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

摘要 本申请表是关于临床科学家导师研究职业发展奖的申请书，标题为 系统性硬化症相关间质肺肌成纤维细胞分化的基因组调控网络 疾病“，由肺科医学助理教授埃莉诺·瓦伦齐博士提交， 匹兹堡大学的过敏和重症护理医学。本意见书中详述的短期目标 旨在帮助申请人实现成为一名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)扩展技术和分析 有效执行翻译ILD研究的工具，以及(3)开发领导技能以获得 学术独立。这项工作将在肺、过敏和重症监护科完成 匹兹堡大学的医学，丰富的研究环境，坚定地致力于并得到证实 促进内科科学家发展的记录。这项研究提案的中心目标是 是为了研究推动系统性硬化症肌成纤维细胞分化的基因调控网络- 相关性间质性肺疾病(SSC-ILD)，因为肌成纤维细胞是纤维化和NO的关键效应细胞 目前存在针对这些细胞的治疗方法。间质性肺疾病是系统性肺炎的主要死亡原因。 硬化症是病死率最高的风湿病。申请人以前发表的作品被确认 SSC-ILD致病肌成纤维细胞在移植肺组织中的转录组 目前尚不清楚驱动成纤维细胞向病理性肌成纤维细胞分化的调控网络。 更多的初步数据表明，常驻肺成纤维细胞是肌成纤维细胞的主要来源， RUNX2/3转录因子可能是肌成纤维细胞分化的正调控因子。这个 主要假设是，RUNX2和RUNX3与特定的复合基序TF伙伴一起促进 SSc-ILD中肌成纤维细胞分化和异常表型。有了这项建议， 申请者将以以下具体目标扩展她之前的工作：(1)定义表观遗传学和 肌成纤维细胞分化所必需的转录程序，(2)重建调控网络 Runx2/3，以及(3)确定Runx2/3调节关键的 肌成纤维细胞功能效应器的细胞外基质表达和收缩功能。这项提案将决定 SSC-ILD肌成纤维细胞RUNX2/3基因调控程序及下游通道的建立 关键转录因子及其调控机制的研究进展 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.