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

基本信息

批准号：
10160822
负责人：
Dong Sung An
金额：
$ 46.67万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
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：
2020-05-07 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10160822
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 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 Lentivirus Vector MGMT gene Macaca nemestrina Malignant - descriptor Malignant Neoplasms Methotrexate Modeling Mus Patients Procedures Prodrugs RNA Interference Reaction Risk Safety T-Lymphocyte Testing Therapeutic Thioguanine TimeLine Toxic effect Transplantation Treatment Efficacy Virus Replication base 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

项目摘要

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和安全的阳性和阴性选择策略来调节抗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(2) 结合安全“杀伤开关”，以消除基因工程抗 HIV-1 免疫效应细胞，以防出现意外的不良反应，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实现 HIV-1 治愈的巨大希望，但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。因此，需要一种安全且可滴定的阳性选择策略来最大限度地提高抗 HIV-1 基因工程免疫细胞的水平，从而在没有危险的强化治疗的情况下治疗 HIV-1 患者。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 免疫效应细胞非常重要。因此，我们将开发阴性选择。作为消除基因工程免疫细胞的安全“杀死开关”的策略，我们将确定最有效和最安全的选择策略：(1) 使用 RNA 敲低次黄嘌呤鸟嘌呤磷酸核糖转移酶 (HPRT) 的表达。使我们能够使用临床上可用的前药 6-硫鸟嘌呤或甲氨蝶呤有效消除或消除抗 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的标记物以及 (3)用于阳性选择的人 O6-甲基鸟嘌呤-DNA-甲基转移酶 (MGMTP140K) 的 P140K 突变形式我们发现，通过严格评估我们提出的抗 HIV 筛选策略，可以开发临床相关、安全且有效的阳性和阴性选择策略。 -1 CAR 和 scFv-Fc bNAb 联合疗法可实现 HIV 疾病的治愈。