Dokument: Lange nicht-kodierende RNAs als Biomarker in Plattenepithelkarzinomen von Kopf und Hals

Titel:	Lange nicht-kodierende RNAs als Biomarker in Plattenepithelkarzinomen von Kopf und Hals
URL für Lesezeichen:	https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=53302
URN (NBN):	urn:nbn:de:hbz:061-20200603-101438-8
Kollektion:	Dissertationen
Sprache:	Deutsch
Dokumententyp:	Wissenschaftliche Abschlussarbeiten » Dissertation
Medientyp:	Text
Autor:	Voß, Madeleine [Autor]
Dateien:	[Dateien anzeigen] Adobe PDF [Details] 4,04 MB in einer Datei [ZIP-Datei erzeugen] Dateien vom 28.05.2020 / geändert 28.05.2020
Beitragende:	Prof. Dr. Schulz, Wolfgang A. [Gutachter] Prof. Dr. med. Sabel, Michael [Gutachter]
Stichwörter:	Biomarker, CASC9, Plattenepithelkarzinom, lncRNA
Dewey Dezimal-Klassifikation:	600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit
Beschreibungen:	Plattenepithelkarzinome des Kopf-Hals-Bereiches (HNSCC) entstehen aus Zellen der Mukosa in Mund, Nase und Schlund. Das HNSCC stellt den sechsthäufigsten Tumor in Industriestaaten dar und gilt als häufigste bösartige Veränderung des oberen Respirations-traktes. Gehäuft tritt diese Tumorerkrankung bei Männern zwischen 50 und 70 Jahren auf; sie wird vor allem durch das Rauchen und erhöhten Alkoholkonsum verursacht. Die Inzidenz in jüngeren Bevölkerungsgruppen steigt, was in ca. 30% aller Fälle mit einer vorliegenden HPV-Infektion assoziiert ist. Die HPV-assoziierten Tumore stellen offenbar eine biologisch und klinisch distinkte Subgruppe dar, die mit einem besseren Therapieansprechen und somit einer besseren Prognose vergesellschaftet ist. In frühen Tumorstadien haben Patienten eine gute Überle-bensprognose, die sich jedoch mit Fortschreiten der Erkrankung drastisch reduziert. Da viele Tumore - aufgrund fehlender Früherkennung - erst im fortgeschrittenen Stadium diagnostiziert werden, sind die Überlebenschancen der Patienten bei Erstdiagnose bereits vermindert. Für die Therapieeinschätzung dieser Tumore ist es wichtig, jenseits von his-topathologischen Parametern und HPV-Status geeignete Biomarker zur Früherkennung, Prädiktion von Therapieansprechen und klinischem Verlauf zu finden. Als potentieller Biomarker werden in dieser Arbeit lange nicht kodierende RNAs, kurz lncRNAs, unter-sucht. Von diesen mindestens 200 bp langen RNAs ist aus verschiedenen Tumorentitäten bekannt, dass sie vielfältige Wirkungen auf Zellen ausüben und auch in der Tumorgenese eine bedeutende Rolle spielen können. Vor diesem Hintergrund wurden mittels Literatur- und Datenbankrecherchen potentiell in HNSCC verändert exprimierte lncRNAs identifiziert und ihre Expression in Gewebesets mit Tumor- und Normalproben gemessen. Hierbei stellten sich die lncRNAs HOTAIR, HOXB-AS3 und CASC9 als signifikant überexprimiert in Tumoren dar. Das Hauptaugenmerk wurde im weiteren Verlauf auf CASC9 gelegt und dessen Expression in einer Serie von 49 Tumoren und 20 Normalge-weben mittels qRT-PCR gemessen. Nach Auswertung der klinischen Patientendaten zeig-te sich CASC9 - im Einklang mit den Datenbankanalysen - signifikant über- ex-primiert, auch in fortgeschrittenen Tumorstadien sowie metastasierten Tumoren. Weitere Datenbankrecherchen ergaben, dass CASC9 signifikant in vielen weiteren Plattenepithelkarzinomen überexprimiert wird, sodass der funktionelle Einfluss dieser lncRNA auf Tumorzellen näher betrachtet wurde. Hierzu wurde in zwei verschiedenen Versuchsansätzen auf der einen Seite die Über- expression von CASC9 mittels lentiviraler Transduktion in Vektoren, sowie auf der ande-ren Seite eine Herabregulation von CASC9 mittels shRNA („Knockdown“) erreicht und die Effekte auf Proliferation, Migration und Invasion verschiedener Zelllinien analysiert. Sowohl die - jeweils verifizierte - Überexpression als auch der Knockdown von CASC9 in Zelllinien lösten jedoch nur geringe zelluläre Effekte im Zellkulturmodell aus. Es