Exploring miR-29 in melanoma progression and prevention

探索 miR-29 在黑色素瘤进展和预防中的作用

基本信息

批准号：
10456977
负责人：
Florian Karreth
金额：
$ 22.64万
依托单位：
H. LEE MOFFITT CANCER CTR & RES INST
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456977
关键词：
Address Alleles Biological Process Biology Cell Line Cell Proliferation Clinical Complement Data Development Disease Down-Regulation Ensure Evaluation Family Gene Expression Genetic Genetic Engineering Genetic Transcription Genetically Engineered Mouse Goals Growth Human Human Cell Line Human Engineering Immunotherapy Impairment In Vitro Individual MAP Kinase Gene MYBL2 gene Maintenance Malignant Neoplasms Mediating Melanoma Cell Messenger RNA MicroRNAs Modeling Mutation Nevi and Melanomas Oncogenic Pathway interactions Persons Pharmacology Play Porifera Prevention Property Regulation Repression Research Personnel Role Signal Transduction Stress TP53 gene Testing Therapeutic Tumor Suppressor Genes Tumor Suppressor Proteins Untranslated RNA base clinically translatable embryonic stem cell experimental study human disease improved in vivo in vivo Model innovation melanocyte melanoma melanomagenesis mouse model novel novel strategies overexpression patient derived xenograft model prevent promoter restoration targeted treatment therapeutic target transcription factor treatment strategy tumor tumor initiation tumorigenic

项目摘要

PROJECT SUMMARY The miR-29 family of microRNAs (miRNAs), consisting of miR-29a, miR-29b1, miR-29b2, and miR-29c, is encoded by two miRNA clusters (miR-29a~b1 and miR-29b2~c) and has tumor suppressive properties in a variety of human cancers. We found that, paradoxically, oncogenic BRAFV600E induces the transcription of both clusters in melanocytes through either MAPK pathway activation (miR-29a~b1) or oncogenic stress-mediated p53 activation (miR-29b2~c). This creates a barrier that prevents melanoma development, and we showed in a genetically engineered mouse model that inactivation of the miR-29 family with a sponge construct promotes melanoma progression. Interestingly, engineered human cell line models and human expression data from nevi and melanomas show that melanoma progression is associated with decreased expression of miR-29b2~c. Based on these findings we hypothesize that de-repression and overexpression of critical miR-29 targets promotes melanoma development and that such targets are vulnerabilities of melanoma that can be exploited for therapy. We have established a high-throughput genetically engineered mouse modeling platform to evaluate the role of miR-29 targets in melanoma and to assess their potential as therapeutic targets. In this proposal, we will extensively use this platform to first test if restoration of miR-29 expression and, thus, repression of all miR- 29 targets impairs melanoma development. We will further evaluate if silencing of a single miR-29 target elicits significant effects on melanomagenesis. We have identified the oncogenic transcription factor MYBL2 as a bona fide miR-29 target whose silencing reduces the aggressiveness of melanoma cell lines in vitro. We will test if de- repressed MYBL2 mediates the effects of miR-29 loss by silencing MYBL2 in a melanoma mouse model driven by miR-29 inactivation. Moreover, we will silence MYBL2 in a model with normal miR-29 activity to determine if MYBL2 is a general vulnerability of melanoma. To ensure relevance to human melanoma, we will complement these genetically engineered mouse models with experiments in human melanoma PDXs. Our proposed studies will use our newly developed mouse modeling platform as a pipeline to systematically evaluate if miR-29 targets in general, and MYBL2 specifically, have the potential to be further explored as therapeutic targets in melanoma.

项目概要 miR-29 的 microRNA (miRNA) 家族由 miR-29a、miR-29b1、miR-29b2 和 miR-29c 组成，是由两个 miRNA 簇（miR-29a~b1 和 miR-29b2~c）编码，具有肿瘤抑制特性多种人类癌症。我们发现，矛盾的是，致癌基因 BRAFV600E 诱导这两种基因的转录通过 MAPK 通路激活 (miR-29a~b1) 或致癌应激介导的黑色素细胞簇 p53 激活 (miR-29b2~c)。这创造了一个阻止黑色素瘤发展的屏障，我们在一项研究中表明基因工程小鼠模型，用海绵结构灭活 miR-29 家族促进黑色素瘤进展。有趣的是，工程化的人类细胞系模型和来自痣的人类表达数据黑色素瘤显示黑色素瘤进展与 miR-29b2~c 表达下降相关。基于这些发现，我们假设关键 miR-29 靶标的去抑制和过度表达促进黑色素瘤的发展，这些目标是黑色素瘤可以利用的弱点用于治疗。我们建立了高通量基因工程小鼠建模平台来评估 miR-29 靶点在黑色素瘤中的作用并评估其作为治疗靶点的潜力。在这个提案中，我们将广泛使用该平台来首先测试是否恢复 miR-29 表达，从而抑制所有 miR-29 29 个目标会损害黑色素瘤的发展。我们将进一步评估单个 miR-29 靶标的沉默是否会引发对黑色素瘤生成有显着影响。我们已经鉴定出致癌转录因子 MYBL2 是一种真正的致癌转录因子。 miR-29 的靶标，其沉默可降低体外黑色素瘤细胞系的侵袭性。我们将测试是否在黑色素瘤小鼠模型中，抑制 MYBL2 通过沉默 MYBL2 介导 miR-29 丢失的影响通过 miR-29 失活。此外，我们将在具有正常 miR-29 活性的模型中沉默 MYBL2，以确定是否 MYBL2 是黑色素瘤的普遍弱点。为了确保与人类黑色素瘤的相关性，我们将补充这些基因工程小鼠模型在人类黑色素瘤 PDX 中进行了实验。我们提出的研究将使用我们新开发的小鼠建模平台作为管道来系统地评估 miR-29 是否靶向一般来说，特别是 MYBL2，有可能作为黑色素瘤的治疗靶点得到进一步探索。