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 局灶节段性肾小球硬化（FSGS）是慢性肾脏病的重要病因， 治疗选择，通常进展为终末期肾病（ESRD）。FSGS具有实质性的 遗传成分。对导致FSGS的基因突变的研究为正常人提供了宝贵的见解。 肾小球功能和FSGS的发病机制。 TRPC 6的突变是FSGS的已知原因。此外，TRPC 6水平通常在更多的人中增加。 常见形式的肾脏疾病，这意味着TRPC 6活性可能影响获得性和遗传性形式的肾脏疾病。 FSGS。TRPC 6是在细胞表面发现的离子通道，使其成为有吸引力的药理学靶点。 然而，关于TRPC 6和FSGS发展的两个关键问题仍然没有答案。是因为 过量的TRPC 6活性对肾小球功能有害，或者TRPC 6像金发姑娘， 还是太少有害？与TRPC 6交叉的多个信号通路中的哪一个与 肾小球病变的发展我们已经创造了新的动物模型-小鼠相当于一个 Trpc 6中的人FSGS突变和具有显性阴性（DN）Trpc 6突变的其他人-为了能够 回答这些问题。我们的实验计划有三个具体目标，目的是提供答案， 上述问题：（1）为了确定Trpc 6中具有FSGS-或DN-突变的小鼠是否发生FSGS， 年龄或需要“第二次打击”来表现疾病。(2)为了表征TRPC 6突变对 足细胞行为已知在FSGS中受到影响，重点是调节其肌动蛋白细胞骨架， 钙调神经磷酸酶-NFAT信号传导和细胞死亡。(3)为了确定突变型TRPC 6诱导的细胞凋亡所必需的基因， 细胞死亡 我们提出的研究有可能显着提高我们对FSGS的理解。他们将帮助我们 了解TRPC 6活性的增加和减少如何改变肾小球对应激的反应，并将澄清 由TRPC 6激活的已知信号通路中的哪一个介导病理。这项研究有 有可能揭示参与调节TRPC 6和促进FSGS的新分子机制。 最终，这些结果将有助于确定阻断TRPC 6活性是否可能是一种潜在的， 需要新的治疗方法来治疗FSGS。

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