti-angiogenesis chemical dosage to a minimum. Existence of optimal solutions is proved and necessary conditions are formulated in terms of the Pontryagin maximum principle. Based on this principle, a so-called sequential quadratic Hamiltonian (SQH) method is discussed and benchmarked with an “interior point optimizer―a mathematical programming language” (IPOPT-AMPL) algorithm. Results of numerical experiments are presented that successfully validate the SQH solution scheme. Further, it is shown how to choose the optimisation weights in order to obtain treatment functions that successfully reduce the tumor volume to zero. KW - cancer KW - therapy KW - optimal control problem Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-177152 VL - 9 IS - 8 ER - TY - JOUR A1 - Haussmann, Alexander A1 - Schmidt, Martina E. A1 - Illmann, Mona L. A1 - Schröter, Marleen A1 - Hielscher, Thomas A1 - Cramer, Holger A1 - Maatouk, Imad A1 - Horneber, Markus A1 - Steindorf, Karen T1 - Meta-analysis of randomized controlled trials on yoga, psychosocial, and mindfulness-based interventions for cancer-related fatigue: What intervention characteristics are related to higher efficacy? JF - Cancers N2 - Cancer-related fatigue (CRF) is a burdensome sequela of cancer treatments. Besides exercise, recommended therapies for CRF include yoga, psychosocial, and mindfulness-based interventions. However, interventions conducted vary widely, and not all show a significant effect. This meta-analysis aimed to explore intervention characteristics related to greater reductions in CRF. We included randomized controlled trials published before October 2021. Standardized mean differences were used to assess intervention efficacy for CRF and multimodel inference to explore intervention characteristics associated with higher efficacy. For the meta-analysis, we included 70 interventions (24 yoga interventions, 31 psychosocial interventions, and 15 mindfulness-based interventions) with 6387 participants. The results showed a significant effect of yoga, psychosocial, and mindfulness-based interventions on CRF but with high heterogeneity between studies. For yoga and mindfulness-based interventions, no particular intervention characteristic was identified to be advantageous for reducing CRF. Regarding psychosocial interventions, a group setting and work on cognition were related to higher intervention effects on CRF. The results of this meta-analysis suggest options to maximize the intervention effects of psychosocial interventions for CRF. The effects of yoga and mindfulness-based interventions for CRF appear to be independent of their design, although the limited number of studies points to the need for further research. KW - fatigue KW - cancer KW - psychosocial KW - mindfulness KW - yoga KW - quality of life KW - patient-reported outcomes Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-270753 SN - 2072-6694 VL - 14 IS - 8 ER - TY - JOUR A1 - Pellegrino, Marsha A1 - Del Bufalo, Francesca A1 - De Angelis, Biagio A1 - Quintarelli, Concetta A1 - Caruana, Ignazio A1 - de Billy, Emmanuel T1 - Manipulating the metabolism to improve the efficacy of CAR T-cell immunotherapy JF - Cells N2 - The adoptive transfer of the chimeric antigen receptor (CAR) expressing T-cells has produced unprecedented successful results in the treatment of B-cell malignancies. However, the use of this technology in other malignancies remains less effective. In the setting of solid neoplasms, CAR T-cell metabolic fitness needs to be optimal to reach the tumor and execute their cytolytic function in an environment often hostile. It is now well established that both tumor and T cell metabolisms play critical roles in controlling the immune response by conditioning the tumor microenvironment and the fate and activity of the T cells. In this review, after a brief description of the tumoral and T cell metabolic reprogramming, we summarize the latest advances and new strategies that have been developed to improve the metabolic fitness and efficacy of CAR T-cell products. KW - cancer KW - metabolic reprogramming KW - combined therapy KW - Chimeric Antigen Receptor T cells KW - immunotherapy Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220140 SN - 2073-4409 VL - 10 IS - 1 ER - TY - JOUR A1 - Hausmann, Stefan A1 - Brandt, Evelyn A1 - Köchel, Carolin A1 - Einsele, Hermann A1 - Bargou, Ralf C. A1 - Seggewiss-Bernhardt, Ruth A1 - Stühmer, Thorsten T1 - Loss of serum and glucocorticoid-regulated kinase 3 (SGK3) does not affect proliferation and survival of multiple myeloma cell lines JF - PLoS ONE N2 - Multiple myeloma (MM) is a generally fatal plasma cell cancer that often shows activation of the phosphoinositide 3-kinase/Akt (PI3K/Akt) pathway. Targeted pharmacologic therapies, however, have not yet progressed beyond the clinical trial stage, and given the complexity of the PI3K/Akt signalling system (e.g. multiple protein isoforms, diverse feedback regulation mechanisms, strong variability between patients) it is mandatory to characterise its ramifications in order to better guide informed decisions about the best therapeutic approaches. Here we explore whether serum and glucocorticoid-regulated kinase 3 (SGK3), a potential downstream effector of PI3K, plays a role in oncogenic signalling in MM cells-either in concert with or independent of Akt. SGK3 was expressed in all MM cell lines and in all primary MM samples tested. Four MM cell lines representing a broad range of intrinsic Akt activation (very strong: MM. 1s, moderate: L 363 and JJN-3, absent: AMO-1) were chosen to test the effects of transient SGK3 knockdown alone and in combination with pharmacological inhibition of Akt, PI3K-p110\(\alpha\), or in the context of serum starvation. Although the electroporation protocol led to strong SGK3 depletion for at least 5 days its absence had no substantial effect on the activation status of potential downstream substrates, or on the survival, viability or proliferation of MM cells in all experimental contexts tested. We conclude that it is unlikely that SGK3 plays a significant role for oncogenic signalling in multiple myeloma. KW - Akt KW - phosphorylation KW - downstream KW - mechanism KW - pathway KW - isoforms KW - activation KW - cancer KW - inhibition KW - phosphatidylinositol 3-kinase/Akt Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148708 VL - 10 IS - 4 ER - TY - JOUR A1 - Altieri, Barbara A1 - Sbiera, Silviu A1 - Della Casa, Silvia A1 - Weigand, Isabel A1 - Wild, Vanessa A1 - Steinhauer, Sonja A1 - Fadda, Guido A1 - Kocot, Arkadius A1 - Bekteshi, Michaela A1 - Mambretti, Egle M. A1 - Rosenwald, Andreas A1 - Pontecorvi, Alfredo A1 - Fassnacht, Martin A1 - Ronchi, Cristina L. T1 - Livin/BIRC7 expression as malignancy marker in