Replication of Human Endogenous Retroviruses in Modern Humans

人内源性逆转录病毒在现代人类中的复制

• 批准号: 8550159
• 负责人: David Michael Markovitz
• 金额: $ 8.26万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-28 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8550159
• 关键词: Address; Antibiotic Resistance; Applications Grants; Autoimmune Diseases; Autoimmunity; Bioinformatics; Blood; Cancer Biology; Cancer Patient; Cell Line; Cells; Cloning; Computing Methodologies; DNA Sequence; Disease; Elements; Endogenous Retroviruses; Gene Expression; Gene Order; Genes; Genetic; Genetic Materials; Genome; Genomics; Green Fluorescent Proteins; HERVs; Health; Human; Human Genetics; Human Genome; Individual; Inherited; Junk DNA; Laboratories; Lead; Length; Life; Link; Lymphoma; Malignant Neoplasms; Methodology; Methods; Molecular; Mutation; Normal tissue morphology; Nuclear Magnetic Resonance; Pathogenesis; Patients; Proteins; RNA; Research Personnel; Role; Safety; Surrogate Markers; System; Techniques; Teratocarcinoma; Testing; Thinking; Tumor Tissue; Vascular blood supply; Virus; cell type; conformational conversion; human DNA sequencing; human disease; in vivo; malignant breast neoplasm; melanoma; nervous system disorder; novel diagnostics; novel therapeutics; transmission process; tumorigenesis

DESCRIPTION (provided by applicant): The functions of the approximately 98% of the human genome that do not encode human cellular proteins remain to be elucidated. Actively replicating endogenous retroviruses entered the human genome millions of years ago and became a stable part of the inherited genetic material, with retroviral elements presently making up approximately 8% of the modern human genome. These viruses subsequently acquired multiple mutations, leading to the widely-held assumption that they are no longer competent to replicate. However, in studying living patients rather than the standard cell lines, we have recently discovered surprising evidence suggesting that in certain patients with cancer HERV-K (HML-2), an endogenous retrovirus that is a relatively recent entrant into the human genome and has been linked to oncogenesis, might still be capable of replication. Replication and transmission of endogenous retroviruses is difficult to prove using standard techniques, however, as these viruses are already present in the genomes of all human cells. Therefore, we have assembled a diverse and expert group of investigators who will test the hypothesis that modern HERV-K (HML-2) can replicate using cutting-edge and complementary techniques. We will use a newly devised molecular system in which antibiotic resistance serves as a surrogate marker to assess whether we can passage virus from the blood of patients in the laboratory. We will also use high-throughput DNA sequencing and Bioinformatics to find full-length functional virus that is present in cancer patients but is not represented in the current draft of the human genome. An infectious clone that is representative of modern HERV-K (HML-2) will be made by using the information gained from the Bioinformatics studies and/or by cloning a full-length virus from the HERV-K RNA found in the blood of patients. This clone will then be tagged with green fluorescent protein and used to directly study replication. Building on our advances using Nuclear Magnetic Resonance (NMR) to visualize spatially correlated dynamics that direct RNA and protein conformational transitions, we will also develop NMR methods to visualize HERV-K (HML-2) replication in vivo. Proof that endogenous retroviruses can still replicate in modern humans will lead to a paradigm shift in thinking about these viruses, and will suggest a role for them in reshaping individual genomes. In addition, as increased expression of chromosomal endogenous retroviral sequences has been linked to cancer and autoimmunity, these findings will be relevant to understanding the pathogenesis of significant diseases. Finally, if replicating, infectious endogenous human retroviruses capable of causing disease can be found in the blood of given individuals, this information will have important implications for the safety of the blood supply.

描述（由申请人提供）：未编码人类细胞蛋白的大约98％人类基因组的功能仍有待阐明。积极复制内源性逆转录病毒进入了数百万年前的人类基因组，并成为遗传材料的稳定部分，目前，逆转录病毒元素占现代人类基因组的8％。这些病毒随后获得了多个突变，导致人们普遍认为它们不再有能力复制。然而，在研究活着的患者而不是标准细胞系时，我们最近发现了令人惊讶的证据，表明在某些癌症HERV-K（HML-2）的患者中，这是一种内源性逆转录病毒，是一种相对较新的参与者进入人类基因组，并且与肿瘤发生有关，可能仍然能够复制。使用标准技术很难证明内源性逆转录病毒的复制和传播，但是，这些病毒已经存在于所有人类细胞的基因组中。因此，我们组建了一个多样化的研究人员组，他们将检验现代HERV-K（HML-2）可以使用尖端和互补技术复制的假设。我们将使用新设计的分子系统，其中抗生素耐药性用作替代标记，以评估我们是否可以从实验室患者的血液中传播病毒。我们还将使用高通量DNA测序和生物信息学来找到癌症患者中存在但在人类基因组目前的草稿中没有表示的全长功能病毒。通过使用从生物信息学研究中获得的信息和/或通过从患者血液中发现的HERV-K RNA克隆全长病毒的信息，将获得代表现代HERV-K（HML-2）的传染性克隆。然后，该克隆将用绿色荧光蛋白标记，并用于直接研究复制。在我们使用核磁共振（NMR）的前进的基础上，我们还将开发NMR方法来可视化HERV-K（HML-2）复制，从而可视化直接RNA和蛋白质构象转变的空间相关动力学。证明内源性逆转录病毒仍然可以在现代人类中复制，这将导致对这些病毒的思考范式转移，并将暗示它们在重塑单个基因组中的作用。此外，随着染色体内源性逆转录病毒序列的表达增加与癌症和自身免疫性有关，这些发现将与理解明显疾病的发病机理有关。最后，如果在给定个体的血液中可以找到能够引起疾病的复制，感染性内源性逆转录病毒，则此信息将对血液供应的安全具有重要意义。