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

摘要 脑、肠道、淋巴等组织中单核/巨噬细胞系细胞中持续的HIV-1感染 组织或脂肪组织形成了一个未被研究的病毒储备库，这将阻止艾滋病毒-1的根除，甚至 如果CD4+T细胞中的储备库能够被解决。然而，除了潜伏感染HIV-1的T细胞，所以 到目前为止，髓系细胞潜伏期的生物分子控制还没有详细说明。我们证明了潜伏的艾滋病毒- 1感染的单核/巨噬细胞从正常的2n染色体组转变为4n染色体状态。这 与之前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4N转换是一种自然机制 巨噬细胞在病原体反应过程中。已知多倍体不仅会增加对许多 挑战，否则将导致细胞死亡，但也触发非线性转录效应，这 干扰适当的细胞信号，我们认为这是HIV-1无法建立活动性感染的部分原因。非- 线性转录效应在全基因组转录签名的比较中变得明显。 不同潜伏感染HIV-1的单核细胞克隆的序列序列(SEQ)，显示出广泛的克隆间异质性。 然而，尽管这种广泛的转录异质性，潜伏感染HIV-1的巨噬细胞共享一小部分 转录组(RNA-seq)和蛋白质组水平(动态组分析)的常见生物分子标记改变。 我们确定磷酸酶PPM1a是这一潜伏期控制信号的关键元件。PPM1a的调制 巨噬细胞表达控制HIV-1活性感染，调节巨噬细胞对HIV-1的易感性 1诱导细胞死亡，改变髓系细胞建立潜伏感染的能力。从而以PPM1a为目标 有可能提供一种新的治疗途径来根除巨噬细胞中的艾滋病毒-1储存库。这 应用将有助于我们基本的分子理解髓系细胞的内在抗病毒反应 细胞被控制，以及PPM1A活性的治疗性调节如何有助于消除 持续的HIV-1储存库一般存在于髓系细胞中，特别是驻留在脑内的小胶质细胞中。

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.