adrenocortical tumors JF - Oncotarget N2 - Livin/BIRC7 is a member of the inhibitors of apoptosis proteins family, which are involved in tumor development through the inhibition of caspases. Aim was to investigate the expression of livin and other members of its pathway in adrenocortical tumors and in the adrenocortical carcinoma (ACC) cell line NCI-H295R. The mRNA expression of livin, its isoforms α and β, XIAP, CASP3 and DIABLO was evaluated by qRT-PCR in 82 fresh-frozen adrenal tissues (34 ACC, 25 adenomas = ACA, 23 normal adrenal glands = NAG). Livin protein expression was assessed by immunohistochemistry in 270 paraffin-embedded tissues (192 ACC, 58 ACA, 20 NAG). Livin, CASP3 and cleaved caspase-3 were evaluated in NCI-H295R after induction of livin overexpression. Relative livin mRNA expression was significantly higher in ACC than in ACA and NAG (0.060 ± 0.116 vs 0.004 ± 0.014 and 0.002 ± 0.009, respectively, p < 0.01), being consistently higher in tumors than in adjacent NAG and isoform β more expressed than α. No significant differences in CASP3, XIAP and DIABLO levels were found among these groups. In immunohistochemistry, livin was localized in both cytoplasm and nuclei. The ratio between cytoplasmic and nuclear staining was significantly higher in ACC (1.51 ± 0.66) than in ACA (0.80 ± 0.35) and NAG (0.88 ± 0.27; p < 0.0001). No significant correlations were observed between livin expression and histopathological parameters or clinical outcome. In NCI-H295R cells, the livin overexpression slightly reduced the activation of CASP3, but did not correlate with cell viability. In conclusion, livin is specifically over-expressed in ACC, suggesting that it might be involved in adrenocortical tumorigenesis and represent a new molecular marker of malignancy. KW - cancer KW - livin KW - BIRC7 KW - adrenocortical cancer KW - adrenal tumor KW - caspase-3 Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-171887 VL - 8 IS - 6 ER - TY - THES A1 - Laufer, Antje T1 - Kombinierte zytogenetische und morphologische Analyse follikulärer Non-Hodgkin Lymphome : Eine neue rekurrente chromosomale Aberration bei prädominant diffusen follikulären Lymphomen T1 - Combined cytogenetic and morphologic analysis of follicular Non Hodgkin lymphomas. A new rekurrent chromosomal aberration with predominant diffuse follicular lymphomas. N2 - Das follikuläre Lymphom ist eines der häufigsten Non-Hodgkin Lymphome und überwiegend eine Erkrankung des erwachsenen Menschen. In der WHO-Klassifikation ist es als ein Lymphom von Keimzentrumszellen definiert, das follikulär und/oder diffus wachsen kann. Zur Subklassifikation follikulärer Lymphome empfiehlt die WHO-Klassifikation eine Unterscheidung der Grade 1, 2 und 3 durch Auszählen der Zentroblasten pro zehn Gesichtsfelder in starker Vergrößerung. Beim Grad 3A liegen neben Zentroblasten auch Zentrozyten vor. FL Grad 3B bestehen ausschließlich aus Zentroblasten. Hinsichtlich der Zytomorphologie, Immunhistologie und Genetik bestehen deutliche Unterschiede zwischen FL Grad 1, 2 und 3A gegenüber FL Grad 3B. In der vorliegenden Arbeit konnte gezeigt werden, dass die weit überwiegende Zahl der follikulären Lymphome Grad 1, 2 und 3A ein prädominant follikuläres Wachstumsmuster aufwies. Ein follikulärer und diffuser Wuchstyp lag seltener vor. Noch seltener war ein überwiegend bzw. „rein“ diffuses Wachstumsmuster. Die mitotische und proliferative Aktivität stieg mit dem Tumorgrad linear an. Hinsichtlich der CD10 Reaktivität, der BCL-2 und p53 Expression sowie des Nachweises einer sekretorischen Differenzierung ergaben sich beim Vergleich der FL Grad 1 bis 3A keine statistisch signifikanten Unterschiede. Die BCL-2 Expression nahm allerdings bei den FL1-3A mit zunehmendem Grad ab. In zytogenetischen Untersuchungen wurden in allen follikulären Lymphomen Grad 1 bis 3A primäre bzw. sekundäre Chromosomenaberrationen gefunden. Unter den rekurrenten chromosomalen Alterationen trat die Translokation t(14;18)(q32;q21) am häufigsten auf und war insbesondere bei follikulären Lymphomen Grad 1 und 2, in etwas geringerem Maße auch bei FL Grad 3A anzutreffen. Diese Translokation scheint also in einem frühen Stadium der B-Zell-Entwicklung aufzutreten und führt primär zu einem höher differenzierten (zentrozytenreichen) Lymphom. Die t(14;18) bedingt zumeist eine Überexpression des BCL-2 Gens, die sich auch immunhistochemisch nachweisen lässt und diagnostische Verwendung findet. Das BCL-2 Protein ist daher von Nutzen für die Unterscheidung neoplastischer von reaktiven Follikeln, nicht aber, um follikuläre von anderen „low grade“ B-Zell Lymphomen zu unterscheiden. Die sekundären Alterationen charakterisieren bestimmte undifferenzierte Stadien mit hohem Blastenanteil und einer hohen mitotischen und proliferativen Aktivität. In zytogenetischen Untersuchungen von überwiegend diffus wachsenden FL konnte keine Translokation t(14;18)(q32;q21) nachgewiesen werden. Die identische Morphologie dieser Lymphome und die identischen Veränderungen auch auf genetischer Ebene deuten auf die nahe Verwandtschaft der überwiegend diffus wachsenden FL mit den typischen Keimzentrums-lymphomen hin. Es handelt sich jedoch um eine eigenständige Identität, die differenzierten Keimzentrumslymphome, die wahrscheinlich aufgrund des Fehlens einer t(14;18)(q32;q21) ein primär und ausgeprägt diffuses Wachstumsmuster aufweisen. N2 - The follicular lymphoma is predominant one the most frequent Non Hodgkin of lymphomas and an illness of the adult humans. In the WHO classification it is defined as a lymphoma of germ center cells, which can grow follicular and/or vaguely. To the Subklassifikation of follicular lymphomas the WHO classification recommends a distinction of the degrees of 1, 2 and 3 by counting out the centroblasten per ten visual field in strong enlargement. With the degree of 3A also centrocyten are present beside centroblasten. Flat steel bars degrees of 3B exclusively consist of centroblasten. Regarding the cytomorphology, immunhistology and genetics clear differences between flat steel bar degrees 1, 2 and 3A exist opposite flat steel bar degree 3B. In the available work it could be shown that the number of the follicular lymphomas degrees of 1, 2 and 3A outweighing far predominant folliculars growth sample exhibited. A follicular and diffuse type of stature was present more rarely. Still more rarely a “diffuse growth sample was predominant and/or „pure. The mitotic and proliferative activity rose with the tumor degree linear. Regarding the CD10 reactivity, the BCL-2 and p53 Expression as well as the proof of a secretoric differentiation did not result in the case of the comparison of the flat steel bars degrees of 1 to 3A statistically significant differences. The BCL-2 Expression removed however