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）是一种致死性疾病，涉及肺部进行性瘢痕形成，每10万美国人中有50人受到影响。其病理包括临时细胞外基质（ECM）蛋白（包括纤维蛋白和纤连蛋白）的异常和慢性沉积，以及瓜氨酸形式的这些蛋白质的大量翻译后修饰。重要的是，瓜氨酸会影响精氨酸残基，包括在纤维蛋白和纤连蛋白中发现的RGD和PHSRN基序中可接近的那些，因此预计此类修饰会影响介导整合素相互作用改变的局部成纤维细胞行为。IPF还与“活化的”成纤维细胞表型相关，包括侵袭性、抗细胞凋亡和分泌过量ECM蛋白等特征。总的假设是，成纤维细胞与这些瓜氨酸化临时ECM蛋白的长期相互作用是导致成纤维细胞改变的原因。 表型将在三个具体目标中探讨这一假设。首先，将在存在或未被瓜氨酸化的纤维蛋白、纤维蛋白原和纤连蛋白的情况下评价成纤维细胞行为，包括细胞附着、扩散、迁移、抗凋亡和分泌特征，包括趋化因子、ECM分子和降解酶。在第二，这些改变的成纤维细胞的机制将通过评估特定整合素的结合能力和激活的变化进行探索。最后，在第三个目标中，将在博来霉素诱导的Thy 1（-/-）IPF模型中评价抑制瓜氨酸化的靶向治疗的体内疗效。这项研究意义重大，因为它探索了IPF成纤维细胞活化的新机制，这可能最终提出一类新的早期介入治疗。因此，它具有巨大的翻译潜力。它也很重要，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性疾病中普遍存在，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理解。该提案是申请人培训计划的中心，使她能够扩展她的科学知识，技术专业知识，沟通技巧和领导能力，为她在生物技术领域具有挑战性的职业生涯做准备。

 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.