MMRRC COVID-19 variant testing in humanized mouse models

MMRRC 在人源化小鼠模型中进行 COVID-19 变异测试

• 批准号: 10412858
• 负责人: KC KENT LLOYD
• 金额: $ 49.88万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10412858
• 关键词: 2019-nCoV; ACE2; Acute; Address; Administrative Supplement; Aging; Alleles; Animal Model; Antiviral Therapy; Archives; Awareness; Binding; Biomedical Research; Body Weight; Breeding; C57BL/6 Mouse; COVID-19; California; Cells; Centers for Disease Control and Prevention (U.S.); Communities; Cryopreservation; Deposition; Development; Disease; Effectiveness; Embryo; Ensure; Environment; Evaluation; Event; FURIN gene; Female; Fertilization in Vitro; Frequencies; Future; Genes; Genetic; Genotype; Grant; Hand; Health; Heart; Human; Immune response; Inbred BALB C Mice; Infection; Inflammatory Response; K-18 conjugate; Kinetics; Knock-in; Knock-in Mouse; Knockout Mice; Long COVID; Lung; Lung Inflammation; Modeling; Morbidity - disease rate; Mus; Mutant Strains Mice; Orthologous Gene; Partner in relationship; Pathogenesis; Pathologic; Pathway interactions; Phenotype; Predisposition; Prevention; Research; Research Personnel; Resources; SARS-CoV-2 B.1.1.7; SARS-CoV-2 infection; SARS-CoV-2 variant; Scientist; TMPRSS2 gene; Testing; Therapeutic; Tissues; Transgenic Organisms; Translational Research; Translations; U-Series Cooperative Agreements; United States National Institutes of Health; Universities; Vaccines; Validation; Viral; Viral Load result; Virus; Virus Diseases; Work; cohort; disorder prevention; experience; experimental study; follow-up; genetic variant; human disease; humanized mouse; in vivo; male; member; mortality; mouse model; novel therapeutic intervention; pandemic disease; pre-clinical; programs; response; screening; therapeutic vaccine; transmission process; validation studies; viral genomics; working group

ABSTRACT & SCOPE OF WORK The Mutant Mouse Resource and Research Center at the University of California, Davis (MMRRC-UC Davis) is pleased to submit this administrative supplement for up to 1 year of support in response to ORIP’s participation in PA-20-272, “Administrative Supplements to Existing NIH Grants and Cooperative Agreements” specifically related to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease (COVID-19). This application addresses a number of the stated objectives of the call to support COVID-19 related research, including to develop and characterize animal models for post-acute sequelae of COVID-19 (PASC) and to study the susceptibility of existing COVID-19 animal models to emerging viral genomic variants. Specifically, this application will generate polygenic humanized mouse models for virus challenge, validation, and post- acute sequellae of COVID-19 (PASC) in both young and aging mice using well-established testing and screening platforms in an ABSL3 environment. This project will build upon our successful efforts to generate monogenic humanized knockin/murine knockout mouse lines for several genes (ACE2, TMPRSS2, and FURIN) involved in SARS-CoV-2 binding, entry, and activation. With this prior experience in hand, we now propose to determine infectivity and transmission in new polygenic humanized mice to assess the extent to which they can be used as suitable models of PASC in humans. Specifically in this project we will 1) use in vitro fertilization (IVF) expansion and intercrossing of our extant monogenic models to rapidly generate polygenic humanized mouse models of hACE2/hTMPRSS2 and hACE2/hTMPRSS2/hFURIN, 2) validate the pathophysiological effects and assess PASC after challenge with currently dominant circulating (B.1.1.7; strain: USA/CA_CDC_5574/2020) in young and aging male and female cohorts of polygenic humanized mice under ABSL3 conditions, and 3) establish breeding colonies and cryopreserved germplasm of humanized mouse models for archiving and distribution to the biomedical research community. Validation studies will involve systematic characterization of viral load and clearance, body weight kinetics, and lung inflammation after SARS-CoV-2 challenge of male and female cohorts of mice; positive results will be communicated to the ACTIV-Preclinical Working Group and others at NIH. In addition, observational and pathological screening of surviving aging mice will be conducted to screen for evidence of PASC; promising findings will be communicated with members of the PASC Initiative and Investigator Consortium (OTA-21-015A and B) and other NIH staff to ensure rapid translation of findings for human studies and functional studies in animal models. Further, we will ensure that our mouse models and testing platform will be made readily available for use by other researchers to swiftly assess the in vivo consequences of not only newly appearing SARS-CoV-2 variants that escape current therapeutic and vaccine strategies but also of future viruses with similarly high- impact pandemic potential. This study is essential to overcome genetic discrepancies between mouse and human and to fill crucial gaps in existing small animal models of COVID-19 that hinder translation of research findings to improvements in human health, including understanding the development, treatment, and prevention of PASC, the effectiveness of antiviral therapies, and the reliability of disease-prevention vaccine strategies.

