Uncovering the genomic regulatory network of myofibroblast differentiation in systemic sclerosis-associated interstitial lung disease

摘要 本申请是为指导临床科学家研究职业发展奖题为“发现 系统性硬化症肺间质成肌纤维细胞分化的基因调控网络 疾病”，由Eleanor Valenzi博士提交，他是肺科医学助理教授， 过敏和重症监护医学在匹兹堡大学。本文件中详细说明的短期目标 旨在帮助申请人实现成为独立医生的长期目标- 自身免疫性间质性肺病（ILD）研究领域的科学家和领导者。这些短期目标 包括（1）提高计算和分子生物学知识;（2）扩大技术和分析 工具，以有效地进行翻译ILD研究，和（3）发展领导技能，以获得 学术独立。这项工作将在肺、过敏和重症监护部门完成 医学在匹兹堡大学，一个丰富的研究环境与坚定的承诺，并证明 促进医生-科学家发展的良好记录。这项研究的主要目的是 是研究系统性硬化症中驱动肌成纤维细胞分化的基因调控网络- 相关间质性肺病（SSc-ILD），因为肌成纤维细胞是纤维化的关键效应细胞， 目前存在靶向这些细胞的治疗。间质性肺病是全身性疾病中死亡的主要原因。 硬化症是死亡率最高的风湿性疾病。申请人先前发表的作品 肺组织中SSc-ILD致病性肌成纤维细胞的转录组，然而， 驱动固有成纤维细胞分化为致病性肌成纤维细胞的调节网络仍然未知。 额外的初步数据表明，肺成纤维细胞是肌成纤维细胞的主要来源， 和RUNX 2/3转录因子（TF）作为肌成纤维细胞分化的假定正调节因子。的 主要假设是RUNX 2和RUNX 3与特定的复合基序TF配偶体相关，促进 肌成纤维细胞分化和SSc-ILD中异常肌成纤维细胞表型。根据这项建议， 申请人将扩展她以前的工作，具体目标如下：（1）定义表观遗传， 转录程序必不可少的肌成纤维细胞分化，（2）重建调控网络， RUNX 2/3在肌成纤维细胞中的作用，以及（3）确定RUNX 2/3调节肌成纤维细胞中关键细胞的机制。 肌成纤维细胞效应子功能的细胞外基质表达和收缩性。该提案将决定 SSc-ILD肌成纤维细胞RUNX 2/3基因调控程序，并建立下游 研究关键的转录因子及其调控机制， 肌成纤维细胞表型

ABSTRACT This application is for a Mentored Clinical Scientist Research Career Development Award entitled “Uncovering the genomic regulatory network of myofibroblast differentiation in systemic sclerosis-associated interstitial lung disease”, submitted by Dr. Eleanor Valenzi, an Assistant Professor of Medicine within the Division of Pulmonary, Allergy and Critical Care Medicine at the University of Pittsburgh. The short-term goals detailed in this submission are designed to help the applicant achieve her long-term objective of becoming an independent physician- scientist and leader in the field of autoimmune interstitial lung disease (ILD) research. These short-term goals include (1) advancing knowledge of computational and molecular biology (2) expansion of technical and analytic tools to effectively perform translational ILD research, and (3) development of leadership skills to obtain academic independence. This work will be completed in the Division of Pulmonary, Allergy and Critical Care Medicine at the University of Pittsburgh, a rich research environment with a strong commitment to and proven track record of fostering the development of physician-scientists. The central objective of this research proposal is to investigate the gene regulatory networks driving myofibroblast differentiation in systemic sclerosis- associated interstitial lung disease (SSc-ILD), as myofibroblasts are the key effector cell in fibrosis and no treatments currently exist to target these cells. Interstitial lung disease is the leading cause of death in systemic sclerosis, the rheumatic disease with the highest case mortality. The applicant’s prior published work identified the transcriptome of the SSc-ILD pathogenic myofibroblasts in explanted lung tissues, however the specific gene regulatory networks driving differentiation of resident fibroblasts to pathogenic myofibroblasts remain unknown. Additional preliminary data implicates resident pulmonary fibroblasts as the primary source of myofibroblasts, and the RUNX2/3 transcription factors (TFs) as putative positive regulators of myofibroblast differentiation. The primary hypothesis is that RUNX2 and RUNX3, in association with specific composite motif TF partners, promote myofibroblast differentiation and the aberrant myofibroblast phenotype in SSc-ILD. With this proposal, the applicant will expand on her prior work with the following specific aims: (1) define the epigenetic and transcriptional programs essential to myofibroblast differentiation, (2) reconstruct the regulatory network of RUNX2/3 in myofibroblasts, and (3) determine the mechanism by which RUNX2/3 regulate the critical myofibroblast effector functions of extracellular matrix expression and contractility. This proposal will determine the SSc-ILD myofibroblast RUNX2/3 gene regulatory program and establish a pipeline for downstream investigation of the critical implicated transcription factors and the mechanisms by which they regulate the myofibroblast phenotype.