SOCS domain-mediated fibronectin matrix degradation on fibrosis reversal

摘要 特发性肺纤维化(IPF)是一种不可逆转的肺部疾病，没有有效的治疗选择。 2001-2011年间IPF发病率的保险索赔数据估计为每10万人中有31-43人 2000年，美国有89,000名IPF患者和34,000例新病例 每年确诊一次。IPF是一种病因不明的疾病，已知有多种环境因素 在易感人群中促进疾病的发展。在纤维化过程中，成纤维细胞分化 转化成肌成纤维细胞，产生大量改变基质的细胞外基质(ECM)蛋白 僵硬程度和肺构型对肺功能的影响。ECM的一个关键组成部分是核心母体 蛋白质纤维连接蛋白(FN)，组装成FN基质，有助于胶原基质的形成和细胞外基质 僵硬。Von Hippel Lindau(VHL)蛋白是FN基质形成所必需的，在肺组织中上调 在IPF患者中。我们已经证明了SOCS结构域(细胞因子信号的抑制物)，一种40个氨基酸的 SOCS蛋白家族中的保守结构域，靶向VHL以被依赖于cullin的泛素降解 连接酶机制。我们假设在肌成纤维细胞中SOCS结构域的过度表达将导致 肌成纤维细胞病理性纤维连接蛋白基质降解及纤维化逆转。我们将执行这些 使用转导SOCS结构域和SOCS结构域突变体的肺成纤维细胞的实验 VHL降解有缺陷。利用RNAseq和磷酸蛋白质组分析，我们将开发一种计算 确定肌成纤维细胞逆转过程中可能的信号介质的管道。我们的体外研究结果， 将在博莱霉素诱导的小鼠体内肺纤维化模型中进行评估，以测试纤维化的有效性 通过腺病毒介导的SOCS域的传递逆转。我们预计肝纤维化标记物在 我们的体外实验在我们的体内模型中显著逆转了纤维化。这些研究的结果 实验将提供新的基于多肽的靶点，并提供可用于以下两种用途的治疗剂 单用或与现有治疗方法联合使用，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.