Expression, hormonal regulation and function of microRNA in leiomyoma

描述（由申请人提供）：平滑肌瘤是良性子宫肿瘤，其生长依赖于卵巢类固醇。通过基因组学和蛋白质组学，我们已经确定了一些差异表达和调控的基因在平滑肌瘤具有不同的生物学功能，包括细胞转化，增殖，凋亡和促炎/促纤维化活动。其中一些基因的表达受卵巢类固醇激素通过ER/PR基因组和非基因组途径调节。microRNAs（miRNAs）是一类新型的非蛋白质编码小RNA，通过抑制和降解来调节靶基因表达的稳定性。我们已经鉴定了许多miRNA在成对的子宫肌层和平滑肌瘤及其分离的平滑肌细胞（MSMC和LSMC）中的表达，包括miR-18 a、21、181 a、142- 5 p和542- 3 p，预测分别靶向ER、PR、GPR 30和促炎/促纤维化基因的表达。此外，基因的表达，共同发挥核心作用，在平滑肌瘤的发病机制。我们的核心假设是，这些miRNAs的表达在平滑肌瘤生长和消退过程中显示出特定的模式，卵巢类固醇对它们的调节代表了一种先于它们对靶基因作用的机制，从而影响了对平滑肌瘤生长至关重要的多种细胞活动的结果。为了检验这一假设，目标#1将评估miR-18 a、21、181 a、142- 5 p和542- 3 p的表达及其预测的编码ER、PR、GPR 30、IL-13、TGF-2、FBLN 4、FMOD、MMP 7和TIMP-3在子宫肌层和平滑肌瘤生长过程中的表达根据种族，可分为月经周期的增殖期和分泌期和消退期（GnRHa治疗）。利用先前获得的成对子宫肌瘤和子宫肌层的基因微阵列谱，我们将识别和确定这些miRNA的总体靶基因的功能注释。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，我们将利用生物化学，分子和细胞生物学方法的组合。我们预计，从这些研究中获得的信息导致鉴定由miRNA指导的新分子机制，导致特定基因的调节，其产物在平滑肌瘤生长和消退中发挥核心作用，从而允许开发新的治疗方法来控制其生长。公共卫生相关性：子宫肌瘤是一种良性的子宫肿瘤，估计70%的女性在生育期间会发生，有症状的肿瘤会引起慢性盆腔疼痛和异常子宫出血。该提案将研究肌瘤如何生长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.