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.

项目摘要 MiR-29 MicroRNA家族（miRNA），由miR-29a，miR-29b1，miR-29b2和miR-29c组成由两个miRNA簇（miR-29a〜b1和miR-29b2〜c）编码，并在A中具有肿瘤抑制特性人类癌症的多样性。我们发现，矛盾的是，致癌Brafv600E引起了两者的转录通过MAPK途径激活（miR-29a〜b1）或致癌应力介导的黑色素细胞中的簇。 p53激活（miR-29b2〜c）。这产生了阻止黑色素瘤发育的障碍，我们在用海绵结构灭活miR-29家族的基因工程小鼠模型会促进黑色素瘤进展。有趣的是，来自NEVI的工程人类细胞系模型和人类表达数据黑色素瘤表明黑色素瘤的进展与miR-29b2〜c的表达降低有关。基于这些发现，我们假设关键miR-29目标的消除抑制和过表达促进黑色素瘤的发育，这种靶标是可以被利用的黑色素瘤的脆弱性用于治疗。我们已经建立了一个高通量基因工程的鼠标建模平台来评估 miR-29靶标在黑色素瘤中的作用并评估其作为治疗靶标的潜力。在这个建议中，我们如果miR-29表达的恢复，并抑制了所有miR- 29个目标会损害黑色素瘤的发育。我们将进一步评估单个miR-29目标引起的沉默是否对黑色素作用的显着影响。我们已经将致癌转录因子mybl2确定为bona FIDE miR-29靶标的沉默在体外降低了黑色素瘤细胞系的攻击性。我们将测试是否de- 抑制MYBL2通过在黑色素瘤小鼠模型驱动的黑色素瘤中介导miR-29损失的影响通过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，以确定是否是否 MYBL2是黑色素瘤的普遍脆弱性。为了确保与人黑色素瘤相关，我们将补充这些基因工程的小鼠模型与人类黑色素瘤PDX的实验。我们提出的研究将使用我们新开发的鼠标建模平台作为管道，以系统地评估miR-29目标是否目标通常，特别是MYBL2，有可能进一步探索黑色素瘤的治疗靶标。