The role and mechanism of RNA m6A modification in the pathogenesis and drug-resistance of prostate cancer

RNA m6A修饰在前列腺癌发病及耐药中的作用及机制

基本信息

批准号：
10638634
负责人：
Jihoon Lee
金额：
$ 38.55万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10638634
关键词：
Address Adenosine Affect Androgen Receptor Androgens Binding Biological Assay Biological Process Cancer Patient Castration Cessation of life ChIP-seq Chromatin Clinical Complex Data Deposition Development Disease Drug resistance Enhancers Gene Expression Genetic Enhancer Element Genetic Transcription Goals Growth Hormones Human Link Malignant Neoplasms Malignant neoplasm of prostate Maps Mediating Messenger RNA Methylation Methyltransferase Modification Molecular Mutation Nitrogen Nuclear Oncogenic Pathogenesis Phase Phenotype Physical condensation Play Positioning Attribute Prostate Prostate Cancer therapy Proteins RNA RNA Splicing Reader Receptor Signaling Regulation Regulator Genes Relapse Reporting Resistance Role Signal Pathway Site-Directed Mutagenesis Testing Therapeutic Transactivation Transcriptional Activation Transcriptional Regulation Translations Treatment Failure United States Untranslated RNA Xenograft procedure abiraterone advanced prostate cancer androgen deprivation therapy antagonist cancer diagnosis cancer survival castration resistant prostate cancer drug relapse drug resistance development enzalutamide experimental study gain of function gene repression genome-wide human disease loss of function methylome mouse model novel therapeutic intervention patient derived xenograft model pharmacologic prostate cancer cell prostate cancer progression receptor binding receptor function receptor-mediated signaling transcriptome transcriptome sequencing tumor

项目摘要

PROJECT TITLE: The role and mechanism of RNA m6A modification in the pathogenesis and drug-resistance of prostate cancer PROJECT SUMMARY (ABSTRACT): Background: N6-methyladenosine (m6A) modification is the most prevalent and abundant internal modification in eukaryotic messenger RNAs (mRNAs) and plays diverse and important roles in normal biological processes. Emerging evidence suggest that m6A and m6A-assicated proteins also play critical roles in human diseases including cancers. Prostate cancer (PCa) is the most commonly diagnosed cancer with the second leading estimated deaths at the United States for more than a decade. Androgens and androgen receptor (AR) regulate normal growth and function of the prostate gland. However, aberrant regulation of androgens and AR signaling pathway are closely associated with PCa pathogenesis and progression. Thus, androgen deprivation therapies (ADT) or targeting AR function have always been the mainstay of therapeutic strategy against advanced prostate cancers. However, the current therapeutic strategies cannot cure most PCa patients due to drug resistance and relapse, and eventually develop metastatic, castration-resistant prostate cancer (mCRPC), a lethal stage of the disease. The acquired resistance to abiraterone and enzalutamide is considered as the main problem for the treatment failure and relapse of PCa. Therefore, it is crucial to elucidate the molecular mechanisms underlying PCa pathogenesis and drug resistance, which may contribute to the development of effective new therapeutic approaches to overcome the limitations of current treatment for PCa. RNA m6A modification is deposited by m6A methyltransferase complex composed of three core subunits METTL3, METTL14 and WTAP and may have functions as an important regulator of gene expression that can affect mRNA transcription, splicing, export, stability, and translation. Previously, we have reported that METTL3-catalyzed m6A modification of target mRNA plays critical oncogenic roles in human cancers including PCa, but its definitive roles in AR signaling activation and especially in drug-resistant AR signaling remain elusive. Our preliminary data imply that m6A modification on enhancer RNAs (eRNAs), which are regulatory RNAs transcribed from non-coding enhancer elements, may also have roles in drug resistance in PCa, which requires further systematic studies. The goal of this proposal is to prove the central hypothesis that METTL3-mediated RNA m6A modification plays essential roles in AR signaling, PCa pathogenesis and drug resistance. The m6A methylome of chromatin-associated regulator RNAs (carRNAs) and nascent RNAs will be mapped, analyzed and correlated with the genome-wide binding of AR and transcriptional machineries. Also, the role of YTHDC1, a nuclear m6A reader protein, will be systemically examined to investigate its relationship with the transactivation of AR and genome-wide distribution, as it has been known to regulate transcriptional activation or repression via binding to m6A-modified RNAs. Loss- and gain-of-function experiments will address the widespread impact of RNA m6A modification in controlling the activation of AR signaling and downstream gene expression, and thus will help uncover the molecular mechanism underlying the role of m6A modification in PCa pathogenesis. Finally, the effects of m6A modification manipulation in the AR signaling regulation with drug-resistant AR mutations will be explored using a xenograft PCa mouse model, as well as a patient-derived xenograft (PDX) models and a panel of assays will be placed to examine the underlying molecular mechanism. Our proposed study will successfully reveal the role of RNA m6A modification in AR signaling and PCa pathogenesis, and help evaluate the clinical potential pharmacological inhibition of m6A modification in PCa, especially metastatic and drug resistant PCa.

