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：
10602472
负责人：
Wenhui Hu
金额：
$ 79.52万
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/10602472
关键词：
Acute Animals Bar Codes Blood - brain barrier anatomy Blood Vessels Brain Brain region CCL2 gene CCR5 gene CRISPR/Cas technology Capsid Cells Central Nervous System Chinese Chronic Phase Clinical Clinical Research Clustered Regularly Interspaced Short Palindromic Repeats Cytoprotection DNA Data Early treatment Epidemic Excision Genes Genome Goals Guide RNA HIV HIV Infections HIV-1 HIV/AIDS Immunotherapy In Vitro Individual Infection Injections Intervention Intravenous Latent virus infection phase Libraries Macaca Macaca mulatta Macrophage Measurement Mediating Microglia Modeling Monitor Monkeys Mus Myelogenous Myeloid Cells Names Outcome Plasma Play Prevention Prevention therapy Proviruses Recommendation Regimen Reporter Residual state Role SIV Serotyping Shock Site Source Specificity System Technology Testing Therapeutic Effect Time Tissues Vero Cells Viral reservoir Viremia Virus Virus Diseases antiretroviral therapy blood-brain barrier penetration brain size cell type efficacy evaluation gene therapy genome editing high risk high risk population humanized mouse in vivo mouse model nervous system disorder neural neuroAIDS neurocognitive disorder neuroinflammation nonhuman primate novel novel strategies prevent promoter treatment guidelines viral detection 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.

项目摘要长寿命的髓样细胞，例如中枢神经系统中的血管周围巨噬细胞和小胶质细胞（CNS）持续怀有艾滋病毒。这些受感染的细胞可能有助于在长期抗逆转录病毒疗法（ART）或在停止艺术时反弹病毒。毫无疑问地紧急需要制定新型策略，以专门针对CNS髓样细胞以消除HIV和治愈。是- 导致广泛研究的“冲击和杀伤”方法的影响可能加剧神经炎症，基因治疗是最佳策略，尤其是先进的CRISPR基因组编辑技术。猿人猕猴的免疫缺陷病毒（SIV）感染是测试新策略的最佳可用模型进行临床研究。我们已经表明，即使在动物上，SIV感染在中枢神经系统中也有广泛的传播。因为该病毒在感染后几天内进入大脑，并建立潜在储层很早就发生了，艺术的倡议应尽早。此外，许多研究表明该CCR5/CCR2在HIV进入和神经炎症中起着重要作用。最重要的是，我们使用了AAV 将CRISPR/CAS基因组编辑器传递到放射性艾滋病毒/SIV病毒中的人性化小鼠和非 - 人类的素数。但是，缺乏高效且可靠地转导髓样的AAV血清型中枢神经系统中的细胞仍然是一个关键挑战。因此，我们假设多个目标基因编辑整个血脑屏障（BBB）的系统可以清除感染的髓样细胞中的SIV病毒，保护细胞针对新的感染并抑制神经炎症。为了检验这一假设，我们将优化一种新型的AAV血清使用BBB渗透和粒细胞特异性翻译（即AAV-BM）类型，以有效提供较小的 CJCAS9具有多个针对4个目标位点（SIV LTR，GAG和HOST CCR5，CCR2）的特异性SGRNA（BMCJ4）整个用于体内HIV/SIV洗脱的中枢神经系统（AIM 1）。我们将评估BMCJ4早期治疗的效率 preventing brain SIV infection or/and excising SIV proviral DNA from brain myeloid cells in acute SIV infection 与早期艺术（AIM 2A）。我们还将用ART确定BMCJ4的治疗作用，然后将其提高持续性脑SIV潜在感染的辐射辐射的另一种AAV-BM（AIM 2B）。我们期望早期或长期重复的BMCJ4 AAV基因疗法将有效地避免急性和潜在感染的HIV病毒并广泛地最大程度地减少脑病毒储存库的大小，以实现消毒或功能治愈艾滋病毒/艾滋病，尤其是神经辅助。