Kinetics and effect of integrin expression on human CD34+ cells during murine leukemia virus-derived retroviral transduction with recombinant fibronectin for stem cell gene therapy

Yasuomi Horiuchi, Masafumi Onodera, Yoshitaka Miyagawa, Ban Sato, Keiko Onda, Yohko U. Katagiri, Hajime Okita, Mayumi Okada, Makoto Otsu, Akihiro Kume, Torayuki Okuyama, Junichiro Fujimoto, Tadatoshi Kuratsuji, Nobutaka Kiyokawa

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2 Citations (Scopus)

Abstract

The CH-296 recombinant fragment of human fibronectin is essential for murine leukemia virus (MLV)-derived retroviral transduction of CD34+ cells for the purpose of stem cell gene therapy. Although the major effect of CH-296 is colocalization of the MLV-derived retrovirus and target cells at specific adhesion domains of CH-296 mediated by integrins expressed on CD34 + cells, the precise roles of the integrins are unclear. We examined the kinetics of integrin expression on CD34+ cells during the course of MLV-derived retrovirus-mediated gene transduction with CH-296. Flow cytometry revealed that the levels of both very late activation protein (VLA)-4 and VLA-5 on CD34+ cells freshly isolated from cord blood were insufficient for effective MLV-derived retroviral transduction. However, increases were achieved during culture for preinduction and MLV-derived retrovirus-mediated gene transduction in the presence of a cocktail of cytokines. In addition, we confirmed by using specific antibodies that inhibition of the cell adhesion mediated by the integrins significantly reduced transduction efficiency, indicating that integrin expression is indeed important for CH-296-based MLV-derived retroviral transduction. Only a few cytokines are capable of inducing integrin expression, and stem cell factor plus thrombopoietin was found to be the minimal combination that was sufficient for effective transduction of an MLV-derived retrovirus based on CH-296. Our findings should be useful for improving the culture conditions for CH-296-based MLV-derived retroviral transduction in stem cell gene therapy.

Original languageEnglish
Pages (from-to)777-783
Number of pages7
JournalHuman Gene Therapy
Volume20
Issue number7
DOIs
Publication statusPublished - 2009 Jul 1
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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    Horiuchi, Y., Onodera, M., Miyagawa, Y., Sato, B., Onda, K., Katagiri, Y. U., Okita, H., Okada, M., Otsu, M., Kume, A., Okuyama, T., Fujimoto, J., Kuratsuji, T., & Kiyokawa, N. (2009). Kinetics and effect of integrin expression on human CD34+ cells during murine leukemia virus-derived retroviral transduction with recombinant fibronectin for stem cell gene therapy. Human Gene Therapy, 20(7), 777-783. https://doi.org/10.1089/hum.2008.159