Mechanism of Long Non-coding RNAs Action in leiomyoma

子宫肌瘤(肌瘤)是一种良性肿瘤，影响着相当数量的生育年龄。 女性，而且没有已知的原因。我们的实验室一直致力于了解miRNAs如何影响 肌瘤中的促炎和促纤维化途径，并鉴定了两个miRNAsmiR-200C和 MIR-29c，主要针对细胞周期、炎症和细胞外基质(ECM)成分 基因分别作为子宫肌瘤发生发展的关键基因。我们使用下一代技术的最新发现 测序已经证明了包括长的非编码RNA在内的一整套非编码RNA (LncRNAs)在肌瘤中调节失调。IncRNAs可以作为miRNAs的海绵，因此可能是 肌瘤基因失调的驱动因素。我们之前的研究表明曲尼司特，一种抗肿瘤药物 炎性药物在日本和韩国被批准用于治疗哮喘，在 通过抑制细胞增殖和刺激两个关键miRNAs的表达来抑制纤维瘤细胞 在肌瘤中表达下调(miR-200c和miR-29c)15、16。这些关键的miRNAs在肌瘤中起着关键作用 靶向细胞周期调节蛋白表达和炎症的发病机制(miR-200C)， 和ECM的组成和重塑(miR-29c)。此外，最近的报告显示，H19 在各种恶性肿瘤中也可作为miR-29和miR-200家族18-20的海绵。在这个项目中，我们 建议确定H19是否可以作为肌瘤miR-29/miR-200家族的海绵，以及曲尼司特是否可以 能降低H19的表达，从而增加miR-29和miR-200的表达 一家人。这个项目显然属于我们的R01资助项目的范围，该项目探索 H19，含miR-29和miR-200家族。我们假设H19海绵miR-29和miR-200家族在 肌瘤和曲尼司特通过抑制miR-29/miR-200家族上调miR-29/miR-200的表达 H19的表达。这一假设将在两个目标上得到检验。 目的1：确定LncRNAH19在肌瘤中是否作为miR-29和miR-200家族的海绵，以及 测定曲尼司特在体内外对H19表达的影响。 目的2：确定曲尼司特对miR-29和miR-200家族表达的影响是否介导 在H19之前。 这些目标的完成不仅将提高我们对长非编码RNA在 肌瘤的发病机制和潜在的靶向治疗方法，但也将提供 我们的候选人昆塔尼拉先生有机会学习一系列新的分子和外科技术 为他在生物医学研究领域的职业生涯做好准备。

Uterine leiomyoma (fibroids) are benign tumors afflicting a significant number of reproductive age women and without a known cause. Our laboratory has focused on understanding how miRNAs impact pro-inflammatory and pro-fibrotic pathways in leiomyoma, and identified two miRNAsmiR-200c and miR-29c which primarily target cell cycle, inflammation and extracellular matrix (ECM) component genes respectively as key in the development of leiomyoma. Our recent findings using next generation sequencing has demonstrated a whole host of non-coding RNAs including long non-coding RNAs (lncRNAs) are dysregulated in fibroids. LncRNAs can act as sponge for miRNAs and therefore may be a driver of gene dysregulation in leiomyomas. Our previous work demonstrated that tranilast, an anti- inflammatory drug approved for the treatment of asthma in Japan and Korea, has potent effects in fibroid cells by inhibiting cell proliferation and stimulating the expression of two key miRNAs that are down regulated in fibroids (miR-200c and miR-29c)15, 16. These key miRNAs play a pivotal role in fibroid pathogenesis by targeting the expression of cell cycle regulatory proteins and inflammation (miR-200c), and composition and remodeling of the ECM (miR-29c). Furthermore, recent reports showed that H19 also can act as sponge for miR-29 and miR-200 family18-20 in various malignancies. In this project we propose to determine if H19 can act as a sponge for miR-29/miR-200 family in fibroids and if tranilast can decrease the expression of H19 and thereby increase the expression of miR-29 and miR-200 family. This project clearly falls within the scope of our R01 funded project exploring the interaction of H19 with miR-29 and miR-200 family. We hypothesize that H19 sponges miR-29 and miR-200 family in fibroids and that tranilast upregulates the expression of miR-29/miR-200 family by inhibiting the expression of H19. This hypothesis will be tested in two Aims. Aim 1: Determine if lncRNA H19 functions as a sponge for miR-29 and miR-200 family in fibroids, and determine the effects of tranilast on the expression of H19 in vitro and in vivo. Aim 2: Determine if the effect of tranilast on the expression of miR-29 and miR-200 family is mediated by H19. Completion of these aims will not only advance our knowledge of the role of long non-coding RNAs in fibroid pathogenesis and potential targeting them as a therapeutic approach but will also provide the opportunity for our candidate Mr. Quintanilla to learn a host of new molecular and surgical techniques and prepare him for a career in the biomedical research.