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

基本信息

批准号：
10374397
负责人：
Jennifer Eileen Adair
金额：
$ 87.79万
依托单位：
FRED HUTCHINSON CANCER CENTER
依托单位国家：
美国
项目类别：
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/10374397
关键词：
Anti-Retroviral Agents Antibodies Antibody Formation Antibody Therapy Antigens Autologous Transplantation B-Lymphocytes Bar Codes Blood Cells Bone Marrow CRISPR/Cas technology Cell Survival Cell surface Cells Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Coupling DNA Data Engineering Ensure Epidemic 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 Time 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 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

项目摘要

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 /摘要自流行病开始以来，对艾滋病毒治愈的追求仍然不完整。抗逆转录病毒药物鸡尾酒可以抑制艾滋病毒感染，但由于副作用而遭受成功和访问和合规性的限制。注射广泛中和抗体（BNAB）防止艾滋病毒反弹取得了一定的成功，但需要定期重新注射多种抗体保持抑制和病毒逃生。那是成本和持续访问仍然存在的局限性。遗传已经提出了患者细胞的工程来克服所有这些缺口，并且可能构成如果成功，则具有终身治疗价值的一次性治疗。在这个建议中，我们利用一本小说贾斯汀·泰勒（Justin Taylor）博士的实验室开发的方法为基因工程B细胞表达BNABS 用于治疗人类免疫缺陷病毒（HIV）。该策略已经习惯了工程师B细胞产生抗体免受影响病毒，呼吸综合病毒的抗体， Epstein-Barr病毒和HIV [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产生，基因工程过程需要10天复杂的EX 体内制造，不可分割。为了克服这些障碍，我们将选择一本小说，詹妮弗·阿黛尔（Jennifer Adair）博士实验室中开发的合成纳米颗粒以提供遗传一个被动的步骤[Shahbazi等人，自然材料，2019年]。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，以遗传刺激的原代人血细胞可以修改以在体内特异性与靶血细胞类型特异性相互作用。我们会在这里开发这种可扩展的纳米成型，作为一种类似于疫苗的体内递送系统，以引导体液在临床上与HIV感染的非人类私有模型中，具有多个BNAB的免疫力。我们将使用这些纳米颗粒直接加密设计天然的原代B细胞亚型，并且造血干细胞和祖细胞，可以提供抗体的终身复制产生B细胞。这项研究不仅将开发针对艾滋病毒的独特工具，而且会提供基因编辑疗法公平分布的变革性进步。