with the FL1-3A with increasing degree. In cytogenetic investigations in all follicular lymphomas degrees of 1 to 3A primary and/or secondary chromosome aberrations were found. Under the recurrent chromosomalen old person rations the translokation t (14 stepped; 18) (q32; q21) most frequently up and was in particular with follicular lymphomas degree of 1 and 2 to find in somewhat smaller measure also with flat steel bar degree of 3A. This translokation seems to arise thus in an early stage of the B-cell-development and leads primarily to a more highly differentiated (centrocyt) lymphoma. The t (14; ) mostly a overexpression of the BCL-2 of gene, which can be proven also immune-histochemical, causes 18 and diagnostic use finds. The BCL-2 protein is from there of use for the distinction more neoplastic of reactive follicles, not however, in order to differentiated follicular from others „low degrees “B-cell lymphomas. The secondary old person rations characterize certain undifferentiated stages with high blowing portion and a high mitotic and proliferativen activity. In cytogenetic investigations of predominantly vaguely growing flat steel bar no Translokation t (14 could; 18) (q32; q21) to be proven. The identical morphology of these lymphomas and the identical changes also on genetic level point growing flat steel bar with the typical germ center lymphomas to the close relationship that predominantly vaguely. However the differentiated germ center lymphomas, those concern probably due to the absence of a t their own identity, (14; 18) (q32; q21) primarily and minted diffuse growth sample exhibit. KW - Lymphdrüse KW - Krebs KW - Tumor KW - Lymphknoten KW - Hodgkin KW - Lymphdrüse KW - Krebs KW - Tumor KW - Lymphknoten KW - Hodgkin KW - lymphoma KW - cancer KW - Hodgkin KW - tumor KW - lymph node Y1 - 2008 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-29701 ER - TY - JOUR A1 - Klement, Rainer A1 - Kämmerer, Ulrike T1 - Is there a role for carbohydrate restriction in the treatment and prevention of cancer? N2 - Over the last years, evidence has accumulated suggesting that by systematically reducing the amount of dietary carbohydrates (CHOs) one could suppress, or at least delay, the emergence of cancer, and that proliferation of already existing tumor cells could be slowed down. This hypothesis is supported by the association between modern chronic diseases like the metabolic syndrome and the risk of developing or dying from cancer. CHOs or glucose, to which more complex carbohydrates are ultimately digested, can have direct and indirect effects on tumor cell proliferation: first, contrary to normal cells, most malignant cells depend on steady glucose availability in the blood for their energy and biomass generating demands and are not able to metabolize significant amounts of fatty acids or ketone bodies due to mitochondrial dysfunction. Second, high insulin and insulin-like growth factor (IGF)-1 levels resulting from chronic ingestion of CHO-rich Western diet meals, can directly promote tumor cell proliferation via the insulin/IGF1 signaling pathway. Third, ketone bodies that are elevated when insulin and blood glucose levels are low, have been found to negatively affect proliferation of different malignant cells in vitro or not to be usable by tumor cells for metabolic demands, and a multitude of mouse models have shown antitumorigenic properties of very low CHO ketogenic diets. In addition, many cancer patients exhibit an altered glucose metabolism characterized by insulin resistance and may profit from an increased protein and fat intake. In this review, we address the possible beneficial effects of low CHO diets on cancer prevention and treatment. Emphasis will be placed on the role of insulin and IGF1 signaling in tumorigenesis as well as altered dietary needs of cancer patients. KW - Medizin KW - Ketogenic diet KW - cancer KW - review KW - low carbohydrate diet KW - cachexia KW - insulin KW - insulin-like growth factor 1 (IGF1) Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-69178 ER - TY - THES A1 - Dejure, Francesca Romana T1 - Investigation of the role of MYC as a stress responsive protein T1 - Untersuchung der Rolle von MYC als stress-reguliertes Protein N2 - The transcription factor MYC is deregulated in over 70% of all human tumors and, in its oncogenic form, plays a major role in the cancer metabolic reprogramming, promoting the uptake of nutrients in order to sustain the biosynthetic needs of cancer cells. The research presented in this work aimed to understand if MYC itself is regulated by nutrient availability, focusing on the two major fuels of cancer cells: glucose and glutamine. Initial observations showed that endogenous MYC protein levels strongly depend on the availability of glutamine, but not of glucose. Subsequent analysis highlighted that the mechanism which accounts for the glutamine-mediated regulation of MYC is dependent on the 3´-untranslated region (3´-UTR) of MYC. Enhanced glutamine utilization by tumors has been shown to be directly linked to MYC oncogenic activity and MYC-dependent apoptosis has been observed under glutamine starvation. Such effect has been described in experimental systems which are mainly based on the use of MYC transgenes that do not contain the 3´-UTR. It was observed in the present study that cells are able to survive under glutamine starvation, which leads to cell cycle arrest and not apoptosis, as previously reported. However, enforced expression of a MYC transgene, which lacks the 3´-UTR, strongly increases the percentage of apoptotic cells upon starvation. Evaluation of glutamine-derived metabolites allowed to identify adenosine nucleotides as the specific stimulus responsible for the glutamine-mediated regulation of MYC, in a 3´-UTR-dependent way. Finally, glutamine-dependent MYC-mediated effects on RNA Polymerase II (RNAPII) function were evaluated, since MYC is involved in different steps of global transcriptional regulation. A global loss of RNAPII recruitment at the transcriptional start site results upon glutamine withdrawal. Such effect is overcome by enforced MYC expression under the same condition. This study shows that the 3´UTR of MYC acts as metabolic sensor and that MYC globally regulates the RNAPII function according to the availability of glutamine. The observations presented in this work underline the importance of considering stress-induced mechanisms impinging on the 3´UTR of MYC. N2 - In über 70% aller Krebserkrankungen ist der Transkriptionsfaktor MYC dereguliert. Dabei spielt onkogenes MYC unter anderem eine wichtige Rolle bei der Umprogrammierung metabolischer Prozesse indem es z.B. die Aufnahme von Nährstoffen wie Glutamin oder Glukose fördert, um den veränderten Bedürfnissen