In vivo CRISPR engineering of B cells to produce anti-HIV broadly neutralizing antibodies using novel nanoparticles

B 细胞体内 CRISPR 工程利用新型纳米粒子产生抗 HIV 广泛中和抗体

• 批准号: 10640843
• 负责人: Jennifer Eileen Adair
• 金额: $ 90.72万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-08 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640843
• 关键词: Anti-Retroviral Agents; Antibodies; Antibody Formation; Antibody Therapy; Antigens; Autologous Transplantation; B-Lymphocytes; Bar Codes; Binding; Blood Cells; Bone Marrow; Cell Survival; Cell surface; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Coupling; DNA; Data; Engineering; Ensure; Epidemic; Equity; Frequencies; Gene Delivery; Genes; Genetic; Genetic Engineering; HIV; HIV Infections; Half-Life; Hematopoietic stem cells; Human; Human Herpesvirus 4; Humoral Immunities; Immune; Immunity; Immunoglobulin G; Immunoglobulin-Secreting Cells; Immunology; Injections; Interruption; Intervention; Laboratories; Life; Liver; Mediating; Memory B-Lymphocyte; Modeling; Molecular Target; Mus; Nature; Patients; Persons; Pharmaceutical Preparations; Plasma Cells; Process; Proliferating; Research; Resources; Respiratory syncytial virus; Risk; Science; Single-Stranded DNA; Source; Surface; System; Technology; Testing; Therapeutic; Tissues; Transplantation; Vaccines; Viral; Viral Vector; Virus Diseases; Work; antibody engineering; cell type; clinically relevant; cost; cost effective; design; experience; fitness; gene therapy; genetically modified cells; immunogenic; improved; in vivo; in vivo evaluation; influenzavirus; lipid nanoparticle; manufacture; mouse model; nanoGold; nanoformulation; nanoparticle; neutralizing antibody; nonhuman primate; novel; novel strategies; preference; prevent; promoter; side effect; simian human immunodeficiency virus; success; synergism; therapeutic genome editing; tool; viral rebound

PROJECT SUMMARY / ABSTRACT The quest for an HIV cure remains incomplete, nearly half a century since the onset of the epidemic. Antiretroviral drug cocktails can suppress HIV infection, but suffer in their success owing to side effects and limitations in access and compliance. Injection of broadly neutralizing antibodies (bNAbs) to prevent HIV rebound has had some success, but requires regular re-injection of multiple antibodies to maintain suppression and viral escape. Thus, cost and continued access remain limitations. Genetic engineering of patient cells has been proposed to overcome all of these shortfalls, and could constitute a one-time treatment with lifelong therapeutic value if successful. In this proposal, we leverage a novel approach developed by Dr. Justin Taylor’s laboratory to genetically engineer B cells to express bNAbs for the treatment of human immunodeficiency virus (HIV). This strategy has already been used to engineer B cells to produce antibodies protective against influenza virus, respiratory syncytial virus, Epstein-barr virus and HIV [Moffett et al., Science Immunology, 2019]. While this approach can ensure protective antibody production, the genetic engineering process required 10 days of complicated ex vivo manufacturing and is not broadly distributable. To overcome these barriers, we will co-opt a novel, synthetic nanoparticle that was developed in Dr. Jennifer Adair’s laboratory to deliver genetic engineering in a single, passive step [Shahbazi et al., Nature Materials, 2019]. We show that this nanoparticle can be assembled in less than a day to genetically engineer unstimulated, primary human blood cells and can be modified to specifically interact with target blood cell types in vivo. Here we will develop this scalable nanoformulation as a vaccine-like in vivo delivery system to direct humoral immunity with multiple bNAbs in a clinically-relevant nonhuman primate model of HIV infection. We will use these nanoparticles to directly genetically engineer native primary B cell subtypes, and hematopoietic stem and progenitor cells, which can provide lifelong replenishment of antibody- producing B cells. This research will not only develop a unique tool set against HIV but will provide transformative advances in equitable distribution of gene editing therapies.

项目摘要/摘要 在艾滋病疫情爆发近半个世纪后，对艾滋病毒治愈方法的探索仍未完成。 抗逆转录病毒药物鸡尾酒可以抑制艾滋病毒感染，但由于副作用，其成功受到影响 以及访问和合规性方面的限制。注射广谱中和抗体(BNAbs) 预防艾滋病毒反弹已取得一定成功，但需要定期重新注射多种抗体 保持抑制和病毒逃逸。因此，费用和持续使用仍然受到限制。遗传 对患者细胞进行工程已被提出以克服所有这些不足，并可能构成 如果成功，一次性治疗将具有终身治疗价值。在这个提案中，我们利用了一本小说 贾斯汀·泰勒博士的实验室开发的基因工程B细胞表达bNAbs的方法 用于治疗人类免疫缺陷病毒(HIV)。这种策略已经被用来 改造B细胞以产生针对流感病毒、呼吸道合胞病毒、 爱泼斯坦-巴尔病毒与艾滋病毒[Moffett等人，科学免疫学，2019年]。虽然这种方法可以确保 保护性抗体的生产，基因工程过程需要10天的复杂实验 VIVO制造，不能广泛分销。为了克服这些障碍，我们将增选一部小说， 由詹妮弗·阿代尔博士的实验室开发的合成纳米颗粒，用于传递基因 一步完成工程设计[Shahbazi等人，《自然材料》，2019]。我们证明了这一点 纳米颗粒可以在不到一天的时间内组装成基因工程，在没有刺激的情况下，原生人类 它可以被改造成与体内的目标血细胞类型发生特异性的相互作用。在这里，我们将 开发这种可伸缩的纳米制剂作为疫苗样的体内递送系统来引导体液 HIV感染的临床相关非人类灵长类动物模型中多种bNAbs的免疫。我们会 使用这些纳米颗粒直接对原始B细胞亚型进行基因工程，以及 造血干细胞和造血祖细胞，可提供终生补充抗体- 产生B细胞。这项研究不仅将开发一种针对艾滋病毒的独特工具，而且将提供 基因编辑疗法公平分配方面的变革性进展。