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

调节组织驻留巨噬细胞谱系的表观遗传编程以影响 HIV-1 感染、维持和持久性。

基本信息

批准号：
10675934
负责人：
DAVID G RUSSELL
金额：
$ 69.52万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-06 至 2028-03-31
项目状态：
未结题

项目摘要

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.

项目概要/摘要直到最近，该领域才意识到某些组织驻留巨噬细胞，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 储存库及其对病毒持久性的贡献。我们已经表明，监管受感染巨噬细胞中的途径，例如促生存途径，可以通过靶向特定的途径来抑制 lncRNA，从而驱动受感染而非未感染巨噬细胞的选择性细胞死亡。这样的观察表明根除 HIV-1 病毒库的基础，然而，lncRNA 等生物制剂并不像小分子抑制剂进入治疗领域。我们在小鼠和人肺的巨噬细胞生物学方面拥有广泛且有记录的专业知识，并且我们一直在维持基于化合物表型筛选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。凭借这些专业知识，我们提出了鉴定和功能表征能够改变宿主巨噬细胞编程以驱动选择性的小分子表观遗传抑制剂诱导特定骨髓细胞谱系的细胞死亡，并探讨潜在机制。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。我们将进行对 HIV-1 感染的 HMDM 和 AM 进行转录分析，以评估细胞的多样性这两个谱系在活跃和潜伏感染状态下对感染的反应。 2. 筛选实验感染中表观遗传编程的小分子抑制剂 HMDM 和 AM，以及 HC69.5 小胶质细胞。我们拥有 735 个小分子表观遗传库抑制剂，并将筛选该化合物集合以对抗 HIV-1 感染的 HMDM、AM 和永生化人类小胶质细胞系 HC69.5，重点是识别驱动的化合物不同巨噬细胞谱系和感染模型的细胞死亡。 3. 通过作用模式研究来确定可作用的化合物，从而取得进展。我们建议 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，评估候选化合物驱动细胞死亡和抑制细胞死亡的能力病毒持久性我们将通过离体药物治疗和细胞存活和病毒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。