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

描述（由申请人提供）：子宫平滑肌瘤在流产、不孕症、早产、月经过多、盆腔疼痛和尿失禁中起重要作用。尽管如此戏剧性的症状，病因的子宫平滑肌瘤是知之甚少。这种有限的理解已经转化为有限的治疗选择。对于希望保持生育能力的子宫平滑肌瘤患者，大手术是目前唯一可行的选择，而且复发并不罕见。因此，这些妇女必须在冒着健康的未出生婴儿死亡的风险或冒着重复手术的风险之间做出选择，而短期和长期发病率和死亡率的可能性日益增加。更好地了解子宫平滑肌瘤的发展将为医学治疗提供新的靶点，从而最大限度地降低子宫平滑肌瘤对妊娠和一般健康的风险，同时消除与手术干预相关的风险。其中一种药物治疗，米非司酮，可以减小肿瘤大小，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平滑肌瘤细胞外基质（ECM）的产生。然而，米非司酮是否调节平滑肌瘤中ECM的形成并能刺激ECM的溶解，以及ECM调节平滑肌瘤细胞表型的机制尚不清楚。为了解决这一缺陷，我们从人类平滑肌瘤和患者匹配的肌层中生产了3-D永生化细胞系，并计划表征米非司酮治疗对ECM形成和降解的影响。我们还将通过描述3-D平滑肌瘤培养中相对于3-D子宫肌瘤培养中ECM产生的特征，进一步扩展我们在ECM产生和通过Rho信号通路信号传导的异常机械转导的发现。我们的两个具体目标是：(1)表征米非司酮对3-D平滑肌瘤培养物中ECM形成和溶解的影响；(2)确认通过形成ECM的机械应力扰动改变Rho/ERK信号并增加异常ECM的形成。通过完成拟议的研究，我们将描述临床有效疗法（米非司酮）对3-D平滑肌瘤ECM的治疗影响，以及Rho信号对ECM成分形成的影响。这个项目的创新之处在于：(1)它使用了唯一一个用于研究ECM的模型系统，(2)它描述了一种临床有效的治疗方法，(3)它使用了一个人类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.