PMID- 35890334 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220731 IS - 1999-4923 (Print) IS - 1999-4923 (Electronic) IS - 1999-4923 (Linking) VI - 14 IP - 7 DP - 2022 Jul 9 TI - Codelivery of HBx-siRNA and Plasmid Encoding IL-12 for Inhibition of Hepatitis B Virus and Reactivation of Antiviral Immunity. LID - 10.3390/pharmaceutics14071439 [doi] LID - 1439 AB - Chronic hepatitis B is a critical cause of many serious liver diseases such as hepatocellular carcinoma (HCC). The main challenges in hepatitis B treatment include the rebound of hepatitis B virus (HBV)-related antigen levels after drug withdrawal and the immunosuppression caused by the virus. Herein, we demonstrate that the HBV-related antigen can be effectively inhibited and antiviral immunity can be successfully reactivated through codelivery of the small interfering RNA (siRNA) targeting HBV X protein (HBx) and the plasmid encoding interleukin 12 (pIL-12) to hepatocytes and immune cells. After being treated by the siRNA/pIL-12 codelivery system, HBx mRNA and hepatitis B surface antigen (HBsAg) are dramatically reduced in HepG2.215 cells. More importantly, the downregulated CD47 and programmed death ligand 1 (PD-L1) and the upregulated interferon-beta promoter stimulator-1 (IPS-1), retinoic acid-inducible gene-1 (RIG-1), CD80, and human leukocyte antigen-1 (HLA-1) in treated HepG2.215 cells indicate that the immunosuppression is reversed by the codelivery system. Furthermore, the codelivery system results in inhibition of extracellular regulated protein kinases (ERK) and phosphoinositide-3-kinase (PI3K)/protein kinase B (Akt) pathways, as well as downregulation of B-cell lymphoma-2 (Bcl-2) and upregulation of p53, implying its potential in preventing the progression of HBV-induced HCC. In addition, J774A.1 macrophages treated by the codelivery system were successfully differentiated into the M1 phenotype and expressed enhanced cytokines with anti-hepatitis B effects such as interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha). Therefore, we believe that codelivery of siRNA and pIL-12 can effectively inhibit hepatitis B virus, reverse virus-induced immunosuppression, reactivate antiviral immunity, and hinder the progression of HBV-induced hepatocellular carcinoma. This investigation provides a promising approach for the synergistic treatment of HBV infection. FAU - Mu, Yan AU - Mu Y AD - National & Local Joint Engineering Research Center of High-Throughput Drug Screening Technology, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Key Laboratory of Biotechnology of Chinese Traditional Medicine of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, College of Life Sciences, Hubei University, Wuhan 430062, China. FAU - Ren, Xiao-He AU - Ren XH AD - Key Laboratory of Biomedical Polymers of Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, China. FAU - Han, Di AU - Han D AD - Key Laboratory of Biomedical Polymers of Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, China. FAU - Guan, Ying-Ying AU - Guan YY AD - National & Local Joint Engineering Research Center of High-Throughput Drug Screening Technology, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Key Laboratory of Biotechnology of Chinese Traditional Medicine of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, College of Life Sciences, Hubei University, Wuhan 430062, China. FAU - Liu, Pei-Ling AU - Liu PL AD - National & Local Joint Engineering Research Center of High-Throughput Drug Screening Technology, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Key Laboratory of Biotechnology of Chinese Traditional Medicine of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, College of Life Sciences, Hubei University, Wuhan 430062, China. FAU - Cheng, Si-Xue AU - Cheng SX AD - Key Laboratory of Biomedical Polymers of Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, China. FAU - Liu, Hong AU - Liu H AUID- ORCID: 0000-0003-1586-2431 AD - National & Local Joint Engineering Research Center of High-Throughput Drug Screening Technology, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Key Laboratory of Biotechnology of Chinese Traditional Medicine of Hubei Province, College of Life Sciences, Hubei University, Wuhan 430062, China. AD - Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, College of Life Sciences, Hubei University, Wuhan 430062, China. LA - eng GR - 20210201/Opening Project of Key Laboratory of Biomedical Polymers of Ministry of Education at Wuhan University/ PT - Journal Article DEP - 20220709 PL - Switzerland TA - Pharmaceutics JT - Pharmaceutics JID - 101534003 PMC - PMC9318813 OTO - NOTNLM OT - hepatitis B treatment OT - immune regulation OT - macrophage polarity OT - plasmid DNA OT - siRNA COIS- The authors declare no conflict of interest. EDAT- 2022/07/28 06:00 MHDA- 2022/07/28 06:01 PMCR- 2022/07/09 CRDT- 2022/07/27 01:39 PHST- 2022/06/06 00:00 [received] PHST- 2022/06/24 00:00 [revised] PHST- 2022/07/04 00:00 [accepted] PHST- 2022/07/27 01:39 [entrez] PHST- 2022/07/28 06:00 [pubmed] PHST- 2022/07/28 06:01 [medline] PHST- 2022/07/09 00:00 [pmc-release] AID - pharmaceutics14071439 [pii] AID - pharmaceutics-14-01439 [pii] AID - 10.3390/pharmaceutics14071439 [doi] PST - epublish SO - Pharmaceutics. 2022 Jul 9;14(7):1439. doi: 10.3390/pharmaceutics14071439.