摘要和工作范围 加州大学戴维斯分校突变小鼠资源和研究中心 (MMRRC-UC Davis) 很高兴提交此行政补充文件，以获得长达 1 年的支持，以响应 ORIP 的要求 参与 PA-20-272，“现有 NIH 拨款和合作协议的行政补充” 与严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 疾病 (COVID-19) 特别相关。 此应用程序解决了支持 COVID-19 相关研究的呼吁的许多既定目标， 包括开发和表征 COVID-19 急性后遗症动物模型 (PASC) 以及 研究现有的 COVID-19 动物模型对新兴病毒基因组变异的敏感性。 该应用程序将生成用于病毒攻击、验证和后处理的多基因人源化小鼠模型 使用完善的测试和方法，在年轻和老年小鼠中发现 COVID-19 (PASC) 的急性后遗症 该项目将建立在我们成功生成的基础上。 多个基因（ACE2、TMPRSS2 和 FURIN）参与 SARS-CoV-2 的结合、进入和激活。有了这些先前的经验，我们现在可以了解。 提议确定新的多基因人源化小鼠的感染性和传播性，以评估感染和传播的程度 它们可以用作人类 PASC 的合适模型，具体来说，在这个项目中，我们将 1) 使用。 体外受精 (IVF) 扩展和现有单基因模型的交叉，以快速生成 hACE2/hTMPRSS2 和 hACE2/hTMPRSS2/hFURIN 多基因人源化小鼠模型，2) 验证 病理生理学影响并评估当前占主导地位的循环攻击后的 PASC（B.1.1.7；菌株： USA/CA_CDC_5574/2020）在多基因人源化小鼠的年轻和老年男性和女性群体中 ABSL3条件，以及3)建立人源化小鼠的繁殖群体和冻存种质 归档和分发给生物医学研究界的模型将涉及验证研究。 病毒载量和清除、体重动力学和肺部炎症的系统特征 雄性和雌性小鼠组的 SARS-CoV-2 攻击阳性结果将传达给 NIH 的 ACTIV 临床前工作组和其他人员还进行了观察和病理筛查。 将对幸存的衰老小鼠进行筛选，以寻找 PASC 的证据； 与 PASC 倡议和研究者联盟 (OTA-21-015A 和 B) 的成员进行了沟通， NIH 的其他工作人员确保快速转化人类研究和动物功能研究的发现 此外，我们将确保我们的鼠标模型和测试平台可供随时使用。 其他研究人员使用它来快速评估新出现的 SARS-CoV-2 的体内后果 逃避当前治疗和疫苗策略的变种，以及未来病毒的类似高病毒变种 这项研究对于克服小鼠和小鼠之间的遗传差异至关重要。 人类并填补现有的 COVID-19 小动物模型中阻碍研究转化的关键空白 改善人类健康的发现，包括了解发展、治疗和 PASC 的预防、抗病毒治疗的有效性以及疾病预防疫苗的可靠性 策略。