Function of Long Non-Coding RNA MD1 in Leiomyoma Pathogenesis

长链非编码RNA MD1在平滑肌瘤发病机制中的作用

• 批准号: 10370413
• 负责人: OMID A. KHORRAM
• 金额: $ 7.71万
• 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: 2021
• 资助国家: 美国
• 起止时间: 2021-03-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370413
• 关键词: Address; African American population; Age; Animal Model; Area; Benign; Biological Assay; Cell Cycle; Cell Proliferation; Cells; Characteristics; Code; Data; Development; Disease; Epigenetic Process; Etiology; Extracellular Matrix; Family; Female Genital Neoplasms; Fibroid Tumor; Fibrosis; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Hemorrhage; High Prevalence; Immunoprecipitation; In Vitro; Inflammation; Investigation; Kidney; Knowledge; Laboratories; Leiomyoma; Lentivirus; Link; MicroRNAs; Modeling; Mus; Nuclear; Pain; Pathogenesis; Pathway interactions; Pelvic Pain; Porifera; Preclinical Testing; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Repression; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Symptoms; Testing; Therapeutic; Time; Tissues; Transplantation; United States National Institutes of Health; Untranslated RNA; Uterine Fibroids; Uterine Neoplasms; Uterine hemorrhage; Woman; Work; angiogenesis; base; beta catenin; capsule; clinically relevant; differential expression; experimental study; in vivo imaging; in vivo monitoring; knock-down; metaplastic cell transformation; myometrium; next generation; next generation sequencing; novel; overexpression; pressure; protein expression; reproductive; response; targeted treatment; transcriptome sequencing; tumor growth; tumorigenesis

Uterine leiomyoma (fibroids) develop during reproductive years in over 70% of women; however, their etiology remains unknown. Our group has identified two miRNAs, miR-200c and miR-29c, which are critical to regulation of genes functionally associated with cell cycle, cellular transformation, inflammation and extracellular matrix accumulation and thus of great significance to fibroid pathogenesis. Our recent findings using next generation sequencing provided a comprehensive profile of non-coding RNAs including long non- coding RNAs (lncRNAs) as well as novel groups of small non-coding RNAs (sncRNAs). Our preliminary findings here have identified aberrant expression of linc-MD1 and miR-135 family in fibroids and thus the impetus for this proposal. Using qRT-PCR we detected markedly reduced levels of linc-MD1 while significantly higher expression of miR-135 family in leiomyomas. The sequences of the linc-MD1 and our preliminary data suggest that linc-MD1 can act as miRNA sponge for miR-135 family (miR-135a/-135b) in leiomyoma smooth muscle cells. Therefore, based on this preliminary data we hypothesize that leiomyoma as compared to myometrium displays reduced expression of linc-MD1 which through a miRNA-guided mechanism involving miR-135 regulates the expression of specific target genes functionally associated with Wnt/β-catenin signaling pathway which is known to be central to leiomyoma pathogenesis. To test our core hypothesis we propose 2 specific aims. In Aim 1 we will determine the interaction between linc-MD1 and miR-135 family by RNA immunoprecipitation and RNA pull-down assay. We will over or under express linc-MD1 in LSMC or MSMC spheroid cells and determine its effect on miR-135 family and its downstream target genes namely GSK3β and APC which are known to regulate β-Catenin degradation. In these experiments β-Catenin, its phosphorylated form and its nuclear localization will be determined in response to overexpression and knockdown studies in vitro. To establish the clinical relevance and therapeutic potential of linc-MD1 in Aim 2 we will determine the effect of linc-MD1 overexpression in leiomyoma cells on fibroid progression in a leiomyoma animal model. This translational proposal addresses a significant gap in knowledge on the role of a novel lncRNA-miRNA network in the pathogenesis of fibroids which is a priority area of investigation for NIH, and could potentially be targeted for therapeutic purposes for fibroids.

