Toyoharu Mine

Publications of Toyoharu Mine

• Multiple components in serum contribute to hepatic transgene expression by lipoplex in mice.

  Authors: Naoki Yoshikawa, Keiko Sakamoto, Sachiyo Mizuno, Junichiro Sakaguchi, Hirotaka Miyamoto, Toyoharu Mine, Hitoshi Sasaki, Shintaro Fumoto, Koyo Nishida

  The journal of gene medicine. 11/2011; 13(11):632-43.

  Interaction of cationic liposome/plasmid DNA complex (lipoplex) with serum was not a limiting factor for in vivo transfection. After intraportal injection of lipoplex, hepatic transgene expressionInteraction of cationic liposome/plasmid DNA complex (lipoplex) with serum was not a limiting factor for in vivo transfection. After intraportal injection of lipoplex, hepatic transgene expression was enhanced by interaction with serum in mice. In the present study, we analyzed the mechanism of enhanced hepatic transgene expression of lipoplex by interaction with serum components. Lipoplexes were incubated with several serum components for 5  min at 37  ° C before administration. Transfection efficiency of lipoplexes was measured 6  h after intraportal injection of lipoplex in mice. Depletion of divalent cation from serum decreased hepatic transgene expression. The addition of calcium ion to divalent cation-depleted serum restored transgene expression. Heat-inactivated serum and bovine serum albumin diminished the enhancing effect of serum on hepatic transgene expression. On the other hand, removal of anionic proteins from serum using an anion-exchanging column was critical for the enhancing effect of serum on transgene expression. Among the serum components tested, fibronectin and complement component C3 enhanced hepatic transgene expression. Hepatic transgene expression by lipoplex was enhanced by interaction with multiple components in serum. Interaction of lipoplex with serum could be an important factor for successful in vivo gene transfer. Hence, the information obtained in the present study is valuable for the future development of effective gene carriers.

• Rubbing gastric serosal surface enhances naked plasmid DNA transfer in rats and mice.

  Authors: Toyoharu Mine, Hiroki Ishii, Sayuri Nakajima, Naoki Yoshikawa, Hirotaka Miyamoto, Mikiro Nakashima, Junzo Nakamura, Shintaro Fumoto, Koyo Nishida

  Biological & pharmaceutical bulletin. 01/2011; 34(9):1514-7.

  We have developed in vivo gene transfer to mesothelial cells on the peritoneal organs, including the stomach. Simple instillation of naked plasmid DNA onto the gastric serosal surface in miceWe have developed in vivo gene transfer to mesothelial cells on the peritoneal organs, including the stomach. Simple instillation of naked plasmid DNA onto the gastric serosal surface in mice resulted in effective but transient transgene expression. Here, we developed a simple method to improve not only the transfection efficiency but also the duration of transgene expression. Rubbing the gastric serosal surface using a medical spoon immediately after instillation of naked plasmid DNA onto the gastric serosal surface resulted in 59-fold higher transgene expression 24 h after administration in rats. Without rubbing, transgene expression decreased under the detection limit 7 d after administration. On the other hand, rubbing the gastric serosal surface with a medical spoon after instillation of plasmid DNA prolonged transgene expression for one month. Mechanistic study in mice revealed that improved transfection should not be due to stimulation of cell function such as macropinocytosis by rubbing because rubbing before instillation of plasmid DNA did not improve transfection. Plasmid DNA should enter effectively into cells during rubbing. These findings are valuable to develop an effective method of in vivo gene transfer into peritoneal organs.