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(摘自申请者的摘要)：老年、营养不良和 伴有血管功能不全的糖尿病患者罹患 与高血压性皮肤损伤相关的无法愈合的皮肤伤口的发展 发病率，有时甚至死亡率。该应用程序的目标是 获得研究皮肤伤口愈合过程的新技能和知识， 在教授他人方面获得经验，并获得必要的专业知识 领导一个独立的研究小组。这些经验和培训在 布里格姆妇女医院/哈佛医学院将把 参与几项有组织的研究活动，最终导致 为科学独立干杯。该实验室占地2500平方英尺 新装修的空间配备了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。这项拟议的研究将考察细胞外细菌的作用。 真皮创面中的基质蛋白、ACLP(主动脉羧肽酶样蛋白) 治愈。他们产生了ACLP缺失的小鼠，这些小鼠会自发地发展为无法愈合 皮肤溃疡。他们推测，ACLP的缺失可能会阻碍骨髓瘤的愈合 通过损害细胞的增殖、分化或迁移而造成的皮肤创伤 真皮成纤维细胞。在目标1中，他们将确定一种 缺乏ACLP导致创面愈合不良，通过组织学分析和 体外试验。在目标2中，他们将识别特定的DNA序列和 调控ACLP基因表达的重要转录因子 皮肤成纤维细胞体外报告基因分析。ACLP启动子LacZ 将产生报告转基因小鼠来评估转录 ACLP在体内皮肤创面愈合过程中的调节作用。在《目标3》中，他们将 研究ACLP的生物学功能及其与其他蛋白的相互作用 细胞外基质蛋白与盘状结构域受体酪氨酸激酶 (解甲返乡)。将产生一系列ACLP和DDR缺失突变体来鉴定 对于这些交互很重要的领域。此外，函数增益 将进行使用产生ACLP的重组腺病毒的实验。