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Evaluation of primitive murine hematopoietic stem and progenitor cell transduction in vitro and in vivo by recombinant adeno-associated virus vector serotypes 1 through 5.
Hum Gene Ther 2006; 17(3):321-33HG

Abstract

Conflicting data exist on hematopoietic cell transduction by AAV serotype 2 (AAV2) vectors, and additional AAV serotype vectors have not been evaluated for their efficacy in hematopoietic stem/progenitor cell transduction. We evaluated the efficacy of conventional, single-stranded AAV serotype vectors 1 through 5 in primitive murine hematopoietic stem/progenitor cells in vitro as well as in vivo. In progenitor cell assays using Sca1+ c-kit+ Lin- hematopoietic cells, 9% of the colonies in cultures infected with AAV1 expressed the transgene. Coinfection of AAV1 with self-complementary AAV vectors carrying the gene for T cell protein tyrosine phosphatase (scAAV-TC-PTP) increased the transduction efficiency to 24%, indicating that viral secondstrand DNA synthesis is a rate-limiting step. This was further corroborated by the use of scAAV vectors, which bypass this requirement. In bone marrow transplantation studies involving lethally irradiated syngeneic mice, Sca1+ c-kit+ Lin- cells coinfected with AAV1 +/- scAAV-TC-PTP vectors led to transgene expression in 2 and 7.5% of peripheral blood (PB) cells, respectively, 6 months posttransplantation. In secondary transplantation experiments, 7% of PB cells and 3% of bone marrow (BM) cells expressed the transgene 6 months posttransplantation. Approximately 21% of BM-derived colonies harbored the proviral DNA sequences in integrated forms. These results document that AAV1 is thus far the most efficient vector in transducing primitive murine hematopoietic stem/progenitor cells. Further studies involving scAAV genomes and hematopoietic cell-specific promoters should further augment the transduction efficiency of AAV1 vectors, which should have implications in the optimal use of these vectors in hematopoietic stem cell gene therapy.

Authors+Show Affiliations

Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32610, USA.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)

Evaluation Studies
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16544981

Citation

Zhong, Li, et al. "Evaluation of Primitive Murine Hematopoietic Stem and Progenitor Cell Transduction in Vitro and in Vivo By Recombinant Adeno-associated Virus Vector Serotypes 1 Through 5." Human Gene Therapy, vol. 17, no. 3, 2006, pp. 321-33.
Zhong L, Li W, Li Y, et al. Evaluation of primitive murine hematopoietic stem and progenitor cell transduction in vitro and in vivo by recombinant adeno-associated virus vector serotypes 1 through 5. Hum Gene Ther. 2006;17(3):321-33.
Zhong, L., Li, W., Li, Y., Zhao, W., Wu, J., Li, B., ... Srivastava, A. (2006). Evaluation of primitive murine hematopoietic stem and progenitor cell transduction in vitro and in vivo by recombinant adeno-associated virus vector serotypes 1 through 5. Human Gene Therapy, 17(3), pp. 321-33.
Zhong L, et al. Evaluation of Primitive Murine Hematopoietic Stem and Progenitor Cell Transduction in Vitro and in Vivo By Recombinant Adeno-associated Virus Vector Serotypes 1 Through 5. Hum Gene Ther. 2006;17(3):321-33. PubMed PMID: 16544981.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of primitive murine hematopoietic stem and progenitor cell transduction in vitro and in vivo by recombinant adeno-associated virus vector serotypes 1 through 5. AU - Zhong,Li, AU - Li,Weiming, AU - Li,Yanjun, AU - Zhao,Weihong, AU - Wu,Jianqing, AU - Li,Baozheng, AU - Maina,Njeri, AU - Bischof,Daniela, AU - Qing,Keyun, AU - Weigel-Kelley,Kirsten A, AU - Zolotukhin,Irene, AU - Warrington,Kenneth H,Jr AU - Li,Xiaomiao, AU - Slayton,William B, AU - Yoder,Mervin C, AU - Srivastava,Arun, PY - 2006/3/21/pubmed PY - 2006/5/5/medline PY - 2006/3/21/entrez SP - 321 EP - 33 JF - Human gene therapy JO - Hum. Gene Ther. VL - 17 IS - 3 N2 - Conflicting data exist on hematopoietic cell transduction by AAV serotype 2 (AAV2) vectors, and additional AAV serotype vectors have not been evaluated for their efficacy in hematopoietic stem/progenitor cell transduction. We evaluated the efficacy of conventional, single-stranded AAV serotype vectors 1 through 5 in primitive murine hematopoietic stem/progenitor cells in vitro as well as in vivo. In progenitor cell assays using Sca1+ c-kit+ Lin- hematopoietic cells, 9% of the colonies in cultures infected with AAV1 expressed the transgene. Coinfection of AAV1 with self-complementary AAV vectors carrying the gene for T cell protein tyrosine phosphatase (scAAV-TC-PTP) increased the transduction efficiency to 24%, indicating that viral secondstrand DNA synthesis is a rate-limiting step. This was further corroborated by the use of scAAV vectors, which bypass this requirement. In bone marrow transplantation studies involving lethally irradiated syngeneic mice, Sca1+ c-kit+ Lin- cells coinfected with AAV1 +/- scAAV-TC-PTP vectors led to transgene expression in 2 and 7.5% of peripheral blood (PB) cells, respectively, 6 months posttransplantation. In secondary transplantation experiments, 7% of PB cells and 3% of bone marrow (BM) cells expressed the transgene 6 months posttransplantation. Approximately 21% of BM-derived colonies harbored the proviral DNA sequences in integrated forms. These results document that AAV1 is thus far the most efficient vector in transducing primitive murine hematopoietic stem/progenitor cells. Further studies involving scAAV genomes and hematopoietic cell-specific promoters should further augment the transduction efficiency of AAV1 vectors, which should have implications in the optimal use of these vectors in hematopoietic stem cell gene therapy. SN - 1043-0342 UR - https://www.unboundmedicine.com/medline/citation/16544981/Evaluation_of_primitive_murine_hematopoietic_stem_and_progenitor_cell_transduction_in_vitro_and_in_vivo_by_recombinant_adeno_associated_virus_vector_serotypes_1_through_5_ L2 - https://www.liebertpub.com/doi/full/10.1089/hum.2006.17.321?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -