Tollip deficiency enhances mitophagy and reduces STING activation in influenza A virus-infected mice.

Toll-interacting protein (Tollip) is an intracellular adaptor protein with diverse functions including regulation of autophagy of mitochondria-mitophagy. Tollip deficiency promotes viral infection, but whether mitophagy is involved remains unclear. We sought to determine if mitophagy and associated signaling such as mitochondrial DNA (mtDNA) release and activation of stimulator of interferon genes (STING) contribute to worsened viral infection due to Tollip deficiency. Wild-type and Tollip knockout (KO) C57/BL6 mice were intranasally infected with influenza A virus (IAV), and then treated with or without a STING agonist 2'3'cGAMP for 4 d. PINK1 (an initiator of mitophagy) KO mouse tracheal epithelial cells (mTECs) or PINK1 KO mice were infected with IAV to reveal the role of mitophagy in viral infection. In IAV-infected mice, Tollip deficiency enhanced lung mitophagy (more PINK1 and BNIP3L, but less p62), and decreased release of mtDNA. Furthermore, Tollip deficiency suppressed STING activation and the antiviral response (eg IFN-β and MX1), and increased viral load. In IAV-infected Tollip KO mice, 2'3'cGAMP activated STING and increased antiviral response coupled with less virus. PINK1-deficient mice increased lung release of mtDNA and augmented STING activation and antiviral responses. PINK1 deficiency in mTECs increased STING activation and significantly decreased the viral load. Our findings suggest that enhanced mitophagy due to Tollip deficiency reduces mtDNA release and STING activation during viral infection, resulting in decreased antiviral responses. Reduction of mitophagy and/or STING activation may open novel avenues for therapeutic intervention in human subjects with Tollip deficiency and viral infection.