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

Li Zhong, Weiming Li, Yanjun Li, Weihong Zhao, Jianqing Wu, Baozheng Li, Njeri Maina, Daniela Bischof, Keyun Qing, Kirsten A. Weigel-Kelley, Irene Zolotukhin, Kenneth H. Warrington, Xiaomiao Li, William B. Slayton, Mervin Yoder, Arun Srivastava

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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 second-strand 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.

Original languageEnglish
Pages (from-to)321-333
Number of pages13
JournalHuman Gene Therapy
Volume17
Issue number3
DOIs
StatePublished - Mar 2006

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Dependovirus
Hematopoietic Stem Cells
Non-Receptor Type 2 Protein Tyrosine Phosphatase
Transgenes
Blood Cells
Cell- and Tissue-Based Therapy
Serogroup
In Vitro Techniques
Bone Marrow Transplantation
Coinfection
Bone Marrow Cells
Genetic Therapy
Genes
Stem Cells
Transplantation
Bone Marrow
Genome
DNA

ASJC Scopus subject areas

  • Genetics

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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. / Zhong, Li; Li, Weiming; Li, Yanjun; Zhao, Weihong; Wu, Jianqing; Li, Baozheng; Maina, Njeri; Bischof, Daniela; Qing, Keyun; Weigel-Kelley, Kirsten A.; Zolotukhin, Irene; Warrington, Kenneth H.; Li, Xiaomiao; Slayton, William B.; Yoder, Mervin; Srivastava, Arun.

In: Human Gene Therapy, Vol. 17, No. 3, 03.2006, p. 321-333.

Research output: Contribution to journalArticle

Zhong, L, Li, W, Li, Y, Zhao, W, Wu, J, Li, B, Maina, N, Bischof, D, Qing, K, Weigel-Kelley, KA, Zolotukhin, I, Warrington, KH, Li, X, Slayton, WB, Yoder, M & 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, vol. 17, no. 3, pp. 321-333. https://doi.org/10.1089/hum.2006.17.321
Zhong, Li ; Li, Weiming ; Li, Yanjun ; Zhao, Weihong ; Wu, Jianqing ; Li, Baozheng ; Maina, Njeri ; Bischof, Daniela ; Qing, Keyun ; Weigel-Kelley, Kirsten A. ; Zolotukhin, Irene ; Warrington, Kenneth H. ; Li, Xiaomiao ; Slayton, William B. ; Yoder, Mervin ; Srivastava, Arun. / 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. In: Human Gene Therapy. 2006 ; Vol. 17, No. 3. pp. 321-333.
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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 second-strand 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.",
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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.

AU - Li, Xiaomiao

AU - Slayton, William B.

AU - Yoder, Mervin

AU - Srivastava, Arun

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AB - 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 second-strand 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.

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