Autologous HIV-1 resistant T cells through accelerated CCR5 gene disruption

通过加速 CCR5 基因破坏产生自体 HIV-1 抗性 T 细胞

基本信息

批准号：
8603742
负责人：
Alexander Astrakhan
金额：
$ 50.77万
依托单位：
PRECISION GENOME ENGINEERING, INC.
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8603742
关键词：
Adverse effects Alleles Anti-Retroviral Agents Autologous Biodistribution Biological Biological Assay Bone Marrow Transplantation CCR5 gene CD4 Positive T Lymphocytes Case Study Cell Separation Cells Characteristics Clinic Clinical Clinical Trials DNA Data Set Dependence Development Dose Electroporation Engineering Engraftment Enhancement Technology Evaluation Event Exonuclease Gene Delivery Gene Targeting Gene-Modified Generations Genes Genetic Genome Genomics HIV HIV Infections HIV-1 Health Heterogeneity Highly Active Antiretroviral Therapy Homing Human Immunodeficient Mouse In Vitro Individual Infection Lead Length Lymphocyte Mediating Messenger RNA Methods Modeling Monitor Mutation Patients Performance Pharmaceutical Preparations Pharmacologic Substance Phase Positioning Attribute Process Production Protocols documentation Qualifying Quality of life Reagent Regimen Resistance Safety Site Small Business Technology Transfer Research Specificity Staging Surface T cell therapy T-Lymphocyte Technology Testing Therapeutic Toxic effect Toxicology Transcription Coactivator Validation Variant Viral Viremia alternative treatment base endonuclease improved in vivo manufacturing process mouse model novel nuclease outcome forecast peripheral blood potency testing pre-clinical preclinical efficacy preclinical safety product development programs public health relevance receptor reconstitution research study success

项目摘要

DESCRIPTION (provided by applicant): Treating patients infected with Human Immunodeficiency Virus-1 (HIV-1) is a substantial clinical challenge with evolving pharmaceutical drug and dosing options. Treatments should improve the health and quality of life for HIV-infected individuals by reducing viremia and preserving immunological function. Currents therapeutic approaches utilize combination antiretroviral drug regiments (cARV, historically referred to highly active antiretroviral therapy, or HAART) to reduce ongoing viral replication and improve the patient's prognosis. However, cARV therapies are not curative and furthermore they are expensive, are associated with high rates of drug-related toxicity, and are highly susceptible to viral evasion through resistance-conferring mutation. Genetic targeting CCR5 is a promising alternative approach to cARV therapy. The CCR5 gene is used by HIV-1 for cellular entry and those individuals who naturally lack CCR5 expression are fully resistant to HIV-1 infection. In an important clinical case study, bone marrow transplantation from a CCR5-deficient donor has produced the first recorded case of functional cure of an ongoing HIV-1 infection. This has prompted therapeutic strategies to create HIV-1 resistant patient-derived T cells using nuclease- based reagents to create disruptive mutations in the CCR5 gene. Though providing critical proof-of- concept in HIV-1 patients, the success of the gene-targeting therapy currently in clinical trials appears to be limited due to low gene disruption activity of these firt-generation nuclease technologies. Pregenen has used its surface display engineering platform to produce highly specific CCR5- targeting LAGLIDADG homing endonucleases (LHE), recently reformatted as extremely powerful MegaTAL nucleases and combined with an additional layer of rate-enhancement technology - exonuclease-accelerated genome editing (X-AGE) developed at SCRI. The proposed therapy will isolated and process patient-derived T-cells by delivering these enhanced reagents using a simplified and scalable mRNA-based method. This manufacturing process will be used to create HIV-resistant, CCR5-deficient autologous CD4+ T cell. Autologous CCR5-disrupted T cells have the potential to durably suppress HIV-1 infection and reconstitute the full functionality of the CD4+ compartment while reducing dependence on antiretroviral drugs. The experiments described in this proposal are part of a comprehensive plan to bring the CCR5-targeting nuclease therapy to the clinic. We plan to profile the safety, biodistribution, and potency of primary human T cells treated with CCR5-targeting reagents. The best-performing reagents and protocols will be used to generate CCR5-deficient human CD4+ T cells to test the capacity of these cells to withstand HIV infection in vitro and in vivo. We will se this data-set to support pivotal preclinical experiments and subsequent IND submission for the continued clinical development of our cellular therapy.

描述（由申请人提供）：治疗感染人类免疫缺陷病毒-1 (HIV-1) 的患者是一项重大的临床挑战，药物和剂量选择不断变化。治疗应通过减少病毒血症和保留免疫功能来改善艾滋病毒感染者的健康和生活质量。目前的治疗方法利用联合抗逆转录病毒药物疗法（cARV，历史上称为高效抗逆转录病毒疗法，或HAART）来减少持续的病毒复制并改善患者的预后。然而，cARV 疗法没有治愈性，而且价格昂贵，与药物相关的毒性发生率很高，并且很容易通过赋予耐药性的突变逃避病毒。基因靶向 CCR5 是一种有前途的 cARV 治疗替代方法。 CCR5 基因被 HIV-1 用于进入细胞，那些天然缺乏 CCR5 表达的个体对 HIV-1 感染具有完全抵抗力。在一项重要的临床案例研究中，来自 CCR5 缺陷供体的骨髓移植产生了第一个有记录的 HIV-1 感染功能性治愈病例。这促使人们采用基于核酸酶的试剂在 CCR5 基因中产生破坏性突变来创建 HIV-1 抗性患者来源的 T 细胞的治疗策略。尽管在 HIV-1 患者中提供了关键的概念验证，但由于这些第一代核酸酶技术的基因破坏活性较低，目前临床试验中的基因靶向治疗的成功似乎受到限制。 Pregenen 利用其表面显示工程平台生产高度特异性的 CCR5 靶向 LAGLIDADG 归巢核酸内切酶 (LHE)，最近重新格式化为极其强大的 MegaTAL 核酸酶，并结合了额外的速率增强技术 - 核酸外切酶加速基因组编辑 (X-AGE) ）在 SCRI 开发。拟议的疗法将通过使用简化且可扩展的基于 mRNA 的方法提供这些增强的试剂来分离和处理患者来源的 T 细胞。该制造工艺将用于制造抗 HIV、CCR5 缺陷的自体 CD4+ T 细胞。自体 CCR5 破坏的 T 细胞有潜力持久抑制 HIV-1 感染并重建 CD4+ 区室的全部功能，同时减少对抗逆转录病毒药物的依赖。该提案中描述的实验是将 CCR5 靶向核酸酶疗法推向临床的综合计划的一部分。我们计划分析用 CCR5 靶向试剂处理的原代人类 T 细胞的安全性、生物分布和效力。性能最佳的试剂和方案将用于生成 CCR5 缺陷型人类 CD4+ T 细胞，以测试这些细胞在体外和体内抵抗 HIV 感染的能力。我们将利用该数据集来支持关键的临床前实验和随后的 IND 提交，以支持我们细胞疗法的持续临床开发。