Hippo-YAP in podocyte health and disease

Hippo-YAP 在足细胞健康和疾病中的作用

• 批准号: 10188524
• 负责人: Kirk N Campbell
• 金额: $ 51.03万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-03 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188524
• 关键词: 3-Dimensional; Actins; Agonist; Architecture; Atomic Force Microscopy; Biological; Biophysics; Calcium; Cells; Cellular Structures; Clinical; Cytoplasmic Protein; Cytoskeleton; Dasatinib; Data; Development; Disease; Disease Progression; Electrophysiology (science); Focal Adhesions; Focal Segmental Glomerulosclerosis; Gene Expression; Genes; Genetic Transcription; Goals; Health; Homeostasis; Homology Modeling; Human; In Vitro; Injury; Kidney; Kidney Diseases; Knowledge; Left; Mediating; Microfilaments; Modeling; Molecular; Mus; Nuclear; Nuclear Protein; Oncogenes; Pathogenesis; Pathogenicity; Pathway interactions; Phosphorylation; Phosphotransferases; Potassium; Potassium Channel; Predisposition; Protein Inhibition; Proteins; Purinoceptor; Renal glomerular disease; Reporter Genes; Research; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Testing; Therapeutic; Therapeutic Agents; Tyrosine Kinase Inhibitor; Up-Regulation; Work; base; biophysical properties; cell injury; clinically relevant; druggable target; glomerular filtration; in vivo; innovation; mechanotransduction; new therapeutic target; novel; novel therapeutics; podocyte; protein activation; protein expression; protein function; restoration; slit diaphragm; small molecule; therapeutic development; therapeutic target; transcription factor; uptake

Project Summary A limited understanding of clinically relevant signaling pathways has limited the development of therapeutic agents for human glomerular disease. Our long-term goal is to enhance the pipeline of putative therapeutic targets available to tackle human glomerular disease by elucidating the details and functional significance of key signaling pathways that regulate podocyte injury and survival. Our preliminary data have identified YAP, the key effector of the Hippo signaling pathway, as an important regulator of podocyte survival. YAP inactivation in podocytes causes FSGS in mice and decreased YAP expression is associated with the development and progression of human glomerular disease. We have detected increased intracellular calcium uptake and marked upregulation of calcium-gated potassium channel expression in YAP silenced podocytes. The overall objective of this application is to define the mechanism by which YAP regulates podocyte survival and test its role as a potential therapeutic target. Our central hypothesis is that YAP is inactivated in podocytes by canonical phosphorylation and cytoplasmic sequestration under the influence of the Hippo kinase LATS. YAP expression and function can also be regulated at the genetic and transcriptional level. Decreased YAP signaling enhances purinergic receptor-mediated calcium uptake in podocytes and calcium-gated potassium channel activation contributing to disruption of the actin cytoskeleton. The rationale for the proposed research is that defining the underlying mechanisms that regulate YAP function will advance understanding of glomerular disease progression as well as the quest for novel therapeutic targets available for clinical use. Our hypothesis will be tested by pursuing two specific aims: Aim 1 will explore the functional significance of YAP phosphorylation and nuclear-cytoplasmic shuttling in podocyte survival. We will determine whether cytoplasmic YAP expression in podocytes enhances injury susceptibility and enhancing nuclear YAP signaling is protective in proteinuric kidney disease. We will also develop a novel YAP agonist and test its role in protecting podocytes from injury. In Aim 2 we will determine the key signaling pathways and cellular structural changes induced by YAP inactivation. Our innovative approach utilizes state of the art microfabricated 3-D chips, electrophysiology and atomic force microscopy to quantify the biophysical properties of podocytes during YAP inhibition and activation under normal and disease conditions. By homology modeling, we will generate novel small molecule YAP agonists that could be protective in proteinuric kidney disease. These contributions are significant because they have the potential to not only advance understanding of the pathogenesis of glomerular disease but could help identify novel therapeutic targets.

项目摘要 对临床相关信号通路的有限理解限制了治疗方法的发展。 治疗人类肾小球疾病的药物。我们的长期目标是加强假定的治疗渠道 可用于治疗人类肾小球疾病的靶点通过阐明其细节和功能意义 调节足细胞损伤和存活的关键信号通路。我们的初步数据已经确认了YAP， 河马信号通路的关键效应者，作为足细胞生存的重要调节因子。耶普 足细胞失活导致小鼠FSGS，YAP表达减少与 人类肾小球疾病的发生发展。我们检测到细胞内钙离子升高 YAP沉默的足细胞摄取并显著上调钙门控钾通道的表达。 本应用的总体目标是确定YAP调节足细胞存活的机制 并测试其作为潜在治疗靶点的作用。我们的中心假设是YAP在足细胞中失活 在河马蛋白激酶LATS的影响下，通过典型的磷酸化和细胞质隔离。 YAP的表达和功能也可以在基因和转录水平上进行调节。减少了YAP 信号转导增强嘌呤能受体介导的足细胞钙摄取和钙门控钾 通道激活导致肌动蛋白细胞骨架的破坏。建议进行这项研究的理由 定义调节YAP功能的潜在机制将促进对 肾小球疾病的进展以及对可用于临床的新的治疗靶点的探索。我们的 假设将通过追求两个具体目标来检验：目标1将探索YAP的功能意义 足细胞存活中的磷酸化和核质穿梭。我们将确定细胞质是否 YAP在足细胞中的表达增强损伤易感性，增强核YAP信号具有保护作用 在蛋白尿性肾病中。我们还将开发一种新型的YAP激动剂，并测试其在保护 足细胞因受伤而死亡。在目标2中，我们将确定关键的信号通路和细胞结构变化 由YAP失活诱导。我们的创新方法利用最先进的微型制造3-D芯片， 电生理学和原子力显微镜对足细胞生物物理特性的定量研究 在正常和疾病条件下的抑制和激活。通过同源建模，我们将生成新颖的 小分子YAP激动剂，可在蛋白尿性肾病中起到保护作用。这些贡献是 意义重大，因为它们不仅有可能促进对糖尿病发病机制的了解 治疗肾小球疾病，但可能有助于确定新的治疗靶点。