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

皮肤伤口愈合有缺陷，导致慢性伤口或纤维增生性疾病，如肥厚性疤痕，是全球医疗保健的重大挑战，市场估计至少为100亿欧元，并注定随着人口老龄化而增加。目前缺乏针对发病机制的治疗方法，这在很大程度上是由于创伤愈合过程中涉及的多个阶段的复杂性，以及缺乏关于不同参与者之间联系的信息。这个项目的目的是确定一个新的金属蛋白酶家族，称为阿司匹林样蛋白酶(Alps)，在正常和病理性伤口愈合中的作用。与主要以组织降解而闻名的基质金属蛋白酶不同，阿司匹林样蛋白酶最近被发现协调组织修复的几个方面。其中骨形态发生蛋白-1/类托洛样蛋白酶(BTPs)和meprinα和meprin beta是在皮肤中发现的。在这方面尤其重要的是细胞因子和生长因子的激活、血管生成和细胞外基质(ECM)的组装。最近在定量蛋白质组学研究复杂样品中的蛋白分解以及针对阿尔卑斯的抑制剂的开发方面的进展使人们有可能采取综合的方法来理解这些似乎具有重叠和互补底物特异性的蛋白酶在组织重建中的作用。该项目将是多学科的，包括i)专门针对单个阿尔卑斯的新型膦酸多肽类抑制剂的表征和应用，ii)使用小鼠模型、人类活检组织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不同阿尔卑斯在皮肤中的表达模式，iii)应用定量蛋白质组学(Tail方法)来鉴定碱性磷酸酶底物，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(抑制物)来提出和测试针对单个alp成员或亚家族的瘢痕形成的新治疗策略。使用上述鼠标模型。来自基尔和弗莱堡的合作伙伴在阿尔卑斯山、蛋白酶活性分析、定量蛋白质组学、小鼠模型、纤维化和伤口愈合方面拥有互补的专业知识。在拟议项目过程中获得的知识将成为制定预防和治疗伤口愈合障碍的新治疗战略的基础。

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.