Functional and mechanistic delineation of HuR-Wisp1 signaling on myofibroblast activity

HuR-Wisp1信号对肌成纤维细胞活性的功能和机制描述

• 批准号: 10736289
• 负责人: Onur Kanisicak
• 金额: $ 71.88万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2023-09-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10736289
• 关键词: Affect; Antigens; Binding; Cardiac; Cardiac Myocytes; Cells; Consensus; Data; Extracellular Matrix; Fibroblasts; Foundations; Gene Expression; Genetic; Goals; Heart; Heart Diseases; Heart failure; Human; In Vitro; Individual; Knowledge; Mechanics; Mediating; Mediator; Modeling; Morphology; Myocardial; Myocardium; Myofibroblast; Outcome; Paracrine Communication; Pathologic; Pathway interactions; Phenotype; Play; Population; Proteomics; RNA-Binding Proteins; Role; Signal Pathway; Signal Transduction; Therapeutic; Transforming Growth Factor beta; Work; aorta constriction; cell type; connective tissue growth factor; coronary fibrosis; heart function; heart preservation; in vivo; mechanical properties; mouse model; periostin; pharmacologic; pressure; response; response to injury; single-cell RNA sequencing; small molecule inhibitor; transcriptomics; translational impact; Affect; Antigens; Binding; Cardiac; Cardiac Myocytes; Cells; Consensus; Data; Extracellular Matrix; Fibroblasts; Foundations; Gene Expression; Genetic; Goals; Heart; Heart Diseases; Heart failure; Human; In Vitro; Individual; Knowledge; Mechanics; Mediating; Mediator; Modeling; Morphology; Myocardial; Myocardium; Myofibroblast; Outcome; Paracrine Communication; Pathologic; Pathway interactions; Phenotype; Play; Population; Proteomics; RNA-Binding Proteins; Role; Signal Pathway; Signal Transduction; Therapeutic; Transforming Growth Factor beta; Work; aorta constriction; cell type; connective tissue growth factor; coronary fibrosis; heart function; heart preservation; in vivo; mechanical properties; mouse model; periostin; pharmacologic; pressure; response; response to injury; single-cell RNA sequencing; small molecule inhibitor; transcriptomics; translational impact

PROJECT SUMMARY: Cardiac fibrosis is regulated by the activation and phenotypic switching of quiescent cardiac fibroblasts (CFs) to active myofibroblasts (MFs). These MFs have extracellular matrix (ECM) remodeling and contractile functions which play a central role in cardiac remodeling in response to injury. As such, a consensus effort in the field is to manipulate fibroblast activity for therapeutic gain. However, a more complete understanding of the signaling pathways that regulate MF activity in cardiac remodeling remains an unmet need. We have previously demonstrated that the RNA binding protein Human antigen R (HuR) directly mediates hypertrophic signaling in cardiac myocytes (CMs), and that CM-specific genetic deletion or pharmacological inhibition of HuR reduces pathological remodeling and preserves cardiac function following transverse aortic constriction (TAC)-induced pressure overload, in part through a reduction in pro-fibrotic gene expression.1,2 In this proposal, we now present new preliminary data showing that HuR (1) expression in cardiac fibroblasts strongly correlates with established markers (periostin, 𝛼SMA, CTGF, etc) of MFs, (2) is necessary for TGFߚ- dependent MF activation and ECM-remodeling in vitro, and (3) CF-specific deletion of HuR preserves cardiac function following TAC. Critically, we have identified Wisp1 (Ccn4) as a downstream HuR-dependent mediator of MF activation, and show that expression of Wisp1 is sufficient to induce MF activity downstream/independent of HuR. The primary goals of this proposal are to determine the functional benefit of targeting HuR deletion in cardiac fibroblasts during pathological cardiac remodeling and the mechanistic impact of HuR deletion in CFs across other cell types in the myocardium (Aim 1), as well as the mechanisms by which HuR mediates Wisp1 expression in MFs, the functional benefit of fibroblast-specific Wisp1 deletion on cardiac remodeling, and the translational impact of these pathways on cardiac remodeling in human heart failure (Aim 2). Our central hypothesis is that HuR-Wisp1 signaling in cardiac fibroblasts is necessary for myofibroblast activity and promotes pathological cardiac remodeling. The specific aims of this proposal are to: (1) Determine the functional role of HuR in CFs in vivo and define its pleiotropic role across cell types during pathological cardiac remodeling. (2) Identify the functional and mechanistic role of HuR-dependent control of Wisp1 expression on MF activity and pathological cardiac remodeling. The expected results of this proposal will provide a mechanistic understanding of how HuR-Wisp1 signaling in cardiac fibroblasts modulates the functional response during pathological cardiac remodeling. Additionally, we will integrate previous translational work using a HuR small molecule inhibitor to better define the transcriptomic and functional changes across cardiac cell types in response to global and CF-specific abrogation of HuR signaling. In all, we expect this work to fill knowledge gaps in the signaling pathways that mediate cardiac fibroblast activation and ECM remodeling in the failing heart and provide a translational foundation for the targeting of HuR-Wisp1 signaling and/or HuR-dependent gene expression for therapeutic gain.

