Abstract Mpox, formerly known as monkeypox, is a zoonotic illness of international concern that can lead to severe disease including neurological sequelae. However, it remains unclear what the neurotropism of monkeypox virus (MPXV) is and how MPXV infection leads to neurological deficits. Here, we determined the neurotropism and neurovirulence of MPXV using human pluripotent stem cell- (hPSC)-derived neural stem cells, astrocytes, cortical neurons, and microglia together with ex vivo human brain tissue. We found that MPXV infects and replicates more efficiently in astrocytes and microglia compared to cortical neurons, which unlike glial cells showed activation of distinct antiviral programs that may confer differential susceptibility to MPXV. Ex vivo infection of human brain tissue confirmed the susceptibility of astrocytes to MPXV infection, which also had the strongest disease-associated changes. Molecular pathway analyses revealed induction of cellular senescence and a senescence-associated secretory phenotype upon MPXV infection in astrocytes. Finally, we demonstrated that antiviral treatment using tecovirimat inhibits MPXV replication and prevents virus-induced senescence in hPSC-derived astrocytes. Altogether, leveraging hPSC-derived brain cells, we reveal MPXV-induced cell type-specific effects at the molecular and cellular level, which provide important insights into the neuropathogenesis of MPXV infection.

Dissecting the neurotropism and neurovirulence of MPXV using human stem cell-based models