Improved transduction efficiencies of adeno-associated virus vectors by synthetic cell-permeable peptides.
Various serotypes of adeno-associated virus (AAV) vectors have been used for gene therapy and as research tools. Among these serotypes, the AAV type 2 vector has been used successfully in human gene therapies. However, the transduction efficiency of AAV2 depends on the cell type, and this poses a problem in the efficacy of gene therapy. To improve the transduction efficiency of AAV2, we designed a small peptide consisting of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor peptide and the HIV-Tat sequence Tat-Y1068. Pre- or co-treatment of CYNOM-K1 cells from cynomolgus monkey embryo skin with Tat-Y1068 increased the transduction efficiencies in a dose-dependent manner and caused p38 phosphorylation. The transduction efficiency of AAV2 into the rat fibroblast cell line RAT-1 highly expressing EGFR was less than the transduction efficiency of AAV2 into CYNOM-K1 cells. Tat-Y1068 increased the transduction efficiency in RAT-1 cells in the same manner as in CYNOM-K1 cells. In conclusion, cell-permeable peptides possessing the EGFR tyrosine kinase inhibitor function might serve as a useful ingredient of AAV2 vector solution for increasing the transduction efficiency of gene therapies.
Laboratory of Visual Neuroscience, Graduate Course in Biological Sciences, Iwate University Division of Science and Engineering, 4-3-5 Ueda, Morioka, Iwate 020-8551, Japan. Electronic address: ktabata@iwate-u.ac.jp.
Kitako
K
Tabata
Laboratory of Visual Neuroscience, Graduate Course in Biological Sciences, Iwate University Division of Science and Engineering, 4-3-5 Ueda, Morioka, Iwate 020-8551, Japan; Soft-Path Engineering Research Center (SPERC), Faculty of Engineering, Iwate University, Morioka, 020-8551, Japan. Electronic address: sseriko@iwate-u.ac.jp.
Eriko
E
Sugano
Laboratory of Visual Neuroscience, Graduate Course in Biological Sciences, Iwate University Division of Science and Engineering, 4-3-5 Ueda, Morioka, Iwate 020-8551, Japan. Electronic address: t0113073@iwate-u.ac.jp.
Fumika
F
Murakami
Soft-Path Engineering Research Center (SPERC), Faculty of Engineering, Iwate University, Morioka, 020-8551, Japan; Department of Biological Chemistry, Iwate University Faculty of Agriculture, Morioka, Japan. Electronic address: yamashit@iwate-u.ac.jp.
Tetsuro
T
Yamashita
Soft-Path Engineering Research Center (SPERC), Faculty of Engineering, Iwate University, Morioka, 020-8551, Japan. Electronic address: tozaki@iwate-u.ac.jp.
Taku
T
Ozaki
Laboratory of Visual Neuroscience, Graduate Course in Biological Sciences, Iwate University Division of Science and Engineering, 4-3-5 Ueda, Morioka, Iwate 020-8551, Japan; Soft-Path Engineering Research Center (SPERC), Faculty of Engineering, Iwate University, Morioka, 020-8551, Japan; Clinical Research, Innovation and Education Center, Tohoku University Hospital, 1-1 Seiryo, Aoba, Sendai, Miyagi 980-8574, Japan. Electronic address: htomita@iwate-u.ac.jp.
Hiroshi
H
Tomita
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D057846:Cell-Penetrating Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D000229:Dependovirus / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D008252:Macaca fascicularis; D008856:Microscopy, Fluorescence; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D015203:Reproducibility of Results; D012867:Skin / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D057846:Cell-Penetrating Peptides; D048051:p38 Mitogen-Activated Protein Kinases
Adeno-associated virus vector; Cell-permeable peptide; Epidermal growth factor receptor; Transduction efficiency