Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection

Saiaditya Badeti; Qingkui Jiang; Alireza Naghizadeh; Hsiang-Chi Tseng; Yuri Bushkin; Salvatore A E Marras; Annuurun Nisa; Sanjay Tyagi; Fei Chen; Peter Romanienko; Ghassan Yehia; Deborah Evans; Moises Lopez-Gonzalez; David Alland; Riccardo Russo; William Gause; Lanbo Shi; Dongfang Liu

doi:10.1186/s13578-022-00822-6

Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection

Cell Biosci. 2022 Jun 11;12(1):88. doi: 10.1186/s13578-022-00822-6.

Authors

Saiaditya Badeti^#^{1

2

3}, Qingkui Jiang^#⁴, Alireza Naghizadeh^{1

3}, Hsiang-Chi Tseng^{1

3}, Yuri Bushkin⁴, Salvatore A E Marras⁴, Annuurun Nisa⁴, Sanjay Tyagi⁴, Fei Chen^{1

3}, Peter Romanienko⁵, Ghassan Yehia⁵, Deborah Evans^{1

3}, Moises Lopez-Gonzalez⁶, David Alland⁴, Riccardo Russo⁴, William Gause^{1

3}, Lanbo Shi⁷, Dongfang Liu^{8

9

10}

Affiliations

¹ Department of Pathology, Immunology and Laboratory Medicine, Center for Immunity and Inflammation, Rutgers-New Jersey Medical School, Rutgers University, 205 S. Orange Ave., CC-H1218, Newark, NJ, 07103, USA.
² School of Graduate Studies, Biomedical and Health Sciences, Rutgers University, Newark, NJ, 07103, USA.
³ Center for Immunity and Inflammation, New Jersey Medical School, Rutgers University, Newark, NJ, 07103, USA.
⁴ Public Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ, 07103, USA.
⁵ Genome Editing Shared Resources, Cancer Institute of New Jersey, Rutgers University, New Brunswick, NJ, 08901, USA.
⁶ Regional Bio-Containment Laboratory, Center for COVID-19 Response and Pandemic Preparedness (CCRP2), Rutgers-New Jersey Medical School, Newark, NJ, 07103, USA.
⁷ Public Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ, 07103, USA. shila@njms.rutgers.edu.
⁸ Department of Pathology, Immunology and Laboratory Medicine, Center for Immunity and Inflammation, Rutgers-New Jersey Medical School, Rutgers University, 205 S. Orange Ave., CC-H1218, Newark, NJ, 07103, USA. dongfang.liu@rutgers.edu.
⁹ School of Graduate Studies, Biomedical and Health Sciences, Rutgers University, Newark, NJ, 07103, USA. dongfang.liu@rutgers.edu.
¹⁰ Center for Immunity and Inflammation, New Jersey Medical School, Rutgers University, Newark, NJ, 07103, USA. dongfang.liu@rutgers.edu.

^# Contributed equally.

Abstract

Background: An animal model that can mimic the SARS-CoV-2 infection in humans is critical to understanding the rapidly evolving SARS-CoV-2 virus and for development of prophylactic and therapeutic strategies to combat emerging mutants. Studies show that the spike proteins of SARS-CoV and SARS-CoV-2 bind to human angiotensin-converting enzyme 2 (hACE2, a well-recognized, functional receptor for SARS-CoV and SARS-CoV-2) to mediate viral entry. Several hACE2 transgenic (hACE2Tg) mouse models are being widely used, which are clearly invaluable. However, the hACE2Tg mouse model cannot fully explain: (1) low expression of ACE2 observed in human lung and heart, but lung or heart failure occurs frequently in severe COVID-19 patients; (2) low expression of ACE2 on immune cells, but lymphocytopenia occurs frequently in COVID-19 patients; and (3) hACE2Tg mice do not mimic the natural course of SARS-CoV-2 infection in humans. Moreover, one of most outstanding features of coronavirus infection is the diversity of receptor usage, which includes the newly proposed human CD147 (hCD147) as a possible co-receptor for SARS-CoV-2 entry. It is still debatable whether CD147 can serve as a functional receptor for SARS-CoV-2 infection or entry.

Results: Here we successfully generated a hCD147 knock-in mouse model (hCD147KI) in the NOD-scid IL2Rgamma^null (NSG) background. In this hCD147KI-NSG mouse model, the hCD147 genetic sequence was placed downstream of the endogenous mouse promoter for mouse CD147 (mCD147), which creates an in vivo model that may better recapitulate physiological expression of hCD147 proteins at the molecular level compared to the existing and well-studied K18-hACE2-B6 (JAX) model. In addition, the hCD147KI-NSG mouse model allows further study of SARS-CoV-2 in the immunodeficiency condition which may assist our understanding of this virus in the context of high-risk populations in immunosuppressed states. Our data show (1) the human CD147 protein is expressed in various organs (including bronchiolar epithelial cells) in hCD147KI-NSG mice by immunohistochemical staining and flow cytometry; (2) hCD147KI-NSG mice are marginally sensitive to SARS-CoV-2 infection compared to WT-NSG littermates characterized by increased viral copies by qRT-PCR and moderate body weight decline compared to baseline; (3) a significant increase in leukocytes in the lungs of hCD147KI-NSG mice, compared to infected WT-NSG mice.

Conclusions: hCD147KI-NSG mice are more sensitive to COVID-19 infection compared to WT-NSG mice. The hCD147KI-NSG mouse model can serve as an additional animal model for further interrogation whether CD147 serve as an independent functional receptor or accessory receptor for SARS-CoV-2 entry and immune responses.

Keywords: Accessory receptor; BSG; Basigin; CD147; COVID-19; Immune responses; Knock-in mouse model; NSG; Physiological expression; SARS-CoV-2; Spike protein; Viral entry; hCD147KI.

Abstract

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