CRISPR for Cure

虽然抗逆转录病毒疗法(Art)极大地降低了艾滋病毒的发病率和死亡率，但它并没有起到作用。 消灭病毒库。中断治疗会导致潜伏病毒的激活和体内的反弹病毒血症 几周。迫切需要新的战略来根除潜伏感染和增强免疫系统 导致在停止抗逆转录病毒疗法后持续、持久地控制病毒反弹。为了回应RFA-AI- 20-035马丁·德莱尼艾滋病毒治疗研究合作中心，我们现在提交题为 “CRISPR for Cure”该计划的首要目标是使用CRISPR中介的基因组编辑来 增强免疫反应，直接消灭艾滋病毒前病毒。我们已经组建了一个协作团队， 成就卓著的基础和翻译科学家与社区利益相关者和 一家小型生物技术公司将开发基于CRISPR的疗法，以直接针对艾滋病毒前病毒并 增强免疫应答。该研究计划由三项高度互动研究组成 Foci(RF)将利用跨学科、创新和与社区协作的研究方法 以及政府的投入。RF1将使用下一代测序和新型条形码病毒来定义HIV 储存库和表观遗传机制对前病毒反弹的影响。在RF2中，我们将增强效应器NK 和CTL细胞的功能，并通过创新的基因组编辑杀死和限制目标细胞的病毒传播 战略。RF3将创建和测试下一代可诱导的、多路CRISPR，并增加 CD4嗜性淋巴AAV9用于根除HIV-1前病毒DNA的特异性、有效性和安全性 在其免疫细胞在RF2中被修饰的动物模型中，评估普遍的和 个体化CRISPR在体内清除复制能力强的病毒。除了共同关注 CRISPR技术，合作实验室将通过 条形码病毒在人源化小鼠中的共享使用 支持 MISTRG人源化小鼠 人类造血干细胞和祖细胞的维持和骨髓生成；恒河猴 感染一种新的SIV条形码病毒；来自具有良好特征的队列的体外临床样本及其用途 腺病毒高效地将CRISPR运送到体内人源化动物模型 患者来源的PBMCs。这一综合性和多学科计划的结果是由CRISPR for 治愈“ 这个 合作，将加快使用基因编辑策略，以根除来自 停止抗逆转录病毒治疗后体内或持续的病毒缓解。 从我们的私营部门合作伙伴那里，我们将为GMP制造的进一步发展做好准备，以及 未来的初步临床研究。

While antiretroviral therapy (ART) has dramatically reduced HIV disease morbidity and mortality, it has failed to eliminate viral reservoirs. Interruption of treatment leads to activation of latent virus and rebound viremia within weeks. Novel strategies are urgently needed to eradicate latent infections and enhance the immune system leading to sustained, durable control of viral rebound following the cessation of ART. In response to RFA-AI- 20-035 Martin Delaney Collaboratories for HIV Cure Research, we now submit the application entitled "CRISPR for Cure." The overarching goal of this program is to use genome editing mediated by CRISPR to enhance immune responses and directly ablate HIV proviruses. We have assembled a collaborative team of highly accomplished basic and translational scientists working in tandem with community stakeholders and a small biotechnology company to develop CRISPR-based therapies to directly target the HIV provirus and to enhance immunological responses. The research program is comprised of three highly interactive research foci (RF) that will utilize interdisciplinary, innovative and collaborative research approaches with community and government input. RF1 will use next generation sequencing and novel barcoded viruses to define the HIV reservoir and the impact of epigenetic mechanisms on proviral rebound. In RF2, we will enhance effector NK and CTL cell function and killing and limit viral spread by target cells using innovative genome editing strategies. RF3 will create and test the next generation of inducible, multiplex CRISPR with increased specificity, potency and safety for delivery by CD4 tropic lymphoid AAV9 for eradication of HIV-1 proviral DNA in animal models whose immune cells are modified in RF2 and assess the possibility of both a universal and personalized CRISPR in eliminating replication competent virus in vivo. In addition to the shared focus on CRISPRs technology, the Collaboratory will undertake a highly integrated experimental agenda through the shared use of barcoded viruses in humanized mice and unique support MISTRG humanized mice that differentially human hematopoietic stem and progenitor cell maintenance and myelopoiesis;rhesus macaques infected with a novel SIV barcoded virus; ex vivo clinical samples from a well characterized cohort and the use of adenoviruses to efficiently deliver CRISPRs to an in vivo humanized animal model carrying cells from patient-derived PBMCs. The outcome of this comprehensive and multidisciplinary program by the “CRISPR for Cure” the Collaboratory, will accelerate the use of gene editing strategies towards eradication of HIV infection from body or sustained viral remission following cessation of antiretroviral therapy.With resources available from our private sector partner, we will be well positioned for further GMP manufacturing development, and future initial clinical investigations.