tetano
Editor, Senior Moderator
Cell Rep
. 2024 Oct 30;43(11):114921.
doi: 10.1016/j.celrep.2024.114921. Online ahead of print. TMEM106B-mediated SARS-CoV-2 infection allows for robust ACE2-independent infection in vitro but not in vivo
Kexin Yan[SUP] 1 [/SUP], Troy Dumenil[SUP] 1 [/SUP], Romal Stewart[SUP] 1 [/SUP], Cameron R Bishop[SUP] 1 [/SUP], Bing Tang[SUP] 1 [/SUP], Wilson Nguyen[SUP] 1 [/SUP], Andreas Suhrbier[SUP] 2 [/SUP], Daniel J Rawle[SUP] 3 [/SUP]
Affiliations
Angiotensin-converting enzyme 2 (ACE2) is the primary entry receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but ACE2-independent entry has been observed in vitro for strains with the spike-E484D substitution. Here, we conduct a whole-genome CRISPR-Cas9 knockout screen using SARS-CoV-2 mouse adapted 1 (SARS-CoV-2[SUB]MA1[/SUB]), which carries spike-E484D, to identify the ACE2-independent entry mechanisms. SARS-CoV-2[SUB]MA1[/SUB] infection in HEK293T cells relies on heparan sulfate and endocytic pathways, with TMEM106B, a transmembrane lysosomal protein, the most significant contributor. While SARS-CoV-2[SUB]MA1[/SUB] productively infects human brain organoids and K18-hACE2 mouse brains, it does not infect C57BL/6J or Ifnar[SUP]-/-[/SUP] mouse brains. This suggests that ACE2-independent entry via TMEM106B, which is predominantly expressed in the brain, does not overtly increase the risk of SARS-CoV-2 neuroinvasiveness in mice with endogenous Ace2 expression. Importantly, SARS-CoV-2[SUB]MA1[/SUB] does not replicate in the Ace2[SUP]-/-[/SUP] mouse respiratory tract. Overall, this suggests that robust ACE2-independent infection by SARS-CoV-2[SUB]MA1[/SUB] is likely an in vitro phenomenon with no apparent implications for infection in vivo.
Keywords: ACE2-independent entry; CP: Immunology; CP: Microbiology; CRISPR-Cas9 screen; SARS-CoV-2; SARS-CoV-2 entry receptors; TMEM106B; brain organoids; mouse models.
. 2024 Oct 30;43(11):114921.
doi: 10.1016/j.celrep.2024.114921. Online ahead of print. TMEM106B-mediated SARS-CoV-2 infection allows for robust ACE2-independent infection in vitro but not in vivo
Kexin Yan[SUP] 1 [/SUP], Troy Dumenil[SUP] 1 [/SUP], Romal Stewart[SUP] 1 [/SUP], Cameron R Bishop[SUP] 1 [/SUP], Bing Tang[SUP] 1 [/SUP], Wilson Nguyen[SUP] 1 [/SUP], Andreas Suhrbier[SUP] 2 [/SUP], Daniel J Rawle[SUP] 3 [/SUP]
Affiliations
- PMID: 39480813
- DOI: 10.1016/j.celrep.2024.114921
Angiotensin-converting enzyme 2 (ACE2) is the primary entry receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but ACE2-independent entry has been observed in vitro for strains with the spike-E484D substitution. Here, we conduct a whole-genome CRISPR-Cas9 knockout screen using SARS-CoV-2 mouse adapted 1 (SARS-CoV-2[SUB]MA1[/SUB]), which carries spike-E484D, to identify the ACE2-independent entry mechanisms. SARS-CoV-2[SUB]MA1[/SUB] infection in HEK293T cells relies on heparan sulfate and endocytic pathways, with TMEM106B, a transmembrane lysosomal protein, the most significant contributor. While SARS-CoV-2[SUB]MA1[/SUB] productively infects human brain organoids and K18-hACE2 mouse brains, it does not infect C57BL/6J or Ifnar[SUP]-/-[/SUP] mouse brains. This suggests that ACE2-independent entry via TMEM106B, which is predominantly expressed in the brain, does not overtly increase the risk of SARS-CoV-2 neuroinvasiveness in mice with endogenous Ace2 expression. Importantly, SARS-CoV-2[SUB]MA1[/SUB] does not replicate in the Ace2[SUP]-/-[/SUP] mouse respiratory tract. Overall, this suggests that robust ACE2-independent infection by SARS-CoV-2[SUB]MA1[/SUB] is likely an in vitro phenomenon with no apparent implications for infection in vivo.
Keywords: ACE2-independent entry; CP: Immunology; CP: Microbiology; CRISPR-Cas9 screen; SARS-CoV-2; SARS-CoV-2 entry receptors; TMEM106B; brain organoids; mouse models.