Elucidating the Role of Microenvironment Mechanics in Regulating Cardiac Myofibroblast Plasticity

阐明微环境力学在调节心脏肌成纤维细胞可塑性中的作用

基本信息

批准号：
10570135
负责人：
Sangkyun Cho
金额：
$ 13.04万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10570135
关键词：
Advisory Committees Affect Animal Model Attenuated Back Biocompatible Materials Biology Biomedical Engineering Cardiac Cardiology Cardiovascular Diseases Cardiovascular system Cells Cellular Assay Characteristics Child Chromatin Committee Members Complement Custom Development Effector Cell Engineering Environment Epigenetic Process Etiology Event Extracellular Matrix Feedback Fibroblasts Fibrosis Genes Genetic Genomics Goals Heart Heart failure Human Hydrogels Hypertrophy Immunoprecipitation Mass Spectrum Analysis Mechanics Mediating Mediator Medicine Mentors Mentorship Modeling Molecular Morbidity - disease rate Mus Myocardial Myocardial Ischemia Myofibroblast Pathologic Pathway interactions Patient-Focused Outcomes Play Population Proteins Proteomics Protocols documentation Regenerative Medicine Regulation Regulatory Element Reporter Research Research Training Role Series Signal Transduction System Testing Therapeutic Time Tissues Training Transforming Growth Factor beta Transposase Universities antifibrotic treatment cardiac tissue engineering career cell type coronary fibrosis drug candidate druggable target effective therapy efficacy testing epigenomics fibrogenesis in vitro Assay in vivo induced pluripotent stem cell inherited cardiomyopathy insight live cell imaging mechanical properties mechanical signal mortality mouse model multiple omics novel novel therapeutic intervention pressure programs response single cell analysis synergism training opportunity transcription factor transdifferentiation virtual

项目摘要

PROJECT SUMMARY Fibrosis underlies a vast number of cardiac pathological conditions, ranging from genetic cardiomyopathies to ischemic heart failure. Although substantial progress has been made in identifying molecular signals that trigger the characteristic activation of quiescent cardiac fibroblasts (CFs) and their transdifferentiation into myofibroblasts (MyoFBs), far less is known about the mechanisms that govern their long-term fate and persistence, which presents major obstacles to the development of effective anti-fibrotic therapies. This K99/R00 application describes a five-year research training plan that proposes to leverage (i) human induced pluripotent stem cell-derived cardiac fibroblasts (iPSC-CFs), (ii) engineered biomaterials with tunable mechanical properties, and (ii) single-cell multiomics platforms to investigate molecular mechanisms that govern MyoFB fate and plasticity. Given the well-established sensitivity of CFs and MyoFBs to extracellular matrix (ECM) stiffness, the applicant Dr. Sangkyun Cho will test the hypothesis that modulation of ECM-mediated mechanical signaling potentiates the de-differentiation of MyoFBs, by synergizing with soluble factors known to regulate major pathways in fibrogenesis. In Aim 1 (K99), Dr. Cho will use reporter iPSC lines (with fluorescently tagged canonical MyoFB ‘marker’ genes, e.g., CFP-TAGLN) and a novel dynamically softening hydrogel system to characterize in real-time the effects of mechanical unloading on MyoFB fate. In Aim 2 (K99), Dr. Cho will investigate the synergy between ECM softening and the TGF-beta pathway in regulating MyoFB states, (i) by examining stiffness-dependent protein interactions among mechanosensitive transcription factors (e.g., yes- associated protein 1 (YAP)), and (ii) by identifying epigenetic regulators downstream of ECM stiffness with single- cell assay for transposase transposase-accessible chromatin (scATAC-seq). In Aim 3 (R00), Dr. Cho will identify potential druggable targets along the cell’s mechanosensory apparatus, and test candidate compounds in engineered heart tissues and a mouse model of pressure-overload induced hypertrophy and heart failure. The proposed studies build upon PI Dr. Sangkyun Cho’s well-suited prior training in biomaterials, proteomics, and ECM mechanobiology, while providing new training opportunities in (i) reporter iPSC-CFs, (ii) single-cell multiomics platforms, and (iii) animal models. Mentor Dr. Joseph Wu is a pioneer in iPSCs and cardiovascular biology, and co-mentor Dr. Sarah Heilshorn is a leading expert in biomaterials and regenerative medicine, whose mentorship complements that of Dr. Wu. Advisory Committee members Drs. Jeffery Molkentin (cardiac fibrosis), Joseph Hill (heart failure models), and Michal Snyder (single-cell genomics) provide additional expertise and guidance. In Summary, the well-tailored research training plan, exceptional mentoring team, and an outstanding Environment at Stanford University are anticipated to help propel Dr. Cho toward his long-term goal of establishing an independent research program at the intersection of bioengineering and cardiovascular stromal biology.

项目概要纤维化是大量心脏病理学病症的基础，从遗传性心肌病到尽管在识别引发缺血性心力衰竭的分子信号方面已经取得了实质性进展。静止心脏成纤维细胞（CF）的特征性激活及其转分化为对于肌成纤维细胞（MyoFBs）的长期命运和控制机制知之甚少。持久性，这对开发有效的抗纤维化疗法构成了主要障碍。该 K99/R00 应用程序描述了一项为期五年的研究培训计划，建议利用 (i) 人力诱导多能干细胞衍生的心脏成纤维细胞（iPSC-CF），（ii）具有可调性的工程生物材料机械特性，以及（ii）单细胞多组学平台来研究控制的分子机制鉴于 CF 和 MyoFB 对细胞外基质 (ECM) 的既定敏感性，MyoFB 的命运和可塑性。刚度方面，申请人 Sangkyun Cho 博士将测试以下假设：ECM 介导的机械调节信号传导通过与已知调节的可溶性因子协同作用，增强 MyoFB 的去分化在目标 1 (K99) 中，Cho 博士将使用报告 iPSC 系（带有荧光标记）。典型的 MyoFB“标记”基因，例如 CFP-TAGLN）和新型动态软化水凝胶系统在 Aim 2 (K99) 中，Cho 博士将实时描述机械卸载对 MyoFB 命运的影响。研究 ECM 软化和 TGF-β 通路在调节 MyoFB 状态中的协同作用，(i) 检查机械敏感转录因子之间的刚度依赖性蛋白质相互作用（例如，是的- 相关蛋白 1 (YAP))，以及 (ii) 通过使用单-鉴定 ECM 硬度下游的表观遗传调节因子在 Aim 3 (R00) 中，Cho 博士将鉴定转座酶可及染色质的细胞分析 (scATAC-seq)。沿着细胞的机械感觉装置潜在的可成药靶点，并测试候选化合物工程心脏组织和压力超负荷引起肥厚和心力衰竭的小鼠模型。拟议的研究建立在 PI 博士 Sangkyun Cho 之前在生物材料、蛋白质组学、和 ECM 机械生物学，同时提供以下方面的新培训机会：(i) 报告 iPSC-CF，(ii) 单细胞多组学平台，以及 (iii) 动物模型导师 Joseph Wu 博士是 iPSC 和心血管领域的先驱。生物学和联合导师 Sarah Heilshorn 博士是生物材料和再生医学领域的专家，其导师与吴博士顾问委员会成员 Jeffery Molkentin 博士（心脏病）相辅相成。纤维化）、Joseph Hill（心力衰竭模型）和 Michal Snyder（单细胞基因组学）提供了额外的专业知识总而言之，精心定制的研究培训计划、出色的指导团队和专业的指导。斯坦福大学出色的环境有望帮助曹博士实现他的长期目标在生物工程和心血管交叉领域建立独立的研究计划基质生物学。