Targeting myofibroblast activation in chronic fibrotic disease

靶向慢性纤维化疾病中的肌成纤维细胞激活

• 批准号: 7536051
• 负责人: ARTHUR ROGER STRAUCH
• 金额: $ 37.5万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-05 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7536051
• 关键词: 1-Phosphatidylinositol 3-Kinase; Affect; Attenuated; Behavior; Biochemical Pathway; COL1A2 gene; Cardiac; Cells; Cessation of life; Chronic; Chronic Disease; Cicatrix; Collagen; Collagen Gene; Collagen Type I; Complex; Connective Tissue Cells; Contractile Proteins; DNA; DNA binding protein B; Deformity; Deposition; Diagnosis; Disease; Exhibits; Extracellular Matrix; Feedback; Fibrosis; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genetic Translation; Growth Factor; Heart; Heart Diseases; Human; Hypertrophic Cicatrix; In Vitro; Individual; Injury; Interleukin-13; Intervention; Ischemia; Lung; Mediating; Mediator of activation protein; Messenger RNA; Modeling; Molecular; Mus; Myofibroblast; Operative Surgical Procedures; Organ Transplantation; Outcome; Output; Participant; Peptides; Pharmacotherapy; Phosphorylation Site; Phosphotransferases; Process; Property; Proteins; Pulmonary Heart Disease; Pur-1 protein; Quality of life; RNA Binding; RNA-Binding Proteins; Relative (related person); Reperfusion Injury; Reperfusion Therapy; Repressor Proteins; Research; Signal Transduction; Smooth Muscle; Solid; Structure of parenchyma of lung; Therapeutic; Thrombin; Tissues; Transcript; Transcription Coactivator; Transcription Regulatory Protein; Transcription Repressor/Corepressor; Translations; Transplant Recipients; Vascular Smooth Muscle; Werdnig-Hoffmann Disease; Wound Healing; alpha Actin; base; design; improved; in vivo; inhibitor/antagonist; interstitial; loss of function; novel; open wound; polypeptide; promoter; research study; small molecule

DESCRIPTION (provided by applicant): Chronic accumulation of myofibroblasts in healing wounds is associated with hypercontractility, excessive deposition of interstitial collagens, and destructive tissue remodeling. Vascular smooth muscle alpha-actin ((-SMA) is a contractile protein transiently expressed by differentiated myofibroblasts for generating tensile force required to close open wounds. In chronic fibrotic disease, myofibroblast differentiation is dysfunctional and we discovered that molecular signaling required for activation of both the (-SMA and type I collagen genes in these cells also provides negative feedback that could potentially limit the recruitment of hyperactive myofibroblasts during wound healing and destructive remodeling. Studies outlined in this proposal are expected to reveal novel forms of functional interplay of the DNA- and mRNA-binding proteins YB-1, Pur (, and Pur ( with the (-SMA and type I collagen promoters and clarify how these proteins are affected by pro- fibrotic agents such as TGF(1 and thrombin that, if unchecked, may cause myofibroblast progression to hypertrophic scarring. Experiments are designed to initiate, amplify, or attenuate myofibroblast differentiation to better understand strategies for controlling (-SMA and type I collagen gene output at the transcriptional and translation levels as well as reveal novel interventional strategies that might be useful for minimizing aberrant wound healing outcomes. Aim 1 will examine TGF(1-regulated interaction of YB-1 and Pur protein repressors with (-SMA and collagen promoter DNA and the transcriptional activators Sp1, SRF, and Smads 2,3, delineate regions of repressor polypeptide chains required for this functional interplay, and attempt to disrupt complex formation and disable pathobiologic myofibroblast differentiation using peptide decoys and small molecule pharmacologic inhibitors. Aim 2 will determine if thrombin potentiates myofibroblast differentiation at the level of translational control thus functioning as a TGF(1 supplement or instead antagonizes this growth factor by blocking transcription and myofibroblast recruitment by inducing the anti-fibrotic transcriptional regulatory protein, Egr-1. Aim 3 studies will explore alternative, Smad-independent mechanisms of myofibroblast differentiation and fibrosis. Loss-of-function approaches based on pharmacologic inhibition of TGF(1/Smad kinase- or phosphatidylinositol-3-kinase (PI3K)/Akt kinase signaling will be used to evaluate their possible suppressive effect on myofibroblast activation in vitro and cardiac fibrosis in mice after ischemia/reperfusion injury. The ability of TGF(1 and thrombin to exploit the unique DNA-, RNA-, and protein-binding properties of YB-1 and Pur proteins adds a new dynamic perspective to control of gene expression during myofibroblast differentiation that may reveal optimum strategies for therapeutic management of chronic fibrotic diseases. Project Narrative: The proposed research investigates cellular and molecular changes in connective tissue cells in heart and lung tissues that are required for normal wound healing after surgery. In transplant patients as well as individuals diagnosed with various forms of chronic cardiopulmonary diseases, these processes are accelerated leading to permanent scar formation, structural deformities in the heart and lung, loss of tissue function, and early death. New research studies outlined in this application will examine the individual biochemical pathways that drive normal wound healing and attempt to identify new targets for drug therapy that will help control destructive wound healing activity and improve the quality of life for individuals suffering from chronic diseases that cause abnormal scar tissue formation in the heart and lung.

