Development of proteomic-based ECM signatures for lung fibrosis

摘要 系统性硬化症（SSc;硬皮病）是一种特发性结缔组织疾病，其特征在于 皮肤和内脏器官（如肺）中胶原蛋白的产生和沉积增加。的 SSc的病因尚不清楚，尽管遗传影响、环境损伤和异常 免疫功能是积极研究课题。间质性肺病（ILD）是SSc的并发症 并且目前是患有这种疾病的患者的主要死亡原因。特发性肺纤维化（IPF）是 也是具有高发病率和死亡率的纤维化疾病。我们有一个独特的资源， 来自正常供体、SSc患者和IPF患者以及匹配的原代肺成纤维细胞。我们 已经使用肺组织来产生组织微阵列（TMA），其包括正常肺、SSc肺和 同一TMA上的IPF肺芯。我们假设胶原蛋白组的改变，包括后- 翻译修饰，代表了肺并发症的一种新的和临床相关的签名， SSc和IPF。我们进一步假设，非常具体的网站胶原羟基化调节的反应， 成人肺原代成纤维细胞我们建议使用一种新的胶原靶向蛋白质组学方法 定位和测量肺组织内的胶原类型和翻译后修饰， 鉴别疾病（SSc vs. IPF）和表型（正常vs.纤维化）变化。事后确认 将合成修饰的胶原肽以测试来自正常、SSc和IPF的成纤维细胞的反应 肺组织对肽的反应我们的发现将促进靶向治疗的发展， 支持肺纤维化新生物标志物的鉴定。

ABSTRACT Systemic sclerosis (SSc; scleroderma) is an idiopathic disorder of connective tissue characterized by increased production and deposition of collagen in the skin and internal organs such as the lungs. The etiology of SSc is unknown, although the role of genetic influences, environmental insults and abnormal immune function are subjects of active investigation. Interstitial lung disease (ILD) is a complication of SSc and is currently the leading cause of death in patients with this disease. Idiopathic pulmonary fibrosis (IPF) is also a fibrotic disease with high morbidity and mortality. We have a unique resource consisting of lung tissues from normal donors, patients with SSc and patients with IPF as well as matching primary lung fibroblasts. We have used the lung tissues to generate tissue microarrays (TMAs) that include normal lung, SSc lung, and IPF lung cores on the same TMA. We hypothesize that alteration of the collagen proteomes, including post- translational modifications, represents a novel and clinically relevant signature of pulmonary complications of SSc and IPF. We further hypothesize that very specific sites of collagen hydroxylation regulate the response of primary adult human lung fibroblasts. We propose to use a novel collagen-targeting proteomic approach to localize and measure collagen types and post- translational modifications within the lung tissues and identify disease (SSc vs. IPF) and phenotype (normal vs. fibrosis) changes. Identified post-translationally modified collagen peptides will be synthesized to test the response of fibroblasts from normal, SSc and IPF lung tissues to the peptides. Our findings will facilitate the development of targeted therapies and will also support the identification of novel biomarkers for pulmonary fibrosis.