基于高通量筛选方案（STARR-seq）的垂体瘤遗传易感基因区域精细定位和初步功能研究

Pituitary adenoma (PA) is one of the most common intracranial and neuroendocrine neoplasms, presenting severe clinical manifestations. However, the pathogenesis of PA remains largely unknown. To explore the somatic and germline mutations of PA is a hot topic in this field. Previously, we performed a genome-wide association study to identify three new genetic susceptibility loci (10p12.31, 10q21.1 and 13q12.13) for PA. They may lead to PA by regulating the expression of NEBL, PCDH15 and CDK8, respectively. Further, for fine mapping the causal single nucleotide polymorphisms (SNPs) in these loci, which can affect gene expression by modulating activities of distal regulatory elements, we will apply the self-transcribing active regulatory region sequencing (STARR-seq) strategy in current study. Moreover, a preliminary study will be carried out on the function of target genes. The finding from the present project would help to provide a solid theoretical basis for developing new methods to prevent and treat PA.

垂体瘤是最常见的神经内分泌肿瘤和颅脑肿瘤，临床危害严重，但发病机制不清。探索其肿瘤突变基因和遗传致病基因，一直是该领域的研究热点。申请人前期通过全基因组关联分析研究，在国际上首次发现三个与中国人垂体瘤发病密切相关的遗传易感基因区域（10p12.31，10q21.1和13q12.13），并表明其可能分别通过调控NEBL、PCDH15及CDK8表达而影响垂体瘤发生发展。现为对该三个易感基因区域开展精细定位，本研究将采用高通量基因转录调控序列鉴定方法（STARR-seq）从中筛选并验证以获得具有基因表达调控功能的单核苷酸多态性（SNP）位点，并对其下游靶基因开展初步的功能研究，以期为开发垂体瘤新的防治方法提供坚实的理论依据。

垂体瘤是最常见的神经内分泌肿瘤和颅脑肿瘤，临床危害严重，但至今其发病机制仍然不清。为进一步探索其肿瘤突变基因和遗传致病基因，我们前期已通过全基因组关联分析研究，发现与中国人垂体瘤发病密切相关的遗传易感基因区域。在本项目中，我们首先采用高通量基因转录调控序列鉴定法（STARR-seq），在易感基因区间内对具有基因表达调控功能的单核苷酸多态性（SNP）位点进行精细定位；再通过Hi-C对垂体瘤染色质进行空间结构测序，以初步明确功能型SNP所调控的下游靶基因；最后通过分子生物学实验，验证两者间参与调控作用的组蛋白乙酰化相关蛋白BRD4可为垂体瘤治疗新靶点。总体而言，本研究使得我们对垂体瘤遗传易感基因的作用机制有了更深入的认识，也为开发垂体瘤新的防治方法提供了坚实的理论依据。