TY - JOUR A1 - Sanges, C. A1 - Scheuermann, C. A1 - Zahedi, R. P. A1 - Sickmann, A. A1 - Lamberti, A. A1 - Migliaccio, N. A1 - Baljuls, A. A1 - Marra, M. A1 - Zappavigna, S. A1 - Rapp, U. A1 - Abbruzzese, A. A1 - Caraglia, M. A1 - Arcari, P. T1 - Raf kinases mediate the phosphorylation of eukaryotic translation elongation factor 1A and regulate its stability in eukaryotic cells JF - Cell Death & Disease N2 - We identified eukaryotic translation elongation factor 1A (eEF1A) Raf-mediated phosphorylation sites and defined their role in the regulation of eEF1A half-life and of apoptosis of human cancer cells. Mass spectrometry identified in vitro S21 and T88 as phosphorylation sites mediated by B-Raf but not C-Raf on eEF1A1 whereas S21 was phosphorylated on eEF1A2 by both B-and C-Raf. Interestingly, S21 belongs to the first eEF1A GTP/GDP-binding consensus sequence. Phosphorylation of S21 was strongly enhanced when both eEF1A isoforms were preincubated prior the assay with C-Raf, suggesting that the eEF1A isoforms can heterodimerize thus increasing the accessibility of S21 to the phosphate. Overexpression of eEF1A1 in COS 7 cells confirmed the phosphorylation of T88 also in vivo. Compared with wt, in COS 7 cells overexpressed phosphodeficient (A) and phospho-mimicking (D) mutants of eEF1A1 (S21A/D and T88A/D) and of eEF1A2 (S21A/D), resulted less stable and more rapidly proteasome degraded. Transfection of S21 A/D eEF1A mutants in H1355 cells increased apoptosis in comparison with the wt isoforms. It indicates that the blockage of S21 interferes with or even supports C-Raf induced apoptosis rather than cell survival. Raf-mediated regulation of this site could be a crucial mechanism involved in the functional switching of eEF1A between its role in protein biosynthesis and its participation in other cellular processes. KW - signal transduction KW - mass spectrometry KW - elongation KW - protein docking KW - factor EEF1A2 KW - cancer-cells KW - lung cancer KW - EF-1A KW - Raf kinases KW - aminoacyl-transfer-RNA KW - tyrosine phosphorylation KW - factor 1-alpha KW - nucleotide exchange KW - polyarcylamide gels KW - chain KW - apoptosis KW - ubiquitin Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-134673 VL - 3 IS - e276 ER - TY - JOUR A1 - Ceteci, Fatih A1 - Ceteci, Semra A1 - Zanucco, Emanuele A1 - Thakur, Chitra A1 - Becker, Matthias A1 - El-Nikhely, Nefertiti A1 - Fink, Ludger A1 - Seeger, Werner A1 - Savai, Rajkumar A1 - Rapp, Ulf R. T1 - E-Cadherin Controls Bronchiolar Progenitor Cells and Onset of Preneoplastic Lesions in Mice JF - Neoplasia N2 - Although progenitor cells of the conducting airway have been spatially localized and some insights have been gained regarding their molecular phenotype, relatively little is known about the mechanisms regulating their maintenance, activation, and differentiation. This study investigates the potential roles of E-cadherin in mouse Clara cells, as these cells were shown to represent the progenitor/stem cells of the conducting airways and have been implicated as the cell of origin of human non-small cell lung cancer. Postnatal inactivation of E-cadherin affected Clara cell differentiation and compromised airway regeneration under injury conditions. In steady-state adult lung, overexpression of the dominant negative E-cadherin led to an expansion of the bronchiolar stem cells and decreased differentiation concomitant with canonical Wnt signaling activation. Expansion of the bronchiolar stem cell pool was associated with an incessant proliferation of neuroepithelial body-associated Clara cells that ultimately gave rise to bronchiolar hyperplasia. Despite progressive hyperplasia, only a minority of the mice developed pulmonary solid tumors, suggesting that the loss of E-cadherin function leads to tumor formation when additional mutations are sustained. The present study reveals that E-cadherin plays a critical role in the regulation of proliferation and homeostasis of the epithelial cells lining the conducting airways. KW - injury KW - lung cancer KW - stem cells KW - clara cell KW - gene expression KW - basal cell KW - in vivo KW - epithelium KW - airway KW - renewal Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-135407 VL - 14 IS - 12 ER - TY - JOUR A1 - Steinmann, Diana A1 - Paelecke-Habermann, Yvonne A1 - Geinitz, Hans A1 - Aschoff, Raimund A1 - Bayerl, Anja A1 - Bölling, Tobias A1 - Bosch, Elisabeth A1 - Bruns, Frank A1 - Eichenseder-Seiss, Ute A1 - Gerstein, Johanna A1 - Gharbi, Nadine A1 - Hagg, Juliane A1 - Hipp, Matthias A1 - Kleff, Irmgard A1 - Müller, Axel A1 - Schäfer, Christof A1 - Schleicher, Ursula A1 - Sehlen, Susanne A1 - Theodorou, Marilena A1 - Wypior, Hans-Joachim A1 - Zehentmayr, Franz A1 - van Oorschot, Birgitt A1 - Vordermark, Dirk T1 - Prospective evaluation of quality of life effects in patients undergoing palliative radiotherapy for brain metastases JF - BMC Cancer N2 - Background: Recently published results of quality of life (QoL) studies indicated different outcomes of palliative radiotherapy for brain metastases. This prospective multi-center QoL study of patients with brain metastases was designed to investigate which QoL domains improve or worsen after palliative radiotherapy and which might provide prognostic information. Methods: From 01/2007-01/2009, n=151 patients with previously untreated brain metastases were recruited at 14 centers in Germany and Austria. Most patients (82 %) received whole-brain radiotherapy. QoL was measured with the EORTC-QLQ-C15-PAL and brain module BN20 before the start of radiotherapy and after 3 months. Results: At 3 months, 88/142 (62 %) survived. Nine patients were not able to be followed up. 62 patients (70.5 % of 3-month survivors) completed the second set of questionnaires. Three months after the start of radiotherapy QoL deteriorated significantly in the areas of global QoL, physical function, fatigue, nausea, pain, appetite loss, hair loss, drowsiness, motor dysfunction, communication deficit and weakness of legs. Although the use of corticosteroid at 3 months could be reduced compared to pre-treatment (63 % vs. 37 %), the score for headaches remained stable. Initial QoL at the start of treatment was better in those alive than in those deceased at 3 months, significantly for physical function, motor dysfunction and the symptom scales fatigue, pain, appetite loss and weakness of legs. In a multivariate model, lower Karnofsky performance score, higher age and higher pain ratings before radiotherapy were prognostic of 3-month survival. Conclusions: Moderate deterioration in several QoL domains was predominantly observed three months after start of palliative radiotherapy for brain metastases. Future studies will need to address the individual subjective benefit or burden from such treatment. Baseline QoL scores before palliative radiotherapy for brain metastases may contain prognostic information. KW - breast cancer KW - brain tumours KW - survival KW - validation KW - symptoms KW - EORTC-QLQ-C15-PAL KW - EORTC-BN20 KW - whole-brain radiotherapy KW - partitioning analysis RPA KW - cancer patients KW - lung cancer KW - prognostic index KW - radiation oncology KW - clinical trials Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-135254 VL - 12 IS - 283 ER - TY - THES A1 - Nguyen, Hoang Duong T1 - Vaccinia virus mediated expression of human erythropoietin in colonized human tumor xenografts results in faster tumor regression and increased red blood cell biogenesis in mice T1 - Expression von humanem Erythropietin in Vaccinia Virus-kolonisierten Tumorxenograftmodellen fördert die Tumorregression und die Biogenese roter Blutzellen N2 - Cancer-related anemia is prevalent in cancer patients. Anemia negatively affects normal mental and physical function capacity with common symptoms s like fatigue, headache, or depression. Human erythropoietin (hEPO), a glycoprotein hormone regulating red blood cell formation, is approved for the treatment of cancer-related anemia. It has shown benefits in correcting anemia, and