REGULATION OF UTERINE FIBROIDS BY CCN5

CCN5 对子宫肌瘤的调节

• 批准号: 6921334
• 负责人: John J Castellot
• 金额: $ 27.74万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-26 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6921334
• 关键词: cell motility; cell proliferation; estrogens; extracellular matrix; female; gene frequency; genetically modified animals; human tissue; immunocytochemistry; laboratory mouse; laboratory rat; leiomyoma; messenger RNA; polymerase chain reaction; smooth muscle; tissue /cell culture; uterus; western blottings; women' s health

DESCRIPTION (provided by applicant): The long-term goal of this project is to elucidate the cellular, molecular, and biochemical mechanisms regulating the proliferation and motility of human uterine smooth muscle cells (UtSMC). UtSMC hyperproliferation is the cause of fibroids, a condition that afflicts 20-25% of all women and 75% of African-American women. Fibroids cause severe pain and bleeding, impair fertility, and result in >200,000 hysterectomies annually in the U.S. There is no known treatment--medical or surgical--that permanently reduces or eliminates fibroids, other than hysterectomy. Clearly, a detailed understanding of the mechanisms and molecules that regulate UtSMC mitogenesis and migration will provide a therapeutic rationale for controlling fibroids, and may provide important insights into the pathophysiologic basis for fibroid formation. Our laboratory has provided strong evidence that CCN5, an estrogen-induced growth-arrest specific gene, inhibits proliferation and motility in cultured UtSMC. Furthermore, we have demonstrated that human leiomyomas have greatly reduced levels of CCN5 mRNA and protein compared to normal myometrium from the same uterus. Based on this evidence the following hypothesis will be tested: CCN5 is an autocrine regulator of UtSMC proliferation and motility in culture and in vivo, and exerts it anti-proliferative and anti-motility effects, at least in part, through regulation of extracellular matrix synthesis and composition. To test this hypothesis, we will: 1) Continue our functional analysis of CCN5 and its regulation by estrogen on proliferation, motility, and extracellular matrix in SMC cultured from matched pairs of normal and fibroid human uterine tissue. To do this we will use adenovirus vectors, recombinant CCN5, and small inhibitory RNA approaches. 2) Examine the physiologic functions and estrogen regulation of CCN5 in animal models, including normal cycling rats, ovariectomized rats, pregnant rats, wild-type mice, and genetically manipulated mice that either under- or over-express CCN5. Quantitative PCR, Western blot analysis, and immunohistochemistry will be used to determine the spatial and temporal expression pattern and estrogen regulation of CCN5 in each of these animal models. We will also explore the possibility that CCN5 gene or protein therapy might be a useful approach for suppressing human fibroids in a novel nude mouse model system. The experiments proposed in this application should provide new and important insights into UtSMC pathophysiology in humans.

描述（由申请人提供）：该项目的长期目标是阐明调节人子宫平滑肌细胞（UTSMC）的增殖和运动的细胞，分子和生化机制。 UTSMC过度增殖是肌瘤的原因，这种情况遭受了所有妇女的20-25％和75％的非裔美国妇女的疾病。肌瘤会导致严重的疼痛和出血，损害生育能力，并在美国每年导致> 20万种子宫切除术，除了子宫切除术以外，没有已知的治疗治疗 - 医疗或外科手术 - 永久减少或消除肌瘤。显然，对调节UTSMC有丝分裂发生和迁移的机制和分子的详细理解将为控制肌瘤提供一种治疗原理，并可能为肌瘤形成的病理生理基础提供重要的见解。我们的实验室提供了有力的证据，表明雌激素诱导的特定基因CCN5抑制了培养的UTSMC中的增殖和运动。此外，我们已经证明，与同一子宫的正常肌层相比，人类平滑肌瘤的CCN5 mRNA和蛋白质水平大大降低。基于此证据，将检验以下假设：CCN5是UTSMC增殖和培养物和体内运动的自分泌调节剂，并通过调节细胞外基质合成和成分的调节至少部分地施加了抗增殖和抗动力效应。为了检验这一假设，我们将：1）继续对CCN5的功能分析及其通过雌激素对从匹配的正常和肌瘤人子宫组织成对培养的SMC中的增殖，运动和细胞外基质进行调节。为此，我们将使用腺病毒载体，重组CCN5和小型抑制性RNA方法。 2）检查动物模型中CCN5的生理功能和雌激素调节，包括正常的循环大鼠，卵巢切除大鼠，怀孕的大鼠，野生型小鼠以及遗传操纵的小鼠，这些小鼠均不超过表达CCN5。定量PCR，Western印迹分析和免疫组织化学将用于确定这些动物模型中CCN5的空间和时间表达模式和雌激素调节。我们还将探讨CCN5基因或蛋白质治疗可能是抑制新型裸小鼠模型系统中人肌瘤的有用方法。本应用程序中提出的实验应为人类的UTSMC病理生理学提供新的重要见解。