Role of ACLP in dermal wound healing

ACLP 在真皮伤口愈合中的作用

• 批准号: 6533022
• 负责人: MATTHEW D LAYNE
• 金额: $ 12.78万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6533022
• 关键词: DNA binding protein; biopsy; carboxypeptidase; cell differentiation; cell migration; cell proliferation; extracellular matrix proteins; fibroblasts; gel mobility shift assay; gene expression; genetic promoter element; genetically modified animals; immunocytochemistry; molecular cloning; muscle cells; northern blottings; nucleic acid sequence; polymerase chain reaction; protein structure function; protein tyrosine kinase; site directed mutagenesis; skin ulcer; statistics /biometry; transcription factor; vascular smooth muscle; wound healing

DESCRIPTION (Taken from the applicant's abstract): Geriatric, malnourished, and diabetic patients with vascular insufficiencies are at increased risk for the development of non-healing dermal wounds that are associated with high morbidity and sometimes mortality. The objectives of the application are to acquire new skills and knowledge investigating dermal wound healing processes, to gain experience in teaching others, and to achieve the necessary expertise to lead an independent research group. These experiences and training at Brigham and Womens Hospital/Harvard Medical School will incorporate the participation in several structured and research activities ultimately leading to scientific independence. The laboratory encompasses 2,500 square feet of newly renovated space equipped with instruments designed for research in cellular and molecular biology, protein chemistry, animal physiology, and histology. The proposed research will examine the role of an extracellular matrix protein, ACLP (aortic carboxypeptidase-like protein) in dermal wound healing. They generated ACLP-null mice, which spontaneously develop non-healing skin ulcers. They hypothesize that an absence of ACLP may prevent healing of skin wounds by impairing the proliferation, differentiation, or migration of dermal fibroblasts. In Aim 1, they will determine the mechanisms by which an absence of ACLP leads to poor wound healing using histological analysis and in vitro assays. In Aim 2, they will identify specific DNA sequences and transcription factors important for regulating the expression of ACLP in dermeal fibroblasts in vitro using reporter gene assays. ACLP promoter LacZ reporter transgenic mice will be generated to evaluate the transcriptional regulation of ACLP during dermal wound healing in vivo. In Aim 3, they will investigate the biological function of ACLP and its interaction with other extracellular matrix proteins and discoidin domain receptor tyrosine kinases (DDR). A series of ACLP and DDR-deletion mutants will be generated to identify domains important for these interactions. In addition, gain-of-function experiments using recombinant adenovirus that produces ACLP will be performed.

描述（摘自申请人的摘要）：老年、营养不良和 患有血管功能不全的糖尿病患者患糖尿病的风险增加 与高相关的不愈合皮肤伤口的发展 发病率，有时甚至是死亡率。该应用程序的目标是 获得研究真皮伤口愈合过程的新技能和知识， 获得教学他人的经验，并获得必要的专业知识 领导一个独立的研究小组。这些经验和培训 布莱根妇女医院/哈佛医学院将纳入 参与多项结构化和研究活动，最终领导 科学独立。该实验室占地 2,500 平方英尺 新装修的空间配备了专为研究而设计的仪器 细胞和分子生物学、蛋白质化学、动物生理学和 组织学。拟议的研究将检查细胞外细胞的作用 真皮伤口中的基质蛋白、ACLP（主动脉羧肽酶样蛋白） 康复。他们培育出 ACLP 缺失的小鼠，这些小鼠会自发地出现不可愈合的症状 皮肤溃疡。他们假设缺乏 ACLP 可能会阻碍愈合 通过损害增殖、分化或迁移来治疗皮肤伤口 真皮成纤维细胞。在目标 1 中，他们将确定机制 根据组织学分析，缺乏 ACLP 会导致伤口愈合不良 体外测定。在目标 2 中，他们将识别特定的 DNA 序列并 转录因子对于调节 ACLP 的表达具有重要意义 使用报告基因测定体外真皮成纤维细胞。 ACLP启动子LacZ 将产生报告基因转基因小鼠来评估转录 ACLP 在体内真皮伤口愈合过程中的调节。在目标 3 中，他们将 研究ACLP的生物学功能及其与其他物质的相互作用 细胞外基质蛋白和盘状结构域受体酪氨酸激酶 （DDR）。将生成一系列 ACLP 和 DDR 删除突变体来识别 对于这些交互很重要的领域。此外，功能获得 将使用产生 ACLP 的重组腺病毒进行实验。