an den Stoffwechsel der Krebszellen Rechnung zu tragen. Die im Rahmen dieser Arbeit erzielten Ergebnisse zeigen, dass auch das MYC-Protein selbst durch die Verfügbarkeit von Nährstoffen in der Zelle reguliert werden kann. Erste Beobachtungen zeigten, dass die endogenen MYC Proteinlevel stark von der Verfügbarkeit von Glutamin, jedoch nicht von Glucose, abhängen. Weiterführende Experimente ergaben außerdem, dass der Mechanismus, der der Glutamin vermittelten Regulation von MYC zugrunde liegt, abhängig von der 3´-untranslatierten Region (3´-UTR) der MYC-mRNA ist. Es konnte bereits gezeigt werden, dass in Tumoren die verstärkte Nutzung von Glutamin in direktem Zusammenhang mit der onkogenen Aktivität von MYC steht und Zellen unter Glutaminentzug MYC-abhängig Apoptose einleiten. Diese Effekte wurden in experimentellen Systemen beschrieben, die auf einer Überexpression eines MYCTransgenes basierten, welches keine 3´-UTR enthält. In dieser Arbeit konnte jedoch beobachtet werden, dass Zellen, die ohne Glutamin kultiviert wurden, in der Lage waren zu überleben, da entgegen den Resultaten vorausgegangener Studien, ein Arrest des Zellzyklus und nicht Apoptose eingeleitet wurde. Die verstärkte Expression eines MYCTransgenes ohne 3´-UTR, erhöhte jedoch auch unter diesen Bedingungen die Anzahl apoptotischer Zellen. Weiterhin war es möglich Adenosin, für dessen Biosynthese Glutamin notwendig ist, als Stimulus zu identifizieren, der für die 3´-UTR abhängige Regulation von MYC verantwortlich ist. Da MYC in verschiedene Schritte der globalen Regulation der Transkription eingebunden ist, wurden abschließend die durch MYC vermittelten Glutaminabhängigen Effekte auf die RNA-Polymerase II (RNAPII) untersucht. Dabei zeigte sich, dass es nach Glutaminentzug zu einem globalen Verlust der Rekrutierung von RNAPII zu den Transkriptionsstartstellen kommt, was durch eine verstärkte MYC-Expression wieder aufgehoben werden kann. Zusammenfassend konnte in dieser Arbeit gezeigt werden, dass die 3´-UTR von MYC als metabolischer Sensor fungiert und dass MYC in Abhängigkeit der Verfügbarkeit von Glutamin global die RNAPII Funktion reguliert. Diese Studie hebt weiterhin die Bedeutung der 3´-UTR von MYC für die Vermittlung stressinduzierter Feedback-Mechanismen hervor. KW - cancer KW - metabolism KW - MYC KW - Myc KW - Stress KW - Metabolismus KW - Genregulation KW - Glutamin Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-158587 ER - TY - THES A1 - Gupta, Rohini T1 - Intracellular self-activation of the TrkB kinase domain causes FAK phosphorylation and disrupts actin filopodia dynamics T1 - Intrazelluläre Selbst-aktivierung der TrkB Kinase induziert FAK Phosphorylierung und verändert die Dynamik von Aktinfilopodien N2 - The tropomysin receptor kinase B (TrkB), the receptor for the neurotrophin brain-derived neurotrophic factor (BDNF), plays an important role in neuronal survival, neuronal differentiation, and cellular plasticity. Conventionally, TrkB activation is induced by binding of BDNF at extracellular sites and subsequent dimerization of receptor monomers. Classical Trk signaling concepts have failed to explain ligand-independent signaling of intracellular TrkB or oncogenic NTRK-fusion proteins. The intracellular activation domain of TrkB consists of a tyrosine kinase core, with three tyrosine (Y) residues at positions 701, 705 and 706, that catalyzes the phosphorylation reaction between ATPγ and tyrosine. The release of cisautoinhibition of the kinase domain activates the kinase domain and tyrosine residues outside of the catalytic domain become phosphorylated. The aim of this study was to find out how ligand-independent activation of TrkB is brought about. With the help of phosphorylation mutants of TrkB, it has been found that a high, local abundance of the receptor is sufficient to activate TrkB in a ligand-independent manner. This self-activation of TrkB was blocked when either the ATP-binding site or Y705 in the core domain was mutated. The vast majority of this self-active TrkB was found at intracellular locations and was preferentially seen in roundish cells, lacking filopodia. Live cell imaging of actin dynamics showed that self-active TrkB changed the cellular morphology by reducing actin filopodia formation. Signaling cascade analysis confirmed that self-active TrkB is a powerful activator of focal adhesion kinase (FAK). This might be the reason why self-active TrkB is able to disrupt actin filopodia formation. The signaling axis from Y705 to FAK could be mimicked by expression of the soluble, cytosolic TrkB kinase domain. However, the signaling pathway was inactive, when the TrkB kinase domain was targeted to the plasmamembrane with the help of artificial myristoylation membrane anchors. A cancer-related intracellular NTRK2-fusion protein (SQSTM1-NTRK2) also underwent constitutive kinase activation. In glioblastoma-like U87MG cells, self-active TrkB kinase reduced cell migration. These constitutive signaling pathways could be fully blocked within minutes by clinically approved, anti-tumorigenic Trk inhibitors. Moreover, this study found evidences for constitutively active, intracellular TrkB in tissue of human grade IV glioblastoma. In conclusion, the data provide an explanation and biological function for selfactive, constitutive TrkB kinase domain signaling, in the absence of a ligand. N2 - Die Rezeptortyrosinkinase TrkB, der Rezeptor für das Neurotrophin brain-derived neurotrophic factor (BDNF), spielt eine wichtige Rolle für das neuronale Überleben, die neuronale Differenzierung und die zelluläre Plastizität. Üblicherweise wird TrkB bei der Bindung von BDNF an extrazellulären Domänen durch Dimerisierung von Rezeptormonomeren aktiviert. Klassische Konzepte der Trk Signalübertragung können jedoch die Liganden-unabhängige Signalübertragung von intrazellulären TrkB- oder Onkogen-aktiven NTRK-Fusionsproteinen nicht erklären. Die intrazelluläre Aktivierungsdomäne von TrkB besitzt eine Tyrosinkinasedomäne mit drei Tyrosin (Y)-Resten an den Positionen 701, 705 und 706. Diese katalysieren die Phosphorylierungsreaktion zwischen ATPγ und Tyrosin. Durch die Enthemmung der cis-Autoinhibition wird die Kinase-Domäne aktiv und Tyrosinreste außerhalb der katalytischen Domäne werden phosphoryliert. Ziel dieser Arbeit war herauszufinden, wie es zur Liganden-unabhängigen Aktivierung von TrkB kommen kann. Mit Hilfe von TrkB-Phosphorylierungsmutanten wurde gefunden, dass eine hohe, lokale Abundanz des Rezeptors ausreicht, um TrkB Liganden-unabhängig zu aktivieren. Diese Selbstaktivierung von TrkB konnte blockiert werden, wenn entweder die ATP-bindende Domäne oder Y705 in der Kinasedomäne mutiert wurden. Die überwiegende Mehrheit dieses selbstaktivierenden TrkB wurde intrazellulär, in rundlichen