TAILIEUCHUNG - Báo cáo sinh học: " In vitro evaluation of a double-stranded selfcomplementary adeno-associated virus type2 vector in bone marrow stromal cells for bone healing"

Tuyển tập các báo cáo nghiên cứu về sinh học được đăng trên tạp chí sinh học Journal of Biology đề tài: In vitro evaluation of a double-stranded selfcomplementary adeno-associated virus type2 vector in bone marrow stromal cells for bone healing | Alaee et al. Genetic Vaccines and Therapy 2011 9 4 http content 9 1 4 GENETIC VACCINES AND THERAPY RESEARCH Open Access In vitro evaluation of a double-stranded self-complementary adeno-associated virus type2 vector in bone marrow stromal cells for bone healing 1 - 1 1 -I- 2 2 1 Farhang Alaee Osamu Sugiyama Mandeep S Virk Ying Tang Bing Wang Jay R Lieberman Abstract Background Both adenoviral and lentiviral vectors have been successfully used to induce bone repair by overexpression of human bone morphogenetic protein 2 BMP-2 in primary rat bone marrow stromal cells in pre-clinical models of ex vivo regional gene therapy. Despite being a very efficient means of gene delivery there are potential safety concerns that may limit the adaptation of these viral vectors for clinical use in humans. Recombinant adeno-associated viral rAAV vector is a promising viral vector without known pathogenicity in humans and has the potential to be an effective gene delivery vehicle to enhance bone repair. In this study we investigated gene transfer in rat and human bone marrow stromal cells in order to evaluate the effectiveness of the self-complementary AAV vector scAAV system which has higher efficiency than the single-stranded AAV vector ssAAV due to its unique viral genome that bypasses the rate-limiting conversion step necessary in ssAAV. Methods Self-complementaryAAV2 encoding GFP and BMP-2 scAAV2-GFP and scAAV2-BMP-2 were used to transduce human and rat bone marrow stromal cells in vitro and subsequently the levels of GFP and BMP-2 expression were assessed 48 hours after treatment. In parallel experiments adenoviral and lentiviral vector mediated over-expression of GFP and BMP-2 were used for comparison. Results Our results demonstrate that the scAAV2 is not capable of inducing significant transgene expression in human and rat bone marrow stromal cells which may be associated with its unique tropism. Conclusions In developing ex vivo gene therapy regimens the

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