项目名称： RNA m6A修饰在前列腺癌发病及耐药中的作用及机制项目概要（摘要）：背景：N6-甲基腺苷（m6A）修饰是最普遍和最丰富的内部修饰真核生物信使 RNA (mRNA) 在正常生物过程中发挥着多种重要作用。新证据表明 m6A 和 m6A 相关蛋白在人类疾病中也发挥着关键作用包括癌症。前列腺癌 (PCa) 是最常诊断的癌症，位居第二位十多年来美国的估计死亡人数。雄激素和雄激素受体 (AR) 调节前列腺的正常生长和功能。然而，雄激素和 AR 信号传导的异常调节途径与 PCa 的发病机制和进展密切相关。因此，雄激素剥夺疗法 (ADT) 或靶向 AR 功能一直是晚期前列腺治疗策略的支柱癌症。然而，由于耐药性和耐药性，目前的治疗策略无法治愈大多数 PCa 患者。复发，并最终发展为转移性去势抵抗性前列腺癌 (mCRPC)，这是前列腺癌的致命阶段疾病。对阿比特龙和恩杂鲁胺的获得性耐药被认为是该病的主要问题。 PCa 治疗失败和复发。因此，阐明其背后的分子机制至关重要 PCa 发病机制和耐药性，可能有助于开发有效的新疗法克服当前 PCa 治疗局限性的方法。 RNA m6A 修饰由 m6A 沉积甲基转移酶复合体由三个核心亚基 METTL3、METTL14 和 WTAP 组成，可能具有作为基因表达的重要调节因子，可以影响 mRNA 转录、剪接、输出、稳定性和翻译。此前，我们曾报道过 METTL3 催化 m6A 修饰靶标 mRNA 在包括 PCa 在内的人类癌症中发挥重要的致癌作用，但其在 AR 信号传导激活中发挥决定性作用尤其是在耐药 AR 信号传导方面仍然难以捉摸。我们的初步数据表明 m6A 修饰增强子 RNA (eRNA) 是从非编码增强子元件转录而来的调节性 RNA，可能也与 PCa 耐药有关，这需要进一步的系统研究。该提案的目标是证明 METTL3 介导的 RNA m6A 修饰在 AR 中发挥重要作用的中心假设信号传导、PCa 发病机制和耐药性。染色质相关调节因子的 m6A 甲基化组 RNA (carRNA) 和新生 RNA 将被绘制、分析并与全基因组结合相关联 AR 和转录机器。此外，YTHDC1（一种核 m6A 阅读器蛋白）的作用将被系统地检查其与 AR 反式激活和全基因组分布的关系，因为它已经已知可通过与 m6A 修饰的 RNA 结合来调节转录激活或抑制。损失-和功能获得实验将解决 RNA m6A 修饰在控制 AR信号传导和下游基因表达的激活，从而有助于揭示分子 m6A 修饰在 PCa 发病机制中的作用机制。最后是m6A修改后的效果将使用异种移植物探索耐药 AR 突变对 AR 信号调节的操纵 PCa 小鼠模型以及患者来源的异种移植 (PDX) 模型和一组检测将被用于检查潜在的分子机制。我们提出的研究将成功揭示 RNA m6A 的作用 AR 信号传导和 PCa 发病机制的修饰，并有助于评估临床潜在药理学抑制 PCa 中的 m6A 修饰，尤其是转移性和耐药性 PCa。