应用外显子测序技术鉴定原发性高尿酸血症及痛风的易感基因

Primary gout and hyperuricemia are metabolic disorders caused by urate overproduction and/or reduced urate excretion. It’s an independent risk factor for cardiovascular disease. Multiple risk factors, including genetics, gender, age, OA and diet may interact and lead to development of gout, however, the genetic basis remains unknown. We conducted whole-exome sequencing towards 8 selected individuals in a 3 generation familial gout and hyperuricemia family for identifying causative genes. By extensive bioinformatic analysis, we identified 4 novel mutations that existed in all examined patients. Further studies will detect the genotype of the loci in all family members to screen pathogenic mutation in this pedigree. After the novel mutation is identified, it will be validated in a large number of cases with family history and sporadic cases as well as the normal population, and genotype-phenotype correlation will be studied. The function of the novel mutation in the pathogenesis of gout and hyperuricemia will be explored by generating mice carrying mutations in causative genes using CRISPR/Cas-Mediated genome engineering. The purpose of this study is quickly learning the causative gene for patients with primary gout and hyperuricemia. It is important to the understanding of the molecular mechanism of gout and hyperuricemia and will lead to the development of new therapeutics.

原发性高尿酸血症及痛风为多基因遗传性疾病，是心血管疾病的独立危险因素。受传统检测方法的限制，原发性高尿酸血症及通风的致病基因并不清楚。我们前期对一个家系内2个原发痛风、4个高尿酸血症患者和2个血尿酸水平正常的直系亲属进行了全外显子测序，经过深入的生物信息学分析得到了4个高尿酸血症易感基因。本课题拟通过对该家系内所有成员候选位点进行基因型检测，发现与疾病共分离的新的致病位点；确定致病突变后，选取大量汉族高尿酸血症及痛风家系患者、散发病例以及正常人群，检测致病突变在患者和正常人群中的分布，进一步探讨致病突变与患者临床表型间的关系；通过CRISPR/Cas9技术制备包含致病突变的小鼠模型，进一步验证易感基因的功能，探索该致病突变在高尿酸血症和痛风发病过程中可能的机制，为高尿酸血症和痛风的提早的防治提供新思路。

原发性高尿酸血症及痛风为多基因遗传病，是心血管疾病的独立危险因素。受传统检测方法限制，原发性高尿酸血症及痛风的致病基因并不清楚。本研究利用外显子捕获结合高通量测序的技术，对两个包含三代患者的大家系中18个给个体进行了全基因组测序，经过深入的生物信息学分析，得到了家系内的候选致病基因及候选致病突变，利用sanger测序在家系内进行验证，在两个家系分别鉴定出2个与疾病共分离的候选致病突变；在300例散发高尿酸血症患者和300例正常对照中，利用飞行质谱技术分析了家系候选致病突变、GWAS报道的8个高尿酸血症相关的SNP位点及2个与高尿酸血症用药相关的位点，与正常对照组有统计学显著差异的为ABCG2基因的Rs2231142，飞行质谱结果显示汉族人群HLA-B5801*阳性的个体超过6%，提示在除此使用别嘌呤醇前需进行HLA-B5801*检测。本研究后收集64例早发心梗病例，通过对早发心更患者的统计分析，显示高尿酸血症是心梗发生的独立危险因素，在男性患者中与发病年龄显示出相关性，全外显子测序结果提示，伴有/不伴有高尿酸血症的早发心梗患者可能包含不同的致病基因及致病突变。