描述(由申请人提供)：愈合伤口中肌成纤维细胞的慢性积聚与过度收缩、间质胶原过度沉积和破坏性组织重塑有关。血管平滑肌α-肌动蛋白(-SMA)是一种收缩蛋白，由分化的肌成纤维细胞瞬时表达，用于产生关闭开放伤口所需的张力。在慢性纤维化疾病中，肌成纤维细胞的分化是功能失调的，我们发现这些细胞中激活(-SMA和I型胶原基因所需的分子信号也提供了负反馈，这可能会在伤口愈合和破坏性重塑过程中限制过度活跃的肌成纤维细胞的招募。这项提案中概述的研究有望揭示DNA和mRNA结合蛋白YB-1、PUR(和PUR()与(-SMA和I型胶原启动子)之间的新形式的功能相互作用，并阐明这些蛋白如何受到促纤维化因子如转化生长因子(1)和凝血酶的影响，如果不加以控制，可能会导致肌成纤维细胞进展为肥厚性瘢痕。实验旨在启动、放大或减弱肌成纤维细胞的分化，以更好地了解在转录和翻译水平控制(-SMA和I型胶原基因输出)的策略，以及揭示可能有助于将异常伤口愈合结果降至最低的新型干预策略。目的1研究YB-1和PUR蛋白抑制物与(-SMA和胶原启动子DNA以及转录激活剂Sp1、SRF和Smads 2，3)的相互作用，描绘这种功能相互作用所需的抑制物多肽链区域，并试图利用多肽诱饵和小分子药物抑制物来破坏复合体的形成和阻止病理生物学的肌成纤维细胞分化。目的2将确定凝血酶是否在翻译控制水平上促进肌成纤维细胞的分化，从而发挥转化生长因子(1)的补充作用，或者相反，通过诱导抗纤维化转录调节蛋白Egr-1来阻断转录和肌成纤维细胞募集来拮抗这种生长因子。目的3研究将探索肌成纤维细胞分化和纤维化的替代的、Smad不依赖的机制。基于药物抑制转化生长因子(1/Smad)或磷脂酰肌醇-3-激酶(PI3K)/Akt信号通路的功能丧失方法将被用来评估它们在体外对小鼠心肌成纤维细胞激活和缺血/再灌注损伤后心肌纤维化的可能抑制作用。转化生长因子(1)和凝血酶能够利用YB-1和PUR蛋白独特的DNA、RNA和蛋白质结合特性，为控制肌成纤维细胞分化过程中的基因表达提供了一个新的动态视角，可能为慢性纤维疾病的治疗提供最佳策略。项目简介：这项拟议的研究调查了心脏和肺组织中结缔组织细胞和分子的变化，这是手术后正常伤口愈合所必需的。在移植患者以及被诊断患有各种形式的慢性心肺疾病的患者中，这些过程会加速，导致永久性疤痕形成、心脏和肺结构畸形、组织功能丧失和过早死亡。本申请中概述的新研究将检查推动正常伤口愈合的个体生化途径，并试图确定药物治疗的新靶点，这将有助于控制破坏性伤口愈合活动，并提高慢性疾病患者的生活质量，这些疾病会导致心脏和肺中异常疤痕组织的形成。