Human iPSC-derived Podocytes to Study APOL1 High-Risk Variants

项目摘要/摘要。。 肾脏疾病状态的特征是肾脏清除多余液体和废物的效率。 肾小球内的足细胞是形成这种滤过所需的屏障所必需的。估计为15% 的美国成年人患有慢性肾脏疾病(CKD)，但没有治愈CKD的治疗选择 患者必须求助于耗时的透析或接受肾脏移植来维持质量 在疾病发展过程中的生活。撒哈拉以南非洲裔美国人的人口是美国人口的3.5倍 与欧洲血统的人群相比，终末期肾病的风险增加。这起事件 差异的部分原因是载脂蛋白L1(APOL1)基因中的两个致病变异体G1和G2。 存在一个高危等位基因的个体对非洲昏睡病具有抵抗力，但存在 两个高危等位基因中的两个显著易患肾脏疾病。 因为载脂蛋白1只在人类和 在一些灵长类动物中，这项工作利用了经过基因工程改造的含有APOL1变体的等基因人类IPSC系 基因分型。APOL1介导的肾脏疾病的机制尚不清楚，但与足细胞有关 人口枯竭。这种消耗被认为与足突的消失有关，足细胞应激。 由细胞骨架重排引起的反应，导致过程简化和扁平化， 导致肾功能下降。这项研究提出高危APOL1足细胞在本质上是不同的 与参考APOL1足字节相比。变异的APOL1 IPSC品系将直接分化为纯 并将用于确定高危APOL1的存在是否干扰足细胞 细胞骨架动力学。此外，这项工作将利用实验设计(DoE)方法来检测 参与足细胞生物学的协同输入的影响，以确定高危APOL1足细胞是否 本质上是不同的。这项研究将评估具有代表性的足细胞生物学中人类特有的方面 和同质的细胞群体，为了解 载脂蛋白1介导的疾病。生成的数据将直接提供可翻译和迫切需要的数据 肾脏疾病损伤和进展的治疗干预。

PROJECT SUMMARY/ABSTRACT. . Kidney disease status is characterized by how efficiently the kidney is able to remove excess fluids and waste. Podocytes within the glomerulus are integral to forming the barrier needed for this filtration. An estimated 15% of adults in the US have chronic kidney disease (CKD), but there are no curative treatment options for CKD and patients must resort to time-consuming dialysis or undergoing a kidney transplant to maintain a quality of life during disease progression. The African American population of sub-Saharan descent has a 3.5-fold increased risk for end-stage kidney disease compared to populations of European descent. This incidence discrepancy is, in part, due to two pathogenic variants G1 and G2 in the apolipoprotein L1 (APOL1) gene. Individuals with the presence of one high-risk allele are resistant to African sleeping sickness, but the presence of two high-risk alleles significantly predisposes to kidney disease. As APOL1 is only found in humans and some primates, this work utilizes isogenic human iPSC lines genetically engineered to contain APOL1 variant genotypes. The mechanism of APOL1-mediated kidney diseases is not known but correlates with podocyte population depletion. This depletion is thought to be linked to foot process effacement, a podocyte stress response dictated by cytoskeletal rearrangements that cause the processes to simplify and flatten to flatten, leading to decreased kidney function. This study proposes high-risk APOL1 podocytes are intrinsically different compared to reference APOL1 podocytes. Variant APOL1 iPSC lines will be directly differentiated into pure podocyte populations and will be used to determine if the presence of high-risk APOL1 interferes with podocyte cytoskeleton dynamics. Further, this work will utilize a Design of Experiment (DoE) approach to detect the effects of synergistic inputs involved in podocyte biology to determine if high-risk APOL1 podocytes are intrinsically different. This research will assess human-specific aspects of podocyte biology in a representative and homogenous cell population, providing an opportunity to gain an understanding of the mechanism of APOL1-mediated diseases. Data generated will provide directly translatable and desperately needed therapeutic intervention to kidney disease injury and progression.