Elucidating the role of somatic 3’UTR mutations in prostate cancer pathogenesis

阐明体细胞 3-UTR 突变在前列腺癌发病机制中的作用

• 批准号: 10580822
• 负责人: Samantha Lee Schuster
• 金额: $ 1.69万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2023-06-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10580822
• 关键词: 3' Untranslated Regions; Adenine; Affect; Apoptosis; Area; Binding; Binding Sites; Biological Assay; CRISPR/Cas technology; Cancer Etiology; Cancer Patient; Cancerous; Cell Line; Cells; Cessation of life; Characteristics; Code; Data; Databases; Disease; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genomic Segment; In Vitro; Individual; Laboratories; Lead; Location; Luciferases; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Mediator; Messenger RNA; MicroRNAs; Modeling; Molecular; Mutate; Mutation; Nucleic Acid Regulatory Sequences; Oncogenes; Oncogenic; Pathogenesis; Pathogenicity; Patients; Phenotype; Play; Post-Transcriptional Regulation; Process; Prognosis; Proliferating; Proteins; Publishing; RNA-Binding Proteins; Regulation; Regulatory Element; Reporter; Reporter Genes; Research; Role; Sampling; Site; Somatic Mutation; Testing; Tissue Sample; Tissues; Transcript; Translational Repression; Translations; United States; Variant; Work; base editor; cancer diagnosis; cancer genomics; cancer risk; castration resistant prostate cancer; cell motility; cohort; design; genome-wide; in vivo; insight; interest; mRNA Decay; mRNA Stability; mRNA Translation; men; mortality; new therapeutic target; novel; overexpression; p21 activated kinase; patient subsets; polysome profiling; posttranscriptional; ribosome profiling; therapeutic target; tumorigenesis

PROJECT SUMMARY Prostate cancer is the second most common cause of cancer mortality in US men due to our poor understanding and treatment of metastatic, castration-resistant prostate cancer (mCRPC). New insights into mechanisms of prostate cancer pathogenesis may be found within historically understudied aspects of gene expression, including post-transcriptional regulation of oncogenes. An important mediator of post-transcriptional gene regulation is the 3’ untranslated region (3’UTR), which plays critical roles in controlling mRNA stability and translation. The 3’UTR harbors a significant mutational burden across cancer patients; however, these mutations have not been extensively studied. Individual 3’UTR variants have been associated with cancer risk and shown to regulate expression in reporter assays, but it is still unclear whether these mutations significantly affect pathogenesis in patients and how widespread this dysregulation may be across cancer. By performing somatic mutation calling on a unique cohort of 230 prostate cancer patients, I have uncovered over 13,000 3’UTR somatic mutations in cancer tissues genome-wide. Many of these are in known cancer-related genes and 3’UTR regulatory motifs, indicating they have potential to affect oncogenic expression and cancer pathogenesis. Furthermore, I have determined via ribosome profiling of a subset of patient samples that ~40% of 3’UTR mutations are associated with changes in post-transcriptional gene regulation. Using individual reporter assays, I have validated that several of these patient-identified 3’UTR mutations in known cancer-related genes significantly alter gene expression. One of these functional mutations is in PAK2, a p21-activated kinase involved in cell motility, hyperactivation of which is oncogenic in several cancers. Based on these preliminary data, I hypothesize that 3’UTR mutations drive oncogenic changes in post-transcriptional gene regulation, such as overexpression of PAK2, that contribute to pathogenesis in prostate cancer patients. I now aim to define the molecular and pathogenic mechanisms by which this PAK2 3’UTR mutation increases PAK2 expression and exacerbates cancerous phenotypes in vitro and in vivo using an endogenous cell line model of the mutation. Additionally, I will investigate the extent of 3’UTR mutation-mediated gene dysregulation in cancer by performing a massively-parallel reporter assay to determine the effects of >6,000 patient-identified 3’UTR mutations on transcript levels and translation efficiency. These studies will establish 3’UTR mutations as important drivers of oncogenic dysregulation, expanding the field of functional cancer genomics and our understanding of prostate cancer pathogenesis, in addition to potentially identifying new therapeutic targets.

项目总结 前列腺癌是美国男性癌症死亡的第二大常见原因，因为我们对此缺乏了解 以及治疗转移性、耐去势前列腺癌(MCRPC)。对致病机制的新见解 前列腺癌的发病机制可以在历史上研究不足的基因表达方面发现， 包括癌基因的转录后调控。转录后基因的重要介体 调控是3‘端非翻译区(3’UTR)，在控制mRNA的稳定性和转录水平方面起着关键作用。 翻译。3‘非编码区在癌症患者中存在显著的突变负担；然而，这些突变 还没有得到广泛的研究。个体3‘非编码区变异与癌症风险有关，并显示 来调节报告分析中的表达，但目前还不清楚这些突变是否会显著影响 患者的发病机制，以及这种调节失调在癌症中的普遍程度。通过执行躯体手术 突变呼唤230名前列腺癌患者的独特队列，我发现了超过13,000个3‘非编码区体细胞 全基因组癌症组织中的突变。其中许多存在于已知的癌症相关基因和3‘非编码区 调控基序，表明它们有可能影响癌基因表达和癌症发病机制。 此外，我已经通过对患者样本子集的核糖体分析确定，~40%的3‘UTR 突变与转录后基因调控的变化有关。使用单独的记者分析， 我已经证实了其中几个患者在已知癌症相关基因中发现了3‘UTR突变 显著改变基因表达。其中一个功能突变是在PAK2中，它是一种p21激活的激酶 在细胞运动中，它的过度激活在几种癌症中是致癌的。根据这些初步数据，我 假设3‘非编码区突变导致转录后基因调控的致癌变化，例如 PAK2的过度表达参与了前列腺癌患者的发病机制。我现在的目标是定义 PAK2 3‘非编码区突变增加PAK2表达和致病机制的分子和致病机制 使用这种突变的内源性细胞系模型，在体外和体内加剧癌症的表型。 此外，我还将研究3‘非编码区突变在癌症中介导的基因失调的程度。 一项大规模平行报告试验，以确定6,000名患者识别的3‘UTR突变对人骨肉瘤细胞的影响 笔录水平和翻译效率。这些研究将建立3‘非编码区突变为重要驱动因素 致癌失调，拓展功能癌基因组学领域和我们对前列腺癌的认识 除了潜在地确定新的治疗靶点外，还可以研究癌症的发病机制。