lässt sich dennoch festhalten, dass CASC9 ein vielversprechender Biomarker-Kandidat zur Identifizierung von Plattenepithelkarzinomen ist. Dabei ist CASC9 offenbar kein Hauptinduktor zellulärer Veränderungen, sondern wirkt vermutlich mit entitätsspezifi-schen Kofaktoren zusammen und könnte so zu einer Dysregulation in der Zelle beitragen. Als weiterführende Fragestellung der Forschung um CASC9 ergibt sich somit die Identi-fizierung der Faktoren, die zur Überexpression von CASC9 in Plattenepithelkarzinomen führen. Squamous carcinoma of the head and neck (HNSCC) is the sixth most common malig-nancy overall and the most common malignancy of the upper respiratory tract. It is caused mostly by cigarette smoking and alcohol consumption and occurs especially in men aged 50-70 years. In recent years, the incidence in the younger generation has increased be-cause of human papillomavirus (HPV) infection. Radiation therapy, surgery, chemothera-py, treatment with EGFR antibodies, immune checkpoint inhibitors or combined treat-ments are applied for primary tumors and recurrent or metastatic disease. Patients with localized HNSCC and low tumor stage have a high chance of cure. The disease recurs in up to 50% of the cases. For high stage, metastatic and recurrent HNSCC treatment options are limited and the outcome is therefore unfavorable. One problem in HNSCC is the belat-ed diagnosis of many tumors which aggravates prognosis. To date, clinically validated prognostic biomarkers for HNSCC are lacking except for HPV positivity, which predicts favorable survival and better response to radio- and chemotherapy. For these reasons new diagnostic and prognostic markers are required. Long noncoding RNAs (lncRNAs) are potential biomarkers, as they are considered in the literature as good candidates for tumor biomarkers and regulators of various neoplastic cell properties. By definition, lncRNAs do not contain substantial open reading frames and consist of more than 200 nucleotides. They can be involved in various cellular processes like prolif-eration, metastasis or apoptosis. LncRNAs are known to be differentially expressed in tumors and may actively contribute to their development and progression. In this thesis differentially expressed lncRNAs in HNSCC were identified by literature and database research. Their expression was then analyzed in two different tissue sets of HNSCC tumor and benign tissue samples. CASC9, HOTAIR and HOXB-AS3 were found to be upregulated in tumors. CASC9 expression was in particular upregulated in advanced tumor stages and metastatic cases. Literature data and a pan-cancer analysis based on data from The Cancer Genome Atlas indicated CASC9 to be overexpressed in other squamous cell cancer types suggesting CASC9 as general biomarker of squamous cell cancers as well. To understand the consequences of CASC9 overexpression in tumors, in vitro experi-ments were performed in HNSCC cell lines to explore the effects of CASC9 expression on cell proliferation, clonogenicity, migration or chemosensitivity. However, upregulation or shRNA-mediated downregulation (knockdown) of CASC9 had no major effects on tumor cells. In conclusion, CASC9 does not appear to be a driving factor in cancers of the oral and oropharyngeal tract but can discriminate well between cancerous and benign samples. Being robustly overexpressed in HNSCC CASC9 is a promising candidate for tumor de-tection, potentially also of squamous cell carcinomas in other organs. An important ques-tion for future work is therefore which factors drive the overexpression of CASC9 in HNSCC and other squamous cell carcinomas.
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Lizenz:	Urheberrechtsschutz
Bezug:	2016-2020
Fachbereich / Einrichtung:	Medizinische Fakultät
Dokument erstellt am:	03.06.2020
Dateien geändert am:	03.06.2020
Promotionsantrag am:	10.09.2019
Datum der Promotion:	26.05.2020