Roles of Endogenous Retroviruses in Cancer and Auto-immune Diseases

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。我们基因组中逆转录病毒的插入数约为4万个，与我们DNA编码的基因总数相同。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，这些基因在发育和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。广泛研究的一组内源性逆转录病毒是那些在小鼠中引起重组小鼠白血病病毒（mulv）的病毒。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。内源性逆转录病毒的转移早在感染后一天就可以观察到，这表明在宿主最初感染的细胞中，包装发生在一个复制周期之后。此外，我们发现其他内源性病毒，不同于多嗜性mulv，可能从感染小鼠中动员。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。

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 and are in the same range as the total number of genes encoded by our DNA. Most endogenous retrovirus elements are defective for replication however several contain one or more viral genes that are expressed during development and 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. An extensively investigated group of endogenous retroviruses are those giving 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. Transfer of endogenous retroviruses can be observed as early as one day after infection indicating that packaging occurs after a single replication cycle in the initially infected cells of the host. In addition, we have found that other endogenous viruses, distinct from polytropic MuLVs, are likely mobilized from infected mice. The mobilization of intact endogenous retroviruses is unprecedented and may have important implications for the involvement of endogenous retroviruses in disease processes. In 2011 we have extended our observations to further characterize the endogenous viruses mobilized after infection by exogenous retroviruses. We have found a striking similarity of the viruses we have characterized with those mouse retroviruses reported to be contaminants in studies of putative mouse retrovirus infection in humans. Exogenous mouse retroviruses as well as some other gamma 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, if any, effect on replication in fibroblastic cell lines. APOBEC proteins have evolved as innate defenses against retroviral infections. HIV encodes the VIF protein to evade human APOBEC3G, however mouse retroviruses do not encode a VIF homologue and it has not been understood how they evade mouse APOBEC3. 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 2011 we have focused on the elucidation of the mechanism by which the gGag protein abrogates the action of APOBEC3. To counteract the human APOBEC3G (hA3g), HIV has evolved the Vif protein, which depletes hA3g from infected cells by facilitating its degradation through the proteosome. We have found that a gGag-containing MuLV does not deplete mA3 from an infected cell. Further, immunoprecipitation and localization experiments suggest that mA3 does not interact with gGag directly in an infected cell. In contrast, both gGag and mA3 are associated with the virion core and are coprecipitated in the context of the mature virions. Both hA3g and mA3 have been reported to use deamination-dependent and -independent mechanisms to inhibit retroviral replication. Inhibition by mA3 on gGag-deficient virions correlates with a decrease in the level of transcripts upon infection of cells and appears largely independent of deamination activity. 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, since endogenous polytropic viruses encode gp70s that are closely related to xenotropic gp70, these viruses could be 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. These studies were extended to show that expression levels of a subclass of polytropic MuLVs, termed, modified polytropic (mPT), are highly elevated in mice which develop systemic lupus erythematosus. This elevated expression appears specific for this class of endogenous viruses and under the control of the Sgp3 locus. We have also examined the influence of TLR7 or TLR9 agonist on the expression of serum gp70s. These results suggest an additional pathogenic role of TLR7 and TLR9 in murine lupus nephritis by promoting the expression of nephritogenic gp70 autoantigen. In 2011 we have extended these results to determine the genetic origin of xenotropic viral sequences expressed in wild-type and two different Sgp congenic C57BL/6 mice. The results of these indicated that Sgp3 and Sgp4 independently regulated the transcription of distinct and restricted sets of xenotropic viral sequences in trans, thereby promoting the production of nephritogenic gp70 autoantigens. Among these endogenous viruses induced by Sgp3 and Sgp4 are two potentially replication competent xenotropic viruses. Although xenotropic viruses cannot infect mouse cells it is possible that these viruses may be complemented by mPT env proteins allowing their spread and and activation of TLR 7. In this regard, the presence of Sgp3 induces an approximately 100-fold increase of mPT viruses. Thus Sgp3 and Sgp4 may contribute to the development of autoimmune responses against serum gp70 through the activation of TLR7.