基于激光捕获微分离、依赖配体蛋白质组芯片技术和生物样本库的IgA肾病评估标志物研究

IgA nephropathy is the most common primary glomerulonephropathy in the world, and is the major cause of uremia in China. Clinical, pathological and prognostic manifestations is very different among patients suffered with IgA nephropathy. Markers widely used for evaluation of IgA nephropathy progress, such as urine protein quantification and glomerular filtration rate, are not sufficiently sensitive. It is still confirmed that the correlation between some biomarkers found by proteomics at present and the seriousness of pathological change, activity, and clinical dynamic change. Most markers for evaluation of IgA nephropathy do not target for the key step of IgA nephropathy, ie glomerular lesion. Therefore, in this research, we will utilize the most standardized biospecimen of kidney disease in China and Aptamer-based proteomics arrays to analyze glomerular protein in renal biopsy specimen of IgA nephropathy extracted by laser capture microdissection and urine protein, establish protein expression profile, screen synchronously changed different protein in glomeruli and urine, discovery new biomarker reflected the progress of IgA nephropathy.

IgA肾病是世界范围内最常见的原发性肾小球疾病，也是我国尿毒症最主要的病因，其临床、病理、预后存在很大差异。目前广泛应用的IgA肾病病情评估标志物如尿蛋白定量和肾小球滤过率仍不够敏感，国内外利用尿液蛋白质组学发现的一些标志物与IgA肾病病理改变严重程度以及临床动态变化的相关性仍待考证，且大部分标志物并未针对IgA肾病进展关键环节"肾小球病变"进行评估。因此，本研究将利用符合国际规范的肾病生物样本库资源，通过激光捕获微切割技术和高分辨依赖配体的蛋白质组芯片技术对从IgA肾病患者肾活检组织的肾小球中提取的蛋白质和尿液蛋白质进行分析，建立表达谱，筛选肾小球和尿液中变化同步的差异蛋白，以期发现反映与临床和病理改变相关的新型标志物。

本研究利用通过激光捕获微切割技术和微量蛋白质组学iTRAQ检测技术对从IgA肾病患者肾活检组织和肾脏肿瘤患者癌旁组织的肾小球中提取的蛋白质和尿液蛋白质进行分析，建立表达谱，筛选肾小球和尿液中变化同步的差异蛋白。所选IgA肾病患者病理分级为Lee分级III级，均具有IgA肾病进展过程中出现的典型病变。uniprot-human库蛋白质定性及定量信息分析显示，IgA肾病患者和瘤旁组织肾小球共检测出161个蛋白，期中差异蛋白为20个。后续将结合尿液蛋白质组学检测结果，通过对比分析发现与IgA肾病肾小球病变同步变化的蛋白质，以期发现IgA肾病新型生物标志物。