糖尿病增殖型视网膜病变外显子组测序研究

摘要

Objective We intended to apply targeted capture and exome sequencing to identify genetic variants involved in proliferative diabetic retinopathy. Methods In this case-control study, all subjects were assessed using ophthalmoscopy and fundus photographs. Fifty subjects with proliferative diabetic retinopathy(with less than 15 years of diabetes duration) and 50 subjects without diabetic retinopathy( with hemoglobin Alc above 7.0% , at least 10 years of type 2 diabetes duration) were enrolled. The total DNA was extracted from the blood samples. Targeted capture and exome sequencing were applied. Results Subjects with proliferative diabetic retinopathy were younger and had shorter duration of diabetes. There were no significant differences in body mass index, blood pressure, hemoglobin Alc, lipid profiles and renal function between the proliferative diabetic retinopathy group and the non-diabetic retinopathy group. The prevalence of diabetic nephropathy in the proliferative diabetic retinopathy group (70.6% ) was higher than that of the control group(21. 6% , P = 0. 000). The average depth was 42 - fold in exome sequencing. An excess of single nucleotide polymorphisms, particularly for nonsynonymous single nucleotide polymorphisms including 14 386 known and 6 444 novel single nucleotide polymorphisms, were identified. Conclusion These single nucleotide polymorphisms may be related with proliferative diabetic retinopathy in type 2 diabetes by applying targeted capture and exome sequencing. Further large-scale and experimental studies are needed to verify these associations.%目的 本研究组拟应用目标区域捕获与外显子测序技术对2型糖尿病增殖型糖尿病视网膜病变患者进行外显子组测序分析.方法 选择于首都医科大学附属北京同仁医院内分泌科就诊的2型糖尿病患者中进行彩色眼底照相及眼科医师检眼镜检查者.50名糖尿病人病史小于15年患增殖型糖尿病视网膜病变的患者作为病例组,50名糖尿病病史10年以上而无糖尿病视网膜病变且糖化血红蛋白大于7.0%患者作为对照组.提取外周血DNA,应用NimbleGen商业化2.1M外显子序列捕获芯片结合高通量测序技术进行外显子组区域的重测序.结果 无视网膜病变组和增殖型视网膜病变组年龄分别为(60.00+7.89)岁、(55.81±7.75)岁(P=0.007)；糖尿病病程分别为(13.67±3.87)年、(10.55±4.16)年(P =0.000 1).2组间体质量指数、血压、糖化血红蛋白、血脂、肾功能差异均无统计学意义.增殖型视网膜病变组较无视网膜病变组有较高的糖尿病肾病患病率,分别为70.6%、21.6%,P=0.000.外显子组测序结果显示目标区域的平均测序深度42×.发现已知非同义单核苷酸多态性14 386个,新的非同义单核苷酸多态性6444个.结论 应用目标区域捕获与外显子测序技术对2型糖尿病增殖型糖尿病视网膜病变进行测序分析显示,大量单核骨酸多态性可能与增殖型糖尿病视网膜病变相关,有待于进一步在大样本研究及实验研究中验证其在增殖型糖尿病视网膜病变发病中的具体作用.