A Gene Drive Therapy for HIV: single-administration intervention for high-risk groups

项目摘要/摘要 对于新冠肺炎来说，注射毒品者(PWID)已被确定为接触的高危人群 对SARS-CoV-2(导致新冠肺炎的病毒)的风险，因为他们无家可归或 监禁--与疾病传播率增加和与艾滋病毒-1混合感染有关的情况。 考虑到SARS-CoV-2疫苗很可能在12-24个月后出现，医疗方面存在严重的未得到满足的需求 可以将新冠肺炎疫情控制在普通人群和这些难以接触到的地区的对策 尤其是像PWID这样的高危人群。此外，我们在认识上存在着根本性的差距 在这些PWID高危人群中，SARS-CoV-2感染和发病机制的关系。有证据表明，SARS-- CoV-2感染并耗尽T淋巴细胞，许多感染艾滋病毒的PWID患者获得抗逆转录病毒的机会有限 治疗，因此表现出先前存在的CD4+T细胞耗竭。因此，SARS-CoV-2感染可能 加速临床进展为艾滋病，或者，HIV+PWID中SARS-CoV-2感染可能加剧 新冠肺炎的临床症状导致死亡风险增加。因此，HIV+PWID可能有更高的风险 死于SARS-CoV-2感染。在这些PWID人群中，减少T细胞耗竭将是非常重要的 有益于阻止临床进展，可能是一个可行的长期治疗目标。的具体目标 这项补充建议是为了重新调整现有技术的用途，以快速开发基因驱动疗法(GDT) SARS-CoV-2候选病毒及其在患者T细胞中的干扰和体外传播广度的量化 来自HIV+PWID。这一努力将在很大程度上建立在我们最近在设计HIV-1 GDT方面的成功基础上(见Parent 和针对寨卡病毒的GDT(ZIKV)，表明GDT的概念可以被重新用于其他 病毒。中心假设--基于对艾滋病毒和寨卡病毒的广泛初步研究--是一个假定的 SARS-CoV-2 Gdt去除了所有致病病毒基因，可以靶向与野生型SARS相同的细胞 CoV-2(包括T淋巴细胞)，竞争细胞内资源，降低SARS CoV-2病毒载量和 发病机制，从而起到单次给药的治疗作用。GDT对策的基本原理 对于SARS，CoV-2是基于人源化小鼠中HIV-1的广泛数据和FDA的阳性会议。我们会 通过两个具体目标实现我们的目标：(I)设计一个SARS-CoV-2 GDT候选病毒(通过调整现有的 生物反应器平台)；以及(Ii)测试SARS CoV-2 GDT候选对患者T细胞的保护作用 墨西哥提华纳的HIV+PWID队列。虽然GDT方法带有固有的风险，但单一管理 治疗将是非常有益的，特别是对于治疗难以接触到的高危PWID人群。 不管GDT在预防T细胞枯竭方面是否成功，这里提出的研究将具有 SARS-CoV-2感染对HIV+T淋巴细胞影响的广泛基础意义 PWID。这些研究还将验证一种新的医学对策，其潜在的 快速部署以应对新的病毒威胁。

PROJECT SUMMARY/ABSTRACT For COVID-19, persons who inject drugs (PWID) have been identified as a population at high-risk of exposure to SARS-CoV-2 (the virus that causes COVID-19) because of their increased risk of homelessness or incarceration—situations linked to increased rates of the disease transmission and co-infection with HIV-1. Given that a SARS-CoV-2 vaccine is likely 12–24 months away, there is a critical unmet medical need for medical countermeasures that could contain COVID-19 outbreaks in the general population and in these difficult-to-reach high-risk populations such as PWIDs in particular. Moreover, there is a fundamental gap in our understanding of SARS-CoV-2 infection and pathogenesis in these at-risk PWID populations. Evidence indicates that SARS- CoV-2 infects and depletes T lymphocytes and many HIV-infected PWID have limited access to antiretroviral therapy and consequently exhibit pre-existing CD4+ T-cell depletion. Hence, SARS-CoV-2 infection could accelerate clinical progression to AIDS, or alternatively, SARS-CoV-2 infection in HIV+ PWID could exacerbate COVID-19 clinical symptoms leading to elevated risk of death. Thus, HIV+ PWID may be at elevated risk of death from SARS-CoV-2 infection. In these PWID populations, reducing T-cell depletion would be highly beneficial to halting clinical progression and may a viable long-term therapeutic goal. The specific objective of this supplement proposal is to repurpose existing technologies to rapidly develop a Gene Drive Therapy (GDT) candidate for SARS-CoV-2 and quantify its breadth of interference and transmission in vitro in patient T-cells from HIV+ PWID. This effort will build heavily off our recent success in engineering an HIV-1 GDT (see Parent Award) and a GDT against Zika Virus (ZIKV), demonstrating that the GDT concept can be repurposed for other viruses. The central hypothesis—based on extensive preliminary studies in HIV and ZIKV—is that a putative SARS-CoV-2 GDT, depleted of all the pathogenic viral genes, could target the same cells as wild-type SARS CoV-2 (including T lymphocytes), compete for intracellular resources, and reduce SARS CoV-2 viral load and pathogenesis, thereby serving as a single-administration therapeutic. The rationale for a GDT countermeasure for SARS CoV-2 is based on extensive data for HIV-1 in humanized mice and positive FDA meetings. We will achieve our objectives via two specific aims: (i) Engineer a SARS-CoV-2 GDT candidate (by adapting the existing Bioreactor platform); and (ii) Test the SARS CoV-2 GDT candidate's protective effect on patient T-cells from an HIV+ PWID cohort in Tijuana Mexico. While the GDT approach carries inherent risks, single-administration therapeutics would be highly beneficial particularly for treating difficult-to-reach, high-risk PWID populations. Regardless of the success of GDTs in protecting against T-cell depletion, the studies proposed here will have broad fundamental significance by assaying how SARS-CoV-2 infection impacts T lymphocytes from HIV+ PWID. These studies would also provide validation of a novel medical countermeasure with the potential to be rapidly deployed against new viral threats.