Brain myeloid cell-targeted multiplexed gene editing for SIV/HIV eradication

用于根除 SIV/HIV 的脑髓细胞靶向多重基因编辑

基本信息

批准号：
10476684
负责人：
Wenhui Hu
金额：
$ 91.54万
依托单位：
TEXAS BIOMEDICAL RESEARCH INSTITUTE
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：
2022-04-05 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10476684
关键词：
AIDS/HIV problem Acute Animals Bar Codes Blood - brain barrier anatomy Brain Brain region CCL2 gene CCR5 gene CRISPR/Cas technology Capsid Cells Chinese Chronic Phase Clinical Clinical Research Clustered Regularly Interspaced Short Palindromic Repeats DNA Data Early treatment Epidemic Excision Genes Genome Goals Guide RNA HIV HIV Infections HIV-1 In Vitro Individual Infection Injections Intervention Intravenous Latent virus infection phase Libraries Macaca Macaca mulatta Measurement Mediating Microglia Modeling Monitor Monkeys Mus Myelogenous Myeloid Cells Names Neuraxis Outcome Plasma Play Prevention Prevention therapy Proviruses Regimen Reporter Residual state Rhesus Role SIV Seeds Serotyping Shock Site Source System Technology Testing Therapeutic Effect Time Tissues Vero Cells Viral reservoir Viremia Virus Virus Diseases antiretroviral therapy base blood-brain barrier penetration brain size cell type efficacy evaluation gene therapy genome editing high risk high risk population humanized mouse in vivo macrophage nervous system disorder neuroAIDS neurocognitive disorder neuroinflammation nonhuman primate novel novel strategies prevent promoter relating to nervous system simian human immunodeficiency virus treatment guidelines viral rebound

项目摘要

Project Summary The long-lived myeloid cells such as perivascular macrophages and microglia in the central nervous system (CNS) persistently harbor HIV. These infected cells could contribute to the source of residual viremia during long-term antiretroviral therapy (ART) or to rebounding virus upon ART cessation. It is undoubtfully and urgently needed to develop novel strategies to specifically target CNS myeloid cells for HIV eradication and a cure. Be- cause the effects of the widely-studied “Shock and Kill” approach could exacerbate neuroinflammation, gene therapy emerges as the optimal strategy, particularly the advanced CRISPR genome editing technology. Simian immunodeficiency virus (SIV) infection of macaques is the best available model for testing novel strategies prior to clinical studies. We have shown that SIV infection has a broad spread in the CNS even in animals on ART. Because the virus enters the brain within a few days after infection and the establishment of the latent reservoir occurs very early, the initiation of ART should be as early as possible. In addition, numerous studies suggest that CCR5/CCR2 play a major role in HIV entry and neuroinflammation. Most importantly, we have used AAV delivery of a CRISPR/Cas genome editor to eradicate HIV/SIV provirus in models of humanized mice and non- human primates. However, the lack of AAV serotypes that are highly effective and reliable to transduce myeloid cells in the CNS remains a key challenge. Therefore, we hypothesize that the multiple-targeting gene editing system across the blood brain barrier (BBB) can remove SIV provirus in infected myeloid cells, protect cells against new infection, and inhibit neuroinflammation. To test this hypothesis, we will optimize a novel AAV sero- type with BBB penetration and myeloid-specific transduction (namely AAV-BM) to effectively deliver the smaller cjCas9 with multiplex sgRNAs (BMCj4) specific for 4 target sites (SIV LTR, gag, and host CCR5, CCR2) into the entire CNS for in vivo HIV/SIV eradication (Aim 1). We will evaluate the efficacy of BMCj4 early treatment in preventing brain SIV infection or/and excising SIV proviral DNA from brain myeloid cells in acute SIV infection with early ART (Aim 2a). We will also determine the therapeutic effect of BMCj4 with ART and then boost it with an alternative AAV-BM for eradication of persistent brain SIV latent infection (Aim 2b). We expect that early or long-term repeated BMCj4 AAV gene therapy will effectively eradicate acute and latently-infected HIV provirus and extensively minimize the size of the brain viral reservoir to achieve a sterilizing or functional cure of HIV/AIDS, particularly NeuroAIDS.

项目概要中枢神经系统中的长寿骨髓细胞，例如血管周围巨噬细胞和小胶质细胞（中枢神经系统）持续携带艾滋病毒，这些受感染的细胞可能是导致残留病毒血症的原因。长期抗逆转录病毒治疗（ART）或ART停止后病毒反弹无疑是紧迫的。需要开发新的策略来专门针对中枢神经系统骨髓细胞来根除和治愈艾滋病毒。导致广泛研究的“电击杀伤”方法的效果可能会加剧神经炎症、基因治疗成为最佳策略，特别是先进的 CRISPR 基因组编辑技术。猕猴的免疫缺陷病毒（SIV）感染是先前测试新策略的最佳可用模型临床研究表明，SIV 感染在中枢神经系统中广泛传播，甚至在接受 ART 的动物中也是如此。因为病毒在感染后几天内就进入大脑并建立潜伏储存库发生得很早，因此应尽早开始 ART 此外，大量研究表明。 CCR5/CCR2 在 HIV 进入和神经炎症中发挥重要作用最重要的是，我们使用了 AAV。交付 CRISPR/Cas 基因组编辑器以根除人源化小鼠和非小鼠模型中的 HIV/SIV 原病毒然而，人类灵长类动物缺乏高效可靠的 AAV 血清型。中枢神经系统细胞仍然是一个关键挑战，因此，我们追求多靶点基因编辑。系统跨越血脑屏障（BBB）可清除受感染骨髓细胞中的SIV原病毒，保护细胞为了检验这一假设，我们将优化一种新型 AAV 血清。具有 BBB 穿透和骨髓特异性转导（即 AAV-BM）的类型，可有效递送较小的 cjCas9 与 4 个靶位点（SIV LTR、gag 和宿主 CCR5、CCR2）特异性的多重 sgRNA (BMCj4) 结合到整个 CNS 体内 HIV/SIV 根除（目标 1）我们将评估 BMCj4 早期治疗的功效。预防脑 SIV 感染或/和在急性 SIV 感染时从脑髓细胞中切除 SIV 前病毒 DNA 早期 ART（目标 2a）我们还将确定 BMCj4 与 ART 的治疗效果，然后通过 ART 增强其疗效。一种用于根除持续性脑 SIV 潜伏感染的替代 AAV-BM（目标 2b）。长期重复BMCj4 AAV基因治疗将有效消除急性和潜伏感染的HIV原病毒并通常最大限度地减少脑病毒库的大小，以实现对脑部病毒的灭菌或功能性治愈艾滋病毒/艾滋病，特别是神经艾滋病。