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

项目名称： RNA m6 A修饰在前列腺癌发病及耐药中的作用及机制 项目总结（摘要）： 背景：N6-甲基腺苷（m6 A）修饰是细胞内最普遍和最丰富的内部修饰， 真核生物的信使RNA（mRNAs）在正常的生物过程中发挥着多种重要的作用。 新的证据表明，m6 A和m6 A相关蛋白在人类疾病中也起着关键作用 包括癌症。前列腺癌（PCa）是最常见的癌症， 估计死亡在美国超过十年。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信号的异常调节， 信号通路与PCa的发病和进展密切相关。因此，雄激素剥夺疗法 (ADT)或靶向AR功能一直是治疗晚期前列腺疾病的主要策略。 癌的然而，目前的治疗策略不能治愈大多数PCa患者，由于耐药性， 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性被认为是本发明的主要问题。 治疗失败和PCa复发。因此，阐明其分子机制至关重要 PCa的发病机制和耐药性，这可能有助于开发有效的新的治疗方法 克服目前PCa治疗局限性的方法。RNA m6 A修饰由m6 A 由三个核心亚基L3、胃L14和WTAP组成的甲基转移酶复合物，并且可能具有 作为基因表达的重要调节因子，可以影响mRNA的转录、剪接、输出， 稳定性和翻译。以前，我们已经报道了胃L3催化m6 A修饰靶mRNA， 在包括PCa在内的人类癌症中起关键的致癌作用，但其在AR信号转导激活中的决定性作用 特别是在抗药性AR信号传导中仍然是难以捉摸的。我们的初步数据表明，m6 A修饰 增强子RNA（eRNA）是从非编码增强子元件转录的调节RNA， 也在前列腺癌耐药中发挥作用，需要进一步系统研究。这项提案的目的是 为了证明核心假设，即胃L3介导的RNA m6 A修饰在AR中起重要作用， 信号传导、PCa发病机制和耐药性。染色质相关调节因子m6 A甲基化 RNA（carRNA）和新生RNA将被映射，分析和与全基因组结合的相关性。 AR和转录机制。此外，YTHDC 1（一种核m6 A阅读蛋白）的作用将被系统性地研究。 研究其与AR反式激活和全基因组分布的关系，因为它具有 已知通过与m6 A修饰的RNA结合来调节转录激活或抑制。损失和 功能获得性实验将解决RNA m6 A修饰在控制细胞凋亡中的广泛影响。 AR信号传导和下游基因表达的激活，从而将有助于揭示 m6 A修饰在PCa发病机制中的作用机制。最后，m6 A修饰的效果 将使用异种移植物探索AR信号调节与耐药AR突变的操纵。 将使用PCa小鼠模型以及患者来源的异种移植物（PDX）模型和一组试验， 研究潜在的分子机制。我们提出的研究将成功揭示RNA m6 A的作用， AR信号转导和PCa发病机制的修饰，并帮助评估临床潜在的药理学作用。 抑制PCa中m6 A修饰，尤其是转移性和耐药性PCa。

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.