Roles of Endogenous Retroviruses in Cancer and Auto-immune Diseases

内源性逆转录病毒在癌症和自身免疫性疾病中的作用

• 批准号: 8946483
• 负责人: LEONARD EVANS
• 金额: $ 25.63万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8946483
• 关键词: Acute-Phase Proteins; Antigen-Antibody Complex; Beryllium; Cell Line; Cells; Congenic Mice; Cytidine; Cytidine Deaminase; Data; Deamination; Defect; Development; Disease; Elements; Endogenous Retroviruses; Evolution; Exclusion; Exhibits; Friend Murine Leukemia Virus; Gagging; Genes; Genetic; Genetic Recombination; Genetic Transcription; Genome; Genomics; Germ Cells; Glycoproteins; Goals; HIV; Hepatocyte; Homologous Gene; Host Defense; Human; Immune System Diseases; In Vitro; Infection; Knockout Mice; Length; Liver; Lupus; Lupus Nephritis; Malignant Neoplasms; Mammals; Methionine; Modeling; Mouse Strains; Murine leukemia virus; Mus; Mutation; Pathologic Processes; Pathology; Physiological; Physiological Processes; Physiology; Play; Point Mutation; Polymerase; Polyproteins; Process; Production; Proteins; Proviruses; Recombinants; Regulation; Retroviridae; Retroviridae Infections; Role; Serum; Site; Source; Structure; Time; Transcript; Translations; Viral; Viral Genes; Viral Genome; Viral Physiology; Virion; Virus; Virus Replication; Wild Type Mouse; base; gag Gene Products; human CEM15 protein; in vivo; insight; member; novel

Approximately 8% of the genomes of mammals, including humans and mice, are comprised of retroviral elements acquired by infection of germ line cells during the course of evolution. Retroviral insertions in our genome number about 40,000. Most endogenous retrovirus elements are defective for replication however several contain one or more viral genes that are expressed during development and during certain physiological or pathological conditions. Little is known about the control of retrovirus expression or the influence of such expression on the physiology or pathology of the host. Polytropic MuLVs are an extensively investigated group of endogenous retroviruses that give rise to recombinant murine leukemia viruses (MuLVs) in mice. Upon infection of mice with exogenous ecotropic MuLVs, members of this group undergo recombination to generate new MuLVs with an altered infectious host range. Recombination requires transcription of the endogenous retroviruses. Although the endogenous polytropic proviruses are transcribed; replication of the endogenous polytropic viruses in the absence of recombination has not been observed. This may, in many cases, reflect defects such as point mutations or deletions in the endogenous viral genome but may also be influenced by the activity of various restriction factors. The fact that exogenous MuLVs are capable of replicating in mice indicates that they have evolved mechanisms to circumvent the activity of at least some of the restriction factors such as the murine APOBEC3. Thus, exogenous retroviruses might facilitate,through complementation, active replication of endogenous retroviruses. We have found that infection of mice by an exogenous virus results in the infectious transfer of complete endogenous proviral genetic sequences. This includes proviruses which are severely defective and possess large deletions as well as proviruses that are full-length. Furthermore, the transferred sequences are transcribed and packaged into virions released from the newly infected cells. At early times after infection with the Friend MuLV, packaging and transfer of intact endogenous retroviruses is much more prevalent than recombination. Endogenous retroviruses are transferred as early as one day after infection. Thus, the transcripts are captured within a single in vivo replication cycle and originate from among the initially infected host cells. The mobilization of intact endogenous retroviruses is unprecedented and may have important implications for the involvement of endogenous retroviruses in disease processes. In 2014 we have extended our observations to further characterize the endogenous viruses mobilized after infection by exogenous retroviruses. We have found that the mobilized endogenous viruses appear to be specifically limited to endogenous polytropic proviruses at the exclusion of other endogenous retroviral elements. Furthermore, structures of the mobilized endogenous viruses exhibit frequent recombination among them indicating their active replication, likely as a result of infection of the host with an exogenous retrovirus. The endogenous polytropic proviruses are comprised of two structural subclasses termed Polytropic (PT) and Modified Polytropic (mPT). We observed a distinct shift in the subclass of proviruses from the mPT subclass to the PT subclass of polytropic proviruses detected during the course of infection. These results suggest the spread of infection to cells expressing different classes of polytropic viruses or an alteration in the expression of the endogenous proviruses in infected cells. Exogenous mouse retroviruses encode a glycosylated gag protein (gGag) originating from an alternate translation start site upstream of the methionine start site of the gag structural polyproteins. The functions of gGag remain unclear, but mutations that eliminate its synthesis severely impede in vivo replication of the virus with little effect on replication in fibroblastic cell lines. APOBEC3 proteins have evolved as innate defenses against retroviral infections. Both mice and humans express APOBEC3 proteins that have cytidine deaminase activity leading to hypeermutation of viral transcripts and inactivation of infecting retroviruses. HIV encodes the VIF protein to evade human APOBEC3G (hA3G), however mouse retroviruses do not encode a VIF homologue and it has not been understood how they evade mouse APOBEC3 (mA3). We have found that a mouse retrovirus utilizes its glycosylated gag protein (gGag) to evade APOBEC3. gGag is critical for infection of in vitro cell lines in the presence of APOBEC3. Furthermore, a gGag-deficient virus restricted for replication in wild-type mice replicates efficiently in APOBEC3 knockout mice implicating a novel role of gGag in circumventing the action of APOBEC3 in vivo. In 2014 we have continued to focus on the elucidation of the mechanism by which the gGag protein abrogates the action of APOBEC3. We have found that mA3 packaged at very high levels in wild-type virions results in a loss of specific infectivity, a loss in the specific enzymatic activity of the viral polymerase accompanied by a substantial increase in the overall mutation rate. The mutagenic activity of mA3 in wild-type as well as gGag-deficient viruses does not include G to A hypermutation and is unlikely to involve cytidine deamination. We have found that the ecotropic MuLV, AKV undergos G to A hypermutation with a target sequence indicative of the action of mA3. However, hypermutation of AKV does not require the augmentation of mA3 in cells. Furthermore, hypermutation occurs in only a portion of the virus transcripts suggesting that a restriction factor is present in only a portion of the virions. These results suggest that AKV is exquisitely susceptible to mA3 or another restriction factor expressed at trace levels in the cells. The endogenous retroviral envelope glycoprotein, gp70 is implicated in murine lupus nephritis. This protein is secreted by hepatocytes as an acute phase protein and has been believed to be a product of an endogenous xenotropic virus. However,we have found that endogenous polytropic viruses provide additional sources of serum gp70. To better understand the genetic basis of the expression of serum gp70, we analyzed the abundance of xenotropic and polytropic gp70 RNAs in livers and the genomic composition of corresponding endogenous proviruses in various strains of mice, including two different Sgp (serum gp70 production) congenic mice (Sgp3 and Sgp4). These studies revealed a significant contribution of polytropic gp70s to serum gp70 and a tight regulation of control by the Sgp3 and Sgp4 genes. Sgp3 regulates the transcription of xenotropic, polytropic and modified polytropic (mPT)viruses, and Sgp4 the transcription of only xenotropic viruses. These studies were extended to examine the effects of a third gene, Sgp5. The Sgp5 locus enhanced the expression of xenotropic and mPT viruses, thereby upregulating the production of serum gp70. These data indicate a distinct action of the Sgp5 locus on the expression of endogenous retroviruses as compared with two other Sgp loci. Moreover, it was found that Sgp3 and Sgp4 acted synergistically to elevate the transcription of a potentially replication-competent Xmv18 provirus and the production of serum gp70. The results of these studies indicate that the combined effect of three different Sgp loci markedly enhance the expression of endogenous retroviruses and their gene product, serum gp70, thereby contributing to the formation of nephritogenic gp70-anti-gp70 immune complexes in murine lupus.