subsequently improving health-related quality of life and/or enhancing radio-, and chemotherapy. Several recent clinical trials have suggested that recombinant hEPO (rhEPO) may promote tumor growth that raises the questions concerning the safety of using rhEPO for cancer treatment. However in others, such effects were not indicated. As of today, the direct functional effect of rhEPO in tumor models remains controversial and needs to be further analyzed. Based on the GLV-1h68 backbone, the hEPO-expressing recombinant VACV strains (EPO-VACVs) GLV-1h210, GLV-1h211, GLV-1h212 and GLV-1h213 were generated by replacing the lacZ expression cassette at the J2R locus with hEPO under the control of different vaccinia promoters p7.5, pSE, pSEL, pSL, respectively. Also, GLV-1h209 was generated, which is similar to GLV-1h210 but expresses a mutated non-functinal EPO (R103A). The EPO-VACV strains were characterized for their oncolytic efficacy in lung (A549) cancer cells in culture and tumor xenografts. Concomitantly, the effects of locally expressed hEPO in tumors on virus replication, host immune infiltration, tumor vascularization and tumor growth were also evaluated. As expected, EPO-VACVs enhanced red blood cell (RBC) formation in xenograft model. The number of RBCs and hemoglobin (Hb) levels were significantly increased in EPO-VACVs-treated mice compared to GLV-1h68-treated or untreated control mice. However, the mean size of RBC or Hb content per RBC remained normal. Furthermore, over-expression of hEPO did not significantly affect numbers of lymphocytes, monocytes, leucocytes or platelets in the peripheral blood stream. The expression of hEPO in colonized tumors of mice treated with EPO-VACVs was demonstrated by immunohistological staining. Interestingly, there were 9 - 10 hEPO isoforms detected either in tumors, cells, or supernatant, while 3-4 basic isoforms were missing in blood serum, where only six hEPO isoforms were found. Tumor-bearing mice after treatment with EPO-VACVs showed enhanced tumor regression compared to GLV-1h68. The virus titers in tumors in EPO-VACVs-treated mice were 3-4 fold higher compared to GLV-1h68-treated mice. Nevertheless, no significant difference in virus titers among EPO-VACVs was found. The blood vessels in tumors were significantly enlarged while the blood vessel density remained unchanged compared to the GLV-1h68 treated mice, indicating that hEPO did not affect endothelial cell proliferation in this model. Meanwhile, rhEPO (Epoetin alfa) alone or in combination with GLV-1h68 did not show any signs of enhanced tumor growth when compared to untreated controls and GLV-1h68 groups, while doses used were clinical relevant (500 U/kg). These findings suggested that hEPO did not promote angiogenesis or tumor growth in the A549 tumor xenograft model. Human EPO has been reported to function as an immune modulator. In this study, however, we did not find any involvement of hEPO in immune cytokine and chemokine expression or innate immune cell infiltration (leucocytes, B cells, macrophages and dendritic cells) into infected tumors. The degree of immune infiltration and cytokine expression was directly correlated to the number of virus particles. Increased virus replication, led to more recruited immune cells and secreted cytokines/chemokines. It was proposed that tumor regression was at least partially mediated through activation of innate immune mechanisms. In conclusion, the novel EPO-VACVs were shown to significantly increase the number of RBCs, Hb levels, and virus replication in tumors as well as to enhance tumor regression in the A549 tumor xenograft model. Moreover, locally expressed hEPO did not promote tumor angiogenesis, tumor growth, and immune infiltration but was shown to causing enlarged tumoral microvessels which facilitated virus spreading. It is conceivable that in a possible clinical application, anemic cancer patients could benefit from the EPO-VACVs, where they could serve as “wellness pills” to decrease anemic symptoms, while