项目摘要： 心脏纤维化受静止心脏成纤维细胞的激活和表型切换来调节 （CFS）到活跃的肌纤维细胞（MFS）。这些MF具有细胞外基质（ECM）重塑和收缩 响应损伤而在心脏重塑中起着核心作用的功能。因此，在 场是为了操纵成纤维细胞活性以获得热增益。但是，对 调节心脏重塑中MF活性的信号通路仍然是未满足的需求。 我们以前已经证明RNA结合蛋白人类抗原R（HUR）直接介导 心肌细胞（CMS）中的肥厚信号传导以及CM特异性遗传缺失或药理 HUR的抑制减少病理重塑并保留横向主动脉后心脏功能 收缩（TAC）诱导的压力超负荷，部分通过促纤维基因表达的降低。1,2在 该提案，我们现在提供新的初步数据，表明HUR（1）在心脏成纤维细胞中的表达 与MFS的已建立标记（骨膜，𝛼SMA，CTGF等）密切相关，（2）对于TGF是必需的。 在体外依赖的MF激活和ECM复制，（3）CF特定的HUR删除可保留心脏 TAC之后的功能。至关重要的是，我们将WISP1（CCN4）确定为下游Hur依赖性调解人 MF激活的激活，并表明WISP1的表达足以诱导下游/独立的MF活性 Hur。 该提案的主要目标是确定针对心脏删除HUR删除的功能益处 病理心脏重塑期间的成纤维细胞以及跨越CFS的HUR缺失的机械影响 心肌中的其他细胞类型（AIM 1）以及HUR介导WISP1表达的机制 在MFS中，成纤维细胞特异性WISP1删除的功能益处在心脏重塑上和翻译 这些途径对人心力衰竭心脏重塑的影响（AIM 2）。我们的中心假设是 心脏成纤维细胞中的HUR-WISP1信号传导对于肌纤维细胞活动是必需的，并促进 该提案的具体目的是： （1）确定HUR在体内CFS中的功能作用，并在细胞类型中定义其多效性作用 病理心脏重塑。 （2）确定对HUR依赖性控制Wisp1表达在MF活性上的功能和机械作用 和病理心脏重塑。 该提案的预期结果将提供对HUR WISP1信号的机械理解 在心脏成纤维细胞中，在病理心脏重塑过程中调节功能反应。另外，我们 将使用HUR小分子抑制剂整合以前的翻译工作，以更好地定义转录组 响应于HUR的全球和CF特定归属率，心脏细胞类型之间的功能变化 信号。总的来说，我们希望这项工作能够填补介导心脏的信号传导途径中的知识空白 成纤维细胞激活和ECM在失败的心脏中重塑，并为 靶向HUR-WISP1信号传导和/或HUR依赖性基因表达以进行治疗增益。