Detection and Monitoring of Podocyturia Using RT-QPCR

肾小球基底膜（GBM）足细胞的缺失代表了肾小球滤过屏障中不可替代的成分的缺失，该成分易使裸露的GBM区域与鲍曼囊的壁上皮形成粘连。这些粘连是粘连的开始，其进展为局灶节段性肾小球硬化的组织病理学特征，许多肾脏疾病最终导致慢性肾功能不全（CRI）或终末期肾病（ESRD）。在成人人群中，糖尿病占ESRD病例的近40%，而在儿童人群中，肾小球肾炎占ESRD病例的30%。拟议的研究将利用非侵入性荧光逆转录酶定量聚合酶链反应（RT-QPCR）定量证明足细胞流失到尿液中。纵向收集肾小球肾病和1型糖尿病患者的尿液样本，在获得知情同意后进行检测。荧光RT-QPCR使用一系列复杂的引物和探针为足细胞特异性蛋白nephrin设计。该项目的目标是纵向监测病例患者的临床过程和实验室检查，并将其与尿中足细胞的定量损失联系起来。此外，该项目将量化开始药物治疗前后的尿足细胞损失，并寻求确定那些继续失去足细胞并具有蛋白尿，CRI或ESRD发展风险最大的患者。

The loss of podocytes from the glomerular basement membrane (GBM) represents the loss of an irreplaceable component of the glomerular filtration barrier that predisposes regions of bare GBM to form adhesions to the parietal epithelium of Bowman's capsule. These adhesions are the beginnings of synechiae, which progress to the histopathology of focal segmental glomerulosclerosis characteristic of many renal disorders culminating in chronic renal insufficiency (CRI) or end stage renal disease (ESRD). In the adult population, diabetes mellitus accounts for nearly 40% of the cases of ESRD, whereas in the pediatric population, glomerulonephritis accounts for 30% of cases of ESRD. The proposed research will quantitatively demonstrate podocyte loss into the urine utilizing a non-invasive fluorescent reverse-transcriptase quantitative polymerase chain reaction (RT-QPCR). Longitudinally collected urine samples from case patients with glomerulonephropathies and Type I diabetes mellitus will be tested after informed consent has been obtained. Fluorescent RT-QPCR is performed using an intricate series of primers and probes designed for the podocyte-specific protein nephrin. The goals of this project are to longitudinally monitor the case patients' clinical courses and laboratory tests and to correlate these with their quantitative loss of podocytes in the urine. Additionally, the project will quantify urinary podocyte loss before and after initiation of medical therapy, and seek to identify those patients who continue to lose podocytes and are at greatest risk for the development of proteinuria, CRI or ESRD.