Branched oligomerization of cell-permeable peptides markedly enhances the transduction efficiency of adenovirus into mesenchymal stem cells

• Published in: Gene therapy Vol. 17; no. 8; pp. 1052 - 1061
• Main Authors: Park, S-H, Doh, J, Park, S I, Lim, J Y, Kim, S M, Youn, J-I, Jin, H-T, Seo, S-H, Song, M-Y, Sung, S Y, Kim, M, Hwang, S J, Choi, J-M, Lee, S-K, Lee, H Y, Lim, C L, Chung, Y J, Yang, D, Kim, H-N, Lee, Z H, Choi, K Y, Jeun, S-S, Sung, Y C
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.08.2010; Nature Publishing Group
• Subjects: 631/1647/2300/1850; 631/378/1687/1825; 631/532/2074; 631/61/201; Adenoviridae - genetics; Adenovirus; Adenoviruses; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Applied cell therapy and gene therapy; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Biotechnology; Bone Diseases - genetics; Bone Diseases - therapy; Bone growth; Bone morphogenetic protein 2; Bone Morphogenetic Protein 2 - genetics; Brain-derived neurotrophic factor; Cell Biology; Cell membranes; Cell-Penetrating Peptides - chemistry; Cellular biology; Cellular signal transduction; Cytotoxicity; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene Therapy; Gene transfer; Gene Transfer Techniques; Genetic aspects; Genetic Therapy - methods; Genetic Vectors; Health aspects; Health. Pharmaceutical industry; Human Genetics; Humans; Industrial applications and implications. Economical aspects; Internalization; Male; Medical sciences; Mesenchymal stem cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Nanotechnology; Nonpermissive cells; Oligomerization; original-article; Osteogenesis; Osteogenesis - genetics; Peptides; Physiological aspects; Rats; Rats, Sprague-Dawley; Recovery of function; Skull - growth & development; Spinal cord injuries; Stem cells; Surface charge; Transduction; Transduction, Genetic - methods; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Viruses; South Korea; mesenchymal stem cell; recombinant adenovirus; BMP2; cell-permeable peptide; oligomerization; BDNF; Adenoviridae; Oligomerization; Peptides; Stem cell; Mesenchyma; Virus; Efficiency; Transduction; Gene therapy; Vector
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2010.58

Cell-permeable peptides (CPPs) promote the transduction of nonpermissive cells by recombinant adenovirus (rAd) to improve the therapeutic efficacy of rAd. In this study, branched oligomerization of CPPs significantly enhanced the transduction of human mesenchymal stem cells (MSCs) by rAd in a CPP type-independent manner. In particular, tetrameric CPPs increased transduction efficiency at 3000–5000-fold lower concentrations than did monomeric CPPs. Although branched oligomerization of CPPs also increases cytotoxicity, optimal concentrations of tetrameric CPPs required for maximum transduction are at least 300–1000-fold lower than those causing 50% cytotoxicity. Furthermore, although only ∼60% of MSCs were maximally transduced at 500 μ M of monomeric CPPs, >95% of MSCs were transduced with 0.1 μ M of tetrameric CPPs. Tetrameric CPPs also significantly increased the formation and net surface charge of CPP/rAd complexes, as well as the binding of rAd to cell membranes at a greater degree than did monomeric CPPs, followed by rapid internalization into MSCs. In a critical-size calvarial defect model, the inclusion of tetrameric CPPs in ex vivo transduction of rAd expressing bone morphogenetic protein 2 into MSCs promoted highly mineralized bone formation. In addition, MSCs that were transduced with rAd expressing brain-derived neurotrophic factor in the presence of tetrameric CPPs improved functional recovery in a spinal cord injury model. These results demonstrated the potential for tetrameric CPPs to provide an innovative tool for MSC-based gene therapy and for in vitro gene delivery to MSCs.