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

人 iPSC 衍生的足细胞用于研究 APOL1 高风险变异体

• 批准号: 10607362
• 负责人: Lauren Elizabeth Haines
• 金额: $ 3.68万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607362
• 关键词: Adult; Affect; African; African American population; African Trypanosomiasis; Alleles; Amino Acids; Animal Model; Apolipoproteins; Belief; Biology; C-terminal; Cell Line; Cell model; Cells; Cessation of life; Chronic Kidney Failure; Consumption; Coupled; Cytoplasm; Cytoskeleton; Data; Development; Dialysis procedure; Disease; Disease Progression; End stage renal failure; Epithelium; European; Exhibits; Experimental Designs; Exposure to; Filtration; Foot Process; Functional disorder; Genes; Genetic Engineering; Genotype; Glomerular Capillary; Goals; HIV; Heterozygote; Human; Hypertension; Impairment; Incidence; Individual; Injury; Injury to Kidney; Kidney; Kidney Diseases; Kidney Transplantation; Knowledge; Link; Liquid substance; Maintenance; Mediating; Modeling; Pathogenicity; Pathology; Patients; Physiological; Point Mutation; Population; Primates; Process; Proteins; Protocols documentation; Quality of life; Renal function; Renal glomerular disease; Research; Resort; Risk; Series; Signal Pathway; Structure; Therapeutic Intervention; Time; United States; Urine; Variant; Virus Diseases; Work; biological adaptation to stress; cell type; curative treatments; cytotoxic; cytotoxicity; directed differentiation; health disparity; high risk; in vivo; induced pluripotent stem cell; injured; kidney allograft; migration; non-diabetic; novel; overexpression; podocyte; resistance allele; response; risk variant; slit diaphragm; stressor; transcriptome sequencing; wasting

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.

项目总结/摘要。. 肾脏疾病状态的特征在于肾脏能够有效地清除多余的液体和废物。 肾小球内的足细胞对于形成这种过滤所需的屏障是不可或缺的。估计有15% 的美国成年人患有慢性肾脏病（CKD），但CKD没有治愈性治疗选择 患者必须采取耗时的透析或接受肾脏移植来维持 疾病进展期间的生活。撒哈拉以南非洲裔美国人的人口比非洲裔美国人多3.5倍。 与欧洲血统的人群相比，终末期肾病的风险增加。该发生率 这种差异部分是由于载脂蛋白L1（APOL 1）基因中的两种致病性变体G1和G2。 具有一个高风险等位基因的个体对非洲昏睡病具有抵抗力，但存在 两个高风险等位基因的突变会显著导致肾脏疾病。 由于APOL 1只存在于人类中， 在某些灵长类动物中，这项工作利用了基因工程改造成含有APOL 1变体的同基因人类iPSC系。 基因型APOL 1介导的肾脏疾病的机制尚不清楚，但与足细胞相关。 人口减少这种消耗被认为与足突消失有关，足细胞应激 由细胞骨架重排决定的反应，导致过程简化和变平， 导致肾功能下降这项研究表明，高风险的APOL 1足细胞本质上是不同的， 与参考APOL 1足细胞相比。变体AP 0 L1 iPSC系将直接分化为纯AP 0 L1 iPSC系。 并将用于确定高风险APOL 1的存在是否干扰足细胞群 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，以确定高风险APOL 1足细胞是否 本质上不同。这项研究将评估人类特定方面的足细胞生物学在一个代表性的 和同质的细胞群，提供了一个机会，以获得了解的机制， APOL 1介导的疾病。生成的数据将提供直接可翻译和迫切需要的 对肾脏疾病损伤和进展的治疗干预。