Provisional Matrix Citrullination as an activator of Fibroblasts in Interstitial Lung Disease

 描述(申请人提供)：特发性肺纤维化(IPF)是一种涉及肺部进行性疤痕的致命疾病，每10万名美国人中就有50人受到影响。其病理包括临时细胞外基质(ECM)蛋白的异常和慢性沉积--包括纤维蛋白和纤维连接蛋白--以及以瓜氨酸化形式对这些蛋白进行显著的翻译后修饰。重要的是，瓜氨酸化作用影响精氨酸残基，包括在纤维蛋白和纤维连接蛋白中发现的RGD和PHSRN基序中可获得的精氨酸残基，因此这种修饰有望对局部成纤维细胞的行为产生影响，从而改变整合素的相互作用。IPF还与一种“激活的”成纤维细胞表型有关，包括侵袭性、抗凋亡和分泌过量的ECM蛋白等特征。总体假设是成纤维细胞与这些瓜氨酸化临时ECM蛋白的长时间相互作用是导致成纤维细胞改变的原因。 表型。这一假设将从三个具体目标来探讨。首先，在纤维蛋白、纤维蛋白原和纤维连接蛋白存在的情况下，将评估成纤维细胞的行为，包括细胞附着、扩散、迁移、抗凋亡和分泌型--包括趋化因子、ECM分子和降解酶。第二，将通过评估特定整合素结合能力和激活的变化来探索这些改变的成纤维细胞的机制。最后，在第三个目标中，将在博莱霉素诱导的Thy1(-/-)IPF模型中评估靶向治疗抑制瓜氨酸的体内疗效。这项研究具有重要意义，因为它探索了IPF中成纤维细胞激活的新机制，最终可能建议一种新的早期介入治疗。因此，它具有巨大的翻译潜力。ALS意义重大，因为它研究了一种基本的细胞-ECM相互作用，这种相互作用在各种慢性炎症条件下普遍存在，包括类风湿性关节炎、恶性肿瘤和多发性硬化症，因此它有可能增强我们对许多不同严重人类疾病的发病和进展的了解。这项计划是申请者培训计划的核心，使她能够在准备迎接充满挑战的生物技术职业生涯时，扩大她的科学知识、技术熟练程度、沟通技能和领导能力。

 DESCRIPTION (provided by applicant): Idiopathic pulmonary fibrosis (IPF) is a lethal condition involving progressive scarring of the lungs that affects 50 in every 100,000 Americans. Its pathologies include aberrant and chronic deposition of provisional extracellular matrix (ECM) proteins-including fibrin and fibronectin- as well as prominent amounts of post-translational modifications to these proteins in the form of citrullination. Importantly, citrullination affects arginine residues, including those accessible in RGD and PHSRN motifs found in fibrin and fibronectin, and thus such modifications are expected to have an influence on local fibroblast behavior mediated to altered integrin interactions. IPF is also associated with an "activated" fibroblast phenotype including characteristics such as invasiveness, resistance to apoptosis, and secretion of excessive ECM proteins. The overall hypothesis is that prolonged interaction of fibroblasts with these citrullinated provisional ECM proteins is responsible for altered fibroblast phenotype. This hypothesis will be explored in three specific aims. In the first, fibroblast cell behaviors including cell attachment, spreading, migration, apoptotic resistance, and secretory profile - including that of chemokines, ECM molecules, and degradation enzymes-will be evaluated in the presence of fibrin, fibrinogen, and fibronectin that has or has not been citrullinated. In the second, the mechanism for these altered fibroblasts will be explored by evaluating changes in specific integrins' binding capacities and activation. Finally, in the third aim, the in vivo efficacy of a targeted treatment that inhibits citrullinated will be evaluated in bleomycin-induced Thy1(-/-) model of IPF. This research is significant because it explores a novel mechanism for fibroblast activation in IPF, which may ultimately suggest a new class of early-stage interventional therapy. It therefore possesses great translational potential. It is als significant because it investigates a fundamental cell-ECM interaction that is prevalent in a wide-variety of chronic inflammatory conditions, including rheumatoid arthritis, malignant cancers, and multiple sclerosis, and it thus has the potential to enhance our understanding of both the onset and progressions many different serious human diseases. This proposal serves at the center of the applicant's training plan allowing her to expand her scientific knowledge, technical proficiencies, communication skills, and leadership capabilities as she prepares for a challenging career in biotechnology.