LINC00842调控MUC16与TERT表达在云南宣威家族肺癌遗传易感中的作用及机制研究

云南省宣威地区肺癌发病高居全国首位，是典型的燃煤空气污染相关肺癌。申请人前期研究(81702274)发现：宣威肺癌发病的家族聚集现象明显，家族肺癌临床特征与遗传背景独特;多组学分析发现家族肺癌患者LINC00842下调显著，在肺癌发生中有重要作用;预实验提示LINC00842可能通过调控MUC16与TERT表达影响肺癌的遗传易感性，但具体机制有待进一步研究。.本研究拟使用CRISPR-Cas9基因编辑技术，解析LINC00842的功能及信号调控通路；通过环境致癌物细胞诱变与动物移植瘤实验，研究LINC00842在肺癌易感中的作用及机制，并探索弱化/逆转细胞恶性表型的可能；评估LINC00842、MUC16与TERT作为易感人群筛查、临床诊断和预后指标的价值。本课题可为肺癌的遗传易感机制与癌细胞演化多样性研究提供新思路，为肺癌高危人群的筛查、预防、风险评估和早期诊断提供科学依据。

Xuanwei area in Yunan Province has the highest lung cancer incidence nationwide, which is considered as typical coal-combustion related lung cancer.Our previous project（81702274） found: lung cancer in Xuanwei usually occur in family clusters (familial lung cancer), and has its unique clinical characteristics, mutation spectrum as well as genetic background. Previously, we selected representative lung cancer patients from typical families for whole genome sequencing and transcriptome, after systematical bioinformatics processing, LINC00842、MUC16、TERT emerged as promising candidates for heritable lung cancer susceptibility. In following experiments, we found that LINC00842 could regulate the expression of MUC16 and TERT..Present study uses CRISPR Cas9 gene editing technology to investigate and verify the significance of selected genes in heritable lung cancer susceptibility. Firstly, we plan to study the carcinogenic / cancer-suppressing function of target genes, moreover, the signal pathway of how LINC00842 regulate the expression of MUC16 and TERT. Based on previous data, different gene-editing strategies would be applied to normal lung epithelial cells to generate a group of cell strains with varied susceptibility, then a series of environmental carcinogens were used to mutate gene-edited cells, then compare and analyze which strain is more vulnerable to carcinogen and which strain has carcinogen-resistant potential. By this step, mutated lung epithelial cells showing precancerous / cancerous features would also be selected. Importantly, we can better evaluate the effect of target gene on increasing / decreasing lung cancer susceptibility. Based on these results, by constructing CRISPR vectors designed to do delicate quantity-regulation of target genes, we decide to explore the possible means which may decrease or reverse the malignant phenotype of previously mutated lung epithelial cells, by in vitro / in vivo experiments. Finally, we would measure target genes’ level in lung cancer families, sporadic lung cancer cases and health population, to estimate if they can be used as biomarkers for prevention, diagnosis and prognosis..In summary, the discoveries would provide essential information for future research on cancer susceptibility and biodiversity of cancer evolution, and may eventually helping people in lung cancer susceptibility screen, risk prediction, and diagnostic＆therapeutic strategy development.