Dokument: Analyse der tumorspezifischen zytotoxischen Wirkung von neuen Klasse-I HDAC-Inhibitoren auf Urothelkarzinom-Zellen
| Titel: | Analyse der tumorspezifischen zytotoxischen Wirkung von neuen Klasse-I HDAC-Inhibitoren auf Urothelkarzinom-Zellen | |||||||
| Weiterer Titel: | Analysis of the tumor-specific cytotoxic effect of new class I HDAC inhibitors on urothelial carcinoma cells | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=71295 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20251124-110325-8 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Deutsch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Tanjaya, Fenny [Autor] | |||||||
| Dateien: |
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| Beitragende: | Priv.-Doz. Dr. rer. nat. Hoffmann, Michèle [Gutachter] Prof. Dr. Kurz, Thomas [Gutachter] | |||||||
| Stichwörter: | Histondeacetylase-Inhibitor, Urothelkarzinom, Epigenetik, Kombinationstherapie, Cisplatin | |||||||
| Dewey Dezimal-Klassifikation: | 600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit | |||||||
| Beschreibungen: | Chemoresistenz ist nach wie vor eine große klinische Herausforderung bei der Behandlung von Patienten mit Urothelkarzinomen (UC). Daher besteht ein dringender Bedarf an neuen Therapieansätzen oder Kombinationspartnern, die die Wirkung von Cisplatin verbessern. Der Einsatz von epigenetischen Inhibitoren zusammen mit Cisplatin als Kombinationstherapie könnte die Wirkung von Cisplatin synergistisch verstärken. In früheren Studien der Arbeitsgruppe Harnblasenkarzinom wurden kommerzielle Histondeacetylasen-Inhibitoren (HDACi) wie Romidepsin aufgrund ihrer Wirkung auf die DNA-Schadensreaktion als vielversprechende Kandidaten identifiziert. In dieser Arbeit wurden neu entwickelte HDACi aus AG Kurz im Institut für Pharmazeutische und Medizinische Chemie auf ihre Eignung für die Behandlung von UC-Zelllinien untersucht, wobei insbesondere Substanzen mit einem vergleichbaren Wirkmechanismus wie Romidepsin, aber mit geringerer normaler Toxizität gesucht wurden.
Nach einem Screening von 235 HDACi in fünf verschiedenen Urothelkarzinom-Zelllinien (UCCs) und gutartigen HBLAK-Zellen wurden die zehn besten Kandidaten durch MTT-Tests validiert, wobei die drei wirksamsten Kandidaten eine ausgeprägtere tumorspezifische Wirkung zeigten. Ihre Auswirkungen auf die Zellzyklusprogression und die Apoptose wurde mit Hilfe von Durchflusszytometrie und Caspase-Tests analysiert. Veränderungen der DNA-Schadensreaktion wurden mittels Western Blot, Immunozytochemie und RT-PCR bestimmt. Die synergistischen Effekte in Kombination mit Cisplatin wurden mit der Chou-Talalay-Methode berechnet, und die Enzymspezifität der Inhibitoren wurde durch Western Blot bestimmt. Die IC50-Werte der neuen HDACi DDK122, KSK43 und YAK40 waren in den UCCs UM-UC-3, J82 und SW-1710 niedriger als in benignen HBLAK-Zellen. Alle HDACi induzierten G2/M-Zellzyklusarrest und Zelltod in einer zelllinienabhängigen Weise, wobei in HBLAK-Zellen kein solcher Effekte beobachtet wurde. Die Behandlung mit DDK122 ergab ähnlich starke Wirkungen wie Romidepsin und beeinflusste auch die Expression von Regulatoren, die an der DNA-Replikation und Zytokinese beteiligt sind. Die Kombination von DDK122 mit Cisplatin führte zu einer synergistischen Reduktion der Zellviabilität in UM-UC-3-Zellen. Es konnten neu entwickelte HDACi identifiziert werden, insbesondere DDK122, die eine ausgeprägtere tumorspezifische Wirkung und Synergie mit Cisplatin zeigten. Dies macht sie zu vielversprechenden Kandidaten für einen innovativen Kombinationstherapieansatz bei Urothelkarzinomen.Chemoresistance remains a major clinical challenge in the treatment of patients with urothelial carcinoma (UC). Therefore, there is an urgent need for new therapeutic approaches or suitable combination partners to improve cisplatin efficacy. Combination of epigenetic inhibitors and cisplatin was reported to demonstrate a synergistic effect enhancing therapeutic efficiency. In previous studies by the bladder cancer research group, commercial histone deacetylase inhibitors (HDACi) such as romidepsin were identified as promising candidates due to their effect on DNA damage response. In this study, newly developed HDACi from AG Kurz of the Department of Pharmaceutical and Medical Chemistry were evaluated for their potential as novel treatments for UC. The focus was on identifying substances that share a similar mechanism of action with romidepsin but offer reduced toxicity to normal cells. After screening 235 HDACi in five different urothelial carcinoma cell lines (UCCs) and benign HBLAK cells, the top ten candidates were validated by MTT assays. Three most potent candidates showing a more pronounced tumor-specific effect were identified. Effects on cell cycle progression and apoptosis were analysed by flow cytometry and caspase assays. Changes in the DNA damage response were determined by Western blot, immunocytochemistry and RT-PCR. Synergistic effects in combination with cisplatin were calculated using the Chou-Talalay method, and the enzyme specificity of the inhibitors was determined by Western blot. The IC50 values of the novel HDACi: DDK122, KSK43 and YAK40 were lower in the UCCs UM-UC-3, J82 and SW-1710 than in benign HBLAK cells. All HDACi induced G2/M cell cycle arrest and cell death in a cell-line-dependent manner, whereas no such effect was observed in HBLAK cells. DDK122 treatment resulted in effects similar to those induced by romidepsin and also affected the expression of regulators involved in DNA replication and cytokinesis. The combination of DDK122 with cisplatin resulted in a synergistic reduction of cell viability in UM-UC-3 cells. Newly developed HDACi, especially DDK122, were identified showing a more pronounced tumor-specific effect and synergy with cisplatin. This makes them promising candidates for an innovative combination therapy approach in UC. | |||||||
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| Lizenz: |  Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz | |||||||
| Fachbereich / Einrichtung: | Medizinische Fakultät | |||||||
| Dokument erstellt am: | 24.11.2025 | |||||||
| Dateien geändert am: | 24.11.2025 | |||||||
| Promotionsantrag am: | 08.07.2025 | |||||||
| Datum der Promotion: | 04.11.2025 |
