Role of ACLP in dermal wound healing

ACLP 在真皮伤口愈合中的作用

• 批准号: 6761788
• 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 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6761788
• 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（摘自申请人摘要）：老年、营养不良和 有血管病变的糖尿病患者， 与高血压相关的不愈合皮肤伤口的发展 发病率，有时甚至死亡率。应用程序的目标是 获得新的技能和知识，研究皮肤伤口愈合过程， 获得教学经验，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/哈佛医学院将纳入 参加了一些结构化的研究活动，最终导致 科学独立。实验室占地2,500平方英尺， 新装修的空间配备了专为研究设计的仪器， 细胞和分子生物学，蛋白质化学，动物生理学， 组织学这项拟议中的研究将检查细胞外的作用， 皮肤创伤基质蛋白 治愈他们培育出了ACLP基因缺失的小鼠， 皮肤溃疡他们假设ACLP的缺失可能会阻止 通过损害细胞的增殖、分化或迁移， 真皮成纤维细胞在目标1中，他们将确定 使用组织学分析，ACLP的缺乏导致伤口愈合不良， 体外测定。在目标2中，他们将确定特定的DNA序列， 转录因子对调节ACLP的表达很重要， 使用报告基因测定体外培养真皮成纤维细胞。ACLP启动子LacZ 将产生报告基因转基因小鼠以评估转录水平。 在体内皮肤伤口愈合期间调节ACLP。在目标3中， 研究ACLP的生物学功能及其与其他 细胞外基质蛋白和盘状结构域受体酪氨酸激酶 （DDR）。将产生一系列ACLP和DDR缺失突变体以鉴定 对这些相互作用很重要的领域。此外，功能增益 使用产生ACLP的重组腺病毒进行实验。