Retrovirus Evolution and Cancer

逆转录病毒进化与癌症

• 批准号: 9201540
• 负责人: JOHN M COFFIN
• 金额: $ 99万
• 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/9201540
• 关键词: Acquired Immunodeficiency Syndrome; Bioinformatics; Biology; Cancer cell line; Cells; Chromatin Structure; DNA; DNA Methylation; Databases; 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; Mining; Mus; Normal Cell; Patients; Pattern; 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; Work; Xenograft procedure; base; cDNA Library; cancer cell; env Gene Products; innovation; interest; man; member; 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.

本申请寻求对我的研究项目的持续支持，该项目由 NCI 资助近 40 年，其中 对逆转录病毒及其作用的生物学和进化做出了许多创新和高影响力的贡献 我们将重点关注实验室正在进行的一些项目： 1. 人类HERV-K (HML-2)多态性 HML-2是最近活跃的内源性逆转录病毒。 人类群体中，有 90 种或原病毒，其中 11 种均非传染性，已知在 为了测试活跃成员是否可能出现频率较低但未被发现，我们 开发了生物信息和实验室工具来识别和表征稀有物质 - 因此是最近的 和潜在的传染性 - 大型序列数据库中的原病毒，例如来自“1000个基因组”的原病毒 我们已经发现了一些新的原病毒，它们在人群中很少见，其中一种似乎是在- 我们将确定其编码传染性病毒的潜力以及在癌症中的可能作用。 2. HML-2 表达和癌症 为了测试 HML-2 原病毒在肿瘤发展中的作用，我们进行了研究。 开发了测序、生物信息和其他工具来研究它们在癌细胞系中的表达并阻止 我的 90 种原病毒中的哪些是通过什么机制表达的：即，前病毒中的差异是否 正常细胞和癌细胞之间的压迫是由于顺式（例如 DNA 甲基化、染色质结构）或反式造成的 （例如，转录因子模式）的影响。我们将确定这种上调是否可能导致重新整合。 新的 HML-2 原病毒的作用，以及它是否对肿瘤的发展和生长有积极的影响。 3. HML-2 表达和 HIV 感染 HIV-1 感染患者表现出 HML-2 表达水平升高。 我们正在开发与 2 中相同的方法来测试其细胞和原病毒特异性。 这些个体中的表达以及它是否在恶性肿瘤的发展中发挥作用。 4. HML-2受体特异性的进化 HML-2已经与灵长类动物共同进化了超过2000万个。 多年来，并可能受到类似于 ALV 的进化力量的影响，我们已经制定了一项策略来识别 根据用 HML-2 Env 蛋白假型化的病毒感染表达细胞的能力，确定其宿主受体 然后，我们将使用灵长类动物基因组中的“化石记录”来研究 cDNA 文库。 这种相互作用的共同进化。 5.逆转录病毒-APOBEC相互作用预防免疫缺陷的人类异种移植物的MLV感染。 小鼠是一个重要的问题，因为目前人们对开发“个性化免疫”小鼠以培养人类的兴趣 我们的策略是使受体小鼠全面表达限制。 因子（人 APOBEC3g）将防止移植的人肿瘤细胞系的生产性感染。 还致力于了解 MLV 对人类和小鼠 APOBEC 蛋白的不同敏感性。