Intracellular complement activation in Lupus Nephritis Podocytes

狼疮性肾炎足细胞的细胞内补体激活

• 批准号: 10214745
• 负责人: Abhigyan Satyam
• 金额: $ 21.88万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-06 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214745
• 关键词: American; Antigen-Antibody Complex; Antigens; Biological; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cathepsins; Cathepsins B; Cause of Death; Cells; Chronic Kidney Failure; Cleaved cell; Complement; Complement Activation; Complement Factor B; Complement component C4; Cues; Cytoplasm; Deposition; Development; Epithelial Cells; Exposure to; Extrahepatic; Family; Fibroblasts; Genetic; Genetically Engineered Mouse; Glomerulonephritis; Graft Rejection; Hemolytic-Uremic Syndrome; Homeostasis; Human; Hypoxia; Hypoxia Inducible Factor; IGA Glomerulonephritis; Immunoglobulin G; Impairment; In Vitro; Inflammatory; Injury; Ischemia; Kidney Diseases; Kidney Failure; Kidney Transplantation; Lead; Lupus; Lupus Nephritis; Maintenance; Mediating; Membrane; Methods; Mus; Pathogenesis; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacology; Process; Production; Proliferative Glomerulonephritis; Protocols documentation; Renal glomerular disease; Research; Role; Specialized Epithelial Cell; Surface; System; T-Lymphocyte; Testing; Tumor-Derived; United States; base; cell injury; cell type; complement pathway; complement system; experimental study; exposed human population; genetically modified cells; glomerular filtration; glomerular function; immune activation; inhibitor/antagonist; injured; insight; intestinal epithelium; intestinal injury; kidney epithelial cell; knock-down; lupus prone mice; monocyte; novel; podocyte; prevent; renal damage; response; targeted delivery

Project Summary Deposition of immune complexes and activation of the complement system mediates glomerular injury and is involved in the inflammatory process in patients with lupus nephritis. Podocytes are highly specialized epithelial cells which are critical for the proper function of the glomerular filtration barrier and are injured in several proteinuric kidney diseases. Podocytes produce a number of complement factors and cathepsin proteases. Preliminary evidence suggests that podocytes exposed to IgG form patients with lupus nephritis produce increased amounts of complement which becomes activated intracellularly. We hypothesize that exposure of podocytes to IgG from patients with lupus nephritis or to hypoxia promotes the production and activation of complement intracellularly. We propose to identify complement components that are produced by podocytes after exposure to IgG from patients with lupus nephritis or hypoxia. Next, we will determine how cathepsin proteases activate complement intracellularly. For our studies we will use IgG from patients with lupus nephritis, a newly establish method to culture podocytes on surfaces coated with decellularized fibroblasts and podocytes from lupus-prone and cathepsin-deficient mice. CRISPR/Cas9 system will be utilized to knockdown C3 and cathepsins (B, L and S) in podocytes. The studies will produce a novel concept that intracellularly activated complement contributes to podocyte and glomerular damage independently of the deposited circulating complement. Our studies will point to the pursuit of avenues to limit increased production and activation of complement in podocytes in patients with lupus nephritis.

项目摘要 免疫复合物的沉积和补体系统的激活介导肾小球损伤和IS 参与狼疮性肾炎患者的炎症过程。足细胞高度专门化 上皮细胞对肾小球滤过屏障的正常功能至关重要，并在 几种蛋白尿性肾病。足细胞产生多种补体因子和组织蛋白酶 蛋白酶。初步证据表明，狼疮性肾炎患者的足细胞暴露于免疫球蛋白。 产生更多的补体，并在细胞内激活。我们假设 狼疮性肾炎患者足细胞暴露于免疫球蛋白或低氧环境可促进其产生和 细胞内补体的激活。我们建议确定所生产的补体成分 狼疮性肾炎或缺氧患者暴露于免疫球蛋白后的足细胞。接下来，我们将确定如何 组织蛋白水解酶激活细胞内补体。在我们的研究中，我们将使用来自 狼疮性肾炎--一种在脱细胞表面培养足细胞的新方法 狼疮倾向和组织蛋白酶缺陷小鼠的成纤维细胞和足细胞。CRISPR/CAS9系统将 用于敲除足细胞中的C3和组织蛋白(B，L和S)。这些研究将产生一个新的概念 细胞内激活的补体对足细胞和肾小球的损害独立于 沉积的循环补体。我们的研究将指向限制增加的途径的追求 狼疮性肾炎患者足细胞补体的产生和激活。