In this study, we demonstrate the safety and utility of CRISPR-Cas9 gene editing technology for in vivo editing of proviral DNA in ART-treated, virally controlled simian immunodeficiency virus (SIV) infected rhesus macaques, an established model for HIV infection. EBT-001 is an AAV9-based vector delivering SaCas9 and dual guide RNAs designed to target multiple regions of the SIV genome: the viral LTRs, and the Gag gene. The results presented here demonstrate that a single IV inoculation of EBT-001 at each of 3 dose levels (1.4 × 10¹², 1.4 × 10¹³ and 1.4 × 10¹⁴ genome copies/kg) resulted in broad and functional biodistribution of AAV9-EBT-001 to known tissue reservoirs of SIV. No off-target effects or abnormal pathology were observed, and animals returned to their normal body weight after receiving EBT-001. Importantly, the macaques that received the 2 highest doses of EBT-001 showed improved absolute lymphocyte counts as compared to antiretroviral-treated controls. Taken together, these results demonstrate safety, biodistribution, and in vivo proviral DNA editing following IV administration of EBT-001, supporting the further development of CRISPR-based gene editing as a potential therapeutic approach for HIV in humans.

在这项研究中，我们展示了 CRISPR-Cas9 基因编辑技术在经 ART 治疗、病毒控制的猿猴免疫缺陷病毒 (SIV) 感染恒河猴（一种已建立的 HIV 感染模型）体内编辑原病毒 DNA 的安全性和实用性。EBT-001 是一种基于 AAV9 的载体，提供 SaCas9 和双引导 RNA，旨在靶向 SIV 基因组的多个区域：病毒 LTR 和 Gag 基因。本文提供的结果表明，EBT-001 在 3 个剂量水平（1.4 × 10 ¹²、1.4 × 10 ¹³和 1.4 × 10 ¹⁴基因组拷贝/kg）中的每一个的单次 IV 接种导致 AAV9-EBT 的广泛和功能性生物分布-001 为已知的 SIV 组织储存库。没有观察到脱靶效应或异常病理，动物在接受 EBT-001 后恢复到正常体重。重要的是，与接受抗逆转录病毒治疗的对照组相比，接受 2 个最高剂量 EBT-001 的猕猴的绝对淋巴细胞计数有所改善。总而言之，这些结果证明了 EBT-001 静脉注射后的安全性、生物分布和体内原病毒 DNA 编辑，支持进一步开发基于 CRISPR 的基因编辑作为人类 HIV 的潜在治疗方法。