Heritable immunization of the white-footed mouse reservoir of Lyme disease

白足小鼠莱姆病储库的遗传免疫

• 批准号: 10674208
• 负责人: Kevin Esvelt
• 金额: $ 23.51万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674208
• 关键词: Adopted; Albumins; Alleles; Animal Model; Animals; Antibodies; Area; B-Lymphocytes; Back; Binding; Borrelia burgdorferi; Cell Line; Cell Surface Proteins; Chromosomal translocation; Communities; Deer; Disadvantaged; Disease; Disease Reservoirs; ES Cell Line; Elements; Engineering; Enterobacteria phage P1 Cre recombinase; Environment; Epitopes; Feedback; Gene Transfer Techniques; Genes; Geography; Goals; Heritability; Immunity; Immunization; Immunize; In Vitro; Incidence; Individual; Intervention; Investments; Light; Link; Liver; Lyme Disease; Membrane Proteins; Methods; Midwestern United States; Mus; Mutation; OspA protein; Play; Population; Prevention; Prevention strategy; Production; Protective Clothing; Reciprocal Translocation; Recombinants; Research Personnel; Resistance; Risk; Role; System; Testing; Tick-Borne Diseases; Ticks; Time; Transgenes; Transgenic Organisms; Vaccinated; White-Footed Mouse; Work; Zoonoses; acaricide; cost; disease transmission; embryonic stem cell; exposed human population; field study; fitness; in vitro activity; in vivo; interest; pathogen; prevent; resistance gene; transmission process; underdominance

Project Summary ​ ​ RFA-AI-19-037 The rising incidence of Lyme disease demands new strategies for prevention. Existing methods such as acaricides, deer reduction, landscaping, and personal protective clothing, are inherently short-term and must be regularly re-applied, maintained and worn. ​The Mice Against Ticks project seeks to develop a durable one-time intervention to disrupt the ecological cycle of Lyme disease transmission for many decades​. The causative agent of Lyme disease ​B. burgdorferi ​is passed back and forth between ticks and their small animal hosts, which serve as zoonotic reservoirs of disease. The white-footed mouse ​P. leucopus​ is widely considered to be the most important reservoir because it is both ubiquitous and extremely efficient at acquiring and transmitting pathogens via ticks. Our overarching goal for this proposed project is to heritably immunize white-footed mice against Lyme by encoding protective ​P​. ​leucopus​ antibodies targeting ​B. burgdorferi ​outer surface protein A (OspA) in the mouse germline. According to our calculations, combining at least four such antibodies should prevent evolutionary escape by ​B. burgdorferi ​because too many simultaneous OspA mutations would be needed. Crucially, even if these mice are less important in some areas than currently thought, immunization will reduce the number of infected ticks, which in turn will infect fewer secondary reservoirs, which will infect fewer ticks, reinforcing a negative feedback spiral anticipated to greatly reduce the local burden of Lyme disease. We have already successfully isolated anti-OspA antibodies from OspA-immunized ​P. leucopus​, derived putative ​P. leucopus embryonic stem cells, and shown that the albumin locus appears suitable for antibody secretion from the liver. We now seek to (1) identify antibodies that bind to at least four different OspA epitopes, (2) establish a stable embryonic stem cell line and perform germline editing, and (3) generate heritably resistant mice that express antibodies from a cisgenic cassette linked to a reciprocal chromosomal translocation, a naturally occurring form of high-threshold gene drive that would enable the reversible and tightly localized engineering of wild ​P. leucopus populations. Our open and community-guided approach has met with apparent enthusiasm by residents of Nantucket and Martha’s Vineyard, indicating that local communities suffering from tick-borne disease throughout the Northeast and Upper Midwest may wish to immunize their own wild mouse populations in order to help prevent Lyme disease for many decades.

项目摘要RFA-AI-19-037 莱姆病的不断增长需要新的预防策略。现有方法 例如，减少鹿，园林绿化和个人保护的衣服等葡萄糖苷，本质上是 短期，必须定期重新应用，维护和磨损。小鼠反对tick虫 项目旨在开发一种持久的一次性干预以破坏生态 莱姆病传播的循环数十年。莱姆的病因 疾病B. Burgdorferi在tick虫和它们的小动物宿主之间来回传递 作为人畜共患病疾病储藏。白脚小鼠P. leucopus广泛 被认为是最重要的水库，因为它既普遍又极端 通过壁虱获得和传输病原体的有效效率。我们为此的总体目标 拟议的项目是通过编码对莱姆的遗传性免疫对莱姆的免疫接种 靶向B. burgdorferi外表面蛋白A（OSPA）的保护性白细胞抗体 小鼠种系。根据我们的计算，将至少四种这样的抗体组合 防止B. burgdorferi的进化逃脱，因为太多简单的OSPA突变 需要。十字形，即使这些小鼠在某些领域的重要性不如当前 思想，免疫会减少受感染壁虱的数量，而感染壁虱的数量会减少 次要水库会感染更少的壁虱，增强负面反馈螺旋 预计会大大减少莱姆病的局部燃烧。我们已经成功 来自OSPA免疫的白叶核，衍生的假定P. leucopus的孤立抗OSPA抗体 胚胎干细胞，并表明白蛋白基因座似乎适合抗体分泌 来自肝脏。现在，我们寻求（1）识别与至少四个不同OSPA结合的抗体 表位，（2）建立稳定的胚胎干细胞系并进行种系编辑，（3） 产生可遗传的抗性小鼠，该小鼠从链接的盒子中表达抗体 相互染色体易位，一种天然存在的高阈值基因驱动器的形式 这将使野生假单胞菌的可逆和局限化工程 人群。通过 Nantucket和Martha的葡萄园居民，表明当地社区患有 tick传播的东北和中西部上部疾病可能希望免疫他们 拥有野生小鼠种群，以帮助预防莱姆病数十年。