Endolysosomal two-pore channels regulate a conserved entry pathway for enteroviruses.

Area of Science:

• Virology
• Cell Biology
• Molecular Biology

Background:

• Enteroviruses are major human pathogens lacking effective antiviral treatments.
• Two-pore channels (TPCs) regulate calcium signaling and endosomal trafficking, but their role in non-enveloped virus infections is unclear.

Purpose of the Study:

• To investigate the role of human TPC1 and TPC2 in enterovirus infection.
• To identify TPCs as potential targets for broad-spectrum antiviral therapies.

Main Methods:

• CRISPR/Cas9 gene editing to knockout TPC isoforms.
• Pharmacological inhibition of NAADP-evoked calcium signaling.
• Live-cell calcium imaging.
• Transcriptomic profiling.
• Enterovirus replication assays.

Main Results:

• Human TPC1 and TPC2 are essential host factors enhancing enterovirus infection.
• Inhibition of TPCs or NAADP signaling suppressed enterovirus 71 replication.
• TPC loss delayed viral progression through endosomal compartments without affecting early entry steps.
• Cepharanthine showed inhibitory effects by modulating TPC-mediated calcium signaling.
• TPC depletion broadly restricted replication of multiple enterovirus serotypes.
• Disruption of TPCs altered gene networks involved in endocytosis and vesicular trafficking.

Conclusions:

• Enteroviruses exploit TPC-dependent calcium signaling for replication.
• Endolysosomal TPCs are critical for enterovirus infection by regulating endosomal trafficking.
• Targeting endolysosomal TPCs presents a promising strategy for broad-spectrum antiviral drug development against enteroviruses.