Control of latent/persistent HIV-1 infection in macrophages/microglia: A key role for the phosphatase PPM1A

摘要 组织中单核细胞/巨噬细胞谱系细胞中的持续HIV-1感染，如脑、肠、淋巴 组织或脂肪组织形成了一个研究不足的病毒库，这将阻止根除HIV-1，甚至 如果CD 4 + T细胞的储存库可以解决的话。然而，除了潜伏的HIV-1感染的T细胞， 到目前为止，还没有详细描述骨髓细胞中潜伏期的生物分子控制。我们证明潜伏的艾滋病毒- 1感染的单核细胞/巨噬细胞从正常的2n染色体组转变为4 n染色体状态。这 与先前的报告一致，该报告描述了最活跃的巨噬细胞对抗 结核分枝杆菌感染到多倍体状态，表明4 n转换是结核分枝杆菌感染的自然机制。 巨噬细胞在其病原体反应期间。多倍性不仅可以增加对许多疾病的抵抗力， 挑战，否则会导致细胞死亡，但也会触发非线性转录组学效应， 干扰正常的细胞信号传导，我们认为这是HIV-1无法建立活动性感染的部分原因。非 线性转录组效应在全基因组转录组标签（RNA- seq），其显示出广泛的克隆间异质性。 然而，尽管有这种广泛的转录组异质性，潜伏的HIV-1感染的巨噬细胞与其他巨噬细胞有一个小的共同点。 在转录组（RNA-seq）和蛋白质组水平（激酶组分析）的常见改变的生物分子特征。 我们确定磷酸酶PPM 1A是这种潜伏期控制特征的关键要素。PPM 1A的调节 表达控制巨噬细胞中的活性HIV-1感染，调节巨噬细胞对HIV的易感性， 1诱导细胞死亡，并改变骨髓细胞建立潜伏感染的能力。因此，针对PPM 1A 有可能提供一种新的治疗途径来根除巨噬细胞中的HIV-1储库。这 应用将有助于我们从分子水平了解骨髓细胞的内在抗病毒反应， 细胞是受控的，以及PPM 1A活性的治疗性调节如何有助于消除 在骨髓细胞中，特别是在脑内的小胶质细胞中，HIV-1的持续储存。

ABSTRACT Persistent HIV-1 infection in cells of the monocyte/macrophage lineage in tissues such as the brain, gut, lymphoid tissues, or adipose tissue forms an under-investigated viral reservoirs that will prevent eradication of HIV-1, even if the reservoir in CD4+ T cells could be addressed. However, other than for latently HIV-1 infected T cells, so far the biomolecular control of latency in myeloid cells has not been detailed. We demonstrate that latently HIV- 1 infected monocyte/macrophages transition from a normal 2n chromosome set to a 4n chromosome state. This is consistent with a previous report that describes the transition of the most active macrophages combating Mycobacterium tuberculosis infection to a polyploid state, indicating that 4n transition is a natural mechanism of macrophages during their pathogen response. Polyploidy is not only known to increase resistance to many challenges that would otherwise result in cell death, but also triggers non-linear transcriptomic effects, which disrupt proper cell signaling and we propose are part of the inability of HIV-1 to establish active infection. Non- linear transcriptomic effects become obvious in the comparison of genome-wide transcriptomic signatures (RNA- seq) of different latently HIV- 1 infected monocyte clones, which show extensive inter-clonal heterogeneity. However, despite this extensive transcriptomic heterogeneity, latently HIV-1 infected macrophages share a small common altered biomolecular signature at the transcriptome (RNA-seq) and proteome level (kinome analysis). We identified the phosphatase PPM1A as a key element of this latency control signature. Modulation of PPM1A expression controlled active HIV-1 infection in macrophages, regulated the susceptibility of macrophages to HIV- 1 induced cell death, and altered the capacity of myeloid cells to establish latent infection. Thus targeting PPM1A has the potential to provide a new therapeutic avenue to eradicate HIV-1 reservoirs in macrophages. This application will contribute to our basic molecular understanding of how the intrinsic antiviral response of myeloid cells is controlled and how therapeutic modulation of PPM1A activity could contribute to the elimination of persistent HIV-1 reservoirs in myeloid cells in general and specifically in brain-resident microglia.