Combination gene editing for local and systemic HIV resistance

局部和全身艾滋病毒耐药性的组合基因编辑

基本信息

批准号：
10409805
负责人：
Paula M Cannon
金额：
$ 49.38万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10409805
关键词：
Adoptive Transfer Antibodies Antigens Autologous B-Lymphocytes Berlin Binding CCR5 gene CD4 Positive T Lymphocytes Cell Line Cell Transplantation Cell physiology Cells Cellular biology Collaborations DNA DNA Repair DNA cassette Elements Engineering Evolution Gene Combinations Generations Genes Genetic Genome engineering HIV HIV Infections HIV resistance Hematopoietic Hematopoietic stem cells Human IgG1 Immune Immune response Immunoglobulin G Immunoglobulin Somatic Hypermutation Immunoglobulins In Vitro Individual Knock-out Macaca Macaca mulatta Mediating Membrane Methods Modification Mus Nonhomologous DNA End Joining Outcome Pathway interactions Patients Peripheral Production Reagent Safety Series Signal Transduction Site T-Cell Depletion Technology Time Transplantation V(D)J Recombination Vaccination Viral Viremia base combinatorial engineered stem cells experience gain of function gain of function mutation gene repair gene therapy humanized mouse improved inhibitor mimetics mouse model nanobodies neutralizing antibody novel nuclease plasma cell differentiation predictive tools repaired targeted nucleases

项目摘要

ABSTRACT Gene editing holds the promise of enhancing the precision and safety of anti-HIV gene therapies based on engineering hematopoietic stem and progenitor cells (HSPC), in order to recreate elements of the Berlin Patient cure. Targeted nuclease technology is already being used to exploit NHEJ-mediated repair of DNA breaks and thereby disrupt the CCR5 gene, which mimics one aspect of his cure. However, extending this treatment to non- cancer patients, receiving engineered autologous HSPC, will likely need to include other approaches. To do this, we are exploiting the alternate pathway of DNA break repair based on homology-directed repair (HDR). This can be used to introduce gain-of-function mutations into cellular restriction factors, or to direct the controlled secretion of soluble anti-viral factors, including the broad entry inhibitor eCD4-Ig. Moreover, the site-specific insertion at eCD4-Ig at the CCR5 locus would combine HIV-resistance with systemic protection and thereby provide a combinatorial anti-HIV approach. In the current proposal we aim to continue to improve the safety and efficacy of gene editing in HSPC, and to apply the technology to provide both local and systemic HIV resistance. We will combine our established anti-HIV approaches with a new direction to engineer production of broadly neutralizing antibodies, and thereby provide a novel synthetic immune capability. Individual strategies and combination approaches will be evaluated in appropriate humanized mouse models, to evaluate their impact on HIV infection and the latent reservoir.

抽象的基因编辑有望提高基于抗 HIV 基因疗法的精确度和安全性对造血干细胞和祖细胞（HSPC）进行改造，以重建柏林病人的元素治愈。靶向核酸酶技术已被用于利用 NHEJ 介导的 DNA 断裂修复和从而破坏 CCR5 基因，这模仿了他的治疗方法的一方面。然而，将这种治疗扩展到非接受工程化自体 HSPC 的癌症患者可能需要采用其他方法。为此，我们正在开发基于同源定向修复（HDR）的 DNA 断裂修复替代途径。这个可以用于将功能获得性突变引入细胞限制因子，或指导受控分泌可溶性抗病毒因子，包括广泛进入抑制剂 eCD4-Ig。此外，位点特异性插入 CCR5 基因座上的 eCD4-Ig 将 HIV 抗性与系统保护结合起来，从而提供组合抗 HIV 方法。在当前的提案中，我们的目标是继续提高安全性和有效性 HSPC 中的基因编辑，并应用该技术提供局部和系统的 HIV 抵抗力。我们将将我们已建立的抗艾滋病毒方法与新的方向相结合，以设计广泛中和的生产抗体，从而提供新的合成免疫能力。个人策略和组合将在适当的人源化小鼠模型中评估方法，以评估其对艾滋病毒感染的影响和潜在的水库。