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）已成为参与其中的杰出参与者大多数（如果不是全部）生物过程的调节。鉴于它们的重要作用，可以想象有助于癌症等疾病的发展。圆形RNA（CIRCRNA）是NCRNA 通过称为后平面的过程对线性PREMRNA的替代剪接形成。 Circrnas是丰富，稳定和进化保守并显示细胞类型的特定表达模式，表明它们可能在细胞稳态，分化，发育和疾病中起重要作用。但是，形成circrnas，他们的调节及其功能的理解很少，部分原因是缺乏工具驱动circrNA的永久异位表达。我们开发了稳定过表达的遗传工具在体外和体内的候选circrnas的大量，我们将作为R21提案的一部分实施。使用这些工具，我们将确定我们假设的两个ciRCRNA的致癌潜力，以促进黑素作用。我们将Circpms1和CircDNAJC2鉴定为候选黑色素瘤肿瘤基因。良性黑素细胞和恶性黑色素瘤细胞中的表达分析。我们将测试是否过表达这些 CircDNA在我们的高通量黑色素瘤中促进体外中黑素细胞转化和肿瘤形成鼠标模型。我们还将开始对Circpms1和 CircDNAJC2致癌活性，重点是肽翻译和miRNA sponging。而且，我们将在人类黑色素瘤细胞的体外和异种移植物中特异性沉默CippMS1和CircDNAJC2 模型检查是否需要ciRCRNA表达来维持转化状态并继续黑色素瘤的生长。我们提出的研究将表明Circpms1或 CircDNAJC2驱动黑色素作和建立一个易于适应的实验框架，可以铺路未来研究致癌性circrnas的方式。