Novel technologies for the detection of podocytes from preeclamptic patients

总结/摘要 先兆子痫是一种妊娠并发症，其特征是血压升高，蛋白尿， 涉及多个器官系统的内皮功能障碍。在世界范围内，先兆子痫是胎儿死亡的最大原因。 以及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治疗和对母亲和胎儿的持续监测，因此， 是一个迫切的和未满足的需要，诊断测试，可以确定这些患者的风险先兆子痫。 当患者的血压测量值≥140/90 mm Hg两次≥4小时时，诊断为先兆子痫 并且存在蛋白尿或以下病症之一：血小板减少症，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， 在疾病发作之前的先兆子痫。最近，研究表明肾小球足细胞脱落到 先兆子痫患者的尿液，并可以使用足细胞特异性抗体可视化。此外， 足细胞可以在妊娠中期检测到，在先兆子痫诊断之前，具有高灵敏度， 的特异性检测足细胞的方法目前依赖于过夜培养和将细胞沉降到 胶原蛋白包被的载玻片或Cytospin，由于污染的红细胞， 细胞（RBC）或管型。在足细胞检测方法和漫长的过夜孵育中受到干扰 易受污染削弱了足细胞作为先兆子痫诊断标志物的效用。本 最后，Affinergy正在开发一种载玻片平台，可以特异性地捕获和保留诊断足细胞， 同时将模糊污染物移动到载玻片的单独区域。这种技术适用于 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。在第一阶段的申请中，我们 将专注于开发最佳的肽修饰载玻片，用于从尿液中分离足细胞内脏上皮细胞 以提高临床足细胞载玻片制备的诊断质量。

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.