Targeted genetic engineering of B cells to induce protective antibody responses to viral pathogens

项目概要/摘要 仅在美国，呼吸道病毒病原性感染如流感就导致数百万人死亡。 提供者访问和数万个工作日的损失，数十万人住院， 死亡据世界卫生组织称，病毒致病性感染的数量仍在继续 在资源有限的国家死亡率更高。虽然一个成功的疫苗策略是高度 这种方法依赖于诱导B细胞产生保护性抗体， 防止病毒进入细胞或将受感染的细胞作为破坏目标。不幸的是，成功 经过几十年的研究，许多病毒的疫苗还没有问世，如呼吸道合胞病毒， 病毒（RSV）。在这个提议中，我们利用了贾斯汀·泰勒博士实验室开发的一种新方法， 对B细胞进行基因工程改造，使其表达抗呼吸道病毒的抗体， RSV和流感。这种策略已经被证明可以产生保护性的 抗流感、RSV和人免疫缺陷病毒感染的抗体[Moffett等，科学 免疫学，2019]。虽然这种方法可以确保保护性抗体的产生，但遗传性抗体的产生可能会增加。 工程过程需要10天的复杂离体制造， 可分配的为了克服这些障碍，我们将选择一种新的合成纳米颗粒， 在詹妮弗·阿代尔博士的实验室里开发的一种基因工程， [Shahbazi等人，Nature Materials，2019].我们发现这种纳米粒子可以组装成 在不到2天的时间内对原代人类血细胞进行基因工程改造， 在体内与靶血细胞类型相互作用。在这里，我们将开发这种可扩展的纳米制剂， 疫苗样体内递送系统，以引导针对呼吸道病毒的免疫应答， RSV。我们将使用这些纳米颗粒直接基因工程最具保护性的原代B细胞 亚型，造血干细胞和祖细胞，可以提供终身补充， 保护性B细胞和抗体。这项研究不仅将开发一种独特的工具， 病原体，但将在基因编辑疗法的公平分配方面提供变革性进展。

PROJECT SUMMARY / ABSTRACT In the United States alone, respiratory viral pathogenic infections such as influenza cause millions of provider visits and tens of thousands of work days lost, hundreds of thousands of hospitalizations and deaths. According to the World Health Organization, the number of viral pathogenic infections continues to rise with higher mortalities in resource limited countries. While a successful vaccine strategy is highly desirable, this approach relies on the induction of B cells to produce protective antibodies that either prevent viruses from entering cells or target infected cells for destruction. Unfortunately, successful vaccines for many viruses are not yet available after decades of research such a respiratory syncytial virus (RSV). In this proposal, we leverage a novel approach developed by Dr. Justin Taylor’s laboratory to genetically engineer B cells to express antibodies protective against respiratory viruses including RSV and influenza. This strategy has already been shown to result in the production of protective antibodies against influenza, RSV, and human immunodeficiency virus infection [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 to genetically engineer primary human blood cells in less than 2 days, 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 immune responses against respiratory viruses such as RSV. We will use these nanoparticles to directly genetically engineer the most protective primary B cell subtypes, and hematopoietic stem and progenitor cells, which can provide lifelong replenishment of protective B cells and antibodies. This research will not only develop a unique tool set against viral pathogens, but will provide transformative advances in equitable distribution of gene editing therapies.