simultaneously destroying tumors. N2 - Blutarmut stellt eine häufige Begleiterscheinung in Krebspatienten dar. Anämie beeinträchtigt die normale mentale und körperliche Funktionsfähigkeit. Menschliches Erythropoetin (hEPO), welches die Bildung roter Blutzellen reguliert, ist klinisch zur Behandlung von Krebs-induzierter Blutarmut zugelassen. Wenn es zur Behandlung von Anämie benutzt wird, verbessert es den Gesundheitszustand sowie Bestrahlungs- und Chemotherapie. Verschiedene klinische zeigten, dass rekombinantes hEPO (rhEPO) das Tumorwachstum anregen kann, was die Frage nach Sicherheit der Anwendung von rhEPO aufbringt. In anderen Studien hingegen, gab es keine Anzeichen für eine Tumorwachstum anregenden Wirkung oder für ein Eingreifen in krebsspezifische Signalwege. Verschiedene hEPO exprimierende rekombinante VACV Stämme (EPO-VACV) wurden hergestellt, GLV-1h210, GLV-1h211 und GLV-1h213, in welchen die lacZ Expressionskassette im J2R Lokus durch das hEPO Gen unter der Kontrolle von verschiedenen Promotoren, p7.5, pSE und pSL, ersetzt wurde. Ebenfalls wurde GLV-1h209 hergestellt, welches ähnlich zu GLV-1h210 ist, jedoch ein mutiertes und nicht-funktionelles EPO Protein (R103A) exprimiert. Alle EPO-VACV Stämme wurden bezüglich ihrer onkolytischen Funktion in Zellkulturexperimenten sowie in in vivo Tumormodellen charakterisiert. Die Expression von zwei Markergene war in Zellkultur sowie in Tumorxenograften für alle EPO-VACV vergleichbar mit der des parentalen GLV-1h68 Virus. Unterschiede in hEPO Transkription und Translation der EPO-VACV war deutlich abhängig von der Promotorstärke und stieg an von p7.5, über pSE und pSL zu pSEL 12 h nach Infektion von Zellen. Darüberhinaus hatte die Insertion von hEPO in das virale Genom keinen Einfluss auf Replikation oder Zytotoxizität aller EPO-VACV in A549 oder NCI-H1299 Zelllinien, obwohl zu frühen Zeitpunkten (24-48 hpi) die Replikation der EPO-VACV etwas höher war, als die des GLV-1h68 Virus. Die A549 Zellen war zugänglicher für virale Infektion durch alle untersuchten Viren als die NCI-H1299 Zellen. Von besonderem Interesse ist, dass hypoxische Bedingungen (2% O2) die Replikation und damit Expression des Markergens gusA, sowie Zytotoxizität für alle untersuchten VACV unabhängig von hEPO Expression verlangsamte. Alle EPO-VACV erhöhen die Bildung von roten Blutzellen (RBC) in Mausmodellen. Anzahl und RBCs sowie Hämoglobin (Hb) Level waren signifikant erhöht im Vergleich zu unbehandelten oder GLV-1h68 behandelten Mäusen. Die Durchschnittsgröße einer RBC sowie der Hämoglobinanteil hingegen waren unverändert. Darüberhinaus hatte die Expression von hEPO keinen signifikanten Einfluss auf Lymphozyten, Monozyten, Leukozyten oder Blutplättchen im peripheren Blut. Die Expression von hEPO in EPO-VACV kolonisierten Tumoren wurde durch immunohistologische Färbungen bestätigt. Interessanterweise konnten 9-10 EPO Isoformen in Tumoren, Zellen oder Zellüberständen gefunden werden, während im Blutserum 3-4 basische Isoformen fehlten und nur 6 Isoformen auftraten. Tumortragende Mäuse, die mit EPO-VACV behandelt wurden, wiesen im Vergleich zu GLV-1h68 behandelten Mäusen eine erhöhte Tumorregression auf. Ausserdem waren virale Titer in EPO-VACV behandleten Tumoren 3-4 fach höher also in denen, die mit GLV-1h68 behandelt wurden. Kein signifikanter Unterschied hingegen wurde zwischen viralen Titern der verschiedenen EPO-VACV in Tumoren gefunden. Tumorale Blutgefäße waren im Vergleich zu GLV-1h68 behandelten Mäusen deutlich vergrößert, wohingegen die Dichte an Blutgefäßen unverändert war, was andeuted, dass keine Proliferation von Endothelzellen angeregt wurde. Rekombinant hergestelltes Epoetin alfa in klinisch relevanten Dosen allein oder in Kombination mit GLV-1h68 hatte keinen Einfluss auf Verbesserung der Tumorregression verglichen mit unbehandelten oder GLV-1h68 behandelten Mäusen. Diese Ergbnisse legen nahe, dass weder Angiogenese noch Tumorwachstum durch hEPO im A549 Tumormodell angeregt wurde. In dieser Studie hingegen wurde kein Einfluss