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Optimized Cationic Lipid-assisted Nanoparticle for Delivering CpG Oligodeoxynucleotides to Treat Hepatitis B Virus Infection

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Abstract

Purpose

Hepatitis B virus (HBV) infection is such a global health problem that hundreds of millions of people are HBV carriers. Current anti-viral agents can inhibit HBV replication, but can hardly eradicate HBV. Cytosine-phosphate-guanosine (CpG) oligodeoxynucleotides (ODNs) are an adjuvant that can activate plasmacytoid dendritic cells (pDCs) and conventional dendritic cells (cDCs) to induce therapeutic immunity for HBV eradication. However, efficient delivery of CpG ODNs into pDCs and cDCs remains a challenge. In this study, we constructed a series of cationic lipid-assisted nanoparticles (CLANs) using different cationic lipids to screen an optimal nanoparticle for delivering CpG ODNs into pDCs and cDCs.

Methods

We constructed different CLANCpG using six cationic lipids and analyzed the cellular uptake of different CLANCpG by pDCs and cDCs in vitro and in vivo, and further analyzed the efficiency of different CLANCpG for activating pDCs and cDCs in both wild type mice and HBV-carrier mice.

Results

We found that CLAN fabricated with 1,2-Dioleoyl-3-trimethylammonium propane (DOTAP) showed the highest efficiency for delivering CpG ODNs into pDCs and cDCs, resulting in strong therapeutic immunity in HBV-carrier mice. By using CLANCpG as an immune adjuvant in combination with the injection of recombinant hepatitis B surface antigen (rHBsAg), HBV was successfully eradicated and the chronic liver inflammation in HBV-carrier mice was reduced.

Conclusion

We screened an optimized CLAN fabricated with DOTAP for efficient delivery of CpG ODNs to pDCs and cDCs, which can act as a therapeutic vaccine adjuvant for treating HBV infection.

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DATA AVAILABILITY

All data generated or analyzed during this study are included in this published article and its supplementary information files, or are available from the corresponding authors on reasonable request.

Abbreviations

AL6:

1,2-Dilauroyl-sn-glycero-3-ethylphosphocholine

AL7:

1,2-Dioleoyl-sn-glycero-3-ethylphosphocholine

ALT:

Alanine aminotransferase

anti-HBs:

Hepatitis B surface antibody

AST:

Aspartate transaminase

BHEM-Chol:

N, N-Bis(2-Hydroxyethyl)-N-methyl-N-(2-cholesteryoxycarbonyl-aminoethyl) ammonium bromide

BMDCs:

Bone marrow-derived dendritic cells

CL3:

Cholesterol lipid # 3, Cholest-5-en-3-ol (3β)-, [2-[bis(2-aminoethyl) amino] ethyl] carbamate

CL7:

Cholesterol lipid # 7, N-[6-(cholest-5-en-3β-yloxycarbonylamino) hexyl]-pyridinium chloride

CLAN:

Cationic lipid-assisted nanoparticle

CLSM:

Confocal laser scanning microscopy

CpG:

Cytosine-phosphate-guanosine

Cy5-CpG:

Cy5-labeled Cytosine-phosphate-guanosine

DCs:

Dendritic cells

DOTAP:

1,2-Dioleoyl-3-trimethylammonium propane

ELISA:

Enzyme-linked immunosorbent assay

FBS:

Fetal bovine serum

H&E:

Hematoxylin and eosin

HBcAg:

Hepatitis B core antigen

HBV:

Hepatitis B virus

HBsAg:

Hepatitis B surface antigen

IFN:

Interferon

MFI:

Mean fluorescence intensity

NK cell:

Natural killer cell

ODNs:

Oligodeoxynucleotides

PBS:

Phosphate buffer saline

pDCs:

Plasmacytoid dendritic cells

PEG5K-b-PLGA10K :

Poly(ethylene glycol)5 K-block-poly(lactic-co-glycolic acid)10 K

PLGA:

Poly(lactic-co-glycolic acid)

rAAV:

Recombinant adeno-associated viruses

rHBsAg:

Recombinant hepatitis B surface antigen

RT-PCR:

Real-time PCR

TBIL:

Total bilirubin

TLR9:

Toll-like receptor 9

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Funding

This study was supported by the National Natural Science Foundation of China (82072048, 81901875, and 32071378), Guangdong Provincial Pearl River Talents Program (2017GC010713 and 2017GC010482), the Science and Technology Program of Guangzhou, China (202103030004), Guangdong Basic and Applied Basic Research Foundation (2022B1515020025, 2019A1515011878), and the Fundamental Research Funds for the Central Universities.

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Authors and Affiliations

  1. School of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou International Campus, Guangzhou, 511442, People’s Republic of China

    Yi-Fang Chen, Yue Wang, Cong-Fei Xu & Jun Wang

  2. School of Medicine, South China University of Technology, Guangzhou, 510006, People’s Republic of China

    Yan Wang, Ying-Li Luo, Zi-Dong Lu & Xiao-Jiao Du

  3. National Engineering Research Center for Tissue Restoration and Reconstruction, South China University of Technology, Guangzhou, 510006, People’s Republic of China

    Yue Wang, Cong-Fei Xu & Jun Wang

  4. Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou, 510006, People’s Republic of China

    Yue Wang

  5. Guangdong Provincial Key Laboratory of Biomedical Engineering, South China University of Technology, Guangzhou, 510006, People’s Republic of China

    Jun Wang

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  1. Yi-Fang Chen
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Contributions

J.W., C.-F.X. and X.-J.D. conceived the project, C.-F.X. and Y.-F.C. designed and conducted the experiments, performed the analysis and interpretations, and wrote the paper. Y.W. and Y.W. assisted the flow cytometry experiments, Y.-L.L. and Z.-D.L. provided help in designing the experiments and editing the manuscript. The manuscript has been read by all authors.

Corresponding authors

Correspondence to Xiao-Jiao Du or Cong-Fei Xu.

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Chen, YF., Wang, Y., Wang, Y. et al. Optimized Cationic Lipid-assisted Nanoparticle for Delivering CpG Oligodeoxynucleotides to Treat Hepatitis B Virus Infection. Pharm Res 40, 145–156 (2023). https://doi.org/10.1007/s11095-022-03307-w

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  • DOI: https://doi.org/10.1007/s11095-022-03307-w

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