Gene delivery into hepatocytes with the preS/liposome/DNA system

Abstract

Gene delivery into human hepatocytes remains a critical issue for the development of liver-directed gene therapy. Gene delivery based on non-viral vectors is an attractive approach relative to viral vectors. In this report, novel delivery system of preS/liposome/DNA virus-like particle (VLP) was developed for gene transfection into hepatocytes in vivo and in vitro. Plasmid pCMVβ, expressing β-galactosidase, was encapsulated with cationic liposome, and then the histidine-tagged preS domain of hepatitis B virus was coated on the surface of liposome/DNA to form preS/liposome/DNA VLP. Transfection efficiencies of preS/liposome/DNA, liposome/DNA, naked DNA and preS were analyzed using several different human cell lines. The highest transfection efficiency was found using preS/liposome/DNA VLP as the transfection reagent in human hepatocyte (HH) cell line. Results show that preS domain of hepatitis B virus coated on liposome/DNA can be used for highly efficient gene transfection into human hepatocytes. Moreover, the target characteristic of preS/liposome/DNA was analyzed in vivo. After preS/liposome/DNA VLP was injected into immunocompromised (Nude) mice via the tail vein, most of β-galactosidase was expressed in the liver; however, no significant target expression was found with the injection of liposome/DNA or naked DNA. Our results show that preS/liposome/DNA VLP can be used as a novel liver-specific gene delivery system.

Ex Vivo Gene Delivery to Hepatocytes: Techniques, Challenges, and Underlying Mechanisms

“…14 Both non-viral vectors and viral vectors have been employed for delivering plasmid DNA and siRNA to hepatocytes in vitro and in vivo in order to either initiate or inhibit the expression of specific proteins for pathway studies and for therapeutic applications. Similar to the methods used in other types of cells or tissues, non-viral gene delivery to hepatocytes includes commercially available transfection agents, 6,36 liposomes, 37 electroporation, 2,36 carbohydrate-based gene delivery vectors, 30 and hydrodynamic injection 12 ; while viral gene delivery vectors include retroviruses 36,41 (especially lentiviruses 24 ), adenoviruses 3 and adeno-associated viruses. 20 It has often been more challenging, however, to transfect primary hepatocytes compared to cell lines or to readily dividing primary cells.…”

Hepatitis B virus pre‐S region: Clinical implications and applications

“…� The pre-S1-9Arg peptide successfully combined and delivered siRNA to the HepG2 cells � The pre-S1-9Arg-siRNA complex significantly decreased HBsAg synthesis without hepatotoxicity in vitro and in vivo Pre-S-conjugated liposome 125 Liposome coated with histidine-tagged pre-S peptide on the surface.…”

“…The construction of hepatocyte-targeted delivery systems is based on biological carriers or synthetic carriers. Several carriers containing pre-S peptides have been developed (Table 2), [120][121][122][123][124][125][126][127][128][129][130][131] and targeting delivery systems based on these carriers have shown good specificity to liver cells. Moreover, there are several targeting systems that can specifically identify abnormal liver cells (e.g., hepatoma cells).…”

Current Progress of Virus-mimicking Nanocarriers for Drug Delivery

“…The synthetic peptide corresponding to the N-terminal region of the HBV envelope L protein (e.g., myristoylated pre-S1(2-47) peptide) interacts with the high-affinity HBV receptor, sodium taurocholate cotransporting polypeptide (NTCP) 108 . When this peptide is conjugated with synthetic nanocarriers, such as LPs 109 , and lipoplex (complex of cationic LPs and plasmid DNA) 110 , they can target human hepatic cells specifically. The tropism of HBV is strictly limited to human hepatocyte probably due to the restricted expression of the HBV receptor NTCP in hepatocytes 108 .…”