von hEPO im Bezug auf Zytokin- oder Chemokinexpression sowie Immunzellinfiltration in Tumore nachgewiesen. Das Ausmass an Immunzellinfiltratrion und Zytokinexpression konnte direkt mit der Anzahl an viralen Partikeln korreliert werden. Es wurde angenommen, dass Tumorregression zumindest teiweise durch eine Aktivierung des angeborenen Immunsystems bedingt ist. Zusammenfassend kann gesagt werden, dass durch die neuartigen EPO-VACV die Bildung von RBC, die Level an Hb und die virale Replikation signifikant angeregt wurden sowie eine erhöhte Tumorregression im Xenograftmodell auftrat. Darüberhinaus leitete lokal exprimiertes hEPO keine Tumorangiogenese oder Tumorwachstum ein, aber führte zu einer Vergrößerung von Tumorblutgefäßen, was die virale Ausbreitung erleichtern könnte. Es ist vorstellbar, dass anämische Patienten von einer möglichen klinischen Anwendung der EPO-Viren profitieren würden. KW - Erythropoietin KW - Lungenkrebs KW - Anämie KW - onkolytische Virotherapie KW - erythropoietin KW - lung cancer KW - anemia KW - oncolytic therapy KW - Onkolyse Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-85383 ER - TY - JOUR A1 - Behr, Daniel S. A1 - Peitsch, Wiebke K. A1 - Hametner, Christian A1 - Lasitschka, Felix A1 - Houben, Roland A1 - Schönhaar, Kathrin A1 - Michel, Julia A1 - Dollt, Claudia A1 - Goebeler, Matthias A1 - Marx, Alexander A1 - Goerdt, Sergij A1 - Schmieder, Astrid T1 - Prognostic value of immune cell infiltration, tertiary lymphoid structures and PD-L1 expression in Merkel cell carcinomas JF - International Journal of Clinical and Experimental Pathology N2 - Merkel cell carcinoma (MCC) is an aggressive, virus-associated, neuroendocrine tumor of the skin mainly affecting immunocompromised patients. Higher intratumoral infiltration with CD3 and CD8 positive T-cells is associated with a better prognosis, highlighting the relevance of the immune system for MCC development and progression. In this study 21 primary MCCs were stained with immune cell markers including CD3, CD4, CD8, CD68, CD20, and S100. Furthermore, tumor-infiltrating neutrophils, tertiary lymphoid structures and PD-L1 expression were analyzed and correlated with overall and recurrence free survival. All MCCs were Merkel Cell Polyomavirus positive. Overall and recurrence-free survival did not correlate with intra-and peritumoral CD3 and CD8 T-cell infiltration. In addition, no significant association regarding prognosis was found for tumor-associated neutrophils, tumor-associated macrophages or PD-L1 positivity in MCCs. Interestingly, the presence of tertiary lymphoid structures (TLS) in the tumor microenvironment significantly correlated with recurrence-free survival (P=0.025). In addition, TLS were significantly associated with a higher CD8/CD4 ratio in the tumor periphery (P=0.032), but not in the center of the tumor (P > 0.999). These results demonstrate for the first time that TLS, easily assessed in paraffin-embedded tissue in the tumor periphery of MCCs, may be a valuable prognostic factor indicating prolonged recurrence free survival. KW - CD8(+) KW - PD-L1 KW - tertiary lymphoid structures KW - immune cell infiltration KW - polymavirus KW - survival KW - lymphocytes KW - responses KW - lung cancer KW - B-cells KW - breast cancer KW - antitumor immunity KW - T-antigens KW - Merkel cell carcinoma Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117720 SN - 1936-2625 VL - 7 IS - 11 ER - TY - JOUR A1 - Kunz, Meik A1 - Wolf, Beat A1 - Schulze, Harald A1 - Atlan, David A1 - Walles, Thorsten A1 - Walles, Heike A1 - Dandekar, Thomas T1 - Non-Coding RNAs in Lung Cancer: Contribution of Bioinformatics Analysis to the Development of Non-Invasive Diagnostic Tools JF - Genes N2 - Lung cancer is currently the leading cause of cancer related mortality due to late diagnosis and limited treatment intervention. Non-coding RNAs are not translated into proteins and have emerged as fundamental regulators of gene expression. Recent studies reported that microRNAs and long non-coding RNAs