Zellen ohne Filopodien, gefunden. Live-Zellbildgebung der Aktindynamik zeigte zudem, dass selbstaktives TrkB die Zellmorphologie veränderte, indem es die Bildung von Aktin-Filopodien reduzierte. Die Analyse von Signalkaskaden bestätigte, dass selbstaktives TrkB ein starker Aktivator der Focal Adhesion Kinase (FAK) ist. Dies kann der Grund sein, warum selbstaktives TrkB die Bildung von Aktin-Filopodien zerstört. Die Signalkaskade von Y705 bis FAK konnte durch Expression der löslichen, zytosolischen TrkB-Kinase-Domäne imitiert werden. Der Signalweg war jedoch inaktiv, wenn die TrkB-Kinase-Domäne durch künstliche Myristoylierung an die Plasmamembran gebunden wurde. Ein intrazelluläres NTRK2-Fusionsprotein (SQSTM1-NTRK) zeigte ebenfalls konstitutive Kinaseaktivierung. In Glioblastom-ähnlichen U87MG-Zellen reduzierte die selbstaktive TrkB-Kinase sogar die Zellwanderung. Die konstitutiven Signalwege konnten durch klinisch zugelassene, anti-tumorale Trk-Inhibitoren innerhalb von Minuten vollständig blockiert werden. Darüber hinaus zeigt diese Studie Beweise für konstitutiv-aktives, intrazelluläres TrkB im Gewebe von humanem Glioblastom Grad IV. Die Daten dieser Arbeit geben somit eine Erklärung und eine biologische Funktion für die selbst-aktive, konstitutive Signalübertragung der TrkB-Kinase-Domäne, in Abwesenheit eines Liganden. KW - TrkB KW - self-activation KW - NTRK fusions KW - tyrosine kinase KW - BDNF KW - phosphorylation KW - cancer Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-233829 ER - TY - JOUR A1 - Dürner, Julia A1 - Reinecker, Hans A1 - Csef, Herbert T1 - Individual quality of life in patients with multiple myeloma JF - SpringerPlus N2 - Objective: The situation of patients with multiple myeloma, whose treatment often implies high-dose chemotherapy and stem cell transplantation that can be associated with severe symptoms and psychological distress, has gained attention in recent psychooncological research. This study followed an idiographic approach in order to identify the areas of life most relevant for the interviewed myeloma patients’ quality of life (QoL) as well as their current satisfaction with these. Methods: 64 patients took part in semi-structured interviews according to the SEIQoL-DW Manual (Schedule for the Evaluation of Individual Quality of Life – Direct Weighting). Visual analogue scales (VAS) were used to gain additional information about a general assessment of the present QoL. Qualitative data evaluation preceded quantitative processing. Groups were compared according to the time elapsed since diagnosis regarding specified areas of life, satisfaction with these and their relative weighting. SEIQoL-DW-indices were correlated to the VAS to reflect on an interindividually comparable parameter. Results: Personal social relationships were mentioned significantly more often as important for QoL than healthrelated aspects, and in direct comparison were weighted significantly stronger. Regarding the change of areas relevant for QoL over the time elapsed since diagnosis, there was a significant difference between groups concerning the area of spirituality. Satisfaction differed significantly between groups for the field of leisure. Conclusion: The results for the interviewed patients with multiple myeloma point out the need to take into account the importance of social and individual aspects when reflecting on QoL. Similar findings have been reported for different samples. The relevance of an individualized approach is illustrated by the fact that individually named areas of life were rated comparatively strongly in their importance for the patients’ QoL. An overall assessment for the current QoL by means of VAS is regarded as an adequate supplement to the SEIQoL-Profile and an alternative to the SEIQoL-DW-Index. KW - multiple myeloma KW - individual quality of life KW - cancer KW - psychooncology Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130529 VL - 2 IS - 397 ER - TY - THES A1 - Peter, Leslie T1 - In-vitro-Analyse der Glutamin-Restriktion im murinen Modellsystem L929 sowie des Einflusses potentieller caloric restriction mimetics auf das Plattenepithelkarzinom HNSCC T1 - In vitro analysis of glutamine restriction in the murine model system L929 and the influence of potential caloric restriction mimetics on squamous cell carcinoma HNSCC N2 - Grundsätzlich sollte in dieser Arbeit der Einfluss der Glutamin-Restriktion als eine Form der Aminosäure-Restriktion auf das Proliferationsverhalten, den Stoffwechsel sowie die Morphologie der murinen Fibroblastenzelllinie L929 untersucht werden. Es sollte dabei auch gezeigt werden, ob sich Indizien für die Induktion eines LEM finden lassen. Weiterhin sollten die Polyamine Spermidin, Spermin, Putrescin und N-Acetylputrescin als CRMs diskutiert und ihr Einfluss auf das Proliferationsverhalten an sieben Zelllinien herausgestellt werden. Der antiproliferative Effekt der Polyamine Spermidin und Spermin auf das Wachstum der untersuchten Zelllinien konnte in dieser Arbeit bestätigt werden, wobei sich Spermin als potenter erwies als Spermidin. Eine durch Spermidin/Spermin induzierte Autophagie konnte in den Western Blots nicht signifikant nachgewiesen werden. Putrescin und N-Acetylputrescin zeigten nur leichte Wirkungen bei hoher Dosis (10 mM). Während Putrescin einen leicht antiproliferativen Effekt hatte, zeigte N-Acetylputrescin eine leicht proliferationssteigernde Wirkung. Die Autophagie-Induktion durch Methionin-Restriktion konnte durch den Nachweis Autophagie-assoziierter Proteine in den durchgeführten Western Blots bestätigt werden. Um regelmäßig von den positiven Effekten der Autophagie zu profitieren, wäre eine pflanzenbasierte Ernährungsweise sinnvoll und gut durchführbar, da Methionin hauptsächlich in tierischen Lebensmitteln und nur geringfügig in pflanzlichen Nahrungsmitteln vorkommt. Ein völliger Verzicht auf Methionin ist nicht empfehlenswert, da die Aminosäure essentiell ist und somit über die Nahrung aufgenommen werden muss. Um das Methionin-Level konstant, aber niedrig zu halten, erscheint eine Bedarfsdeckung über pflanzliche Nahrung als ideal. Die an den Zelllinien L929 und HeLa durchgeführten Proliferationsstudien zeigten, dass die Zellen selbst nach fünftägiger Glutamin- Restriktion geringfügig weiter proliferierten und keinerlei Merkmale des Zelltodes aufwiesen. Die massenspektrometrische Analyse der Modellzelllinie L929, welche über einen Zeitraum von fünf Tagen einer Glutamin-Restriktion ausgesetzt war, zeigte deutliche metabolische Veränderungen bei den über 150 untersuchten Metaboliten, die auf die Induktion eines LEM schließen lassen. Es konnte ein metabolischer Fingerabdruck nach 48 h für die Zelllinie L929 unter Glutamin-Restriktion definiert werden, der zukünftig als Referenz