Antibody Mediated Glomerular Injury

抗体介导的肾小球损伤

• 批准号: 7188957
• 负责人: DAVID J SALANT
• 金额: $ 32.05万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7188957
• 关键词: Adult; Affect; Antibodies; Binding; Cell Nucleus; Cells; Clinical; Coculture Techniques; Complement; Complement Membrane Attack Complex; Consensus; Dose; Electron Microscopy; Epithelial Cells; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Glomerulonephritis; Goals; Histopathology; Immune; In Vitro; Injury; Joint Dislocation; Ligands; Location; Measurement; Measures; Mediating; Modeling; Monitor; Morphology; Mus; Nephrotoxic; Nuclear; Pathology; Pathway interactions; Patients; Phenotype; Predisposition; Promoter Regions; Proteins; Proteinuria; RNA; Rattus; Recovery; Regulation; Research Personnel; Rodent; Role; Serum; Stress; Structure; Time; Western Blotting; cell injury; comparative; complement C5b; design; gamma secretase; glomerulosclerosis; in vitro Model; in vivo; in vivo Model; male; mouse model; nephrin; notch protein; podocyte; programs; response; secretase; structural biology; transmission process; young adult

DESCRIPTION (provided by applicant): The primary goal of this proposal is to explore the hypothesis that Notch4 contributes to the integrity of the mature podocyte. This will be accomplished through an analysis of in vitro and in vivo models of antibody-mediated podocyte injury and Notch4-deficient (N4-/-) mice. We found that Notch4 is present in the podocytes of mature rats and mice and that the podocytes of N4-/- mice are focally effaced with increased expression, clustering and dislocation of nephrin. Moreover, Notch4 expression is abundant and total nephrin is reduced coincident with the onset of proteinuria in rats with passive Heymann nephritis, which suggests that Notch4 might regulate podocyte gene transcription. Thus, we will examine the role of Notch4 in the mature podocyte according to four specific aims. In the first specific aim we will use an in vitro model of antibody- and complement-mediated cell injury to determine if Notch4 is activated by gamma-secretase and translocates to the nucleus to bind and activate its nuclear target, CSL. In the second specific aim we will examine N4-/- mice over time to determine if Notch4 deficiency leads to proteinuria and alterations in podocyte morphology and podocyte-associated proteins. The third specific aim is to determine if Notch4 influences the susceptibility to and/or recovery from immune glomerular injury. N4+/+ and N-/- littermates will be studied using various models of antibody-mediated podocyte injury. Their susceptibility to injury and time to recovery will be monitored, and the effect of gamma-secretase inhibition will be examined. The fourth specific aim will utilize a comparative transcriptome analysis of glomerular RNA derived from young male N4+/+ and N-/- littermates before and after antibody-mediated podocyte injury to identify genes that Notch4 regulates in mature glomeruli. Differentially expressed genes will be examined for appropriate location and expression in glomeruli and regulation by Notch4. The results of these studies may disclose mechanisms underlying premature podocyte degeneration and glomerular sclerosis and the variable clinical and pathological response of patients with antibody-mediated podocyte injury.

描述（由申请人提供）：该提案的主要目标是探讨Notch4有助于成熟足细胞的完整性的假设。这将通过分析抗体介导的足细胞损伤和Notch4缺陷型（N4 - / - ）小鼠的体外和体内模型来实现。我们发现，成熟大鼠和小鼠的足细胞中存在Notch4，并且N4 - / - 小鼠的足细胞随着表达，凝聚和肾上腺素的脱位的增加而局部排出。此外，Notch4表达丰富，总肾素与被动Heymann肾炎大鼠的蛋白尿发作相一致，这表明Notch4可能调节足细胞基因转录。因此，我们将根据四个特定目的检查Notch4在成熟的足细胞中的作用。在第一个特定目的中，我们将使用抗体和补体介导的细胞损伤的体外模型来确定Notch4是否被γ-分泌酶激活，并转移到细胞核中以结合并激活其核靶标CSL。在第二个特定目的中，我们将随着时间的推移检查N4 - / - 小鼠，以确定Notch4缺乏是否导致蛋白尿和足细胞形态和足细胞相关蛋白的改变。第三个具体目的是确定Notch4是否影响免疫肾小球损伤的敏感性和/或恢复。将使用各种抗体介导的足细胞损伤模型研究N4+/+和N - / - 窝窝。将监测它们对伤害和恢复时间的敏感性，并将检查γ-分泌酶抑制的作用。第四个特定目的将利用源自年轻男性N4+/+和N - / - 同窝的肾小球RNA的比较转录组分析，并在抗体介导的足细胞损伤之前和之后识别Notch4在成熟肾小球中调节的基因。差异表达的基因将在肾小球中的适当位置和表达检查，并通过Notch4进行调节。这些研究的结果可能会揭示出早熟足细胞变性和肾小球硬化的机制，以及抗体介导的足细胞损伤患者的临床和病理反应可变的临床和病理反应。