are involved in lung cancer development and progression. Moreover, they appear as new promising non-invasive biomarkers for early lung cancer diagnosis. Here, we highlight their potential as biomarker in lung cancer and present how bioinformatics can contribute to the development of non-invasive diagnostic tools. For this, we discuss several bioinformatics algorithms and software tools for a comprehensive understanding and functional characterization of microRNAs and long non-coding RNAs. KW - lung cancer KW - non-invasive biomarkers KW - miRNAs KW - lncRNAs KW - bioinformatics KW - early diagnosis KW - algorithm Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-147990 VL - 8 IS - 1 ER - TY - JOUR A1 - Meder, Lydia A1 - König, Katharina A1 - Ozretić, Luka A1 - Schultheis, Anne M. A1 - Ueckeroth, Frank A1 - Ade, Carsten P. A1 - Albus, Kerstin A1 - Boehm, Diana A1 - Rommerscheidt-Fuss, Ursula A1 - Florin, Alexandra A1 - Buhl, Theresa A1 - Hartmann, Wolfgang A1 - Wolf, Jürgen A1 - Merkelbach-Bruse, Sabine A1 - Eilers, Martin A1 - Perner, Sven A1 - Heukamp, Lukas C. A1 - Buettner, Reinhard T1 - NOTCH, ASCL1, p53 and RB alterations define an alternative pathway driving neuroendocrine and small cell lung carcinomas JF - International Journal of Cancer N2 - Small cell lung cancers (SCLCs) and extrapulmonary small cell cancers (SCCs) are very aggressive tumors arising de novo as primary small cell cancer with characteristic genetic lesions in RB1 and TP53. Based on murine models, neuroendocrine stem cells of the terminal bronchioli have been postulated as the cellular origin of primary SCLC. However, both in lung and many other organs, combined small cell/non-small cell tumors and secondary transitions from non-small cell carcinomas upon cancer therapy to neuroendocrine and small cell tumors occur. We define features of "small cell-ness" based on neuroendocrine markers, characteristic RB1 and TP53 mutations and small cell morphology. Furthermore, here we identify a pathway driving the pathogenesis of secondary SCLC involving inactivating NOTCH mutations, activation of the NOTCH target ASCL1 and canonical WNT-signaling in the context of mutual bi-allelic RB1 and TP53 lesions. Additionaly, we explored ASCL1 dependent RB inactivation by phosphorylation, which is reversible by CDK5 inhibition. We experimentally verify the NOTCH-ASCL1-RB-p53 signaling axis in vitro and validate its activation by genetic alterations in vivo. We analyzed clinical tumor samples including SCLC, SCC and pulmonary large cell neuroendocrine carcinomas and adenocarcinomas using amplicon-based Next Generation Sequencing, immunohistochemistry and fluorescence in situ hybridization. In conclusion, we identified a novel pathway underlying rare secondary SCLC which may drive small cell carcinomas in organs other than lung, as well. KW - lung cancer KW - small cell lung cancer KW - achaete-scute homolog 1 KW - neurogenic locus notch homolog KW - retinoblastoma protein Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-190853 VL - 138 IS - 4 ER - TY - THES A1 - Iltzsche, Fabian T1 - The Role of DREAM/MMB-mediated mitotic gene expression downstream of mutated K-Ras in lung cancer T1 - Die Rolle DREAM/MMB-vermittelter mitotischer Genexpression unterhalb von mutiertem K-Ras in Lungenkrebs N2 - The evolutionary conserved Myb-MuvB (MMB) multiprotein complex has an essential role in transcriptional activation of mitotic genes. MMB target genes as well as the MMB associated transcription factor B-Myb and FoxM1 are highly expressed in a range of different cancer types. The elevated expression of these genes correlates with an advanced tumor state and a poor prognosis. This suggests that MMB could contribute to tumorigenesis by mediating overexpression of mitotic genes. Although MMB has been extensively characterized biochemically, the requirement for MMB to tumorigenesis in vivo remains largely unknown and has not been tested directly so far. In this study, conditional knockout of the MMB core member Lin9 