TNFalpha和NOD通路基因与糖尿病向冠心病演化的关联性及过渡分子标记鉴定

The diabetes-induced complication with coronary artery disease (CAD) is the major cause for the mortality and disability in the patients with type 2 diabetes mellitus (T2D). It is well known that some dysfunctions in lipid metabolism and inflammation are the characteristics shared by the two disorders. By integrating the results from our candidate-gene-based pathway analysis and a previously published pathway analysis using genome-wide SNP data, we select TNFalpha? signaling pathway (lipid metabolism related) and NOD-like receptor signaling pathway (inflammation related) as targets to be systematically investigated. Thus, in this project, we propose (1) to continue to genotyp 43 tagSNPs in NOD-like receptor signaling pathway, by following our previously finished work for 26 tagSNPs in TNFalpha signaling pathway, in order to have a comprehensive and systematic study of the associations of the genetic polymorphisms in the abovementioned two related pathways (involving a total of 40 genes) with T2D or CAD in Chinese Han population. The aim is to identify the SNPs and functional modules shared by two diseases; 2) then, to verify that the shared SNPs (and functional modules) are also associated with CAD in T2D by using our collected samples, which are considered to be the biomarkers related to the disease progressing (from T2D to CAD); (3) next, to screen the genetic variations in the region within 1 LD (linkage disequilibrium) block from the biomarkers, by using DNA direct sequencing, in order to positionally clone their functional loci; and (4) finally, to evaluate the potential of these biomarkers and functional loci for early medical warning or as drug target(s) by using a follow-up clinical study and experimental studies of these functional loci. This project will significantly advance our understanding of the underlying molecular mechanism(s) for the disease progressing, and provide a theoretical foundation for prevention and drug development.

糖尿病引起的冠心病并发症是其致死、致残的主要原因；脂代谢异常和炎症反应是两种疾病的共同特点。综合基于候选基因和全基因组SNP的通路分析结果，选定TNFalpha信号通路(脂质代谢相关)和NOD样受体子信号通路(炎性相关)开展系统研究。本项目拟1）在前期已完成TNFalpha通路基因26个SNP分型的基础上，继续完成NOD样受体信号通路43个SNP的分型工作，全面系统地研究上述通路40个基因的遗传多态性与糖尿病或冠心病的关联性，发掘两病共享的基因位点和功能模块；2）利用已收集的糖尿病并发冠心病样本对上述共享结果进行验证,确定糖尿病向冠心病演化的过渡分子标记；3）利用DNA测序技术筛查位于标记1个LD区块内的基因变异，定位克隆其功能位点；4）最后通过临床验证和基因功能学分析，研究其作为疾病预警和药物研发分子靶标的潜能。本研究将深化对糖尿病向冠心病演化分子机制的认识，为其预防和药物开发提供依据。

本项目系一年期小额资助项目，主要目标是系统地评估脂肪因子信号通路子通路TNFalpha通路的SNP遗传变异与中国汉族人群糖尿病和冠心病的关联性，确定糖尿病和冠心病共享的分子标记。为了更加全面地解析脂肪因子信号通路基因多态性与糖尿病和冠心病的关系，我们在项目初期对计划书做了如下调整：除原计划TNFalpha子通路的分析工作，对脂肪因子信号通路其他两个子通路多态性也进行了分析。本项目主要完成了以下工作：1）累计投入60多万元，利用1000对病例对照设计，对该通路43个基因共79个标签SNP进行了基因型分析和遗传统计分析。评估了脂肪因子信号通路及其3个子通路的疾病风险，确立了核心易感基因；2）由本课题组牵头，联合广东医学院附属医院、广东省茂名市人民医院、广东省深圳市西乡人民医院、中山大学第一附属医院心内科、广东省东莞市厚街人民医院、石龙博爱人民医院和深圳市观澜人民医院、西乡人民医院，建立了中国南方人群遗传资源收集网络。目前，我们的遗传标本资源库已储存了1万多例不同疾病的DNA标本和详细的临床和流行病学资料，包括冠心病病例3000余例、健康对照约2600例和其他疾病（糖尿病等）5000余例。为进一步开展深入的遗传流行病学研究奠定了基础；3）配合本项目通路分析的工作，开展了系列生物信息学方法学研究工作，包括：①建立了基于通路的复杂疾病遗传异质性分析方法；②开展了“基于蛋白质互作知识的生物学通路扩充新方法学”等多项拓展性研究内容；4）最后，做为本项目的姊妹课题，利用本项目现场调查数据，开发和评估了冠心病生存质量评估量表（QLICD-CHD）在广东人群中的适用性和测量学属性。项目研究期间共发表标注本基金号（81373085）学术期刊论文9篇，其中SCI论文4篇。