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Characterization of the oncogenic potential of circRNAs in melanoma

黑色素瘤中 circRNA 致癌潜力的表征

基本信息

批准号：
10355644
负责人：
Florian Karreth
金额：
$ 23.63万
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：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10355644
关键词：
Affect Alternative Splicing Award Back Benign Biological Biological Assay Biological Process Biology Cell Culture Techniques Cell Line Cells Development Disease Ectopic Expression Exhibits Future Generations Genes Genetic Genetic Transcription Genetically Engineered Mouse Growth Homeostasis Human In Vitro Maintenance Malignant Neoplasms Melanoma Cell Messenger RNA MicroRNAs Modeling Molecular Mus Oncogenes Oncogenic PMS1 gene Pattern Peptides Play Porifera Positioning Attribute Process Property Proteins RNA RNA Interference RNA Splicing Regulation Role Series Testing Time Tissues Translating Translations Untranslated RNA Work Xenograft Model Xenograft procedure base cancer type cell growth cell type circular RNA covalent bond differential expression experimental study in vivo mRNA Precursor melanocyte melanoma melanomagenesis mouse model mutant novel novel therapeutic intervention overexpression small hairpin RNA tool transcriptome sequencing tumor tumorigenesis tumorigenic vector

项目摘要

SUMMARY Over the last decade noncoding RNAs (ncRNAs) have emerged as prominent players that are involved in the regulation of most, if not all, biological processes. Given their important roles, it is conceivable that ncRNAs contribute to the development of diseases such as cancer. Circular RNAs (circRNAs) are ncRNAs that are formed through alternative splicing of linear pre-mRNAs via a process termed backsplicing. circRNAs are abundant, stable, and evolutionarily conserved and display cell-type specific expression patterns, indicating that they may play important roles in cell homeostasis, differentiation, development, and disease. However, the formation of circRNAs, their regulation, and their functions are poorly understood, partially due to the lack of tools to drive permanent ectopic expression of circRNAs. We have developed genetic tools for stable overexpression of candidate circRNAs in vitro and in vivo, which we will implement as part of this R21 proposal. Using these tools, we will determine the oncogenic potential of two circRNAs we hypothesize to promote melanomagenesis. We identified circPMS1 and circDNAJC2 as candidate melanoma oncogenes by a comprehensive circRNA expression analysis in benign melanocytes and malignant melanoma cells. We will test if overexpression of these circDNAs promotes melanocyte transformation in vitro and tumor formation in our high-throughput melanoma mouse models. We will also begin the molecular characterization of the functions underlying circPMS1 and circDNAJC2 oncogenic activity, with an emphasis on peptide translation and miRNA sponging. Moreover, we will specifically silence circPMS1 and circDNAJC2 in human melanoma cells in vitro as well as in xenograft models to examine if circRNA expression is required for maintaining the transformed state and continued melanoma growth, respectively. Our proposed studies will demonstrate that overexpression of circPMS1 or circDNAJC2 drives melanomagenesis and establish an easily adaptable experimental framework that will pave the way for future studies of oncogenic circRNAs.

概括在过去的十年中，非编码 RNA (ncRNA) 已成为参与以下领域的重要参与者：大多数（如果不是全部）生物过程的调节。鉴于 ncRNA 的重要作用，可以想象有助于癌症等疾病的发展。环状 RNA (circRNA) 是 ncRNA，线性前体 mRNA 通过称为反向剪接的过程进行选择性剪接而形成。 circRNA 是丰富、稳定、进化保守，并显示细胞类型特异性表达模式，表明它们可能在细胞稳态、分化、发育和疾病中发挥重要作用。然而，人们对 circRNA 的形成、调控和功能知之甚少，部分原因是缺乏工具驱动 circRNA 的永久异位表达。我们开发了用于稳定过度表达的遗传工具我们将在 R21 提案中实施候选 circRNA 的体外和体内研究。使用这些工具，我们将确定我们假设促进黑色素瘤生成的两种 circRNA 的致癌潜力。我们通过综合 circRNA 将 circPMS1 和 circDNAJC2 鉴定为候选黑色素瘤癌基因良性黑色素细胞和恶性黑色素瘤细胞中的表达分析。我们将测试这些是否过度表达 circDNA 促进体外黑色素细胞转化和高通量黑色素瘤中的肿瘤形成鼠标模型。我们还将开始对 circPMS1 和 circDNAJC2 致癌活性，重点是肽翻译和 miRNA 海绵作用。此外，我们将在体外和异种移植物中特异性沉默人黑色素瘤细胞中的 circPMS1 和 circDNAJC2 模型来检查 circRNA 表达是否需要维持转化状态并继续黑色素瘤生长，分别。我们提出的研究将证明 circPMS1 或 circDNAJC2 驱动黑色素瘤生成并建立一个易于适应的实验框架，为未来研究致癌circRNA的方法。