Modulation of epigenetic programming of tissue resident macrophage lineages to impact HIV-1 infection, maintenance, and persistence.

项目总结/摘要 直到最近，该领域才意识到某些组织中的巨噬细胞，包括肺泡巨噬细胞， 巨噬细胞和小胶质细胞是胎儿干细胞衍生的谱系，其行为与巨噬细胞和小胶质细胞显著不同。 血液单核细胞衍生的巨噬细胞。有了这些知识，我们需要重新审视组织驻留者的作用， 巨噬细胞作为HIV-1储库及其对病毒持久性的贡献。我们已经证明，监管 感染巨噬细胞中的通路，如促存活通路，可以通过靶向特异性 lncRNA，从而驱动感染而非未感染的巨噬细胞中的选择性细胞死亡。这些观察结果 然而，根除HIV-1宿主的基础，生物制剂，如lncRNA，并不像 小分子抑制剂发展成为治疗剂。 我们在小鼠和人类肺部巨噬细胞生物学方面拥有广泛的、有文件记录的专业知识， 我们一直在进行一项以化合物的表型筛选为基础的富有成效的抗结核药物发现计划 在受感染的巨噬细胞中活跃。有了这些专业知识，我们提出了鉴定和功能表征 小分子表观遗传抑制剂能够改变宿主巨噬细胞编程， 在特定的骨髓细胞谱系中诱导细胞死亡，并探索潜在的机制。 我们的具体目标是： 1.通过转录分析对HIV-1感染的HMDM和AM进行表型分析。我们会进行 对HIV-1感染的HMDM和AM进行转录谱分析，以评估 在两个谱系中，在活动和潜伏感染状态下对感染的反应。 2.实验性感染中表观遗传编程小分子抑制剂的筛选 HMDM和AM以及HC 69.5小胶质细胞中。我们有一个735个小分子表观遗传的文库， 抑制剂，并将筛选这种化合物收集针对HIV-1感染的HMDM，AM，并针对 永生化的人小胶质细胞系HC69.5，重点是识别驱动细胞增殖的化合物。 不同巨噬细胞谱系和感染模型中的细胞死亡。 3.通过作用模式研究来识别可操作的化合物。我们提出 HIV-1/巨噬细胞生物学分析，优先考虑诱导HIV-1感染者细胞死亡的化合物 巨噬细胞最后，为了评估候选化合物驱动细胞死亡和抑制细胞凋亡的能力， 我们将通过离体药物治疗和细胞存活以及病毒持久性来评估活性， 来自马拉维病毒血症HIV-1阳性人类捐赠者的AM的产物。

Project Summary / Abstract It is only recently that the field became aware that certain tissue resident macrophages, including alveolar macrophages and microglial cells, are fetal stem cell derived lineages that behave markedly differently from blood monocyte derived macrophages. With such knowledge we need revisit the role of tissue resident macrophages as HIV-1 reservoirs and their contribution to viral persistence. We have shown that regulatory pathways in infected macrophages, such as pro-survival pathways, can be inhibited by targeting specific lncRNAs, thus driving selective cell death in infected but not uninfected macrophages. Such observations lay the groundwork for eradication of HIV-1 reservoirs, however, biologics, such as lncRNAs, are not as tractable as small molecule inhibitors to progress into therapeutics. We have extensive, documented expertise in macrophage biology in both mouse and human lung, and we have maintained a productive anti-TB drug discovery program based on phenotypic screening for compounds active in infected macrophages. With this expertise we propose the identification and functional characterization of small molecule epigenetic inhibitors capable of modifying host macrophage programming to drive selective induction of cell death in specific myeloid cell lineages, and probing the underlying mechanism(s). Our Specific Aims are: 1. Phenotypic Profiling HIV-1 infected HMDMs and AMs by transcriptional analysis. We will conduct transcriptional profiling on HIV-1 infected HMDMs and AMs to assess the diversity of the cellular responses to infection in both active and latent infection states in the two lineages. 2. Screening small molecule inhibitors of epigenetic programming in experimental infection in HMDMs and AMs, and in HC69.5 microglial cells. We have a library of 735 small molecule epigenetic inhibitors and will screen this compound collection against HIV-1 infected HMDMs, AMs, and against the immortalized human microglial cell line HC69.5, with the emphasis on identifying compounds that drive cell death across the different macrophage lineages and infection models. 3. Progressing hits through mode-of-action studies to identify actionable compounds. We propose analysis of HIV-1/macrophage biology prioritizing compounds that induce cell death in HIV-1 infected macrophages. Finally, to evaluate candidate compounds for their ability to drive cell death and suppress viral persistence we will assess activity through ex vivo drug treatment and cell survival and viral outgrowth from AMs from viremic HIV-1 positive human donors in Malawi.