Benzodiazepine mediated mechanisms of transcriptional semi-quiescence in discrete myeloid populations

苯二氮卓介导离散骨髓细胞群转录半静止机制

基本信息

批准号：
10573380
负责人：
Peter Jesse Gaskill
金额：
$ 69.51万
依托单位：
DREXEL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10573380
关键词：
Address Affect American Anatomy Atlases Awareness Benzodiazepines Brain Cells Chromatin Clinical Complex DNA Data Data Correlations Development Disease Dose Epigenetic Process Failure Gene Expression Profile Genetic Genetic Transcription HIV HIV Infections HIV-1 Human Image Individual Infection Infection prevention Mediating Microglia Myelogenous Myeloid Cells Neuraxis Neurocognitive Neuropathogenesis Outcome Penetrance Persons Pharmaceutical Preparations Population Process Production Protocols documentation Proviruses Publishing RUNX1 gene Recrudescences Regulation Risk Role Small Interfering RNA Source Substance Use Disorder Symptoms Testing Therapeutic Transcription Process Transcriptional Regulation Viral Viral reservoir Virion Virus Virus Replication Vulnerable Populations Xanax abuse liability antiretroviral therapy benzodiazepine misuse cell type chromatin immunoprecipitation chronic infection drug misuse epigenetic regulation gag Gene Products induced pluripotent stem cell knock-down macrophage memory CD4 T lymphocyte monocyte mutant neuroAIDS neuroinflammation novel therapeutics programs reactivation from latency recruit reduce symptoms viral DNA viral RNA

项目摘要

One of the principal obstacles to curing HIV is the existence of viral reservoirs in distinct compartments and cell types that are unaffected by antiretroviral therapy (ART). In the central nervous system (CNS), these reservoirs are comprised of myeloid cells, such as perivascular macrophages and microglia. Critically, these reservoirs are generally established prior to the initiation of ART and the presence of ART in the CNS does not affect the established reservoir in long-lived myeloid populations. Thus, even in individuals on suppressive ART, persistently infected CNS macrophages and microglia can drive ongoing neuroinflammation and provide a source of HIV for viral recrudescence during medication hiatus or failure. Our ability to eliminate these reservoirs is severely limited by the lack of understanding of the transcriptional and epigenetic regulation of HIV in myeloid cells. To effectively target and eliminate CNS reservoirs in people living with HIV (PLWH), it is critical to define the transcriptional mechanisms mediating persistent infection in distinct CNS myeloid populations. The impact of HIV infection in the CNS is heightened in PLWH who have substance use disorders, including those who take and/or misuse therapeutic drugs such as benzodiazepines (BDZ). Benzodiazepines are used by more than 30 million Americans, have high abuse liability and misuse of these drugs accounts for 15 – 20% of their use. Benzodiazepines are prescribed to a growing number of PLWH and are associated with an increased risk of neurocognitive symptoms in this population. Little is known about the mechanisms by which BDZ impact HIV neuropathogenesis. Our published and preliminary data indicate that BDZ can alter the transcriptional regulation of HIV infection. To better understand and treat the development of neuroHIV in PLWH who use and/or misuse BDZ, it is vital to determine who benzodiazepines affect the transcriptional programs in myeloid cells. We hypothesize that both macrophages and microglia enter a semi-quiescent transcriptional state during ART exposure and that benzodiazepines disrupt this state and increase transcriptional activity and viral replication. Our data show that the BDZ Xanax, as well as some latency reactivation agents, mediate a dose-dependent increase in p24 production in infected, ART-treated cells. Our preliminary studies suggest these effects may result from BDZ interactions with the epigenetic regulator RUNX1. We posit that these changes in HIV replication in myeloid cells reflect the epigenetic state of the provirus. Thus, the growing population of PLWH who use BDZ is at greater risk of more severe neuropathogenesis. It also suggests that BDZ may provide the basis for novel therapeutics to manipulate HIV transcription in myeloid cells. Therefore, we will evaluate the transcriptional and epigenetic mechanisms sustaining the semi-quiescent state of HIV transcription in ART- treated human macrophages (Aim 1), define the differences in transcriptional and epigenetic processes in different types of myeloid cells using human syngeneic iPSC-derived macrophages and microglia (Aim 2), and examine the effect of benzodiazepines on transcription HIV-infected, ART-treated myeloid cells (Aim 3).

治愈艾滋病毒的主要障碍之一是不同区室中存在病毒储存库，在中枢神经系统 (CNS) 中，不受抗逆转录病毒治疗 (ART) 影响的细胞类型。储库由骨髓细胞组成，例如血管周围巨噬细胞和小胶质细胞。病毒库通常在开始 ART 之前就已经建立，而中枢神经系统中 ART 的存在并不影响因此，即使在接受抑制性 ART 的个体中，持续感染的中枢神经系统巨噬细胞和小胶质细胞可以驱动持续的神经炎症并提供药物中断或失败期间病毒复发的艾滋病毒来源。我们消除这些储存库的能力。由于缺乏对骨髓中 HIV 转录和表观遗传调控的了解而受到严重限制为了有效地瞄准和消除艾滋病毒感染者（PLWH）中的中枢神经系统储存库，定义至关重要。介导不同中枢神经系统骨髓群体持续感染的转录机制。 HIV 感染对中枢神经系统的影响是呼吸有物质使用障碍的感染者，包括服用和/或滥用苯二氮卓类药物 (BDZ) 等治疗药物的人。超过 3000 万美国人有很高的滥用倾向，滥用这些药物的人占 15-20% 越来越多的感染者使用了苯二氮卓类药物，并且与增加有关。对于该人群出现神经认知症状的风险知之甚少。 HIV 神经发病机制。我们发表的初步数据表明 BDZ 可以改变转录。 HIV 感染的调节为了更好地了解和治疗使用和/或使用的 PLWH 中神经 HIV 的发展。误用 BDZ 后，确定哪些苯二氮卓类药物影响骨髓细胞的转录程序至关重要。我们追求巨噬细胞和小胶质细胞都进入半静止转录状态在 ART 暴露期间，苯二氮卓类药物会破坏这种状态并增加转录活性我们的数据表明 BDZ Xanax 以及一些潜伏再激活剂可以介导。我们的初步研究表明，经 ART 处理的受感染细胞中 p24 的产量呈剂量依赖性增加。这些影响可能是由于 BDZ 与表观遗传调节因子 RUNX1 的相互作用造成的。 HIV在骨髓细胞中的复制反映了原病毒的表观遗传状态。使用 BDZ 的 PLWH 发生更严重神经病变的风险更大，这也表明 BDZ 可能会提供更严重的神经病变。操纵骨髓细胞中 HIV 转录的新疗法的基础因此，我们将评估 ART-中维持HIV转录半静止状态的转录和表观遗传机制人类巨噬细胞（目标 1），定义了转录和表观遗传过程的差异使用人类同源 iPSC 衍生的巨噬细胞和小胶质细胞研究不同类型的骨髓细胞（目标 2），以及检查苯二氮卓类药物对 HIV 感染、ART 处理的骨髓细胞转录的影响（目标 3）。