Neuronally Active Proteins in IPF

项目摘要 特发性肺纤维化是一种不治之症，其特征是疤痕逐渐堆积。 成人肺中的组织。它通常会在确诊后大约三年内导致死亡，虽然 几种药物已经显示出在延缓疾病进展方面的一些益处，这些药物的效果 干预措施是有限的、不同的，并伴随着毒性。因此，对该事件的进一步调查 推动纤维化反应的机制仍然是一个重要的研究领域。当前肺病的研究范式 纤维化认为这一过程是由于上皮细胞损伤和过度成纤维细胞之间的不匹配造成的 巨噬细胞表型异常可能放大的修复反应。了解所有这些 纤维化的一些方面在IPF中特别重要，在IPF中，患者表现为既有的和经常的 进行性疾病。神经导向蛋白的研究是组织损伤领域的一个新兴领域 并进行修复。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的关联。在紧张中 系统中，SEMA 7a通过-1--1整合素和跨膜蛋白的竞争效应调节神经元生长 蛋白质丛蛋白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和 Sema 7a的刺激作用通过整合素介导的相反过程而发生的炎症 由Plexin C1.我们在这一领域的进一步工作表明，层粘连蛋白样蛋白Netrin-1在 整合素介导的过程。Netrin-1(NTN-1)是一种分泌型神经导向蛋白，可刺激细胞 结直肠癌DCC-1(DCC-1)通过与其吸引力受体结合而产生吸引力，而细胞排斥 侵袭是通过与其排斥性受体非协调5a(UNC5a)的相互作用来驱动的。相比之下，Plexin C_1(PLXNC_1)通过两条途径抑制Sema 7a驱动的过程，即失活Harvey大鼠 肉瘤癌基因rras与LIM激酶-2的磷酸化对细胞功能的调节 (LIMK2)。我们小组成员在这种疾病和其他疾病方面发表的和初步的工作表明 网织蛋白-1‘S刺激成分在特发性肺间质纤维化及多种动物模型中过度激活 肺纤维化，而PLXNc1的S保护功能被抑制。机制和潜力 IPF的治疗益处仍未确定。这项拨款提出了一种翻译方法，将 基于ART质量细胞术的主要人类生物样本的分析，基于生物工程的体外模型， 和复杂的小鼠模型来评估这一假说。在目标1中，我们将确定表达的位置 耶鲁大学IPF队列中血液和肺中NTN-1和PLXNC1途径成分的研究。在《目标2》中我们将 确定NTN-1刺激实验性肺纤维化的机制，并在Aim 3中 将确定PLXNC1信号通路在实验诱导的糖尿病发展中的作用 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.