bei der Testung potentieller CRMs herangezogen werden kann. N2 - The basic aim of this study was to investigate the influence of glutamine restriction as a form of amino acid restriction on the proliferation behavior, metabolism and morphology of the murine fibroblast cell line L929. It should also be shown whether there is evidence for the induction of LEM. Furthermore, the polyamines spermidine, spermine, putrescine and N-acetylputrescine should be discussed as CRMs and their influence on the proliferation behavior of seven cell lines should be highlighted. The antiproliferative effect of the polyamines spermidine and spermine on the growth of the cell lines was confirmed in this study, with spermine proving to be more potent than spermidine. Autophagy induced by spermidine/spermine could not be significantly detected in the Western blots. Putrescine and N-acetylputrescine showed only slight effects at high doses (10 mM). While putrescine had a slight antiproliferative effect, N-acetylputrescine showed a slight proliferation-enhancing effect. The induction of autophagy by methionine restriction was confirmed by the detection of autophagy-associated proteins in the Western blots. In order to benefit regularly from the positive effects of autophagy, a plant-based diet would be sensible and easy to implement, as methionine is mainly found in animal foods and only slightly in plant foods. Completely avoiding methionine is not recommended, as the amino acid is essential and must therefore be obtained from food. In order to keep the methionine level constant but low, it seems ideal to cover the requirement via plant food. The proliferation studies carried out on the cell lines L929 and HeLa showed that the cells continued to proliferate slightly even after five days of glutamine restriction and did not exhibit any signs of cell death. The mass spectrometric analysis of the model cell line L929, which was exposed to glutamine restriction over a period of five days, showed clear metabolic changes in the more than 150 metabolites examined, suggesting the induction of LEM. A metabolic fingerprint could be defined after 48 h for the cell line L929 under glutamine restriction, which can be used as a reference for testing potential CRMs in the future. KW - Plattenepithelcarcinom KW - Massenspektrometrie KW - Glutamin KW - Spermidine KW - HNSCC KW - glutamine KW - methionine KW - LC-MS KW - Glutamin-Restriktion KW - Autophagy KW - restriction KW - cancer Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-349627 ER - TY - JOUR A1 - Philipp-Abbrederis, Kathrin A1 - Herrmann, Ken A1 - Knop, Stefan A1 - Schottelius, Margret A1 - Eiber, Matthias A1 - Lückerath, Katharina A1 - Pietschmann, Elke A1 - Habringer, Stefan A1 - Gerngroß, Carlos A1 - Franke, Katharina A1 - Rudelius, Martina A1 - Schirbel, Andreas A1 - Lapa, Constantin A1 - Schwamborn, Kristina A1 - Steidle, Sabine A1 - Hartmann, Elena A1 - Rosenwald, Andreas A1 - Kropf, Saskia A1 - Beer, Ambros J A1 - Peschel, Christian A1 - Einsele, Hermann A1 - Buck, Andreas K A1 - Schwaiger, Markus A1 - Götze, Katharina A1 - Wester, Hans-Jürgen A1 - Keller, Ulrich T1 - In vivo molecular imaging of chemokine receptor CXCR4 expression in patients with advanced multiple myeloma JF - EMBO Molecular Medicine N2 - CXCR4 is a G-protein-coupled receptor that mediates recruitment of blood cells toward its ligand SDF-1. In cancer, high CXCR4 expression is frequently associated with tumor dissemination andpoor prognosis. We evaluated the novel CXCR4 probe [\(^{68}\)Ga]Pentixafor for invivo mapping of CXCR4 expression density in mice xenografted with human CXCR4-positive MM cell lines and patients with advanced MM by means of positron emission tomography (PET). [\(^{68}\)Ga]Pentixafor PET provided images with excellent specificity and contrast. In 10 of 14 patients with advanced MM [\(^{68}\)Ga]Pentixafor PET/CT scans revealed MM manifestations, whereas only nine of 14 standard [\(^{18}\)F]fluorodeoxyglucose PET/CT scans were rated visually positive. Assessment of blood counts and standard CD34\(^{+}\) flow cytometry did not reveal significant blood count changes associated with tracer application. Based on these highly encouraging data on clinical PET imaging of CXCR4 expression in a cohort of MM patients, we conclude that [\(^{68}\)Ga]Pentixafor PET opens a broad field for clinical investigations on CXCR4 expression and for CXCR4-directed therapeutic approaches in MM and other diseases. KW - FDG PET/CT KW - cells KW - CXCR4/SDF-1 KW - CXCR4 KW - multiple myeloma KW - positron emission tomography KW - chemokine receptor KW - in vivo imaging KW - malignancies KW - involvement KW - microenvironment KW - survival KW - cancer KW - autologous transplantation KW - bone disease Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148738 VL - 7 IS - 4 ER - TY - JOUR A1 - Leikam, C A1 - Hufnagel, AL A1 - Otto, C A1 - Murphy, DJ A1 - Mühling, B A1 - Kneitz, S A1 - Nanda, I A1 - Schmid, M A1 - Wagner, TU A1 - Haferkamp, S A1 - Bröcker, E-B A1 - Schartl, M A1 - Meierjohann, S T1 - In vitro evidence for senescent multinucleated melanocytes as a source for tumor-initiating cells JF - Cell Death and Disease N2 - Oncogenic signaling in melanocytes results in oncogene-induced senescence (OIS), a stable cell-cycle arrest frequently characterized by a bi-or multinuclear phenotype that is considered as a barrier to cancer progression. However, the long-sustained conviction that senescence is a truly irreversible process has recently been challenged. Still, it is not known whether cells driven into OIS can progress to cancer and thereby pose a potential threat. Here, we show that prolonged expression of the melanoma oncogene N-RAS\(^{61K}\) in pigment cells overcomes OIS by triggering the emergence of tumor-initiating mononucleated stem-like cells from senescent cells. This progeny is dedifferentiated, highly proliferative, anoikis-resistant and induces fast growing, metastatic tumors. Our data describe that differentiated cells, which are driven into senescence by an oncogene, use this senescence state as trigger for tumor transformation, giving rise to highly aggressive tumor-initiating cells. These observations provide the first experimental in vitro evidence for the evasion of OIS on the cellular level and ensuing transformation. KW - reactive oxygen KW - human melanoma KW - MITF KW - cancer KW - skin KW - DNA damage KW - kappa-B KW - oncogene-induced senescence KW - cellular senescence Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-148718 VL - 6 IS - e1711 ER - TY - THES A1 - Koderer, Corinna T1 - In Vitro Analyse der Methionin-Restriktion im murinen Modellsystem L929 sowie im Platteneptihelkarzinom - HNSCC T1 - In vitro analysis of methionine restriction