Role of astacin-like proteinases in physiological wound healing and scarring

皮肤伤口愈合缺陷，导致慢性伤口或纤维增生性疾病，如肥厚性疤痕，是世界范围内的主要医疗保健挑战，市场估计至少为100亿欧元，并注定会随着人口老龄化而增加。目前缺乏致病特异性治疗主要是由于伤口愈合涉及多个阶段的复杂性，以及缺乏有关不同参与者之间连通性的信息。该项目旨在确定一个新兴的金属蛋白酶家族，称为astacin样蛋白酶（ALPs），在正常和病理伤口愈合中的作用。基质金属蛋白酶主要以其在组织降解中的作用而闻名，与之不同的是，在皮肤中发现的骨形态发生蛋白-1/ tolloidlike蛋白酶（BTPs）和meprin α和meprin β等astastin样蛋白酶最近被证明在组织修复的几个方面起着协调作用。在此背景下，细胞因子和生长因子激活、血管生成和细胞外基质（ECM）组装尤为重要。最近在定量蛋白质组学研究复杂样品中的蛋白质水解和针对ALPs抑制剂的开发方面取得的进展，使得采取综合方法来理解这些蛋白酶在组织重塑中的作用成为可能，这些蛋白酶似乎具有重叠和互补的底物特异性。该项目将是一个多学科的项目，包括：1)专门针对单个ALPs的新型磷酸肽型抑制剂的表征和应用；2)使用小鼠模型、人体活检和皮肤原代细胞来表征正常和纤维增生性伤口愈合过程中皮肤中不同ALPs的表达模式；3)应用定量蛋白质组学（TAILS方法）来鉴定ALP底物。iv)在体外和体内验证这些底物，以及v)使用项目中提供的数据（表达谱，底物）和工具（抑制剂），使用上述小鼠模型提出和测试针对单个ALP成员或亚家族的新的疤痕治疗策略。Kiel和Freiburg的合作伙伴在ALPs、蛋白酶活性测定、定量蛋白质组学、小鼠模型、纤维化和伤口愈合方面具有互补的专业知识。在拟议的项目过程中获得的知识将为预防和治疗伤口愈合障碍的新治疗策略的发展奠定基础。

Defective wound healing in skin, leading to chronic wounds or fibroproliferative disorders such as hypertrophic scarring, is a major healthcare challenge worldwide for which the market is estimated to be at least 10 billion euros and is destined to increase with the ageing population. The current lack of pathogenesis-specific therapies is largely due to the complexity of the multiple stages involved in wound healing and to the lack of information about the connectivity between the different players involved. This project aims to identify the roles of an emerging family of metalloproteinases, called astacin-like proteinases (ALPs), in normal and pathological wound healing.Unlike the matrix metalloproteinases, known largely for their roles in tissue degradation, the astacin-like proteinases, among which the bone morphogenetic protein-1/tolloid-like proteinases (BTPs) and meprin alpha and meprin beta are those found in skin, have recently been shown to orchestrate several aspects of tissue repair. Particularly important in this context are cytokine and growth factor activation, angiogenesis and extracellular matrix (ECM) assembly. Recent advances in quantitative proteomics to study proteolysis in complex samples and in the development of inhibitors directed against ALPs have made it possible to take an integrative approach to understanding the roles of these proteinases, which appear to have both overlapping and complementary substrate specificities, in tissue remodeling. The project will be multidisciplinary, including i) the characterization and application of new phosphinic peptide-type inhibitors specifically targeting individual ALPs, ii) the use of mouse models, human biopsies and skin primary cells to characterize the expression patterns of different ALPs in skin during the course of normal and fibroproliferative wound healing, iii) the application of quantitative proteomics (TAILS method) for the identification of ALP substrates, iv) the validation of these substrates in vitro and in vivo, and v) the use of the data (expression profiles, substrates) and tools (inhibitors) made available in the project to propose and test novel therapeutic strategies for scarring targeting either individual ALP members or subfamilies, using the above mouse models. The partners from Kiel and Freiburg have complementary expertise in ALPs, protease activity assays, quantitative proteomics, mouse models, fibrosis, and wound healing. The knowledge acquired during the course of the proposed project will form the basis for the development of new therapeutic strategies for the prevention and treatment of wound healing disorders.