Role of ACLP in dermal wound healing

ACLP 在真皮伤口愈合中的作用

• 批准号: 6653248
• 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/6653248
• 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（取自申请人的摘要）：老年医学，营养不良和 血管不足的糖尿病患者的风险增加 与高有关的非愈合皮肤伤口的发展 发病率，有时是死亡率。应用程序的目标是 获取新技能和知识调查皮肤伤口愈合过程， 获得教别人教的经验，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 Brigham和Womens Hospital/Harvard医学院将合并 参与几项结构化和研究活动最终领导 科学独立。实验室包括2500平方英尺 新装修的空间配备了用于研究的工具 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的作用 皮肤伤口中的基质蛋白，ACLP（主动脉羧肽样蛋白） 康复。他们产生了ACLP-NULL小鼠，该小鼠自发发展非治疗 皮肤溃疡。他们假设缺乏ACLP可能会阻止 皮肤伤口通过损害增殖，分化或迁移 真皮成纤维细胞。在AIM 1中，他们将确定一个机制 使用组织学分析和在 体外测定。在AIM 2中，他们将确定特定的DNA序列，并且 转录因子对于调节ACLP的表达很重要 使用报告基因测定法在体外进行皮肤成纤维细胞。 ACLP启动子LACZ 将生成记者转基因小鼠以评估转录 在体内皮肤伤口愈合期间ACLP调节。在AIM 3中，他们将 研究ACLP的生物学功能及其与其他的相互作用 细胞外基质蛋白和盘状蛋白域受体酪氨酸激酶 （DDR）。将生成一系列ACLP和DDR-DRETION突变体以识别 这些相互作用很重要的领域。另外，功能收益 将使用产生ACLP的重组腺病毒的实验。