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

项目摘要/摘要 仅在美国，流感等呼吸道病毒病原体感染就导致数百万人 提供者就诊和数万个工作日的损失，数十万人住院和 死亡。根据世界卫生组织的数据，病毒致病感染的数量仍在继续 在资源有限的国家，随着死亡率的升高而上升。虽然一个成功的疫苗策略是高度 这种方法是可取的，它依赖于诱导B细胞产生保护性抗体，这种抗体 防止病毒进入细胞或以受感染细胞为目标进行破坏。不幸的是，成功了 经过几十年的研究，许多病毒的疫苗还没有问世，比如呼吸道合胞病毒 病毒(RSV)。在这项提案中，我们利用了贾斯汀·泰勒博士的实验室开发的一种新方法 通过基因工程使B细胞表达对呼吸道病毒具有保护作用的抗体，包括 呼吸道合胞病毒和流感。这一战略已被证明会导致生产保护性产品 抗流感、呼吸道合胞病毒和人类免疫缺陷病毒感染的抗体[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，但基因 工程过程需要10天复杂的体外制造，并且不是广泛的 可分发的。为了克服这些障碍，我们将加入一种新的、合成的纳米颗粒 由詹妮弗·阿代尔博士的实验室开发，以一种被动的单一步骤提供基因工程 [Shahbazi等人，《自然材料》，2019]。我们证明了这种纳米颗粒可以组装成 在不到2天的时间内对原代人类血细胞进行基因工程，并可以被修改为特定的 与体内的目标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配方作为 类似疫苗的体内递送系统，可针对呼吸道病毒(如 RSV。我们将使用这些纳米颗粒直接基因工程最具保护性的初级B细胞 亚型，以及造血干细胞和造血祖细胞，可以提供终生补充 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来对抗病毒 但将在基因编辑疗法的公平分配方面取得变革性的进展。

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.