in the murine model system L929 and in squamous cell carcinoma - HNSCC N2 - Die Krebstherapie und Behandlung von Tumoren stellt für die moderne Medizin auch in Zukunft eine enorme Herausforderung dar. Trotz intensiver Forschung konnten in den letzten Jahrzehnten zwar zunehmend Fortschritte erzielt werden, allerdings muss das Spektrum an neuen Therapieformen und Möglichkeiten kontinuierlich erweitert werden. In den letzten Jahren haben die Kalorienrestriktion sowie die Aminosäuren- und Proteinrestriktion zunehmend an Bedeutung gewonnen, da sie einen erheblichen positiven Einfluss auf die Entstehung von altersassoziierten Erkrankungen wie z.B. Krebs haben. Allen Formen gemeinsam ist die Induktion eines Low-Energy-Metabolismus, der die Zellen in einen antiproliferativen und selbst-regenerierenden Zustand versetzt. In dieser Arbeit sollte untersucht werden, ob die Methionin-Restriktion als eine Form der Aminosäurerestriktion sich grundsätzlich als Therapieform im Plattenepithelkarzinom (HNSCC) eignet. Zusätzlich sollte ein einfaches zelluläres Modellsystem etabliert werden, das auf metaboler Ebene die Charakterisierung und Analyse des Low-Energy-Metabolismus ermöglicht. Es konnte aufgezeigt werden, dass die Methionin-Restriktion eine effektive Methode ist, um die Proliferation ausgesuchter Zelllinien des HNSCC zu inhibieren. Des Weiteren konnte aufgezeigt werden, dass der Einsatz von Aminosäure-Analoga eine weitere Möglichkeit darstellt, auf die Proliferation von Tumorzellen Einfluss zu nehmen. Die massenspektrometrische Analyse der murinen Zelllinie L929 mittels LC/MS lieferte über einen Zeitraum von 5 Tagen ein detailliertes Bild des Stoffwechsels von mehr als 150 Metaboliten unter Methionin-Restriktion. Durch die Definition eines charakteristischen Fingerabdrucks nach 48 h und eines nur wenige Metabolite umfassenden Fußabdrucks konnte ein murines Modellsystem etabliert werden, dass die Analyse von potentiellen Wirkstoffen, u.a. sogenannten caloric restriction mimetics, ermöglicht. N2 - Cancer therapy and the treatment of tumors will continue to represent an enormous challenge for modern medicine in the future. Despite intensive research, increasing progress has been made in recent decades, but the spectrum of new forms of therapy and options must be continuously expanded. In recent years, caloric restriction as well as amino acid and protein restriction have become increasingly important as they have a significant positive influence on the development of age-associated diseases such as cancer. Common to all forms is the induction of low-energy metabolism, which places cells in an antiproliferative and self-regenerative state. This work aimed to investigate whether methionine restriction, as a form of amino acid restriction, is suitable in principle as a form of therapy in squamous cell carcinoma (HNSCC). In addition, we aimed to establish a simple cellular model system that would allow characterization and analysis of low-energy metabolism at the metabolic level. It could be shown that methionine restriction is an effective method to inhibit proliferation of selected cell lines of HNSCC. Furthermore, it could be shown that the use of amino acid analogues represents another possibility to influence the proliferation of tumor cells. Mass spectrometric analysis of murine cell line L929 by LC/MS provided a detailed picture of the metabolism of more than 150 metabolites under methionine restriction over a period of 5 days. By defining a characteristic fingerprint after 48 h and a footprint comprising only a few metabolites, a murine model system could be established that allows the analysis of potential active compounds, including so-called caloric restriction mimetics. KW - Plattenepithelkarzinom KW - methionine KW - cancer KW - hnscc KW - restriction Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-271897 ER - TY - JOUR A1 - Kumar, Praveen A1 - Naumann, Ulrike A1 - Aigner, Ludwig A1 - Wischhusen, Joerg A1 - Beier, Christoph P A1 - Beier, Dagmar T1 - Impaired TGF-β induced growth inhibition contributes to the increased proliferation rate of neural stem cells harboring mutant p53 JF - American Journal of Cancer Research N2 - Gliomas have been classified according to their histological properties. However, their respective cells of origin are still unknown. Neural progenitor cells (NPC) from the subventricular zone (SVZ) can initiate tumors in murine models of glioma and are likely cells of origin in the human disease. In both, p53 signaling is often functionally impaired which may contribute to tumor formation. Also, TGF-beta, which under physiological conditions exerts a strong control on the proliferation of NPCs in the SVZ, is a potent mitogen on glioma cells. Here, we approach on the crosstalk between p53 and TGF-beta by loss of function experiments using NPCs derived from p53 mutant mice, as well as pharmacological inhibition of TGF-beta signaling using TGF-beta receptor inhibitors. NPC derived from p53 mutant mice showed increased clonogenicity and more rapid proliferation than their wildtype counterparts. Further, NPC derived from p53\(^{mut/mut}\) mice were insensitive to TGF-beta induced growth arrest. Still, the canonical TGF-beta signaling pathway remained functional in the absence of p53 signaling and expression of key proteins as well as phosphorylation and nuclear translocation of SMAD2 were unaltered. TGF-beta-induced p21 expression could, in contrast, only be detected in p53\(^{wt/wt}\) but not in p53\(^{mut/mut}\) NPC. Conversely, inhibition of TGF-beta signaling using SB431542 increased proliferation of p53\(^{wt/wt}\) but not of p53\(^{mut/mut}\) NPC. In conclusion, our data suggest that the TGF-beta induced growth arrest in NPC depends on functional p53. Mutational inactivation of p53 hence contributes to increased proliferation of NPC and likely to the formation of hyperplasia of the SVZ observed in p53 deficient mice in vivo. KW - mouse brain KW - tumors KW - cancer KW - TGF-beta KW - glioblastoma stem cell KW - pathways KW - expression KW - astrocytoma KW - glioblastoma KW - transforming growth factor-beta-1 KW - neurogenesis KW - gliomas KW - neural stem cell KW - p53 KW - subventricular zone KW - premalignant lesion Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144262 VL - 5 IS - 11 ER - TY - THES A1 - Kaymak, Irem T1 - Identification of metabolic liabilities in 3D models of cancer T1 - Identifikation metabolischer Abhängigkeiten in 3D Tumormodellen N2 - Inefficient vascularisation of solid tumours leads to the formation of oxygen and nutrient gradients. In order to mimic this specific feature of the tumour microenvironment, a multicellular tumour spheroid (SPH) culture system was used. These experiments were implemented in p53 isogenic