inhibits tumor formation in vivo in a mouse model of lung cancer driven by oncogenic K-Ras and loss of p53. The incomplete recombination observed within tumors points towards an enormous selection pressure against the complete loss of Lin9. RNA interference (RNAi)-mediated depletion of Lin9 or the MMB associated subunit B-Myb provides evidence that MMB is required for the expression of mitotic genes in lung cancer cells. Moreover, it was demonstrated that proliferation of lung cancer cells strongly depends on MMB. Furthermore, in this study, the relationship of MMB to the p53 tumor suppressor was investigated in a primary lung cancer cell line with restorable p53 function. Expression analysis revealed that mitotic genes are downregulated after p53 re-expression. Moreover, activation of p53 induces formation of the repressive DREAM complex and results in enrichment of DREAM at mitotic gene promoters. Conversely, MMB is displaced at these promoters. Based on these findings the following model is proposed: In p53-negative cells, mitogenic stimuli foster the switch from DREAM to MMB. Thus, mitotic genes are overexpressed and may promote chromosomal instability and tumorigenesis. This study provides evidence that MMB contributes to the upregulation of G2/M phase-specific genes in p53-negative cells and suggests that inhibition of MMB (or its target genes) might be a strategy for treatment of lung cancer. N2 - Der evolutionär konservierte Myb-MuvB (MMB) Multiproteinkomplex hat eine wesentliche Rolle in der transkriptionellen Aktivierung mitotischer Gene. Zielgene des MMB sowie die MMB assoziierten Transkriptionsfaktoren B-Myb und FoxM1 sind hoch exprimiert in einer Bandbreite verschiedener Krebsarten. Die erhöhte Expression dieser Gene korreliert mit einem fortgeschrittenen Tumorstadium und einer geringen Prognose. Das weißt auf darauf hin, dass MMB an der Tumorentstehung beteiligt sein könnte indem es die Überexpression mitotischer Gene fördert. Obwohl MMB biochemisch eingehend untersucht wurde, ist die Erfordernis von MMB zur Tumorentstehung in vivo weitestgehend unbekannt und wurde bisher nicht direkt getestet. In dieser Studie hemmt der konditionale Knockout der MMB Kerneinheit Lin9 die Tumorbildung in vivo in einem Lungenkrebs-Mausmodell angetrieben durch onkogenes K-Ras und den Verlust von p53. Die unvollständige Rekombination welche in Tumoren beobachtet wurde deutet auf einen starken Selektionsdruck gegen den kompletten Verlust von Lin9 hin. Die Verminderung von Lin9 und der MMB- assoziierten Untereinheit B-Myb durch RNAi-Interferenz (RNAi) liefert Beweise dafür, dass MMB für die Expression mitotischer Gene in Lungenkrebszellen notwendig ist. Zudem wurde gezeigt, dass das Zellwachstum von Lungenkrebszellen stark von MMB abhängig ist. Weiterhin wurde der Zusammenhang zwischen MMB und dem p53-Tumorsuppressor in einer primären Lungenkrebszelllinie mit wiederherstellbarer p53-Funktion untersucht. Expressionsanalysen zeigen, dass mitotische Gene nach Re-expression von p53 runterreguliert werden. Außerdem induziert die Aktivierung von p53 die Bildung des repressiven DREAM-Komplexes und führt zu einer Anreicherung von DREAM an Promotoren mitotischer Gene. Im Gegenzug wird MMB an den Promotoren verdrängt. Basierend auf den Ergebnissen wird das folgende Model vorgeschlagen: In p53- negativen Zellen begünstigen mitogene Reize den Wechsel von DREAM zu MMB. Dadurch werden mitotische Gene überexprimiert und können so chromosomale Instabilität und Tumorentstehung fördern Diese Studie liefert Hinweise, dass MMB an der Hochregulation G2/M- Phasenspezifischer Gene in p53-negativen Zellen beteiligt ist und dass die Hemmung von MMB (oder seiner Zielgene) eine Strategie zur Behandlung von Lungenkrebs sein könnte. KW - Nicht-kleinzelliges Bronchialkarzinom (NSCLC) KW - Lungenkrebs KW - lung cancer KW - DREAM complex KW - MMB KW - K-Ras KW - mitotic gene expression KW - Mitose Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-154108 ER - TY - JOUR A1 - Richter, Anne A1 - Weick, Stefan