Novel technologies for the detection of podocytes from preeclamptic patients

摘要/摘要 先兆子痫是妊娠的一种并发症，其特征是血压升高、蛋白尿和 涉及多个器官系统的内皮功能障碍。在世界范围内，先兆子痫是胎儿的最大原因 和孕产妇发病率和死亡率，仅在美国就占孕产妇死亡的近16% 每年。目前，还没有得到批准的诊断测试来预测哪些患者会发展为先兆子痫。 先兆子痫患者需要住院并持续监测母亲和胎儿，因此， 迫切需要一种诊断测试来识别那些有先兆子痫风险的患者，这一需求尚未得到满足。 当患者的血压测量≥140/90毫米汞柱两次≥4小时时，即可诊断为先兆子痫 出现蛋白尿或下列情况之一：血小板减少，肾功能不全， 肝功能受损、肺水肿或视力障碍。正在进行重大努力，以检测 发病前先兆子痫。最近有研究表明，肾小球足细胞被排入 子痫前期患者的尿液，并可使用足细胞特异性抗体进行可视化。此外，棚改 足细胞可以在妊娠中期，在诊断先兆子痫之前被检测到，具有很高的敏感性和 专一性。目前检测足细胞的方法依赖于过夜培养和细胞在 胶原蛋白涂层玻片或细胞旋体，由于污染红血而容易出现高假阳性率 细胞(红细胞)或管型。足细胞检测方法的不一致和漫长的过夜孵化 易受污染的足细胞作为先兆子痫的诊断标记物的效用减弱。对这件事 最后，AffinEnergy正在开发一种幻灯片平台，可以专门捕获和保留诊断足细胞， 同时将模糊的污染物移动到载玻片的单独区域。这种技术是可以接受的 下游免疫细胞化学，将提高足细胞捕获的诊断质量，并导致 通过最大限度地减少时间和对昂贵设备的需求，显著节省成本。在此第一阶段应用程序中，我们 将专注于开发从尿液中分离足细胞内脏上皮细胞的最佳多肽修饰玻片 目的：提高临床足细胞玻片制剂的诊断质量。

SUMMARY/ABSTRACT Preeclampsia is a complication of pregnancy characterized by elevated blood pressure, proteinuria, and endothelial dysfunction involving multiple organ systems. Worldwide, preeclampsia is the largest cause of fetal and maternal morbidity and mortality, and accounts for nearly 16% of maternal deaths in the United States alone each year. Currently, there is no approved diagnostic test to predict which patients will develop preeclampsia. Patients with preeclampsia require hospitalization and continuous monitoring of the mother and fetus, thus, there is an urgent and unmet need for a diagnostic test that can identify those patients at risk for preeclampsia. Preeclampsia is diagnosed when a patient’s blood pressure measures ≥140/90 mm Hg twice ≥4 hours apart and either proteinuria or one of the following conditions is present: thrombocytopenia, renal insufficiency, impaired liver function, pulmonary edema or visual disturbances. Significant efforts are underway to detect preeclampsia prior to disease onset. Recently, it was demonstrated that glomerular podocytes are shed into the urine of preeclamptic patients and can be visualized using podocyte-specific antibodies. Moreover, shed podocytes can be detected in the second trimester, prior to preeclampsia diagnosis, with high sensitivity and specificity. Methods to detect podocytes currently rely on overnight culture and sedimentation of cells onto collagen-coated slides or Cytospin, which is prone to high false-positive rates due to contaminating red blood cells (RBCs) or casts. Inconsistencies in the method of podocyte detection and lengthy, overnight incubations susceptible to contamination weaken the utility of podocytes as a diagnostic marker of preeclampsia. To this end, Affinergy is developing a slide platform that can specifically capture and retain the diagnostic podocytes, while moving the obscuring contaminants to a separate region of the slide. Such a technology is amenable to downstream immunocytochemistry, will improve the diagnostic quality of podocyte capture, and lead to significant cost savings by minimizing time and need for expensive equipment. In this Phase I application, we will focus on developing optimal peptide-modified slides for isolating podocyte visceral epithelial cells from urine to improve the diagnostic quality of clinical podocyte slide preparations.