哺乳动物（包括人类和小鼠）大约8%的基因组是由在进化过程中通过生殖系细胞感染获得的逆转录病毒元素组成的。在我们的基因组中，逆转录病毒的插入数大约是4万个。大多数内源性逆转录病毒元件在复制方面存在缺陷，但也有一些含有一个或多个病毒基因，这些病毒基因在发育过程中以及某些生理或病理条件下表达。关于逆转录病毒表达的控制以及逆转录病毒表达对宿主生理病理的影响，我们所知甚少。多嗜性mulv是一组被广泛研究的内源性逆转录病毒，可在小鼠体内产生重组小鼠白血病病毒（mulv）。在外源性嗜生态mulv感染小鼠后，该组成员进行重组以产生具有改变的感染宿主范围的新mulv。重组需要内源性逆转录病毒的转录。虽然内源性多性原病毒被转录；没有观察到在没有重组的情况下内源性多性病毒的复制。在许多情况下，这可能反映了内源性病毒基因组中的缺陷，如点突变或缺失，但也可能受到各种限制性因子活性的影响。外源性mulv能够在小鼠体内复制的事实表明，它们已经进化出了规避至少一些限制性因子（如小鼠APOBEC3）活性的机制。因此，外源性逆转录病毒可能通过互补促进内源性逆转录病毒的主动复制。我们发现外源性病毒感染小鼠导致完整的内源性原病毒基因序列的感染性转移。这包括严重缺陷的原病毒和具有大缺失的原病毒以及全长的原病毒。此外，转移的序列被转录并包装成从新感染细胞释放的病毒粒子。在Friend MuLV感染后的早期，包装和转移完整的内源性逆转录病毒比重组更为普遍。内源性逆转录病毒最早在感染后一天转移。因此，转录本在一个体内复制周期内被捕获，并起源于最初感染的宿主细胞。完整内源性逆转录病毒的动员是前所未有的，可能对内源性逆转录病毒参与疾病过程具有重要意义。