Understanding the mechanisms of TRPC6 mediated FSGS

了解 TRPC6 介导的 FSGS 机制

• 批准号: 10063516
• 负责人: JOHANNES S SCHLONDORFF
• 金额: $ 39.38万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-06 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063516
• 关键词: Actins; Address; Affect; Age; Agonist; Angiotensins; Animal Model; Animals; Apoptosis; Behavior; Biochemical; Biological; Biological Assay; Biology; CRISPR screen; CRISPR/Cas technology; Calcineurin; Cations; Cause of Death; Cell Death; Cell Line; Cell model; Cell physiology; Cell surface; Cells; Chronic Kidney Failure; Clustered Regularly Interspaced Short Palindromic Repeats; Cytoskeleton; Data; Development; Disease; Disease model; Dominant-Negative Mutation; Electrophysiology (science); End stage renal failure; Focal Segmental Glomerulosclerosis; Gene Mutation; Genes; Genetic; Genetic Diseases; Genetic Screening; Genotype; Goals; Histologic; Human; Human Genetics; In Vitro; Infusion procedures; Injury; Ion Channel; Kidney; Kidney Diseases; Knock-in Mouse; Lead; Mediating; Methodology; Modeling; Molecular; Molecular Target; Mus; Mutate; Mutation; Nephritis; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Pharmacology; Phenotype; Predisposition; Prevention; Property; Proteinuria; Regulation; Renal glomerular disease; Reporting; Research; Role; Serum; Signal Pathway; Signal Transduction; Small Interfering RNA; Stress; Therapeutic Agents; base; cell immortalization; cytotoxicity; effective therapy; gain of function; gain of function mutation; glomerular function; in vivo; inhibitor/antagonist; insight; knock-down; knockin animal; loss of function mutation; mouse model; mutant; nephrotoxicity; novel; novel strategies; novel therapeutic intervention; overexpression; podocyte; response; response to injury; stressor; therapeutic target

PROJECT SUMMARY Focal segmental glomerulosclerosis (FSGS) is a significant cause of chronic kidney disease, has few effective treatment options, and usually progresses to end-stage renal disease (ESRD). FSGS has a substantive genetic component. Study of gene mutations that cause FSGS has provided invaluable insight into normal glomerular function and the disease pathogenesis of FSGS. Mutations in TRPC6 are a known cause of FSGS. In addition, TRPC6 levels are often increased in more common forms of kidney disease, implying that TRPC6 activity may impact both acquired and genetic forms of FSGS. TRPC6 is an ion channel found on the surface of cells, making it an attractive pharmacologic target. However, two key questions about TRPC6 and the development of FSGS remain unanswered. Is it simply that excess TRPC6 activity is detrimental to glomerular function, or is TRPC6 like Goldilocks, with either too much or too little detrimental? Which of the multiple signaling pathways intersecting with TRPC6 are relevant to the development of glomerular pathology? We have created novel animal models - mice with the equivalent of a human FSGS mutation in Trpc6 and others with a dominant negative (DN) Trpc6 mutation - to be able to address these questions. Our experimental plan has three Specific Aims with the goal of providing answers to the above questions: (1) To ascertain whether mice with FSGS- or DN-mutations in Trpc6 develop FSGS with age or require a “second hit” to manifest disease. (2) To characterize the effect of TRPC6 mutations on podocyte behaviors known to be affected in FSGS, focusing on regulation of their actin cytoskeleton, calcineurin-NFAT signaling, and cell death. (3) To determine the genes necessary for mutant TRPC6-induced cell death. Our proposed studies have the potential to significantly enhance our understanding of FSGS. They will help us understand how increased and decreased TRPC6 activity alters glomerular response to stress, and will clarify which of the known signaling pathways activated by TRPC6 mediate pathology. The research has the potential to uncover novel molecular mechanisms involved in regulating TRPC6 and promoting FSGS. Ultimately, the results will help determine whether blocking TRPC6 activity might be a potential, and much needed, new therapeutic approach for the treatment of FSGS.

项目摘要 局灶性节段性肾小球硬化症（FSGS）是慢性肾脏疾病的重要原因，很少有效 治疗选择，通常会发展为终末期肾脏疾病（ESRD）。 FSG有实质性 遗传成分。研究引起FSG的基因突变已提供了对正常的宝贵洞察力 肾小球功能和FSG的疾病发病机理。 TRPC6中的突变是FSG的已知原因。另外，TRPC6水平通常会增加 肾脏疾病的常见形式，这意味着TRPC6活性可能会影响获得的和遗传形式 FSG。 TRPC6是在细胞表面发现的一个离子通道，使其成为有吸引力的药理靶标。 但是，有关TRPC6和FSG的发展的两个关键问题仍未得到解答。只是简单 多余的TRPC6活性对肾小球功能有害，或者像Goldilocks一样是TRPC6，任一 还是很少的决心？与TRPC6相交的多个信号通路中的哪个与 肾小球病理的发展？我们创建了新颖的动物模型 - 相当于 TRPC6中的人类FSG突变和其他具有显性负（DN）TRPC6突变的突变 - 能够 解决这些问题。我们的实验计划具有三个具体目标，目的是提供答案 以上问题：（1）确定TRPC6中使用FSGS或DN-突变的小鼠是否会与 年龄或需要“第二击”才能表现出疾病。 （2）表征TRPC6突变对 已知在FSG中受到影响的足细胞行为，重点是调节其肌动蛋白细胞骨架， 钙调神经酶-NFAT信号传导和细胞死亡。 （3）确定突变体TRPC6诱导所需的基因 细胞死亡。 我们提出的研究有可能显着增强我们对FSG的理解。他们会帮助我们 了解TRPC6活性的增加和降低如何改变对压力的肾小球反应，并将澄清 TRPC6激活的哪种已知信号通路介导了病理学。这项研究有 揭示参与调节TRPC6并促进FSG的新型分子机制的潜力。 最终，结果将有助于确定阻止TRPC6活动是否可能是潜力，并且很大 需要新的治疗方法来治疗FSG。

项目成果

Trpc6 gain-of-function disease mutation enhances phosphatidylserine exposure in murine platelets.

• DOI: 10.1371/journal.pone.0270431
• 发表时间: 2022
• 期刊: PLOS ONE
• 影响因子: 3.7
• 作者: Boekell, Kimber L.;Brown, Brittney J.;Talbot, Brianna E.;Schlondorff, Johannes S.
• 通讯作者: Schlondorff, Johannes S.

My, oh, MYO9A! Just how complex can regulation of the podocyte actin cytoskeleton get?

我的，哦，MYO9A！

• DOI: 10.1016/j.kint.2021.01.006
• 发表时间: 2021
• 期刊: Kidney international
• 影响因子: 19.6
• 作者: Schlöndorff,JohannesS