COLLAGENOLYSIS AS FUNCTION OF MMPS IN INCONTINENT WOMEN

失禁女性中 MMPS 功能的胶原蛋白溶解

• 批准号: 6362236
• 负责人: MARY L. POLAN
• 金额: $ 19.55万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-15 至 2004-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6362236
• 关键词: clinical research; collagen; collagen disorder; collagenase; cytokine; enzyme activity; female; fibroblasts; gene expression; growth factor; human subject; human tissue; messenger RNA; metalloendopeptidases; polymerase chain reaction; protein metabolism; proteolysis; stromelysin; tissue /cell culture; tissue inhibitor of metalloproteinases; urinary incontinence; western blottings; women's health

DESCRIPTION (Adapted from the Applicant's Abstract): Urinary incontinence is a major public health problem in the United States, resulting in an 11% lifetime risk of surgery for women with incontinence and/or prolapse. The basic molecular pathophysiology of urinary incontinence is poorly understood and, therefore, the development of medical prophylaxis or therapy for this growing area of disability for older women has lagged. The investigators hypothesize that stress urinary incontinence (SUI) results from abnormal pelvic collagen metabolism as a function of differential expression of the matrix metalloproteinases (MMPs) known to degrade extracellular matrix collagen and the tissue inhibitors of metalloproteinases (TIMPs) which specifically inhibit MMP proteolytic activity. Thus, variations in the expression of MMPs and TIMPs govern the amount and type of collagen in the supporting pelvic structures by controlling proteolysis. They propose to investigate MMP and TIMP expression in vaginal cuff tissue isolated from women with SUI and continent women before and after menopause to delineate differences in proteolytic activity and pelvic collagen content by, first, measuring mRNA and protein expression of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 by quantitative, competitive reverse transcription polymerase chain reaction, Western blot analysis, and zymography. Second, they will culture tissue fibroblasts from vaginal cuff samples from control and incontinent women and determine the in vitro mRNA and protein expression of MMPs and TIMPs in response to gonadal steroid, cytokine, and growth factor modulation. Third, to confirm differences in collagenolysis between control and incontinent women, they will measure and compare total collagen content, ratio of type I/type III collagen, amount of the carboxy-terminal epitope, COL2-3/4Cshort, generated by the initial collagen cleavage, and level of pyridinoline crosslinks from both in vivo isolated vaginal cuff tissue and in vitro cultured fibroblasts from control and incontinent patients. Two-thirds of the burden of urinary incontinence is borne by women with prevalence rates of 14 to 41%. The investigators do not believe that any medical therapy for SUI is available. Their fourth goal is to determine the ability of a clinically relevant MMP inhibitor (RS113,456; Roche Bioscience, Inc.) to ablate in vivo MMP proteolysis in fibroblast cultures from women with SUI and control women. Such MMP inhibitors may offer a therapeutic intervention to reverse the pathophysiologc degradation of collagen in women as they age, which results in incontinence.

描述（改编自申请人的摘要）：尿失禁是一种 美国的主要公共卫生问题，导致11%的人一生 失禁和/或脱垂女性的手术风险。 基本 尿失禁的分子病理生理学知之甚少， 因此，针对这种增长的医学预防或治疗的发展 老年妇女的残疾领域滞后。 研究人员假设压力性尿失禁（SUI）的结果 由于盆腔胶原蛋白代谢异常， 基质金属蛋白酶（MMPs）的表达， 细胞外基质胶原和金属蛋白酶组织抑制剂 特异性抑制MMP蛋白水解活性的TIMP。 因此， 在MMPs和TIMPs的表达中， 通过控制蛋白水解来支撑骨盆结构。 他们提议 MMP和TIMP在女性阴道断端组织中表达研究 与SUI和大陆妇女在绝经前后， 蛋白水解活性和盆腔胶原蛋白含量的差异，首先， 测定MMP-1、MMP-2、MMP-9、TIMP-1、TIMP-2的mRNA和蛋白表达， 和TIMP-3的定量竞争性逆转录聚合酶链 反应、Western印迹分析和酶谱分析。 第二，文化 来自对照和失禁妇女的阴道断端样品的组织成纤维细胞 并在体外测定MMPs和TIMPs的mRNA和蛋白表达。 对性腺类固醇、细胞因子和生长因子调节的反应。 三是 证实了对照组和失禁妇女之间胶原溶解的差异， 他们将测量和比较总胶原蛋白含量，I型/III型胶原蛋白的比例， 胶原蛋白，羧基末端表位的量，COL 2 -3/4Cshort，由 最初的胶原蛋白裂解，以及来自两者的吡啶啉交联水平 体内分离的阴道断端组织和体外培养的成纤维细胞， 控制和失禁患者。 三分之二的尿失禁负担是由女性承担的， 患病率为14%至41%。 调查人员认为， SUI的药物治疗是可用的。 他们的第四个目标是确定 临床相关MMP抑制剂的能力（RS 113，456; Roche Bioscience， 公司）消除来自患有乳腺癌的女性的成纤维细胞培养物中的体内MMP蛋白水解， SUI和对照组女性。 这样的MMP抑制剂可以提供治疗性 干预，以扭转妇女胶原蛋白的病理生理降解， 它们会变老，导致失禁。