Recombinant Sendai virus‐mediated gene transfer to vasculature: a new class of efficient gene transfer vector to the vascular system

Abstract

Unsolved issues in clinical gene therapy for cardiovascular diseases include gene transfer efficiency and the requirement of a longer exposure time. We developed a novel mononegavirus vector, recombinant Sendai virus (SeV), and tested whether it can overcome the present hurdles. SeV showed dose‐dependent and persistent gene expression in either proliferating or arrested cells, suggesting stability of RNA genome of the vector. An outstanding feature of the SeV‐mediated gene transfer was that even a brief exposure provided nearly peak gene expression in both culture cells and human veins ex vivo, as well as rabbit carotid arteries in vivo. Gene transfer to human great saphenous veins showed high efficacy in luminal and vasa vasoral endothelial cells and in adventitial fibroblasts via both intraluminal delivery and simple floating; however, only scattered cells were transfected in both neointima and media, regardless of the infusion pressure. Veins with a dissected neointima showed a clear transfection to medial cells, suggesting that the barrier in neointima reduces SeV‐mediated gene transfer to tunica media, similar to the case with adenoviruses. Although the fibromuscular neointima is a common obstacle, these findings suggest that SeV may overcome other limitations of current vectors. SeV may be an important new vector in treating subjects with vascular disorders.

Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved

“…(i) SeV vectors can infect nondividing, quiescent cells as well as dividing cells unlike oncoretroviral vectors. [7][8][9][10][11] Thus, they can be used to infect cells that are terminally differentiated as well as at various stages of differentiation, whether they are dividing or not.…”

“…[3][4][5] We have previously developed Sendai virus (SeV) vectors that replicate in the form of negative-sense single-stranded RNA in the cytoplasm of infected cells and do not go through a DNA phase. 6 SeV vectors can efficiently introduce foreign genes without toxicity into airway epithelial cells, 7 vascular tissue, 8 skeletal muscle, 9 synovial cells, 10 retinal tissue, 11 and hematopoietic progenitor cells. 12 Here we report that the SeV-mediated gene transfer into primate ES cells is very efficient and stable even after the terminal differentiation of the cells.…”

Recombinant Sendai virus vectors for activated T lymphocytes

“…[5][6][7][8][30][31][32][33] However, in our systems, excellent gene transfer could be achieved by the simple addition of SeV vector solution without specific reagents or centrifugation. Moreover, optimal SeV-mediated gene transfer to activated T cells could be performed during a relatively brief exposure time (less than 30 min, Figure 4d), with representative results seen in nasal mucosa, 20 in the vasculature, 21 in retinal tissue, 24 as well as in human umbilical cord bloodderived CD34-positive cells. 45 In addition, rare syncytium formation and few cytopathic effects were observed at the MOI used in this study (data not shown).…”

“…14 Using this vector, we demonstrated markedly superior gene transfer efficiency compared to findings with currently available vectors for use in various organs. [19][20][21][22][23][24][25] In contrast to systems involving retroviridae, replication and transcription of SeV are seen in the cytoplasm accompanying cellular tubulin, 26 and nuclear import and genomic integration are not required, 14 thereby suggesting the elimination of potential risks, including unexpected events related to chromosomal injury.…”

Reduced toxicity of F-deficient Sendai virus vector in the mouse fetus

“…7 Also recombinant virus designed to encode the gp120 glycoprotein resulted in extremely high levels of transgene expression in CV-1 monkey kidney cells. 8 Interestingly, although HVJ liposomes containing Sendai-derived haemaglutinin protein complexed with plasmid DNA carrying the b-galactosidase gene have been applied prenatally by intra-amniotic delivery, which resulted in low and transient expression in rat skin, 9 no previous studies have described the use of recombinant SeV particles in a prenatal context probably due to its intrinsic cytotoxic nature and transient gene expression.…”