colon cancer cell lines (HCT116 p53 +/+ and HCT116 p53-/-) since Tp53 has important regulatory functions in tumour metabolism. First, the characteristics of the cells cultured as monolayers and as spheroids were investigated by using RNA sequencing and metabolomics to compare gene expression and metabolic features of cells grown in different conditions. This analysis showed that certain features of gene expression found in tumours are also present in spheroids but not in monolayer cultures, including reduced proliferation and induction of hypoxia related genes. Moreover, comparison between the different genotypes revealed that the expression of genes involved in cholesterol homeostasis is induced in p53 deficient cells compared to p53 wild type cells and this difference was only detected in spheroids and tumour samples but not in monolayer cultures. In addition, it was established that loss of p53 leads to the induction of enzymes of the mevalonate pathway via activation of the transcription factor SREBP2, resulting in a metabolic rewiring that supports the generation of ubiquinone (coenzyme Q10). An adequate supply of ubiquinone was essential to support mitochondrial electron transport and pyrimidine biosynthesis in p53 deficient cancer cells under conditions of metabolic stress. Moreover, inhibition of the mevalonate pathway using statins selectively induced oxidative stress and apoptosis in p53 deficient colon cancer cells exposed to oxygen and nutrient deprivation. This was caused by ubiquinone being required for electron transfer by dihydroorotate dehydrogenase, an essential enzyme of the pyrimidine nucleotide biosynthesis pathway. Supplementation with exogenous nucleosides relieved the demand for electron transfer and restored viability of p53 deficient cancer cells under metabolic stress. Moreover, the mevalonate pathway was also essential for the synthesis of ubiquinone for nucleotide biosynthesis to support growth of intestinal tumour organoids. Together, these findings highlight the importance of the mevalonate pathway in cancer cells and provide molecular evidence for an enhanced sensitivity towards the inhibition of mitochondrial electron transfer in tumour-like metabolic environments. N2 - In soliden Tumoren führt die ineffiziente Bildung von Blutgefäßen (Vaskularisierung) zu einem Nährstoff- und Sauerstoffgradienten im gesamten Tumor, welches eine spezifische Tumormikroumgebung schafft. Um diese Tumorumgebung nachzuahmen, wurde ein spezielles multi-zelluläres Tumorsphäroid (SPH) Zellkultursystem verwendet. Da Tp53 wichtige regulatorische Funktionen im Tumormetabolismus hat, wurde zur Generierung von Sphäroiden p53 isogene Darmkrebs-Zelllinen HCT116 (p53 +/+ und p53 -/-) verwendet. Zunächst wurden die Sphäroide mittels RNA Sequenzierung und Metabolomik charakterisiert, um die Genexpression und metabolischen Eigenschaften in verschiedenen Zellkulturbedingungen zu vergleichen. Diese Analyse hat gezeigt, dass gewisse Genexpressionsmuster in Tumoren wie beispielsweise Proliferations- und Hypoxia verwandte Gene in Sphäroiden übereinstimmen, nicht jedoch in Monolayer-Kulturen. Vergleicht man die zwei unterschiedlichen Genotypen miteinander, so sind Gene, die in der Cholesterinhomöostase involviert sind, in p53 defizienten Zellen induziert, nicht jedoch in p53 wildtypischen Zellen. Dieser Unterschied ist in Sphäroiden vorhanden, nicht jedoch in Monolayer-Kulturen. Verlust von p53 führt über die Aktivierung des Transkriptionsfaktors SREBP2 zur Induktion von Enzymen des Mevalonat-Synthesewegs und zudem zu einer neuen metabolischen Vernetzung, die die Generierung von Ubichinon (Coenzym Q10) unterstützt. Eine ausreichende Ubichinon-Versorgung ist wichtig, um den mitochondrialen Elektronentransport und die Pyrimidin-Biosynthese in p53-defizienten Krebszellen unter metabolischen Stressbedingungen zu unterstützen. Darüber hinaus induziert die Inhibition des Mevalonat-Synthesewegs durch Statine in p53-defizienten Darmkrebszellen, die Sauerstoff und Nährstoffmangel ausgesetzt sind, selektiv oxidativen Stress und Apoptose. Verursacht wird dies durch einen Mangel an Ubichinon, welches für den Elektronentransfer der Dihydroorotatdehydrogenase, einem essentiellen Enzym der Pyrimidinnukleotid-Biosynthese, notwendig ist. Gabe von exogenen Nukleosiden entlastete die Nachfrage an Elektronentransfer und stellte die Lebensfähigkeit von p53-defizienten Krebszellen unter metabolischem Stress wieder her. Darüber hinaus konnte gezeigt werden, dass der Mevalonat-Syntheseweg auch für die Synthese von Ubichinon für die Pyrimidinnukleotid-Biosynthese unerlässlich ist, um das Wachstum von Darmtumor-Organoiden zu unterstützen. Zusammengenommen interstreichen diese Ergebnisse die Bedeutung des Mevalonat-Syntheseweg in Krebszellen und liefern den molekularen Mechanismus für die erhöhte Empfindlichkeit von Tumorzellen gegenüber der Hemmung des mitochondrialen Elektronentransfers in einer Tumor-ähnlichen Stoffwechselumgebung. KW - p53 KW - cancer KW - CoQ10 KW - Tumor KW - Modell KW - Stoffwechsel Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-181544 ER - TY - JOUR A1 - Wolfahrt, Sonja A1 - Herman, Sandra A1 - Scholz, Claus-Jürgen A1 - Sauer, Georg A1 - Deissler, Helmut T1 - Identification of alternative transcripts of rat CD9 expressed by tumorigenic neural cell lines and in normal tissues JF - Genetics and Molecular Biology N2 - CD9 is the best-studied member of the tetraspanin family of transmembrane proteins. It is involved in various fundamental cellular processes and its altered expression is a characteristic of malignant cells of different origins. Despite numerous investigations confirming its fundamental role, the heterogeneity of CD9 or other tetraspanin proteins was considered only to be caused by posttranslational modification, rather than alternative splicing. Here we describe the first identification of CD9 transcript variants expressed by cell lines derived from fetal rat brain cells. Variant mRNA-B lacks a potential translation initiation codon in the alternative exon 1 and seems to be characteristic of the tumorigenic BT cell lines. In contrast, variant mRNA-C can be translated from a functional initiation codon located in its extended exon 2, and substantial amounts of this form detected in various tissues suggest a contribution to CD9 functions. From the alternative sequence of variant C, a different membrane topology ( 5 transmembrane domains) and a deviating spectrum of functions can be expected. KW - tetraspanin KW - CD9 KW - antigen KW - cancer KW - noncoding RNAs KW - nervous system KW - poor prognosis KW - tetraspanin protein KW - transcript KW - splice variant KW - membrane topology Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-131801 VL - 36 IS - 2 ER -