Yang Bo, Liu Chao, Ju Xiaohui, Wu Bingbing, Wang Zhuangfei, Dong Fucheng, Yu Yanying, Hou Xiaohui, Fang Min, Gao Fei, Guo Xuejiang, Gui Yaoting, Ding Qiang, Li Wei
Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Shenzhen PKU-HKUST Medical Center, Shenzhen, Guangdong, China.
Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, Guangdong, China.
Cell Discov. 2023 Apr 20;9(1):43. doi: 10.1038/s41421-023-00536-0.
Animal models play crucial roles in the rapid development of vaccines/drugs for the prevention and therapy of COVID-19, but current models have some deficits when studying the pathogenesis of SARS-CoV-2 on some special tissues or organs. Here, we generated a human ACE2 and SARS-CoV-2 N knockin mouse line that constitutively expresses human ACE2 and specifically expresses SARS-CoV-2 N gene induced by Cre-recombinase. By crossing with Cre transgenic lines allowing for lung-specific and constitutive expression, we generated lung-specific (Sftpc-hACE2-N) and constitutive SARS-CoV-2 N (EIIa-hACE2-N) expressing mice. Upon intranasal infection with a SARS-CoV-2 GFP/ΔN strain which can only replicate in SARS-CoV-2 N expressed cells, we demonstrated that both the Sftpc-hACE2-N and EIIa-hACE2-N mice support viral replication. Consistent with our design, viral replication was limited to the lung tissues in Sftpc-hACE2-N mice, while the EIIa-hACE2-N mice developed infections in multiple tissues. Furthermore, our model supports different SARS-CoV-2 variants infection, and it can be successfully used to evaluate the effects of therapeutic monoclonal antibodies (Ab1F11) and antiviral drugs (Molnupiravir). Finally, to test the effect of SARS-CoV-2 infection on male reproduction, we generated Sertoli cell-specific SARS-CoV-2 N expressed mice by crossing with AMH-Cre transgenic line. We found that SARS-CoV-2 GFP/ΔN strain could infect Sertoli cells, led to spermatogenic defects due to the destruction of blood-testis barrier. Overall, combining with different tissue-specific Cre transgenic lines, the human ACE2 and SARS-CoV-2 N line enables us to evaluate antivirals in vivo and study the pathogenesis of SARS-CoV-2 on some special tissues or organs.
动物模型在用于预防和治疗新型冠状病毒肺炎的疫苗/药物的快速研发中发挥着关键作用，但目前的模型在研究严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在某些特殊组织或器官的发病机制时存在一些不足。在此，我们构建了一种人血管紧张素转换酶2（ACE2）和SARS-CoV-2 N基因敲入小鼠品系，其组成性表达人ACE2，并特异性表达由Cre重组酶诱导的SARS-CoV-2 N基因。通过与允许肺特异性和组成性表达的Cre转基因品系杂交，我们构建了肺特异性（Sftpc-hACE2-N）和组成性表达SARS-CoV-2 N（EIIa-hACE2-N）的小鼠。在用仅能在表达SARS-CoV-2 N的细胞中复制的SARS-CoV-2 GFP/ΔN毒株进行鼻内感染后，我们证明Sftpc-hACE2-N和EIIa-hACE2-N小鼠均支持病毒复制。与我们的设计一致，Sftpc-hACE2-N小鼠的病毒复制局限于肺组织，而EIIa-hACE2-N小鼠在多个组织中发生感染。此外，我们的模型支持不同的SARS-CoV-2变异株感染，并且它可以成功用于评估治疗性单克隆抗体（Ab1F11）和抗病毒药物（莫努匹拉韦）的效果。最后，为了测试SARS-CoV-2感染对雄性生殖的影响，我们通过与抗苗勒管激素（AMH）-Cre转基因品系杂交构建了支持细胞特异性表达SARS-CoV-2 N的小鼠。我们发现SARS-CoV-2 GFP/ΔN毒株可以感染支持细胞，由于血睾屏障的破坏导致生精缺陷。总体而言，结合不同的组织特异性Cre转基因品系，人ACE2和SARS-CoV-2 N品系使我们能够在体内评估抗病毒药物，并研究SARS-CoV-2在某些特殊组织或器官的发病机制。
