HSV-1 amplicon viral vector-mediated gene transfer to human bone marrow-derived mesenchymal stem cells

Abstract

Human bone marrow-derived mesenchymal stem cells (BM-hMSCs) are nonhematopoietic stem cells that have the potential to differentiate into adipocytes, osteocytes and chondrocytes. Because of its propensity to migrate to the sites of injury and the ability to expand them rapidly, BM-hMSCs have been exploited as potential gene transfer vehicles to deliver therapeutic genes. Herein, we evaluated the feasibility of employing herpes simplex virus type I (HSV-1) amplicon viral vector as a gene delivery vector to BM-hMSCs. High transduction efficiencies were consistently observed in different isolates of BM-hMSCs following infection with HSV-1 amplicon viral vectors. Furthermore, we demonstrated that transduction with HSV-1 amplicon viral vector did not alter the intrinsic properties of the BM-hMSCs. The morphology and cellular proliferation of the transduced BM-hMSCs were not altered. Chromosomal stability, as confirmed by karyotyping and soft agar colony assays, of the transduced BM-hMSCs was not affected. Similarly, transduction with HSV-1 amplicon viral vectors has no effect on the pluripotent differentiation potential and the tumor tropism of BM-hMSCs. Taken together, these results demonstrated that BM-hMSCs could be transduced efficiently by HSV-1 amplicon viral vector in an 'inert' manner and thus enable strategies to express potential therapeutic genes in BM-hMSCs.

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“…A confluent monolayer culture was observed 7-10 days following initial plating. MSC were then characterized based on the presence of cellular markers CD13, CD44, CD73, CD90, and CD105, as well as its ability to differentiate to osteogenic and adipogenic lineage as previously described [25].…”

“…DGli36 cells (kindly provided by Dr. M. Esteves, University of Massachusetts) and 2-2 cells (kindly provided by Dr. R. SandriGoldin, University of California Los Angeles) were cultured as previously described [25]. Human glioma cell line, U87MG, was purchased from American Type Culture Collection (Rockville, MD) and was cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Invitrogen, Grand Island, NY).…”

“…All sequences were verified by DNA sequencing (1st BASE, Singapore). Purification of HSV-1 amplicon viral vectors and amplicon viral transduction were performed as previously described [25].…”

Progress and prospects: Biological properties and technological advances of herpes simplex virus type 1-based amplicon vectors

“…Furthermore, infection with helper-free amplicons did not alter the intrinsic properties of these cells, including morphology, proliferation and chromosomal stability. Most important, amplicon infection did not alter the pluripotent differentiation potential and the tumor tropism of BM-hMSCs, 25 nor integrate into cellular chromosomes, therefore avoiding any genotoxic effect. Although the efficiency of migration of BM-hMSCs to the sites of injury has been reported to be low, this study illustrates a potentially very useful application of HSV-1-based amplicon vectors.…”

“…Earlier studies had shown that amplicons expressing FGF-2 efficiently promoted neuron generation on delivery into mouse embryonic stem cells. 24 In a very interesting recent study, Lam and coworkers 25 used amplicon vectors to deliver GFP to human bonemarrow-derived mesenchymal stem cells (BM-hMSCs), non-hematopoietic stem cells that have the potential to differentiate into adipocytes, osteocytes and chondrocytes. As these cells can migrate to sites of injury, where they expand rapidly, BM-hMSCs can be exploited as gene transfer vehicles to deliver therapeutic genes.…”