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在不同的隔间中存在病毒储层不受抗逆转录病毒疗法影响的细胞类型（ART）。在中枢神经系统（CNS）中，这些储层是髓样细胞积累的，例如周围巨噬细胞和小胶质细胞。批判性，这些通常在艺术计划之前建立水库，而在中枢神经系统中的艺术品没有在长寿髓样人群中影响已建立的储层。即使在抑制艺术的个人中，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正在进行的神经炎症并提供在药物裂变或衰竭期间，HIV的HIV来源。我们消除这些水库的能力由于缺乏对髓样中HIV的转录和表观遗传调节的了解，受到严重限制细胞。为了有效地靶向和消除艾滋病毒（PLWH）的人的中枢神经系统水库，定义至关重要转录机制介导了不同的中枢神经系统种群中的持续感染。在患有药物使用障碍的PLWH中，艾滋病毒感染在中枢神经系统中的影响得到了增强，包括那些服用和/或滥用治疗药物（例如苯二氮卓类药物（BDZ））的人。苯二氮卓类是由超过3000万美国人，负有高昂的虐待责任，对这些毒品的遗漏占15-20％他们的使用。苯二氮卓类药物的处方越来越多，与增加有关该人群中神经认知症状的风险。关于BDZ影响的机制知之甚少 HIV神经病发生。我们已发布的初步数据，表明BDZ可以改变转录调节HIV感染。更好地理解和治疗使用和/或使用和/或滥用BDZ，确定谁苯二氮卓类药物影响髓样细胞中的转录程序至关重要。我们假设巨噬细胞和小胶质细胞都进入半季度的转录状态在艺术暴露期间，苯二氮卓类药物破坏了这种状态并增加了转录活动和病毒复制。我们的数据表明，BDZ Xanax以及某些潜伏重新激活剂，培养基受感染的ART处理细胞中P24产生的剂量依赖性增加。我们的初步研究表明这些影响可能是由于BDZ与表观遗传调节剂Runx1的相互作用而引起的。我们指出这些变化在髓样细胞中的HIV复制中反映了病毒的表观遗传态。那，人口不断增长使用BDZ的PLWH面临更严重的神经病发生的更大风险。这也表明BDZ可能会提供在髓样细胞中操纵HIV转录的新型治疗的基础。因此，我们将评估在艺术中维持HIV转录半频率状态的转录和表观遗传机制处理过的人类巨噬细胞（AIM 1），定义了转录和表观遗传过程的差异使用人类同性IPSC衍生的巨噬细胞和小胶质细胞（AIM 2）和检查苯二氮卓类药物对转录感染的HIV，艺术处理的髓样细胞的影响（AIM 3）。