大骨节病甲基化数量性状基因座鉴定及其对基因表达影响的分子机制研究

Kashin-Beck Disease (KBD) is an endemic osteochondral disease, the etiology and pathogenesis underlying the disease are unknown. Currently, the interaction between environment and gene has been becoming the research hotpot. Some aberrant gene expression and SNPs related to KBD prevalence were reported, but the underlying mechanism is not clear. It is to assume that selenium deficiency and T-2 poisoning are likely to regulate gene transcription by the interaction between genetics and epigenetics. The present study focuses on the methylation quantitative trait loci (meQTLs) in KBD patients and researches the molecular mechanism on gene expression from different aspects including the population, animal, cell, and gene by using high-throughput sequencing and molecular biological techniques, combining epidemiological investigation and laboratory work. Primarily, the meQTLs are located and identified in KBD patients. Subsequently, we explore that the effects of interplay between genetics and epigenetics on gene expression and identify a several of transcription factors and related signaling pathways. Moreover, the differential meQTLs for KBD patients are verified in another larger population, and some biological function experiments are conducted, and it is also explored that effects of selenium deficiency and T-2 poisoning on the interaction between genetics and epigenetics and new targets. This study aims to confirm the regulation of gene expression by meQTLs and its molecular mechanisms in order to illuminate the effects of the interaction between genetics and epigenetics on the etiology and pathogenesis of KBD, identify potential biological markers and regulatory networks, which will provide new concept for the etiology, pathogenesis and treatment of KBD.

大骨节病是一种地方性骨关节病，病因及机制不明，环境与基因相互作用已成为研究热点，研究发现了一些大骨节病风险SNPs及基因表达改变，但其机制不明，推测低硒和T-2毒素可能通过遗传和表观遗传相互作用影响基因表达。本项目拟以现场调查与实验室研究相结合，应用高通量测序及分子生物学技术，从人群、动物、细胞和基因等不同层面，研究大骨节病甲基化数量性状基因座（meQTLs）及其影响基因表达的分子机制。通过meQTL定位，鉴定大骨节病差异性meQTLs；研究遗传与表观遗传相互作用对基因表达的影响，鉴定关键转录因子及相关信号通路；进行人群验证和生物学验证，研究低硒和T-2毒素对遗传与表观遗传相互作用的影响及新靶标。通过该项目实施探索meQTLs调节基因表达的分子机制，阐明遗传与表观遗传相互作用在大骨节病病因和发病机制中的作用，确定大骨节病生物标识物和调控网络，为大骨节病病因、发病机制及防治提供新线索。

项目背景.大骨节病遗传与表观遗传改变之间的关系及其相互作用不明，揭示大骨节病差异性meQTLs的遗传和表观遗传相互作用机制，将为大骨节病环境与基因相互作用研究提供新的视野。.主要研究内容.进行大骨节病全基因组测序，筛选大骨节病差异性meQTLs；研究大骨节病差异性meQTLs影响基因表达的遗传和表观遗传学机制；鉴定影响大骨节病差异性meQTLs关键因子及其通路；探索低硒和T-2毒素中毒导致大骨节病软骨损伤的分子机制，鉴定防治大骨节病新靶标。.重要结果及关键数据.1.获得了大骨节病全基因组转录谱，发现大骨节病患者3194个mRNAs、4103个lncRNAs和1550个miRNAs呈差异表达，提示大骨节病转录谱异常。.2.MGMTrs16906252，SIRT1rs1589064475、rs1313023791、rs568432780、rs550317521、rs1296308235和rs1469169899，ALDH1A2rs3204689，MGMTrs16906252，SEPS1rs34713741和rs4965814，SEP15rs5859，SEPS1G-105A，DIO2rs955849187为大骨节病差异性meQTLs，其相关基因表达异常与大骨节病发病风险显著相关，提示环境反应基因启动子区meQTLs变异是大骨节病的风险因子。.3.meQTLs遗传和表观遗传学改变是大骨节病软骨损伤的重要机制，基因启动子区CpG-SNPs变异可诱导启动子区CpG位点甲基化改变从而影响基因表达。.4.低硒、T-2毒素可介导meQTLs的遗传和表观遗传改变，在大骨节病软骨凋亡和DNA损伤中发挥作用。.科学意义.该项目阐明了大骨节病meQTLs的遗传与表观遗传学作用，揭示了低硒和T-2毒素通过启动子区CpG-SNPs和CpG位点甲基化改变影响基因转录，从而导致大骨节病软骨损伤，为大骨节病预警、诊断及治疗提供了新线索。项目共发表论文32篇，其中SCI收录16篇，中文期刊8篇，会议论文8篇(EI收录3篇)，参编专著2部；获陕西省科技进步二等奖1项和陕西省高等学校科学技术一等奖1项；发明专利2项；主办会议4次，参加国际会议13人次，国内会议42人次，大会特邀报告9人次；邀请外籍专家来华3人次，出国交流2人次；培养博硕士研究生18名，毕业6名。