Tags

Type your tag names separated by a space and hit enter

Sustained transgene expression by human cord blood derived CD34+ cells transduced with simian immunodeficiency virus agmTYO1-based vectors carrying the human coagulation factor VIII gene in NOD/SCID mice.
J Gene Med. 2004 Oct; 6(10):1049-60.JG

Abstract

BACKGROUND

Gene therapy is being studied as the next generation therapy for hemophilia and several clinical trials have been carried out, albeit with limited success. To explore the possibility of utilizing autologous bone marrow transplantation of genetically modified hematopoietic stem cells for hemophilia gene therapy, we investigated the efficacy of genetically engineered CD34+ cell transplantation to NOD/SCID mice for expression of human factor VIII (hFVIII).

METHODS

CD34+ cells were transduced with a simian immunodeficiency virus agmTYO1 (SIV)-based lentiviral vector carrying the enhanced green fluorescent protein (eGFP) gene (SIVeGFP) or the hFVIII gene (SIVhFVIII). CD34+ cells transduced with SIV vectors were transplanted to NOD/SCID mice. Engraftment of transduced CD34+ cells and expression of transgenes were studied.

RESULTS

We could efficiently transduce CD34+ cells using the SIVeGFP vector in a dose-dependent manner, reaching a maximum (99.6 +/- 0.1%) at MOI of 5 x 10(3) vector genome/cell. After transducing CD34+ cells with SIVhFVIII, hFVIII was produced (274.3 +/- 20.1 ng) from 10(6) CD34+ cells during 24 h in vitro incubation. Transplantation of SIVhFVIII-transduced CD34+ cells (5-10 x 10(5)) at a multiplicity of infection (MOI) of 50 vector genome/cell into NOD/SCID mice resulted in successful engraftment of CD34+ cells and production of hFVIII (minimum 1.2 +/- 0.9 ng/mL, maximum 3.6 +/- 0.8 ng/mL) for at least 60 days in vivo. Transcripts of the hFVIII gene and the hFVIII antigen were also detected in the murine bone marrow cells.

CONCLUSIONS

Transplantation of ex vivo transduced hematopoietic stem cells by non-pathogenic SIVhFVIII without exposure of subjects to viral vectors is safe and potentially applicable for gene therapy of hemophilia A patients.

Authors+Show Affiliations

Division of Cell and Molecular Medicine, The Center for Molecular Medicine, Jichi Medical School, Tochigi-ken 329-0498, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15386735

Citation

Kikuchi, Jiro, et al. "Sustained Transgene Expression By Human Cord Blood Derived CD34+ Cells Transduced With Simian Immunodeficiency Virus agmTYO1-based Vectors Carrying the Human Coagulation Factor VIII Gene in NOD/SCID Mice." The Journal of Gene Medicine, vol. 6, no. 10, 2004, pp. 1049-60.
Kikuchi J, Mimuro J, Ogata K, et al. Sustained transgene expression by human cord blood derived CD34+ cells transduced with simian immunodeficiency virus agmTYO1-based vectors carrying the human coagulation factor VIII gene in NOD/SCID mice. J Gene Med. 2004;6(10):1049-60.
Kikuchi, J., Mimuro, J., Ogata, K., Tabata, T., Ueda, Y., Ishiwata, A., Kimura, K., Kimura, K., Takano, K., Madoiwa, S., Mizukami, H., Hanazono, Y., Kume, A., Hasegawa, M., Ozawa, K., & Sakata, Y. (2004). Sustained transgene expression by human cord blood derived CD34+ cells transduced with simian immunodeficiency virus agmTYO1-based vectors carrying the human coagulation factor VIII gene in NOD/SCID mice. The Journal of Gene Medicine, 6(10), 1049-60.
Kikuchi J, et al. Sustained Transgene Expression By Human Cord Blood Derived CD34+ Cells Transduced With Simian Immunodeficiency Virus agmTYO1-based Vectors Carrying the Human Coagulation Factor VIII Gene in NOD/SCID Mice. J Gene Med. 2004;6(10):1049-60. PubMed PMID: 15386735.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sustained transgene expression by human cord blood derived CD34+ cells transduced with simian immunodeficiency virus agmTYO1-based vectors carrying the human coagulation factor VIII gene in NOD/SCID mice. AU - Kikuchi,Jiro, AU - Mimuro,Jun, AU - Ogata,Kyoichi, AU - Tabata,Toshiaki, AU - Ueda,Yasuji, AU - Ishiwata,Akira, AU - Kimura,Kouzoh, AU - Kimura,Konzoh, AU - Takano,Katsuhiro, AU - Madoiwa,Seiji, AU - Mizukami,Hiroaki, AU - Hanazono,Yutaka, AU - Kume,Akihiro, AU - Hasegawa,Mamoru, AU - Ozawa,Keiya, AU - Sakata,Yoichi, PY - 2004/9/24/pubmed PY - 2005/3/3/medline PY - 2004/9/24/entrez SP - 1049 EP - 60 JF - The journal of gene medicine JO - J Gene Med VL - 6 IS - 10 N2 - BACKGROUND: Gene therapy is being studied as the next generation therapy for hemophilia and several clinical trials have been carried out, albeit with limited success. To explore the possibility of utilizing autologous bone marrow transplantation of genetically modified hematopoietic stem cells for hemophilia gene therapy, we investigated the efficacy of genetically engineered CD34+ cell transplantation to NOD/SCID mice for expression of human factor VIII (hFVIII). METHODS: CD34+ cells were transduced with a simian immunodeficiency virus agmTYO1 (SIV)-based lentiviral vector carrying the enhanced green fluorescent protein (eGFP) gene (SIVeGFP) or the hFVIII gene (SIVhFVIII). CD34+ cells transduced with SIV vectors were transplanted to NOD/SCID mice. Engraftment of transduced CD34+ cells and expression of transgenes were studied. RESULTS: We could efficiently transduce CD34+ cells using the SIVeGFP vector in a dose-dependent manner, reaching a maximum (99.6 +/- 0.1%) at MOI of 5 x 10(3) vector genome/cell. After transducing CD34+ cells with SIVhFVIII, hFVIII was produced (274.3 +/- 20.1 ng) from 10(6) CD34+ cells during 24 h in vitro incubation. Transplantation of SIVhFVIII-transduced CD34+ cells (5-10 x 10(5)) at a multiplicity of infection (MOI) of 50 vector genome/cell into NOD/SCID mice resulted in successful engraftment of CD34+ cells and production of hFVIII (minimum 1.2 +/- 0.9 ng/mL, maximum 3.6 +/- 0.8 ng/mL) for at least 60 days in vivo. Transcripts of the hFVIII gene and the hFVIII antigen were also detected in the murine bone marrow cells. CONCLUSIONS: Transplantation of ex vivo transduced hematopoietic stem cells by non-pathogenic SIVhFVIII without exposure of subjects to viral vectors is safe and potentially applicable for gene therapy of hemophilia A patients. SN - 1099-498X UR - https://www.unboundmedicine.com/medline/citation/15386735/Sustained_transgene_expression_by_human_cord_blood_derived_CD34+_cells_transduced_with_simian_immunodeficiency_virus_agmTYO1_based_vectors_carrying_the_human_coagulation_factor_VIII_gene_in_NOD/SCID_mice_ L2 - https://doi.org/10.1002/jgm.609 DB - PRIME DP - Unbound Medicine ER -