Mechanism of Long Non-coding RNAs Action in leiomyoma

长链非编码RNA在平滑肌瘤中的作用机制

• 批准号: 10053201
• 负责人: OMID A. KHORRAM
• 金额: $ 40.65万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-08 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10053201
• 关键词: Achievement; Address; African American; Age; Animal Model; Area; Benign; Bioinformatics; Cell Cycle; Cell Cycle Progression; Cell Proliferation; Cells; Characteristics; Code; Complex; Data; Development; Disease; Down-Regulation; Enantone; Epigenetic Process; Epithelial; Epithelium; Estrogens; Ethnic group; Expression Profiling; Extracellular Matrix; Female Genital Neoplasms; Fibroid Tumor; Fibrosis; Genes; Genetic Transcription; Gonadal Steroid Hormones; Growth; High Prevalence; Human Genome; Immunoprecipitation; Inflammation; Inflammatory; Investigation; Kidney; Knock-in; Knowledge; Laboratories; Leiomyoma; Luteal Phase; Mediator of activation protein; Mesenchymal; Messenger RNA; Modeling; Modification; Mus; Mutation; Ovarian; Pain; Pathogenesis; Pathway interactions; Patients; Phase; Porifera; Post-Transcriptional Regulation; Preclinical Testing; Progesterone; Proteins; Publishing; RNA; Race; Regulation; Rest; Role; Severities; Small Interfering RNA; Small RNA; Smooth Muscle Myocytes; Sp1 Transcription Factor; Steroids; Subfamily lentivirinae; Symptoms; Testing; Therapeutic; Tissues; Transplantation; United States National Institutes of Health; Untranslated RNA; Up-Regulation; Uterine Fibroids; Uterine Neoplasms; Uterine hemorrhage; Woman; Work; base; capsule; cdc Genes; chromosomal location; clinically relevant; experimental study; fibrogenesis; in vivo imaging; in vivo monitoring; knock-down; mRNA Expression; myometrium; new therapeutic target; next generation; next generation sequencing; overexpression; pressure; reproductive; therapeutic target; transcriptome; transcriptome sequencing; tumor; tumor growth; tumorigenesis

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. Based on the level of expression and functional association with tumorigenesis and tissue fibrosis we selected a group of lncRNAs to further characterize. Using QRT-PCR we detected marked overexpression of XIST and MIAT in leiomyomas. The sequences of these lncRNAs and preliminary data suggest they can act as miRNA sponges for miR-29c and miR-200c. Therefore, based on our published and preliminary data we hypothesize that leiomyoma as compared to myometrium display an altered expression of a number of lncRNAs that are transcriptionally regulated and independently or through miRNA-guided mechanisms regulate the expression of specific target genes functionally associated with cell cycle progression, inflammation, fibrosis and epigenetic modification all of which are central to leiomyoma pathogenesis. To test our core hypothesis we propose 3 specific aims. In Aim 1 we will build on our preliminary RNA sequencing study and provide a comprehensive profile of lncRNAs, miRNAs, with concurrent mRNAs expression in large sets of paired leiomyoma and myometrium from different race/ethnic groups derived from proliferative and secretory phase of the menstrual cycle. Through bioinformatic analysis we will identify lncRNAs subtypes, their chromosomal locations at loci often rearranged in leiomyoma and identify lncRNAs which could potentially act as miRNA sponges, or competing endogenous RNAs (ceRNAs). We will also determine if presence of MED12 mutations has any impact on the profiles of these ncRNAs. Aim 2 will address the regulation and function of MIAT and XIST in leiomyoma. Using RNA immunoprecipitation studies we will determine if these lncRNAs will interact with miR-29c and miR-200c known to be important in fibrogenesis. At the cellular level we will assess the role of ovarian steroids on their expression and through knockdown and knock-in strategies identify their effects on expression of ECM and cell cycle genes. To establish the clinical relevance and therapeutic potential of lncRNAs in Aim 3 we will determine the effect of altering the expression of XIST and MIAT which are highly overexpressed in fibroids on fibroid progression and/or establishment in a leiomyoma animal model. This translational proposal addresses a significant gap in knowledge in the pathogenesis of fibroids which is a priority area of investigation for NIH, and could have potential therapeutic applications for treatment of fibroids.

子宫平滑肌瘤（肌瘤）是良性肿瘤，影响大量复制的女性和 没有已知原因。我们的实验室专注于了解miRNA如何影响促炎性 和平滑肌中的促纤维化途径，并确定了两个mirnas-Mir-200c和miR-29c 目标细胞周期，注射和细胞外基质（ECM）成分基因作为关键 平滑肌瘤的发展。我们最近使用下一代测序的发现证明了整个 在肌瘤中，包括长非编码RNA（LNCRNA）在内的非编码RNA宿主在内。 lncrnas 可以充当miRNA的赞助商，因此可能是平滑肌瘤基因失调的驱动力。基于 与肿瘤发生和组织纤维化的表达水平和功能关联，我们选择了一组 lncrnas进一步表征。使用QRT-PCR，我们检测到在 平滑肌瘤。这些LNCRNA和初步数据的序列表明它们可以充当miRNA海绵 用于miR-29c和miR-200c。因此，根据我们已发布的初步数据，我们假设 与肌层相比，平滑肌瘤表现出许多LNCRNA的表达改变 通过转录调节，独立或​​通过miRNA指导的机制调节 特定靶基因在功能上与细胞周期进程，炎症， 纤维化和表观遗传修饰所有这些都是平滑肌瘤发病机理的中心。测试我们的 核心假设我们提出了3个具体目标。在AIM 1中，我们将基于我们的初步RNA测序研究 并提供LNCRNA，miRNA的全面概况，并以大量集合以同时的mRNA表达 来自不同种族/族裔的配对平滑肌瘤和肌层来自增殖和分泌 通过生物信息学分析，我们将确定LNCRNA亚型，它们 在平滑肌瘤中经常重排的局部染色体位置，并鉴定有可能行动的lncRNA 如miRNA海绵或竞争性内源性RNA（CERNAS）。我们还将确定是否存在Med12 突变对这些NCRNA的特征有任何影响。 AIM 2将解决 MIAT和XIST在平滑肌瘤中。使用RNA免疫沉淀研究，我们将确定这些LNCRNA是否会 与MiR-29c和MiR-200C相互作用，已知在纤维发生中很重要。在细胞级别，我们将评估 卵巢立体素在其表达中的作用以及通过敲低和敲击策略确定 对ECM和细胞周期基因表达的影响。建立临床相关性和治疗潜力 AIM 3中的lncRNA的lncrNA，我们将确定改变高度的Xist和MIAT表达的效果 在平滑肌瘤动物模型中，在肌瘤进展和/或建立的肌瘤中过表达。这 翻译提案解决了在肌瘤发病机理中的一个显着差距，这是一个 NIH的优先投资领域，并且可能具有用于治疗肌瘤的潜在治疗应用。