Expression, hormonal regulation and function of microRNA in leiomyoma

描述（由申请人提供）：平滑肌瘤是依赖卵巢类固醇生长的良性子宫肿瘤。通过基因组学和蛋白质组学，我们已经确定了平滑肌瘤中具有多种生物学功能的差异表达和调控基因，包括细胞转化、增殖、凋亡和促炎/促纤维化活性。卵巢类固醇通过ER/PR基因组和非基因组途径调控其中一些基因的表达。MicroRNAs （miRNAs）是一类新型的小非蛋白编码rna，通过抑制和降解来调节靶基因表达的稳定性。我们已经在成对的肌层和平滑肌瘤及其分离的平滑肌细胞（MSMC和LSMC）中鉴定了一些mirna的表达，包括miR-18a、21、181a、142-5p和542-3p，预计它们分别靶向er、PR、GPR30和促炎/促纤维化基因的表达。此外，基因的表达，这些基因共同在平滑肌瘤的发病机制中起着核心作用。我们的核心假设是，这些mirna的表达在平滑肌瘤的生长和消退过程中表现出一种特定的模式，卵巢类固醇对它们的调节代表了一种作用于靶基因之前的机制，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。为了验证这一假设，Aim #1将评估miR-18a、21、181a、142-5p和542-3p的表达及其预测的靶基因编码er、PR、GPR30、IL-13、TGF-2、FBLN4、FMOD、MMP7和TIMP-3在配对子宫肌瘤和平滑肌瘤中基于种族的生长（月经周期的增殖和分泌期）和消退（GnRHa治疗）。利用先前获得的成对平滑肌瘤和子宫肌层的基因微阵列谱，我们将识别并确定这些mirna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将导致鉴定出一种由mirna指导的新的分子机制，该机制导致特定基因的调节，其产物在平滑肌瘤的生长和消退中起着核心作用，从而允许开发一种新的治疗方法来控制其生长。公共卫生相关性：子宫肌瘤是良性子宫肿瘤，估计在70%的育龄妇女中发展，有症状的肿瘤引起慢性盆腔疼痛和子宫异常出血。本提案将探讨肌瘤如何生长和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.