Stable integration of recombinant adeno-associated virus vector genomes after transduction of murine hematopoietic stem cells

Zongchao Han, Li Zhong, Njeri Maina, Zhongbo Hu, Xiaomiao Li, Nitin S. Chouthai, Daniela Bischof, Kirsten A. Weigel-Van Aken, William B. Slayton, Mervin C. Yoder, Arun Srivasta

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31 Scopus citations

Abstract

We previously reported that among single-stranded adeno-associated virus (ssAAV) vectors, serotypes 1 through 5, ssAAV1 is the most efficient in transducing murine hematopoietic stem cells (HSCs), but viral second-strand DNA synthesis remains a rate-limiting step. Subsequently, using double-stranded, self-complementary AAV (scAAV) vectors, serotypes 7 through 10, we observed that scAAV7 vectors also transduce murine HSCs efficiently. In the present study, we used scAAV1 and scAAV7 shuttle vectors to transduce HSCs in a murine bone marrow serial transplant model in vivo, which allowed examination of the AAV proviral integration pattern in the mouse genome, as well as recovery and nucleotide sequence analyses of AAV-HSC DNA junction fragments. The proviral genomes were stably integrated, and integration sites were localized to different mouse chromosomes. None of the integration sites was found to be in a transcribed gene, or near a cellular oncogene. None of the animals, monitored for up to 1 year, exhibited pathological abnormalities. Thus, AAV proviral integration-induced risk of oncogenesis was not found in our study, which provides functional confirmation of stable transduction of self-renewing multipotential HSCs by scAAV vectors as well as promise for the use of these vectors in the potential treatment of disorders of the hematopoietic system.

Original languageEnglish (US)
Pages (from-to)267-278
Number of pages12
JournalHuman gene therapy
Volume19
Issue number3
DOIs
StatePublished - Mar 1 2008

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ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Han, Z., Zhong, L., Maina, N., Hu, Z., Li, X., Chouthai, N. S., Bischof, D., Weigel-Van Aken, K. A., Slayton, W. B., Yoder, M. C., & Srivasta, A. (2008). Stable integration of recombinant adeno-associated virus vector genomes after transduction of murine hematopoietic stem cells. Human gene therapy, 19(3), 267-278. https://doi.org/10.1089/hum.2007.161