Modulation of repopulation of anti HIV-1 gene-modified cells to enhance efficacy and safety

调节抗 HIV-1 基因修饰细胞的再增殖以提高功效和安全性

• 批准号: 10614651
• 负责人: Dong Sung An
• 金额: $ 45.36万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-07 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614651
• 关键词: Adverse effects; Antigens; Autoimmune; B-Lymphocytes; Benefits and Risks; Bone Marrow Purging; CAR T cell therapy; CD19 gene; Cell Count; Cell surface; Cells; Cellular Immunity; Cetuximab; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Comparative Study; Dangerousness; Disease; Effector Cell; Engineered Gene; Engraftment; Epidermal Growth Factor Receptor; Erbitux; FDA approved; Gene Delivery; Gene Modified; Genetic Engineering; Goals; HIV; HIV-1; Hematopoietic stem cells; Human; Humoral Immunities; Hypoxanthine Phosphoribosyltransferase; Immune; In Vitro; Investigational New Drug Application; Learning; Lentivirus Vector; MGMT gene; Macaca nemestrina; Malignant - descriptor; Malignant Neoplasms; Methotrexate; Modeling; Mus; Patients; Population; Procedures; Prodrugs; RNA Interference; Reaction; Safety; T-Lymphocyte; Testing; Therapeutic; Thioguanine; Toxic effect; Transplantation; Treatment Efficacy; Virus Replication; cancer immunotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; clinically relevant; cytokine release syndrome; gene therapy; immunoengineering; immunogenicity; in vivo; knock-down; mutant; neutralizing antibody; nonhuman primate; preclinical development; programs; timeline

Project 3: Summary/Abstract The overall goal of Project 3 is to modulate the levels of anti-HIV-1 chimeric antigen receptor (CAR) and broadly neutralizing antibodies (bNAb) modified immune cells by developing the most effective and safe positive and negative selection strategy to (1) achieve a therapeutic level of repopulation and (2) incorporate a safety “kill-switch” to eliminate the genetically engineered anti-HIV-1 immune effector cells in cases of unexpected adverse effects, such as cytokine storm, autoimmune reaction and malignant transformation. The hematopoietic stem cell-based gene therapy approach has shown great promise to achieve an HIV-1 cure. However, one of the major limitations has been the difficulty of achieving the engraftment levels sufficient to provide therapeutic efficacy, in particular for HIV-1 infected patients where intensive myeloablative conditionings would be an unfavorable risk-benefit. Thus, a safe and titratable positive selection strategy is highly desirable to maximize the level of anti-HIV-1 gene engineered immune cells to treat patients with HIV-1 without dangerous intensive myeloablation. Furthermore, it is important to incorporate a safety “kill-switch” procedure to eliminate the genetically engineered anti-HIV-1 immune effector cells based on lessons learned from severe adverse effects in cancer immunotherapy. Therefore, we will develop a negative selection strategy as a safety “kill-switch” to eliminate genetically engineered immune cells. We will identify the most effective and safe selection strategy from (1) knocking down hypoxanthine-guanine phosphoribosyltransferase (HPRT) expression using RNA interference that enables us to effectively enrich or eliminate anti-HIV-1 gene-modified HSPC using clinically available prodrug 6-thioguanine or methotrexate, (2) co-expressing truncated non-functional human epidermal growth factor receptor (huEGFRt), a cell surface marker for a rapid ex vivo positive selection and in vivo negative selection by an FDA-approved anti-EGFR monoclonal antibody Cetuximab (Erbitux) and (3) the P140K mutant form of human O6-methylguanine-DNA-methyltransferase (MGMTP140K) for a positive selection. We hypothesize that a clinically relevant, safe and effective positive and negative selection strategy can be developed by rigorously evaluating our proposed selection strategies for our anti-HIV-1 CAR and scFv-Fc bNAb combining therapies to achieve a cure of HIV disease.

项目 3：总结/摘要 项目3的总体目标是通过开发最有效、最安全的正向和负向选择策略来调节抗HIV-1嵌合抗原受体（CAR）和广泛中和抗体（bNAb）修饰的免疫细胞的水平，以（1）实现治疗水平的再增殖和（2）结合安全“杀死开关”，以在出现意外不良反应的情况下消除基因工程抗HIV-1免疫效应细胞，例如 如细胞因子风暴、自身免疫反应和恶性转化。基于造血干细胞的基因治疗方法已显示出治愈 HIV-1 的巨大希望。然而，主要限制之一是难以达到足以提供治疗效果的植入水平，特别是对于 HIV-1 感染患者，在这些患者中，强化清髓调理将是不利的风险收益。因此，非常需要一种安全且可滴定的阳性选择策略，以最大限度地提高抗 HIV-1 基因工程免疫细胞的水平，以治疗 HIV-1 患者，而无需危险的强化清髓术。此外，根据癌症免疫治疗中严重不良反应的经验教训，采用安全的“杀死开关”程序来消除基因工程的抗 HIV-1 免疫效应细胞也很重要。因此，我们将开发一种负选择策略作为消除基因工程免疫细胞的安全“杀死开关”。我们将确定最有效和安全的选择策略：(1) 使用 RNA 干扰敲低次黄嘌呤鸟嘌呤磷酸核糖转移酶 (HPRT) 表达，使我们能够使用临床可用的前药 6-硫鸟嘌呤或甲氨蝶呤有效富集或消除抗 HIV-1 基因修饰的 HSPC，(2) 共表达截短的非功能性人表皮生长因子受体 (huEGFRt)，一种细胞表面标记物，用于通过 FDA 批准的抗 EGFR 单克隆抗体西妥昔单抗 (Erbitux) 进行快速离体阳性选择和体内阴性选择，以及 (3) 用于阳性选择的人 O6-甲基鸟嘌呤-DNA-甲基转移酶 (MGMTP140K) 的 P140K 突变形式。我们假设，通过严格评估我们提出的抗 HIV-1 CAR 和 scFv-Fc bNAb 联合疗法的选择策略，可以制定临床相关、安全且有效的阳性和阴性选择策略，以实现 HIV 疾病的治愈。