Expression, hormonal regulation and function of microRNA in leiomyoma

描述(申请人提供)：子宫肌瘤是一种良性子宫肿瘤，依靠卵巢类固醇生长。通过基因组学和蛋白质组学，我们已经发现了一些在肌瘤中差异表达和调控的基因，这些基因具有不同的生物学功能，包括细胞转化、增殖、凋亡和促炎/促纤维化活性。其中一些基因的表达受卵巢类固醇通过ER/PR基因组和非基因组途径调节。MicroRNAs(MiRNAs)是一类新的非蛋白质编码的小RNA，通过抑制和降解来调节靶基因表达的稳定性。我们已经确定了一些miRNAs在成对的子宫肌层和肌瘤及其分离的平滑肌细胞(MSMC和LSMC)中的表达，包括miR-18a、21、181a、142-5p和542-3p，预计分别针对ER、PR、GPR30和促炎/促纤维化基因的表达。此外，基因的表达在肌瘤的发病机制中起着核心作用。我们的核心假设是，这些miRNAs的表达在肌瘤的生长和消退过程中表现出特定的模式，卵巢类固醇对它们的调节代表着一种先于它们对靶基因作用的机制，从而影响对肌瘤生长至关重要的多种细胞活动的结果。为了验证这一假设，目标1将评估miR-18a、21、181a、142-5p和542-3p及其编码ER、PR、GPR30、IL-13、转化生长因子-2、FBLN4、FMOD、MMP7和TIMP-3的预测靶基因在成对肌层和肌瘤生长(月经周期的增殖期和分泌期)和基于种族的消退(GnRHa疗法)中的表达。利用先前获得的成对子宫肌瘤和肌层的基因芯片图谱，我们将识别和确定这些miRNAs整体目标基因的功能注释。目的#2将分别评估卵巢激动剂的调节功能。为了实现我们的目标，我们将结合使用生化、分子和细胞生物学方法。我们预计，从这些研究中获得的信息将导致识别一种新的分子机制，该机制由miRNAs指导，导致对特定基因的调控，这些基因的产物在肌瘤的生长和消退中发挥核心作用，从而允许开发一种新的治疗方法来控制其生长。与公共卫生相关：子宫肌瘤是一种良性子宫肿瘤，估计70%的女性会在生殖年中发生，有症状的肿瘤会导致慢性盆腔疼痛和异常子宫出血。这项提案将调查肌瘤是如何生长的，以及预防其症状和生长的方法。

DESCRIPTION (provided by applicant): Leiomyomas are benign uterine tumors dependent on ovarian steroids for their growth. Through genomics and proteomics we have identified a number of differentially expressed and regulated genes in leiomyomas with diverse biological functions, including cellular transformation, proliferation, apoptosis and proinflammatory/pro-fibrotic activities. The expression of some of these genes is regulated by ovarian steroids through ER/PR genomic and non-genomic pathways. MicroRNAs (miRNAs) are novel class of small non- protein coding RNAs which regulate the stability of target gene expression through repression and degradation. We have identified the expression of a number of miRNAs in paired myometrium and leiomyomas and their isolated smooth muscle cells (MSMC and LSMC), including miR-18a, 21, 181a,142-5p and 542-3p, predicted to target the expression of ERs, PR, GPR30 and proinflammatory/profibrotic genes, respectively. Furthermore, the expression ofgenes, which collectively play a central role in pathogenesis of leiomyoma. Our core hypothesis is that the expression of these miRNAs displays a specific pattern during leiomyomas growth and regression, and their regulation by ovarian steroids represents a mechanism that precedes their actions on target genes, thus influencing the outcome of multiple cellular activities critical to leiomyomas growth. To test this hypothesis Aim #1 will assess the expression of miR-18a, 21, 181a, 142-5p and 542-3p and their predicted target genes encoding ERs, PR, GPR30, IL-13, TGF-2, FBLN4, FMOD, MMP7 and TIMP-3 in paired myometrium and leiomyoma during growth (proliferative and secretory phases of the menstrual cycle) and regression (GnRHa therapy) based on ethnicity. Utilizing gene microarray profiles previously obtained for paired leiomyoma and myometrium, we will identify and determine functional annotation of these miRNAs overall target genes. Aim #2 will assess the regulatory function of ovariagonists, respectively. To achieve our aims, we will utilize a combination of biochemical, molecular and cell biological approaches. We anticipate that the information gained from these studies leads to identification of a novel molecular mechanism directed by miRNAs resulting in regulation of specific genes whose products play a central role in leiomyoma growth and regression, permitting work toward development of a novel therapeutic to control their growth. PUBLIC HEALTH RELEVANCE: Fibroids are benign uterine tumors estimated to develop in 70% of women during their reproductive years, with symptomatic tumors causing chronic pelvic pain and abnormal uterine bleeding. This proposal will investigate how fibroids grow and ways to prevent their symptoms and growth.