Retrovirus Evolution and Cancer

逆转录病毒进化与癌症

• 批准号: 10246396
• 负责人: JOHN M COFFIN
• 金额: $ 79.98万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-20 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246396
• 关键词: APOCEC3G gene; Acquired Immunodeficiency Syndrome; Bioinformatics; Biology; Cancer cell line; Cells; Chromatin Structure; DNA; DNA Methylation; Development; Endogenous Retroviruses; Evolution; Exhibits; Fossils; Frequencies; Funding; Genetic Polymorphism; Genome; Goals; Growth and Development function; HIV Infections; HIV-1; Human; Human Genome; Immune; Immunodeficient Mouse; Individual; Infection; Laboratories; Lead; Malignant Neoplasms; Mus; Normal Cell; Patients; Pattern; Peripheral Blood Mononuclear Cell; Play; Population; Prevention; Primates; Principal Investigator; Proteins; Proviruses; Research; Research Personnel; Research Support; Retroviridae; Role; Specificity; Testing; Therapeutic; Tissues; Tumor Cell Line; Up-Regulation; Virus; Xenograft procedure; base; cDNA Library; cancer cell; env Gene Products; innovation; interest; man; member; molecular sequence database; mouse genome; prevent; programs; receptor; tool; transcription factor; tumor

This application seeks continued support for my research program, funded by the NCI for almost 40 years, with numerous innovative and high-impact contributions to the biology and evolution of retroviruses and their roles in AIDS and cancer. We will focus on a number of ongoing projects in the laboratory: 1. HERV-K (HML-2) Polymorphism in humans. HML-2 is the most recently active endogenous retrovirus group in humans, with 90 or proviruses, of which 11, all noninfectious, were known to be polymorphic in the population. To test whether active members may be present at low frequency, but remain undetected, we have developed bioinformatic and laboratory tools to identify and characterize rare - and therefore more recent and potentially infectious - proviruses in large sequence databases like those from the the “1000 genomes” project. We have identified a number of new proviruses, rare in the population, one of which appears to be in- tact. We will determine its potential to encode infectious virus and possible role in cancer. 2. HML-2 Expression and cancer. To test for a role of HML-2 proviruses in tumor development, we have developed sequencing, bioinformatic and other tools to study their expression in cancer cell lines and deter- mine which of the 90 proviruses are expressed and by what mechanism: i.e., whether the differences in ex- pression between normal and cancer cells are due to cis (e.g., DNA methylation, chromatin structure) or trans (e.g., transcription factor patterns) effects. We will determine whether this upregulation might lead to reintegra- tion of new HML-2 proviruses, and whether it has a positive effect on tumor development and growth. 3. HML-2 expression and HIV infection. HIV-1 infected patients exhibit elevated levels of HML-2 expres- sion in PBMCs. We are developing the same approaches as in 2 to test the cell and provirus specificity of this expression in these individuals and whether it plays any role in the development of malignancy. 4. Evolution of HML-2 receptor specificity. HML-2 has been coevolving with primates for over 20 million years and has likely been subject to evolutionary forces similar to ALV. We have developed a strategy to iden- tify its host receptor based on the ability of viruses pseudotyped with HML-2 Env protein to infect cells express- ing a cDNA library from permissive cells. We will then use the “fossil record” in primate genomes to study the coevolution of this interaction. 5. Retrovirus-APOBEC interaction. Prevention of MLV infection of human xenografts in immunodeficient mice is an issue of importance due the current interest in developing “personalized immune” mice to grow hu- man tissues for therapeutic purposes. Our strategy is to make the recipient mice globally express a restriction factor (human APOBEC3g) that will prevent productive infection of the grafted human tumor cell line. We are also working to understand the differential sensitivity of MLV to human and murine APOBEC proteins.

该申请寻求继续支持我的研究计划，由NCI资助了近40年， 逆转录病毒及其角色的生物学和进化的许多创新和高影响力的贡献 在艾滋病和癌症中。我们将专注于实验室中的许多正在进行的项目： 1。人类中的HERV-K（HML-2）多态性。 HML-2是最近活跃的内源性逆转录病毒 人类中的组，有90个或病毒病毒，其中11个（所有非感染性）在该人中是多态性的 人口。要测试是否可能以低频存在活跃成员，但仍未发现，我们 已经开发了生物信息学和实验室工具来识别和表征稀有的特征 - 因此 以及潜在的传染病 - 大序列数据库中的病毒，例如“ 1000个基因组” 项目。我们已经确定了许多新的病毒病毒，在人口中很少见，其中之一似乎是 机智。我们将确定其编码传染病和可能在癌症中的作用的潜力。 2。HML-2表达和癌症。为了测试HML-2病毒在肿瘤发育中的作用，我们有 开发了测序，生物信息学和其他工具，以研究其在癌细胞系中的表达并确定 - 矿山表达了90个预科病毒中的哪种以及通过什么机制： 正常细胞和癌细胞之间的压力是由于顺式（例如DNA甲基化，染色质结构）或反式 （例如，转录因子模式）效应。我们将确定这种上调是否可能导致重新集成 新的HML-2病毒及其是否对肿瘤发育和生长有积极影响。 3。HML-2表达和HIV感染。 HIV-1感染患者的HML-2表达水平升高 - PBMC中的sion。我们正在开发与2中相同的方法来测试此细胞的特异性 这些人的表达以及它是否在恶性发展中起任何作用。 4。HML-2受体特异性的进化。 HML-2与初级共​​同发展超过2000万 几年，可能受到类似ALV的进化力量的影响。我们已经制定了一种策略来识别 根据HML-2 Env蛋白对感染细胞进行伪型的病毒的能力来对其宿主受体进行表达 - 从宽松细胞中获得cDNA文库。然后，我们将使用灵长类动物基因组中的“化石记录”来研究 这种相互作用的协同进化。 5。逆转录病毒-Apobec相互作用。预防免疫缺陷中人类异种移植物的MLV感染 小鼠是一个重要的问题，这是由于目前有利于开发“个性化免疫”小鼠来增加hu-的兴趣 人体组织用于治疗目的。我们的策略是使接收者小鼠在全球范围内表达限制 因子（人apobec3g）将防止移植的人类肿瘤细胞系的生产性感染。我们是 还致力于了解MLV对人和鼠Apobec蛋白的差异敏感性。