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

Research output: Contribution to journalArticle

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

Fingerprint

Murine Leukemia Viruses
Cell- and Tissue-Based Therapy
Fibronectins
Integrins
Genetic Therapy
Stem Cells
Retroviridae
Cytokines
Thrombopoietin
Stem Cell Factor
Fetal Blood
Cell Adhesion
Genes
Flow Cytometry
Proteins
Antibodies

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

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

In: Human Gene Therapy, Vol. 20, No. 7, 01.07.2009, p. 777-783.

Research output: Contribution to journalArticle

Horiuchi, Y, Onodera, M, Miyagawa, Y, Sato, B, Onda, K, Katagiri, YU, 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, vol. 20, no. 7, pp. 777-783. https://doi.org/10.1089/hum.2008.159
Horiuchi, Yasuomi ; Onodera, Masafumi ; Miyagawa, Yoshitaka ; Sato, Ban ; Onda, Keiko ; Katagiri, Yohko U. ; Okita, Hajime ; Okada, Mayumi ; Otsu, Makoto ; Kume, Akihiro ; Okuyama, Torayuki ; Fujimoto, Junichiro ; Kuratsuji, Tadatoshi ; Kiyokawa, Nobutaka. / 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. In: Human Gene Therapy. 2009 ; Vol. 20, No. 7. pp. 777-783.
@article{0c76719351b245fcac18e0db5aab924f,
title = "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",
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.",
author = "Yasuomi Horiuchi and Masafumi Onodera and Yoshitaka Miyagawa and Ban Sato and Keiko Onda and Katagiri, {Yohko U.} and Hajime Okita and Mayumi Okada and Makoto Otsu and Akihiro Kume and Torayuki Okuyama and Junichiro Fujimoto and Tadatoshi Kuratsuji and Nobutaka Kiyokawa",
year = "2009",
month = "7",
day = "1",
doi = "10.1089/hum.2008.159",
language = "English",
volume = "20",
pages = "777--783",
journal = "Human Gene Therapy",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "7",

}

TY - JOUR

T1 - 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

AU - Horiuchi, Yasuomi

AU - Onodera, Masafumi

AU - Miyagawa, Yoshitaka

AU - Sato, Ban

AU - Onda, Keiko

AU - Katagiri, Yohko U.

AU - Okita, Hajime

AU - Okada, Mayumi

AU - Otsu, Makoto

AU - Kume, Akihiro

AU - Okuyama, Torayuki

AU - Fujimoto, Junichiro

AU - Kuratsuji, Tadatoshi

AU - Kiyokawa, Nobutaka

PY - 2009/7/1

Y1 - 2009/7/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=68949204235&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68949204235&partnerID=8YFLogxK

U2 - 10.1089/hum.2008.159

DO - 10.1089/hum.2008.159

M3 - Article

C2 - 19284246

AN - SCOPUS:68949204235

VL - 20

SP - 777

EP - 783

JO - Human Gene Therapy

JF - Human Gene Therapy

SN - 1043-0342

IS - 7

ER -