Mechanisms of translational output control in pancreatic cancer

项目摘要 我的长期职业目标是领导一个富有成效的学术研究小组，通过 开展有影响力的癌症生物学研究，并指导下一代敬业的科学家。我是 特别感兴趣的是胰腺导管癌的发展机制和对治疗的抵抗， 腺癌（PDAC）。PDAC是最致命的疾病之一，其生存率尚未提高 在过去的25年里，目前对PDAC患者没有有效的治疗方法。NCI，在 根据顽固性癌症研究法，建立了一个科学框架，其中最高 优先事项是开发靶向治疗和疗法，以克服目前对 可用的代理人。我们提出的研究计划通过识别认识不足的非- 组蛋白赖氨酸甲基转移酶胃L10作为eEF 1A（真核细胞延伸因子1）关键调节因子 α）mRNA翻译机制的基本非核糖体组分。失调 蛋白质的产生是癌症的标志，与异常的细胞增殖、存活和细胞周期的改变有关。 免疫反应和癌症能量学。K99/R 00提案的首要目标是 eEF 1A的赖氨酸甲基化调节蛋白质合成的速率，这是最消耗能量的 在细胞中的过程，并在人类癌症生长中起着关键作用。 目的1的目的是阐明胃L10在致癌KRAS驱动的胰腺癌中的作用。 我们将检验这一假设，即胃L10通过其甲基化活性与KRAS信号传导合作， 使用胰腺的小鼠模型促进癌细胞在体内的无限扩增，其中KRAS 通道经常被激活。我们还将研究胃L10在人体组织中的致瘤作用 使用患者来源的异种移植物（PDX）模型。接下来，我们将研究甲基化在特定的 在PDAC小鼠模型中使用核糖体分析技术进行蛋白质生产。最后，我们将探讨 在临床前模型中，胃L10消融联合MAP激酶抑制剂的潜在协同作用 胰腺癌在目的2中，我们将表征胃L10的生理催化活性及其在体外的生物活性。 在体外调控eEF 1A活性和细胞中mRNA翻译生物学中的分子功能。我们将 还研究了胃L10和甲基化eEF 1A相互作用的伴侣，以及这些途径如何交叉， 影响癌细胞表型。 K99/R 00培训奖将使我能够执行这个变革性项目，进一步发展我的 目前的小鼠遗传学，mRNA翻译生物学技能，学习体外和体内的新技术 核糖体分析同时也让我获得了胰腺癌临床方面的知识 以及生化信号和综合系统生物学方面的新专业知识。

PROJECT SUMMARY My long-term career goal is to lead a productive academic research group, promoting science by conducting impactful cancer biology research and mentoring the next generation of dedicated scientists. I am particularly interested in the mechanisms of cancer development and resistance to therapy in pancreatic ductal adenocarcinoma (PDAC). PDAC is one of the deadliest disease for which survival has not improved substantially over the past 25 years. There is currently no effective treatment for PDAC patients. The NCI, in accordance to the Recalcitrant Cancer Research Act, established a scientific framework in which the top priorities are the development of targeted therapeutics and therapies to overcome resistance to currently available agents. Our proposed research plan addresses these priorities by identifying poorly recognized non- histone lysine methyltransferase METTL10 as a critical regulator of eEF1A (eukaryotic elongation factor 1 alpha) a fundamental, non-ribosomal component of the mRNA translational machinery. Dysregulation of protein production is a hallmark of cancer and is linked to aberrant cell proliferation, survival, and alterations in both immune responses and cancer energetics. An overarching goal of this K99/R00 proposal focuses on the idea that lysine methylation of eEF1A regulates the rate of protein synthesis, the most energy-consuming process in the cell, and plays a critical role in human cancer growth. The goal of Aim 1 is to elucidate the role of METTL10 in pancreatic cancer driven by oncogenic KRAS. We will test the hypothesis that METTL10, via its methylation activity, cooperates with KRAS signaling to promote the unlimited expansion of cancer cells in vivo using mouse models of pancreas, in which KRAS pathway is frequently activated. We will also investigate the tumorigenic role of METTL10 in human tissue using patient-derived xenograft (PDX) models. Next, we will investigate the role of methylation on specific protein production using ribosome profiling techniques in mouse models of PDAC. Finally, we will explore potential synergies of METTL10 ablation in combination with inhibitors of MAP-kinases in pre-clinical models of pancreatic cancers. In Aim 2 we will characterize the physiologic catalytic activity of the METTL10 and its molecular functions in the regulation of eEF1A activity in vitro and mRNA translation biology in cells. We will also investigate the METTL10 and methylated eEF1A interacting partners and how these pathways intersect to influence cancer cell phenotypes. A K99/R00 training award will allow me to carry out this transformative project, further developing my current skills in mouse genetics, mRNA translation biology, learn new techniques for in vitro and in vivo ribosome profiling analysis while also allowing me to acquire knowledge in clinical aspects of pancreatic cancer and new expertise in biochemical signaling and integrative systems biology.