Detection and Monitoring of Podocyturia Using RT-QPCR

足细胞从肾小球基底膜（GBM）的损失代表肾小球滤过屏障的不可替代的组分的损失，所述肾小球滤过屏障的不可替代的组分使裸露GBM的区域易于与鲍曼氏囊的壁上皮形成粘连。这些粘连是粘连的开始，粘连进展为局灶节段性肾小球硬化的组织病理学特征，这是许多肾脏疾病的特征，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%，而在儿科人群中，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）定量证明足细胞损失到尿液中。在获得知情同意后，将对从肾小球肾病和I型糖尿病病例患者中纵向采集的尿液样本进行检测。使用针对足细胞特异性蛋白nephrin设计的一系列复杂的引物和探针进行荧光RT-QPCR。本项目的目标是纵向监测病例患者的临床病程和实验室检查，并将其与尿液中足细胞的定量损失相关联。此外，该项目将量化药物治疗开始前后的尿足细胞损失，并寻求确定那些继续损失足细胞的患者，以及发生蛋白尿，CRI或ESRD的风险最大的患者。

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.