Discovering Extracellular Modulators of Lung Fibrogenesis by Profiling Newly Synthesized Extracellular Matrix

通过分析新合成的细胞外基质发现肺纤维生成的细胞外调节剂

• 批准号: 10683787
• 负责人: Xi Ren
• 金额: $ 59.56万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10683787
• 关键词: Address; Adenoviruses; Algorithms; Biocompatible Materials; Biology; Biomimetics; Bleomycin; Cells; Chemistry; Cicatrix; Complement; Deposition; Detection; Disease; Drug Targeting; Endothelial Cells; Endothelium; Engineering; Enzymes; Epithelial; Epithelial Cells; Evaluation; Extracellular Matrix; Extracellular Matrix Proteins; Feedback; Fibroblasts; Fibrosis; Human; Immobilization; In Vitro; Interstitial Lung Diseases; Knowledge; Label; Link; Lung; Lung fibrogenesis; Mass Spectrum Analysis; Mediating; Modeling; Monosaccharides; Mus; Pathogenesis; Pathogenicity; Pathologic; Patients; Perfusion; Physiological; Polysaccharides; Probability; Process; Production; Proteins; Proteomics; Pulmonary Fibrosis; Pulmonary Pathology; Research; Resolution; Sialic Acids; Signal Transduction; Slice; Structure of parenchyma of lung; System; Technology; Therapeutic Intervention; Time; Tissue Engineering; Tissue Model; cell type; cellular targeting; cytokine; extracellular; fibrogenesis; fibrotic interstitial lung disease; fibrotic lung; fibrous protein; glycosylation; idiopathic pulmonary fibrosis; in vivo; innovation; lung health; macrophage; mouse model; multidisciplinary; multiple omics; new technology; new therapeutic target; preference; respiratory; scaffold; targeted treatment; technology development; temporal measurement; therapeutic development; tool; transcriptomics

Project Summary Interstitial lung diseases (ILD) are devastating disorders causing progressive scarring (i.e. fibrosis) of lung tissue, and result from reciprocal interactions of cellular abnormalities and extracellular matrix (ECM) dysregulation. Current ILD treatment primarily targets cellular signaling and remains unable to halt or reverse fibrogenesis. Despite being a key pathological hallmark of ILD, the ECM has rarely been directly targeted for therapeutic intervention. Fundamentally, the extracellular mechanism underlying lung fibrosis progression remains elusive. To bridge this gap, our objective is to develop an innovative approach for selective profiling of newly synthesized ECM (newsECM) along lung fibrogenesis. The proposed technology will selectively label newsECM produced over defined short time spans by incorporating chemoselective azido-tags via post- translational glycosylation. This will enable enrichment of newsECM free from the pre-existing ECM, and thereby enables sensitive proteomic detection of the dynamic ECM synthesis with unprecedented daily temporal resolution, irrespective of the abundant pre-existing ECM background. Our proposed approach has an inherent preference to ECM proteins, the majority of which are glycosylated. The proposed newsECM profiling will address a major technical barrier in conventional, non-selective mass spectrometry analysis, which suffers from limited sensitivity in detecting the dynamic new ECM deposition that is usually in low abundance. The proposed newsECM profiling technology is versatile and will be implemented in three lung fibrosis models, including in vivo mouse fibrosis models, an ex vivo donor lung perfusion model, and an in vitro synthetic fibrosis model. We intend to pursue the following specific aims. Aim 1 will track newsECM dynamics in vivo during the progression and resolution of lung fibrogenesis, which is expected to reveal pro- and anti-fibrotic ECM factors. Aim 2 will establish the correlation between the in vivo and ex vivo newsECM profiles using murine lungs as a model, and apply the resulting optimized newsECM profiling condition and algorithm to the ex vivo lung perfusion (EVLP) of donor human lungs bearing idiopathic pulmonary fibrosis (IPF), the most common form of ILD, to reveal human-specific pathogenic ECM mechanism. Finally, Aim 3 will establish a synthetic lung fibrosis model tissue-engineered combining fibroblast, epithelium, endothelium and macrophage within decellularized native lung ECM scaffold, and use it as an experimentally tractable system to further decode fibrogenic lung cell-ECM interaction. Furthermore, combining newsECM labeling and native ECM biomaterial engineering, we will offer a chemoselective platform for effective functional evaluation of candidate fibrosis-modulating ECM factors in a biomimetic, ECM-associated manner. In summary, this research will facilitate a paradigm shift in ILD treatment by promoting ECM-targeted therapeutic development and by enabling combined therapeutic interventions aiming at both cellular and extracellular targets to bring new hope for the impacted patients.

项目概要 间质性肺疾病 (ILD) 是一种破坏性疾病，会导致肺组织进行性疤痕形成（即纤维化）， 是细胞异常和细胞外基质 (ECM) 失调相互作用的结果。 目前的 ILD 治疗主要针对细胞信号传导，但仍然无法阻止或逆转纤维形成。 尽管 ECM 是 ILD 的一个关键病理标志，但很少直接针对治疗 干涉。从根本上说，肺纤维化进展的细胞外机制仍然难以捉摸。 为了弥补这一差距，我们的目标是开发一种创新方法来选择性分析新的 沿着肺纤维发生合成了 ECM (newsECM)。所提出的技术将选择性地标记 newsECM 通过后置化学选择性叠氮标签在规定的短时间内产生 翻译糖基化。这将能够丰富 newsECM，而无需使用预先存在的 ECM，从而 能够以前所未有的每日时间对动态 ECM 合成进行灵敏的蛋白质组学检测 分辨率，无论现有的 ECM 背景如何。我们提出的方法有一个固有的 优先选择 ECM 蛋白，其中大多数是糖基化的。拟议的 newsECM 分析将 解决了传统非选择性质谱分析中的一个主要技术障碍 由于检测通常丰度较低的动态新 ECM 沉积的灵敏度有限。这 提议的新闻ECM分析技术具有多种用途，将在三种肺纤维化模型中实施， 包括体内小鼠纤维化模型、离体供体肺灌注模型和体外合成纤维化模型 模型。我们打算追求以下具体目标。 Aim 1将跟踪新闻ECM体内动态 肺纤维化的进展和解决，有望揭示促纤维化和抗纤维化的 ECM 因子。 目标 2 将使用小鼠肺作为模型建立体内和离体新闻 ECM 曲线之间的相关性 模型，并将所得优化的 newsECM 分析条件和算法应用于离体肺灌注 (EVLP) 患有特发性肺纤维化 (IPF)（最常见的 ILD 形式）的供体人肺 揭示人类特有的致病ECM机制。最后，目标3将建立合成肺纤维化模型 组织工程将成纤维细胞、上皮细胞、内皮细胞和巨噬细胞结合在脱细胞的天然细胞内 肺 ECM 支架，并将其用作实验上易于处理的系统，以进一步解码纤维化肺细胞 ECM 相互作用。此外，结合 newsECM 标签和原生 ECM 生物材料工程，我们将提供 用于有效功能评估候选纤维化调节 ECM 因子的化学选择性平台 仿生、ECM 相关的方式。总之，这项研究将促进 ILD 的范式转变 通过促进 ECM 靶向治疗的开发和实现联合治疗来治疗 针对细胞和细胞外目标的干预措施为受影响的患者带来新的希望。