SOCS domain-mediated fibronectin matrix degradation on fibrosis reversal

摘要 特发性肺纤维化（IPF）是一种不可逆的肺部疾病，没有有效的治疗选择。 2001-2011年IPF发生率的保险索赔数据估计为每100，000人31-43人， 2000年，美国有89，000例IPF患者，34，000例新发病例 每年诊断。IPF是一种病因不明的疾病，已知多种环境因素 有助于易感个体的疾病发展。在纤维化过程中，成纤维细胞 转化成肌成纤维细胞，产生大量的细胞外基质（ECM）蛋白， 刚性和肺结构影响肺功能。ECM的一个关键组成部分是核心基质体 蛋白质纤连蛋白（FN），其组装成有助于胶原基质形成和ECM的FN基质 刚度Von Hippel Lindau（VHL）蛋白是FN基质形成所必需的，并在肺中上调 IPF患者。我们已经证明，SOCS结构域（细胞因子信号转导抑制因子），一个40个氨基酸， SOCS蛋白家族中的一个保守结构域，靶向VHL，通过cullin依赖性泛素降解 连接酶机制我们假设肌成纤维细胞中SOCS结构域的过度表达将导致 肌成纤维细胞中病理性FN基质的降解和纤维化逆转。我们将执行这些 使用用SOCS结构域和SOCS结构域突变体转导的肺成纤维细胞的实验， VHL降解缺陷。使用RNAseq和磷酸化蛋白质组分析，我们将开发一个计算的 管道，以确定在肌成纤维细胞逆转的过程中可能的信号介质。我们的体外研究结果， 将在博来霉素诱导的肺纤维化的体内小鼠模型中进行评价，以测试纤维化的效率 通过腺病毒介导的SOCS结构域的递送逆转。我们预计， 我们的体外实验和我们的体内模型中纤维化的显著逆转。结果从这些 实验将提供新的基于肽的靶点，并提供可用于以下任一种的治疗剂： 单独使用或与现有疗法联合使用以逆转基质形成和纤维化。

ABSTRACT Idiopathic Pulmonary fibrosis (IPF) is an irreversible lung disease with no effective treatment options. Insurance claims data on IPF incidence between years 2001-2011 was estimated at 31-43 per 100,000 per year, translating to 89,000 patients living with IPF in the United States in the year 2000 and 34,000 new cases diagnosed per year. IPF is a disease of unknown etiology and myriad environmental factors are known to contribute to the development of the disease in susceptible individuals. During fibrosis, fibroblasts differentiate into myofibroblast cells that produce vast amounts of extracellular matrix (ECM) proteins that alter matrix rigidity and lung architecture impacting lung function. A critical constituent of the ECM is the core matrisome protein fibronectin (FN) that assembles into a FN matrix contributing to Collagen matrix formation and ECM rigidity. Von Hippel Lindau (VHL) protein is required for the FN matrix formation and is upregulated in the lungs of IPF patients. We have shown that the SOCS domain (Suppressors Of Cytokine Signaling), a 40 amino acid conserved domain in the SOCS family of proteins, targets VHL for degradation by a cullin-dependent ubiquitin ligase mechanism. We hypothesize that SOCS domain overexpression in myofibroblasts will result in degradation of the pathologic FN matrix in myofibroblasts and fibrosis reversal. We will perform these experiments using lung fibroblasts transduced with the SOCS domain and the SOCS domain mutant that is defective in VHL degradation. Using RNAseq and phosphoproteome analysis, we will develop a computational pipeline to identify probable signaling mediators in the process of myofibroblast reversal. Our in vitro findings, will be evaluated in a Bleomycin-induced in vivo mouse model of lung fibrosis to test the efficiency of fibrosis reversal by adenoviral-mediated delivery of the SOCS domain. We expect a reduction of fibrosis markers in our in vitro experiments and significant reversal of fibrosis in our in vivo model. The results from these experiments will offer new peptide based targets as well as provide therapeutic agents that can be used either on its own or in combination with existing therapies to reverse matrix formation and fibrosis.