MiR-182 overexpression in early tumorigenesis of high grade serous carcinoma

MiR-182在高级别浆液性癌早期肿瘤发生中的过表达

• 批准号: 8606832
• 负责人: Jian-Jun Wei
• 金额: $ 16.3万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8606832
• 关键词: Age; BRCA1 Mutation; BRCA1 gene; BRCA2 Mutation; BRCA2 gene; Carcinoma; Carcinoma in Situ; Cell physiology; Cells; Clinical Trials; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; Defect; Defense Mechanisms; Diagnostic tests; Early Diagnosis; Early identification; Effectiveness; Epigenetic Process; Epithelial; Epithelial Cells; Epithelial ovarian cancer; Event; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Human; In Vitro; Ionizing radiation; Kinetics; Lead; Malignant neoplasm of ovary; Mammalian Oviducts; Mediating; Methylation; MicroRNAs; Molecular; Molecular Target; Mus; Mutation; Neoplasm Metastasis; Neoplasms; Nude Mice; Ovarian; Ovarian Carcinoma; Ovarian Serous Adenocarcinoma; Pathogenesis; Pathway interactions; Patients; Population; Prevention; Radiation Induced DNA Damage; Regulation; Repression; Risk; Risk Factors; Role; Serous; Serous Lesion; Somatic Mutation; Staging; Stress; TP53 gene; Testing; Therapeutic Effect; Transcriptional Regulation; Tumor Burden; Tumor Suppressor Genes; Woman; Xenograft procedure; cancer cell; cell behavior; cell growth; design; effective therapy; homologous recombination; in vivo; mutant; neoplastic cell; novel strategies; ovarian neoplasm; overexpression; public health relevance; repaired; research study; response; tumor; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): High-grade serous ovarian carcinoma (HGSOC) is an aggressive and deadly form of ovarian cancer, yet its pathogenesis is poorly understood. BRCA1 and BRCA2 mutations (identified in 10-40% of patients) are important risk factor for HGSOC. If inactivation of BRCA1/2 is critical in the early transformation of fallopian tube secretory epithelial (FTSE) cells, then other regulatory mechanisms must block BRCA1/2 in the remaining 60% of HGSOC cases. Recent identification of miR-182 repression of BRCA1 expression suggested a role for microRNA in the regulation of BRCA1 expression. MiR-182 is one of a few microRNAs overexpressed in cells with DNA damage and abnormal overexpression early and later stages of PSC. We hypothesize that miR-182 overexpression is an early genetic event and an important trigger of tumorigenesis in HGSOC through dysregulation of HGSOC associated genes. To test our hypothesis, we propose the following specific aims: AIM 1: Investigate the cellular and molecular mechanisms of miR-182-mediated tumorigenesis in FTSE cells in vitro. We propose that impaired DNA damage repair mechanisms and early tumorigenesis in FTSE cells is due to a dysregulation of miR-182 and its target genes. To test our hypothesis, we will examine 1) how miR-182 responds to DNA damage in FTSE cells and whether IR-induced miR-182 overexpression requires mutant P53; and 2) the kinetics of DNA damage repair in FTSE cells in the presence and absence of miR-182. We will also investigate whether miR-182- mediated tumorigenesis can be enhanced by negative regulation of its predicted target genes BNC2 and MTSS1. AIM 2: Test if miR-182 overexpression in FTSE results in tumor formation in xenografts and examine if knockdown of miR-182 expression reduce tumor burden and metastasis in ovarian cancer in vivo. Introducing miR-182 overexpression can result in tumor transformation and enhance invasion and metastasis. We observed that silencing of miR-182 expression in vitro significantly reduced aggressive cell behavior in normal FTSE and ovarian cancer cells. In this study, we will examine ovarian tumor cell growth, invasion, and metastasis in xenografts of nude mice by controlling miR-182 expression. The effectiveness of anti-miR-182 in treated tumors will be evaluated by examination of miR-182 and its target gene expression. If the proposed aims are achieved, we will design experiments for testing the therapeutic effects of conditional activation of miR-182 overexpression in mice, which mimics to early molecular changes in human HGSOC. The ultimate goals are to develop clinical trials in early prevention and a sensitive diagnostic test i detecting early STIC.

描述（由申请人提供）：高级浆液卵巢癌（HGSOC）是卵巢癌的一种侵略性和致命形式，但其发病机理知之甚少。 BRCA1和BRCA2突变（在10-40％的患者中鉴定）是HGSOC的重要危险因素。如果BRCA1/2的失活对于输卵管分泌上皮（FTSE）细胞的早期转化至关重要，则其他调节机制必须阻止其余60％​​的HGSOC病例中的BRCA1/2。 miR-182抑制BRCA1表达的最新鉴定表明microRNA在调节BRCA1表达中的作用。 miR-182是在PSC的早期和更高阶段具有DNA损伤和异常过表达的细胞中过表达的几个microRNA之一。我们假设miR-182的过表达是早期的遗传事件，并且是通过HGSOC相关基因失调而在HGSOC中的重要触发触发因素。为了检验我们的假设，我们提出了以下特定目的：目标1：研究MiR-182介导的FTSE细胞中miR-182介导的肿瘤发生的细胞和分子机制。我们认为，FTSE细胞中DNA损伤修复机制受损和早期肿瘤发生是由于miR-182及其靶基因的失调。为了检验我们的假设，我们将检查1）miR-182如何响应FTSE细胞中的DNA损伤，以及IR诱导的miR-182过表达是否需要突变体p53； 2）在存在和不存在miR-182的情况下，FTSE细胞中DNA损伤修复的动力学。我们还将通过负调控其预测的靶基因BNC2和MTSS1来调查miR-182-介导的肿瘤发生。 AIM 2：测试MiR-182 FTSE中的过表达是否会导致异种移植物中肿瘤形成，并检查miR-182表达的敲低是否减少了体内卵巢癌的肿瘤负担和转移。引入miR-182过表达会导致肿瘤转化并增强侵袭和转移。我们观察到，在体外表达miR-182的沉默会显着降低正常FTSE和卵巢癌细胞中的攻击性细胞行为。在这项研究中，我们将通过控制miR-182表达来检查裸鼠异种移植物中卵巢肿瘤细胞的生长，侵袭和转移。抗MIR-182在治疗肿瘤中的有效性将通过检查miR-182及其靶基因表达来评估。如果实现了所提出的目标，我们将设计实验，以测试小鼠中miR-182过表达的有条件激活的治疗作用，这模仿了人类HGSOC的早期分子变化。最终的目标是在早期预防和敏感的诊断测试中开发临床试验，我检测到了早期的STIC。