A1 - Krieger, Thomas A1 - Exner, Florian A1 - Kellner, Sonja A1 - Polat, Bülent A1 - Flentje, Michael T1 - Evaluation of a software module for adaptive treatment planning and re-irradiation JF - Radiation Oncology N2 - Background: The aim of this work is to validate the Dynamic Planning Module in terms of usability and acceptance in the treatment planning workflow. Methods: The Dynamic Planning Module was used for decision making whether a plan adaptation was necessary within one course of radiation therapy. The Module was also used for patients scheduled for re-irradiation to estimate the dose in the pretreated region and calculate the accumulated dose to critical organs at risk. During one year, 370 patients were scheduled for plan adaptation or re-irradiation. All patient cases were classified according to their treated body region. For a sub-group of 20 patients treated with RT for lung cancer, the dosimetric effect of plan adaptation during the main treatment course was evaluated in detail. Changes in tumor volume, frequency of re-planning and the time interval between treatment start and plan adaptation were assessed. Results: The Dynamic Planning Tool was used in 20% of treated patients per year for both approaches nearly equally (42% plan adaptation and 58% re-irradiation). Most cases were assessed for the thoracic body region (51%) followed by pelvis (21%) and head and neck cases (10%). The sub-group evaluation showed that unintended plan adaptation was performed in 38% of the scheduled cases. A median time span between first day of treatment and necessity of adaptation of 17 days (range 4–35 days) was observed. PTV changed by 12 ± 12% on average (maximum change 42%). PTV decreased in 18 of 20 cases due to tumor shrinkage and increased in 2 of 20 cases. Re-planning resulted in a reduction of the mean lung dose of the ipsilateral side in 15 of 20 cases. Conclusion: The experience of one year showed high acceptance of the Dynamic Planning Module in our department for both physicians and medical physicists. The re-planning can potentially reduce the accumulated dose to the organs at risk and ensure a better target volume coverage. In the re-irradiation situation, the Dynamic Planning Tool was used to consider the pretreatment dose, to adapt the actual treatment schema more specifically and to review the accumulated dose. KW - re-irradiation KW - lung cancer KW - adaptation KW - re-planning Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-158711 VL - 12 IS - 205 ER - TY - JOUR A1 - Kunz, Meik A1 - Göttlich, Claudia A1 - Walles, Thorsten A1 - Nietzer, Sarah A1 - Dandekar, Gudrun A1 - Dandekar, Thomas T1 - MicroRNA-21 versus microRNA-34: Lung cancer promoting and inhibitory microRNAs analysed in silico and in vitro and their clinical impact JF - Tumor Biology N2 - MicroRNAs are well-known strong RNA regulators modulating whole functional units in complex signaling networks. Regarding clinical application, they have potential as biomarkers for prognosis, diagnosis, and therapy. In this review, we focus on two microRNAs centrally involved in lung cancer progression. MicroRNA-21 promotes and microRNA-34 inhibits cancer progression. We elucidate here involved pathways and imbed these antagonistic microRNAs in a network of interactions, stressing their cancer microRNA biology, followed by experimental and bioinformatics analysis of such microRNAs and their targets. This background is then illuminated from a clinical perspective on microRNA-21 and microRNA-34 as general examples for the complex microRNA biology in lung cancer and its diagnostic value. Moreover, we discuss the immense potential that microRNAs such as microRNA-21 and microRNA-34 imply by their broad regulatory effects. These should be explored for novel therapeutic strategies in the clinic. KW - biomarker KW - microRNA–target interaction KW - microRNAs KW - lung cancer KW - therapeutic strategy KW - bioinformatics Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-158399 VL - 39 IS - 7 ER -