Neuronally Active Proteins in IPF

项目摘要 特发性肺纤维化是一种无法治愈的疾病，其特征是瘢痕的进行性积累 成人肺部的组织它通常会导致在诊断后大约三年内死亡， 几种药物在延缓疾病进展方面显示出一定的益处， 干预措施是有限的，异质的，并伴有毒性。因此，进一步调查 驱动纤维化反应的机制仍然是重要的研究领域。当前肺疾病的研究范式 纤维化提出这一过程是由于上皮细胞损伤和过量成纤维细胞之间不匹配造成的 修复反应可能会因巨噬细胞表型异常而放大。了解所有这些 纤维化方面在IPF中特别重要，其中患者存在已确定的并且通常 进行性疾病神经元导向蛋白的研究是组织损伤领域的一个新兴领域 和修复。我们的实验室是第一个在人类肺部疾病的背景下研究这类蛋白质的实验室， 我们确定了GPI锚定的膜蛋白Semaphorin 7a与IPF的相关性。在神经 Sema 7a通过与整合素β 1 β 1和跨膜转运蛋白β 1 β 1的竞争作用调节神经元的生长。 蛋白丛状蛋白C1。我们已经证明这种机制在几种形式的哺乳动物肺纤维化中是活跃的， 炎症，其中Sema 7a的刺激作用通过整合素介导的过程产生， Plexin C1。我们在这一领域的额外工作表明层粘连蛋白样蛋白Netrin-1在细胞凋亡中的新作用。 整合素介导的过程。Netrin-1（NTN-1）是一种分泌的神经元导向蛋白， 通过结合其吸引受体，大肠癌-1（DCC-1）的吸引，而细胞排斥 而入侵是由与其排斥受体Uncoordinated-5a（UNC 5a）的相互作用驱动的。相比之下，Plexin C1（PLXNC 1）通过两种途径抑制Sema 7a驱动的过程;即， 肉瘤癌基因Rras和通过Lim激酶-2磷酸化调节细胞功能 （LimK2）。我们小组成员在这方面和其他疾病方面发表的和初步的工作表明， Netrin-1的刺激成分在IPF和几种实验性IPF模型中过度活化， 肺纤维化，而PLXNC 1的保护功能受到抑制。机制和潜力 IPF的治疗获益仍不明确。这项拨款提出了一种翻译方法， 现有技术基于质谱细胞计数的原始人生物样本的分析，基于新生物工程的离体模型， 和复杂的小鼠模型来评估这一假设。在目标1中，我们将确定表达的位点 NTN-1和PLXNC 1通路组分在Yale IPF队列的血液和肺中的浓度。在目标2中， 确定NTN-1刺激实验诱导的肺纤维化的机制，在目标3中， 将定义PLXNC 1信号通路对实验诱导的 肺纤维化

Project Summary Idiopathic pulmonary fibrosis is an incurable condition characterized by the progressive accumulation of scar tissue in the adult human lung. It typically leads to death within approximately three years of diagnosis and while several pharmacologic agents have shown some benefit in delaying disease progression, the effects of these interventions are limited, heterogeneous, and accompanied by toxicity. Thus, further investigation of the mechanisms driving fibrotic responses remains an important area of study. Current paradigms of pulmonary fibrosis propose this process to result from a mismatch between epithelial cell injury and excessive fibroblast repair responses that may be amplified by abnormalities in macrophage phenotypes. Understanding all of these aspects of fibrosis is of particular importance in IPF, where patients present with established and often progressive disease. The study of neuronal guidance proteins is an emerging area in the field of tissue injury and repair. Our laboratory was the first to study this class of proteins in the context of human lung disease when we defined the association of the GPI-anchored membrane protein Semaphorin 7a with IPF. In the nervous system, Sema 7a regulates neuronal growth via the competing effects of 11 integrin and the transmembrane protein Plexin C1. We have shown this mechanism to be active in several forms of mammalian lung fibrosis and inflammation where Sema 7a's stimulatory effects are enacted via an integrin-mediated process that is opposed by Plexin C1. Our additional work in this area indicates a novel role for the laminin-like protein Netrin-1 in the integrin-mediated processes. Netrin-1 (NTN-1), a secreted neuronal guidance protein, stimulates cellular attraction via binding to its attractive receptor, Deleted in Colorectal Cancer-1 (DCC-1) while cellular repulsion and invasion is driven by interactions with its repulsive receptor, Uncoordinated-5a (UNC5a). In contrast, Plexin C1 (PLXNC1) inhibits Sema 7a-driven processes via two pathways; namely, the inactivation of the Harvey rat sarcoma oncogene, Rras, and the modulation of cellular function via the phosphorylation of the Lim kinase-2 (LimK2). Published and preliminary work by members of our group in this and other diseases indicate that the Netrin-1's stimulatory components are excessively activated in IPF and in several experimental models of pulmonary fibrosis, whereas PLXNC1's protective functions are suppressed. The mechanisms and potential therapeutic benefit to IPF remain undefined. This grant proposes a translational approach combining state of the art mass cytometry based analysis of primary human biospecimens, novel bioengineering based ex vivo models, and sophisticated murine modeling to evaluate this hypothesis. In aim 1 we will determine the site of expression of NTN-1 and PLXNC1 pathway components in the blood and lungs of the Yale IPF cohort. In aim 2 we will determine the mechanism through which NTN-1 stimulates experimentally induced lung fibrosis and in aim 3 we will define the contribution of PLXNC1 signaling pathway to the development of experimentally induced pulmonary fibrosis.