Reverse transcriptase inhibitor effects on the mobilome of colon cancer

项目摘要 RNA测序的大规模转录分析揭示了重复序列的丰富表达。 与正常组织相比，癌症中的RNA元素。此外，这些重复RNA中的许多能够 通过逆转录（RT）逆转录转座，这导致癌症基因组的改变。我们 临床前细胞系和小鼠模型已经证明了这些内源性RT被阻断的能力， 核苷逆转录酶抑制剂（NRTI）通常用于病毒感染。有趣的是， 在3D肿瘤球体中生长的多种结直肠癌（CRC）细胞系中发现了NRTI的抗癌作用， 在小鼠体内异种移植物中。这导致了NRTI 3TC在转移性CRC中的II期临床试验的开始 获得治疗前和治疗后活检的患者用于相关科学研究。 Ting实验室在细胞系和动物模型方面具有专业知识，可以研究NRTI在CRC中的作用， RNA原位杂交方法来定量这些系统中的重复元件。格林鲍姆实验室 开发了计算工具来表征重复RNA表达及其对先天免疫的影响。的 帕克实验室开创了研究癌症基因组中DNA逆转录转座事件的工具。将合并的 Ting、Greenbaum和Park实验室研究重复RNA的专业知识和工具， 让我们实现这个项目的目标。 我们计划： 1)了解3TC对CRC模型系统中细胞应答和重复RNA表达的影响 2)表征3TC对异种移植模型中重复逆转录转座的影响 3)评估患者的重复RNA表达、逆转录转座和肿瘤免疫微环境 从用3TC治疗的CRC患者获得的活检

PROJECT SUMMARY Large scale transcriptional analysis with RNA-sequencing has revealed the abundant expression of repetitive element RNAs in cancers compared to normal tissues. Moreover, many of these repeat RNAs are capable of retrotransposition through reverse transcription (RT), which leads to alterations in the cancer genome. Our preclinical cell line and mouse models have demonstrated the ability of these endogenous RT to be blocked by nucleoside reverse transcriptase inhibitors (NRTIs) commonly used for viral infections. Interestingly, we have found anti-cancer effects of NRTIs in multiple colorectal cancer (CRC) cell lines grown in 3D tumor spheres and in mouse xenografts in vivo. This led to the initial of a Phase II clinical trial of the NRTI 3TC in metastatic CRC patients where pre- and post-treatment biopsies are being obtained for correlative science research. The Ting lab has expertise in cell line and animal models to study NRTI effects in CRC and the sequencing and RNA in situ hybridization methods to quantify repeat elements in these systems. The Greenbaum lab has developed computational tools to characterize repeat RNA expression and their effects on innate immunity. The Park lab has pioneered the tools to study DNA retrotransposition events in cancer genomes. The combined expertise and tools generated to study repeat RNAs by the Ting, Greenbaum, and Park labs uniquely positions us to execute the goals of this project. We plan to: 1) Understand the effects of 3TC on cellular response and repeat RNA expression in CRC model systems 2) Characterize the effect of 3TC on repeat retrotransposition in xenograft models 3) Evaluate repeat RNA expression, retrotransposition, and the tumor immune microenvironment from patient biopsies obtained from CRC patients treated with 3TC