Combined Molecular Excision Therapy (CMET) for Eliminating HIV-1

摘要 人类免疫缺陷病毒(HIV)从其中枢神经系统(CNS)和 外围储集层是治病的必要条件。据我们所知，我们是第一个实现 这一目标是在有限数量的受感染的人源化小鼠身上进行的测试中实现的。来验证这些早期的成功 我们建议建立一个四步梯子，最终达到根除潜伏病毒的顶峰。第一，一种新开发的 人源化的小鼠脑淋巴组织神经艾滋病模型将识别高效感染的血管周围和 脑膜巨噬细胞与实质细胞的限制性感染。这个啮齿动物模型最能反映 人类大脑疾病，由强大的分子、病毒学和神经免疫学测试证明。第二, 长效、缓释、有效的抗逆转录病毒疗法(激光抗逆转录病毒疗法)现在也在我们的 实验室现在可以帮助精确定位潜伏感染的脑部病毒库。第三，病毒式传播 将采用切除策略来消除残留病毒并防止艾滋病毒重新激活。基因编辑 天普大学医学中心研究人员开发的CRISPR/Cas9系统，包括CCR5和Virus 切除策略将减少然后消除任何持续感染和感染的整合前病毒DNA 细胞。CRISPR/Cas9构建物将使用特定的 腺相关病毒的血清型。这将使人源化小鼠在没有艾滋病毒的情况下永久根除艾滋病毒 病毒重新激活，从而防止任何持续的脑感染和随后的神经损伤。第四，在 为了证明该治疗策略对脑和外周淋巴组织病毒(包括 肠道相关淋巴组织、淋巴结、脾和泌尿生殖系统)我们将停止ART 提供交叉验证病毒证据的管理和以下时间段(以月为单位) 通过措施反弹根除。考虑到艾滋病毒在中枢神经系统重新激活的相关风险，这种方法 必须显示其针对潜伏病毒的能力的有效性。综上所述，该提案寻求支持 使用激光抗逆转录病毒疗法和有效的分子病毒和免疫方案相结合来消除 病毒库。总体前提是开发永久消除 在中枢神经系统和外周感染性蓄水池中检测到病毒。

Abstract The elimination of the human immunodeficiency virus (HIV) from its central nervous system (CNS) and peripheral reservoirs is a requirement for a disease-cure. To our knowledge, we are the first to have achieved this goal in tests performed in a limited number of infected humanized mice. To validate such early successes we propose to build a four-step ladder with a final crest of latent virus eradication. First, a newly developed humanized mouse brain-lymphoid model of neuroAIDS will identify productively infected perivascular and meningeal macrophages and restricted infection in parenchymal cells. This rodent model most closely reflects human brain disease as demonstrated by robust molecular, virologic and neuroimmunologic tests. Second, long acting slow effective release antiretroviral therapy (LASER ART) also now fully developed in our laboratories can now facilitate a pinpoint localization of the latently infected viral brain reservoir. Third, viral excision strategies will be employed to eliminate residual virus and preclude HIV reactivation. The gene editing CRISPR/Cas9 system developed by Temple University Medical Center investigators including CCR5 and viral excision strategies will reduce then eliminate any ongoing infection and integrated proviral DNA from infected cells. The CRISPR/Cas9 constructs will deliver its cargo to brain and peripheral tissue sites using specific serotypes of adeno-associated virus. This will enable permanent HIV eradication in humanized mice without viral reactivation and as such preclude any ongoing brain infection and subsequent neural damage. Fourth, in order to prove the therapeutic strategy effective both for brain and peripheral lymphoid tissue virus (including the gut-associated lymphoid tissue, lymph node, spleen and genitourinary system) we will cease ART administrations and following time periods measured in months to provide cross validating evidence for viral eradication by measure rebound. Given the risks associated with HIV reactivation in the CNS this approach must show effectiveness for its abilities to target latent virus. Taken together, the proposal seeks support to employ combination LASER ART and potent molecular viral and immune-based regimens for elimination of viral depots. The overall premise is to develop the “state of the art” tools required to permanently eliminate virus detected in the CNS and peripheral infectious reservoirs.