Lipid nanoparticle‐mediated delivery of IL‐21‐encoding mRNA induces viral clearance in mouse models of hepatitis B virus persistence

Abstract Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), the transcription template for all viral mRNAs, is highly stable and current treatment options cannot effectively induce its clearance. Previously, we established an HBV persistence mouse model based on a clinical isolate (termed BPS) and identified interleukin‐21 (IL‐21) as a potent inducer of HBV clearance. Lipid nanoparticle (LNP) mediated delivery of mRNA has proven to be a highly safe and effective delivery platform. This work explored the applicability and effectiveness of the mRNA‐LNP platform in IL‐21‐based HBV therapies. First, LNP‐encapsulated murine IL‐21 mRNA (LNP‐IL‐21) was prepared, characterized, and demonstrated to engender IL‐21 expression in vitro and in vivo. Next, LNP‐IL‐21 was shown to induce clearance of both serum and intrahepatic HBV antigen and DNA in two HBV persistence mouse models based on BPS and recombinant cccDNA (rcccDNA), respectively, which was associated with HBV‐specific humoral and cellular immune responses. Furthermore, peripheral blood mononuclear cells from BPS persistence mice treated ex vivo with LNP‐IL‐21 and HBV surface antigen (HBsAg) could induce similar HBV clearance upon infusion into recipient mice. These findings indicated that IL‐21 combined with mRNA‐LNP platform represents a valid and promising strategy for developing novel therapeutics against chronic HBV infection.