Extra / Intra-cellular Signaling in Novel 3-Dimensional Human Leiomyoma Cultures

描述(申请人提供)：子宫肌瘤在流产、不孕、早产、月经过多、盆腔疼痛和尿失禁中起重要作用。尽管有如此戏剧性的症状，子宫肌瘤的病因仍鲜为人知。这种有限的理解已经转化为有限的治疗选择。对于患有子宫肌瘤并希望保持生育能力的女性来说，大手术是目前唯一可用的选择，而且复发并不少见。因此，这些妇女必须在冒着本来健康的未出生婴儿死亡的风险和冒着再次手术的风险之间做出选择，短期和长期发病率和死亡率的可能性不断增加。更好地了解子宫肌瘤的发展将为医学治疗提供新的靶点，可以将子宫肌瘤对怀孕和一般健康的风险降至最低，同时消除与手术干预相关的风险。米非司酮就是这样一种药物，可以缩小肿瘤大小，尽管其机制尚不清楚。在初步研究中，我们已经证明米非司酮调节细胞内肌瘤细胞外基质(ECM)的产生。然而，尚不清楚的是，米非司酮是否调节子宫肌瘤细胞外基质的形成和刺激细胞外基质的溶解，以及细胞外基质调节肌瘤细胞表型的机制。为了解决这一缺陷，我们从人子宫肌瘤和患者匹配的子宫肌层中建立了三维永生化细胞系，并计划表征米非司酮治疗对细胞外基质形成和降解的影响。我们还将扩展我们的发现，通过Rho信号通路异常的机械转导对ECM的产生和信号传递，通过表征相对于三维子宫肌层培养的三维肌瘤培养的ECM产生。我们的两个具体目标是：(1)研究米非司酮对三维子宫肌瘤细胞外基质形成和溶解的影响，以及(2)证实机械应力扰动通过形成的细胞外基质改变Rho/ERK信号转导和增加异常细胞外基质的形成。随着拟议研究的完成，我们将表征临床有效的治疗方法(米非司酮)对三维肌瘤细胞外基质的治疗效果，以及Rho信号对细胞外基质成分形成的影响。这个项目的创新之处在于：(1)它使用了为研究ECM而设计的唯一模型系统，(2)它表征了一种临床有效的治疗方法，(3)它使用了一个人体模型来研究一种流行的人类疾病。

DESCRIPTION (provided by applicant): Uterine leiomyoma play a significant role in miscarriage, infertility, preterm labor, menorrhagia, pelvic pain, and urinary incontinence. Despite such dramatic symptoms, the etiology of uterine leiomyomas is marginally understood. This limited understanding has translated to limited therapeutic options. For women suffering from uterine leiomyomas who desire to maintain their fertility, major surgery is the only current option available, and recurrence is not uncommon. As a result, these women must choose between risking the demise of an otherwise healthy unborn child or risking repeat surgery, with ever-increasing likelihood of short- and long-term morbidity and mortality. Better understanding of uterine leiomyoma development would provide novel targets for medical therapy that could minimize the risk of uterine leiomyomas on pregnancy and general health while at the same time eliminating the risks associated with surgical intervention. One such medical therapy, mifepristone, decreases tumor size, although the mechanism is unclear. In preliminary studies, we have demonstrated that mifepristone regulated leiomyoma extracellular matrix (ECM) production within the cell. What is unknown, however, is whether mifepristone regulates ECM formation and can stimulate ECM dissolution in leiomyomas, and the mechanism by which ECM regulates the leiomyoma cellular phenotype. In order to address this deficit, we have produced 3-D immortalized cell lines from human leiomyoma and patient-matched myometrium and plan to characterize the impact of mifepristone treatment on ECM formation and degradation. We will also expand upon our findings of aberrant mechanotransduction on ECM production and signaling via the Rho signaling pathway by characterizing ECM production in 3-D leiomyoma cultures relative to 3-D myometrial cultures. Our two specific aims are: (1) to characterize the impact of mifepristone on ECM formation and dissolution on 3-D leiomyoma cultures, and (2) to confirm that perturbations in mechanical stress through formed ECM alter Rho/ERK signaling and increase aberrant ECM formation. By the completion of the proposed studies, we will have characterized the therapeutic impact of a clinically effective therapy (mifepristone) on 3-D leiomyoma ECM, and the impact of Rho signaling on ECM component formation. This project is innovative in that (1) it uses the only model system designed to study ECM, (2) it characterizes a clinically effective therapy, and (3) it uses a human model to study a prevalent human disease.