在生殖年中，超过70％的女性在生殖年中发展了子宫平滑肌瘤（纤维）；但是，他们的病因 仍然未知。我们的小组已经确定了两个miRNA，miR-200C和miR-29c，这对于 调节基因在功能上与细胞周期，细胞转化，注射和 细胞外基质的积累，因此对纤维性发病机理具有重要意义。我们最近的发现 使用下一代测序提供了非编码RNA的全面概况，包括长期非编码RNA 编码RNA（LNCRNA）以及新型的小型非编码RNA（SNCRNA）。我们的初步 这里的发现确定了肌瘤中linc-md1和miR-135家族的异常表达，因此 推动此提案。使用QRT-PCR，我们检测到LINC-MD1的水平显着降低 miR-135家族在平滑肌瘤中的表达明显更高。 linc-md1和我们的序列 初步数据表明，LINC-MD1可以充当miRNA赞助商MiRNA赞助商（miR-135a/-135b） 平滑肌平滑肌细胞。因此，基于此初步数据，我们假设平滑肌瘤 与子宫肌层相比显示通过miRNA引导的linc-md1的表达降低 涉及miR-135的机制在功能上调节特定靶基因的表达 与Wnt/β-catenin信号通路相关，该途径已知是平滑肌瘤的核心 发病。为了检验我们的核心假设，我们提出了2个具体目标。在AIM 1中，我们将确定 通过RNA免疫沉淀和RNA下拉测定法，LINC-MD1和miR-135家族之间的相互作用。我们 将在LSMC或MSMC球体细胞中超过或以下linc-MD1，并确定其对miR-135的影响 家族及其下游靶基因，即GSK3β和APC，已知可以调节β-catenin 降解。在这些实验β-catenin中，其磷酸化形式及其核定位将是 根据体外的过表达和敲低研究的响应。建立临床相关性 AIM 2中LINC-MD1的治疗潜力，我们将确定LINC-MD1过表达的影响 平滑肌瘤细胞在动物平滑肌瘤模型中的纤维化进展。这个翻译的提案解决了 了解新型lncRNA-mIRNA网络在肌瘤发病机理中的作用的显着差距 这是NIH投资的优先领域，有可能以治疗目的为目标 肌瘤。

项目成果

The Influence of Race/Ethnicity on the Transcriptomic Landscape of Uterine Fibroids.

• DOI: 10.3390/ijms241713441
• 发表时间: 2023-08-30
• 期刊: International journal of molecular sciences
• 影响因子: 5.6

Tryptophan catabolism is dysregulated in leiomyomas.

• DOI: 10.1016/j.fertnstert.2021.05.081
• 发表时间: 2021-10
• 期刊: Fertility and sterility
• 影响因子: 6.7
• 作者: Chuang TD;Quintanilla D;Boos D;Khorram O
• 通讯作者: Khorram O

Therapeutic Effects of Long-Term Administration of Tranilast in an Animal Model for the Treatment of Fibroids.

• DOI: 10.3390/ijms241310465
• 发表时间: 2023-06-21
• 期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
• 影响因子: 5.6
• 作者: Chuang, Tsai-Der;Munoz, Leslie;Quintanilla, Derek;Boos, Drake;Khorram, Omid
• 通讯作者: Khorram, Omid

Differential Expression of Super-Enhancer-Associated Long Non-coding RNAs in Uterine Leiomyomas.

• DOI: 10.1007/s43032-022-00981-4
• 发表时间: 2022-10
• 期刊: Reproductive sciences (Thousand Oaks, Calif.)

Therapeutic effects of in vivo administration of an inhibitor of tryptophan 2,3-dioxygenase (680c91) for the treatment of fibroids: a preclinical study.

体内给予色氨酸 2,3-双加氧酶抑制剂 (680c91) 治疗肌瘤的治疗效果：临床前研究。

• DOI: 10.1016/j.fertnstert.2023.12.006
• 发表时间: 2024
• 期刊: Fertility and sterility
• 影响因子: 6.7
• 作者: Chuang,Tsai-Der;Ton,Nhu;Rysling,Shawn;Quintanilla,Derek;Boos,Drake;Khorram,Omid
• 通讯作者: Khorram,Omid