Regulation and Function of Viral and Endogenous Circular RNA in Cancer

单链环状RNA(CircRNA)在癌症中含量丰富，但其调控和生物学功能 大多数CircRNA的结构尚不清楚。我们的长期目标是理解人类和人类 在感染性和肿瘤性疾病中，乳头瘤病毒(HPV)来源的和内源性的CircRNA。这个 这项建议的具体目标是更全面地了解其职能和 调节头颈部鳞状细胞癌(HNSCC)中存在的CircRNA。我们的中央 假设许多CircRNA编码的RNA在调节和功能上与它们的 相应的全长RNA，促进HNSCC的发展。在初步研究中， 我们已经开发了创新的CircRNA-Seq和Polyome RNA-Seq协议，并发现许多 具有编码潜能的CircRNA在HNSCC肿瘤中较癌旁粘膜富含。在……里面 此外，我们发现了HPV16环状E7 RNA(CircE7)，它被翻译成E7癌蛋白，提供了 这是了解CircRNA形成和功能的机制和生理调节的新工具。 有了这些创新的工具和支持可行性的大量初步证据，我们建议扩大 我们对HNSCC中CircRNA的理解有三个相关的目的。第一，全面转录本 将生成多达50个HPV+和50个HPV-HNSCC肿瘤和邻近组织的简档，包括 丰富CircRNA的RNA-Seq方法。此外，在这一转录图谱中发现的CircRNA 和CircE7作为预后生物标记物的潜力将在存档的验证队列中进行评估 HNSCC。其次，我们将利用HPV16eccE7基因座，确定调节HPV16基因的顺式和反式元件 HNSCC中CircRNA的形成，并确定它们是如何被生理调节的。第三，我们将结合 HNSCC CircRNA-Seq数据集与最近完成的多聚体RNA-Seq鉴定内源CircRNA 很可能会被翻译成中文。这些优先候选人将通过以下方式进行验证和表征 严格控制过表达和敲除实验，包括使用CircRNA来源的 特异性多肽抗体。调查小组包括CircRNA新陈代谢、HPV生物学、 病理学、统计遗传学、头颈癌和流行病学，从而确保我们有技能 以及执行提案所需的资源。提案的完成将提供一份全球和详细的 了解HNSCC中CircRNA及其调控。

Single-stranded circular RNAs (circRNA) are enriched in cancers, yet the regulation and biological function of most circRNA remains unclear. Our long-term goal is to understand the role of both human papillomavirus (HPV)-derived and endogenous circRNAs in both infectious and neoplastic diseases. The specific goal of this proposal is to generate a more comprehensive understanding of the function and regulation of circRNA that are present in head and neck squamous cell carcinoma (HNSCC). Our central hypothesis is that many circRNA are coding RNA that differ in regulation and function from their corresponding full-length RNAs in ways that promote the development of HNSCC. In preliminary studies, we have developed innovative circRNA-Seq and Polysome RNA-Seq protocols and found that many circRNA with coding potential are enriched in HNSCC tumors compared to adjacent non-tumor mucosa. In addition, our discovery of HPV16 circular E7 RNA (circE7), which is translated to the E7 oncoprotein, offers a novel tool to understand the mechanistic and physiological regulation of circRNA formation and function. With these innovative tools and extensive preliminary evidence supporting feasibility, we propose to expand our understanding of circRNAs in HNSCC in three related aims. First, a comprehensive transcriptomic profile from up to 50 HPV+ and 50 HPV- HNSCC tumors and adjacent tissue will be generated, including RNA-Seq approaches that enrich for circRNAs. Moreover, circRNA identified in this transcriptomic profiling and circE7 will be assessed for their potential as prognostic biomarkers in a validation cohort of archived HNSCC. Second, using the HPV16 circE7 locus, we will determine the cis and trans elements that regulate circRNA formation in HNSCC and determine how they are physiologically regulated. Third, we will combine HNSCC circRNA-Seq datasets with recently completed polysome RNA-Seq to identify endogenous circRNA that are likely to be translated. These prioritized candidates will be validated and characterized through rigorously controlled overexpression and knockdown experiments including the use of circRNA-derived specific peptide antibodies. The investigative team includes experts in circRNA metabolism, HPV biology, pathology, statistical genetics, head and neck cancer, and epidemiology, thus ensuring we have the skills and resources to execute the proposal. Completion of the proposal will provide both a global and detailed understanding of circRNA and their regulation in HNSCC.