Matrix Metalloproteinase Regulation of Leukocyte Infiltration during Wound Repair

伤口修复过程中基质金属蛋白酶对白细胞浸润的调节

• 批准号: 7987568
• 负责人: LISA Joy MCCAWLEY
• 金额: $ 30.26万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7987568
• 关键词: Ablation; Adherence; Adhesions; Affect; Animals; Biological Assay; Biological Models; Bioreactors; Blood capillaries; Cell Line; Cell physiology; Chemotaxis; Clinical; Connective Tissue; Cytoskeleton; Endothelial Cells; Endothelium; Environment; Epidermis; Extravasation; Family; Family member; Fibroblasts; Genes; Goals; Growth Factor; Healed; Health; Host Defense; Hydrolysis; Immune; In Vitro; Individual; Infiltration; Inflammation; Leukocytes; MMP3 gene; MMP9 gene; MMPI; Maintenance; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Microfluidics; Modeling; Mus; Optics; Pattern; Peptide Hydrolases; Play; Population; Process; Regulation; Role; Site; Skin; Squamous cell carcinoma; Staging; System; Testing; Tissues; Transgenic Mice; Water; Wound Healing; adhesion receptor; base; capillary; cell type; defense response; design; fibrogenesis; healing; in vivo; in vivo Model; insight; keratinocyte; macrophage; member; migration; monocyte; monolayer; mouse model; neovascularization; neutrophil; novel; overexpression; prevent; public health relevance; response; tumor progression; wound

DESCRIPTION (provided by applicant): Upon loss of existing tissue integrity, wound closure is critical for maintenance of the protective epidermal barrier that shields the host from environmental insult and prevents water loss. An essential, initial response during wound repair is the influx of circulating leukocytes that cleanse the wound and also provide activators and growth factors to stimulate fibrogenesis and neovascularization. Members of the matrix metalloproteinase (MMP) family, such as MMP3, are potential key regulators of leukocyte infiltration. MMP3 is upregulated in the epidermis during processes undergoing tissue remodeling. Mice which are null for MMP3 have delayed closure of excisional wounds and demonstrate reduced infiltration of leukocytes in a number of model systems. Furthermore, we find that MMP3-/- animals have an increased sensitivity to SCC progression that is associated with a reduction in neutrophil and macrophage infiltration evident throughout all stages of tumor progression. Based upon these observations, we propose that MMP3 regulates a protective host defense response during wound repair that is reflective of a protective, antitumorigenic response during SCC. A number of molecules known to affect immune cell function can be directly targeted for hydrolysis and release by MMP3 or matrix proteinases which are activated by MMP3. We hypothesize that MMP3, in combination with other matrix proteinases, directly enhances leukocyte extravasation that subsequently influences the wound repair process. Leukocyte infiltration into sites of inflammation requires the coordinate regulation of multiple steps including arrest on endothelium, migration through the endothelial barrier and directed migration through connective tissue towards the site of inflammation. MMP3 is expressed by a variety of cell types including keratinocytes, fibroblasts, endothelial cells and monocytes and can activate other matrix proteinase classes; thus, MMP3 and other MMPs may modulate multiple key steps required for optimal leukocyte extravasation. The goal of this project is to elucidate the functional role of MMP3 in leukocyte extravasation during wound healing. Our specific aims will test our hypothesis as follows: Specific Aim 1: To identity the cellular matrix proteinase contributions of leukocytes, fibroblasts and keratinocytes to MMP3 dependent leukocyte infiltration into the wound environment using in vivo mouse models that are null for or have targeted overexpression of individual MMP family members. Specific Aim 2. To test the hypothesis that MMP3 activity regulates activation of subsequent MMP family members in wounded epidermis. Specific Aim 3: To identity the cellular mechanisms regulated during MMP3 and matrix protease dependent transendothelial migration in vitro. These studies will provide novel insights into the role of MMP3 and other matrix proteinases during leukocyte transendothelial migration and provide an in vivo model system for testing MMP inhibitors in a clinical setting. PUBLIC HEALTH RELEVANCE: Wounding of the skin requires healing to maintain normal health. An essential part of wound healing is inflammation. Matrix Metalloproteinases such as MMP3 may play a key role in regulating inflammation during wound healing.

描述(由申请人提供)： 一旦失去现有的组织完整性，伤口闭合对于维持保护表皮屏障至关重要，该屏障保护宿主免受环境的侮辱，并防止水分流失。在伤口修复过程中，一个基本的初始反应是循环中的白细胞的涌入，它们清洁伤口，并提供激活剂和生长因子来刺激纤维化和新生血管的形成。基质金属蛋白3(MMP3)是基质金属蛋白酶(MMP3)家族中的一员，是白细胞浸润的重要调节因子。在组织重塑过程中，MMP3在表皮中表达上调。在许多模型系统中，MMP3基因缺失的小鼠延迟了切除伤口的愈合，并显示出白细胞渗透的减少。此外，我们发现MMP3-/-动物对鳞癌进展的敏感性增加，这与在肿瘤进展的所有阶段明显减少中性粒细胞和巨噬细胞的浸润有关。基于这些观察，我们认为MMP3调节伤口修复过程中的保护性宿主防御反应，这反映了SCC过程中的保护性、抗肿瘤反应。许多已知的影响免疫细胞功能的分子可以直接被MMP3或由MMP3激活的基质蛋白酶水解和释放。我们推测，MMP3与其他基质蛋白结合，可直接促进白细胞外渗，进而影响伤口修复过程。白细胞进入炎症部位需要多个步骤的协调调节，包括内皮细胞的抑制、通过内皮屏障的迁移和通过结缔组织向炎症部位的定向迁移。MMP3由多种细胞表达，包括角质形成细胞、成纤维细胞、内皮细胞和单核细胞，并可激活其他基质蛋白类；因此，MMP3和其他MMPs可能调节最佳白细胞外渗所需的多个关键步骤。本项目的目的是阐明MMP3在伤口愈合过程中白细胞渗出中的功能作用。我们的特定目标将验证我们的假设如下：特定目标1：使用单个MMP3家族成员缺失或靶向过表达的体内小鼠模型，鉴定白细胞、成纤维细胞和角质形成细胞在依赖MMP3的白细胞渗入伤口环境中的细胞基质蛋白水解酶的贡献。具体目的2.验证MMP3活性调节创伤表皮中后续的基质金属蛋白酶家族成员激活的假说。具体目的3：明确MMP3和基质蛋白依赖的内皮细胞体外迁移过程中的细胞调控机制。这些研究将为MMP3和其他基质蛋白在白细胞内皮细胞迁移过程中的作用提供新的见解，并为在临床环境下测试基质金属蛋白酶抑制剂提供体内模型系统。 公共卫生相关性： 皮肤受伤需要愈合才能保持正常健康。伤口愈合的一个重要部分是炎症。基质金属蛋白酶，如MMP3，在伤口愈合过程中可能在调节炎症反应中发挥关键作用。