Mutant Surfactant - Induced TGF-beta Secretion in Lung Fibrosis

描述（由申请人提供）：编码表面活性蛋白A2（SP-A2）的基因突变导致家族性特发性肺纤维化（IPF）和肺癌，这两种疾病都是治疗选择有限的致死性疾病。突变型表面活性蛋白的分子机制尚未得到很好的理解，并且是影响儿童和成人的一系列肺部疾病的基础。我们最近发现，肺上皮细胞中某些突变表面活性蛋白的表达导致潜在TGF-β（一种有效的促纤维化和促癌细胞因子）的表达和分泌增加。待检验的具体假设是，肺泡上皮细胞中突变表面活性蛋白的表达通过TGF-β的自分泌和旁分泌效应导致肺纤维化，TGF-β的分泌通过ER应激依赖性和ER应激非依赖性途径诱导。我们提出这项工作的理由是，它将提供增强的理解突变的表面活性蛋白在引起进行性肺纤维化的功能获得的影响。第一个目标将集中在体外研究这种机制，通过确定：哪些变体表面活性蛋白具有诱导潜在TGF-β分泌增加的功能获得效应，以及介导这种机制的ER应激信号的相对贡献是什么？潜在途径的分子机制将使用蛋白质组学和转录组学方法以及化学抑制剂的组合进行探测。第二个目标将定义II型肺泡上皮细胞的损伤或“重编程”， 由突变表面活性蛋白的急性或慢性表达以及影响TGF-β信号传导的细胞环境的各种途径的参与引起的细胞。在第三个也是最后一个目标中， 我们将使用一种新的转基因小鼠模型阐明突变SP-A2蛋白介导的疾病的体内分子机制。我们将特别探讨TGF-β信号传导和端粒酶途径在博莱霉素诱导的肺纤维化中的遗传作用。虽然TGF-β下游的信号传导途径已被很好地表征，但导致其表达和分泌的刺激事件还没有。这些目标的成功完成将阐明突变型表面活性剂诱导潜伏性TGF-β分泌的新途径。这项工作将深入了解连接蛋白质错误折叠，ER应激，EMT，细胞分化和肺纤维化增殖的途径，并将有助于确定药物开发的独特靶点。

DESCRIPTION (provided by applicant): Mutations in the gene encoding surfactant protein A2 (SP-A2) cause familial idiopathic pulmonary fibrosis (IPF) and lung cancer, both lethal diseases with limited treatment options. The molecular mechanism of mutant surfactant proteins are not well understood and underlie a spectrum of lung diseases affecting children and adults. We have recently discovered that expression of certain mutant surfactant proteins in lung epithelial cells leads to increased expression and secretion of latent TGF-ß, a potent pro-fibrotic and pro-oncogenic cytokine. The specific hypothesis to be tested is that expression of mutant surfactant proteins in alveolar epithelial cells leads to pulmonary fibrosis through the autocrine and paracrine effects of TGF-ß, whose secretion is induced through ER stress-dependent and ER stress-independent pathways. Our rationale for the proposed work is that it will provide enhanced understanding of the gain-of-function effects of mutant surfactant proteins in causing progressive pulmonary fibrosis. The first aim will focus on studying this mechanism in vitro by determining: Which variant surfactant proteins have a gain-of-function effect of inducing increased latent TGF-ß secretion and what is the relative contribution of ER stress signaling that mediates this mechanism? The molecular mechanism of the underlying pathways will be probed using a combination of proteomic and transcriptomic approaches as well as chemical inhibitors. The second aim will define the injury or "reprogramming" of type II alveolar epithelial cells caused by acute or chronic expression of mutant surfactant proteins and the involvement of various pathways that affect the cellular context of TGF-ß signaling. In the third and last aim, we will elucidate the in vivo molecular mechanism of mutant SP-A2 protein-mediated disease using a novel transgenic mouse model. We will specifically explore the genetic contributions of the TGF-ß signaling and telomerase pathways in contributing to bleomycin-induced lung fibrosis. While the signaling pathways downstream of TGF-ß are well characterized, the inciting events that lead to its expression and secretion are not. Successful completion of these aims will elucidate this novel pathway of mutant surfactant induced latent TGF-ß secretion. This work will generate insight into the pathways linking protein misfolding, ER stress, EMT, cellular differentiation and proliferation in lung fibrosis and will assist in the identification of unique targets for drug development.