Negative-strand RNA viral vectors: intravenous application of sendai virus vectors for the systemic delivery of therapeutic genes

Abstract

Treatment by gene replacement is critical in the field of gene therapy. Suitable vectors for the delivery of therapeutic genes have to be generated and tested in preclinical settings. Recently, extraordinary features for a local gene delivery by Sendai virus vectors (SeVV) have been reported for different tissues. Here we show that direct intravenous application of SeVV in mice is not only feasible and safe, but it results in the secretion of therapeutic proteins to the circulation, for example, human clotting Factor IX (hFIX). In vitro characterization of first-generation SeVV demonstrated that secreted amounts of hFIX were at least comparable to published results for retroviral or adeno-associated viral vectors. Furthermore, as a consideration for application in humans, SeVV transduction led to efficient hFIX synthesis in primary human hepatocytes, and SeVV-encoded hFIX proteins could be shown to be functionally active in the human clotting cascade. In conclusion, our investigations demonstrate for the first time that intravenous administration of negative-strand RNA viral vectors may become a useful tool for the wide area of gene replacement requirements.

Characterization of retinoic acid–inducible gene-I expression in primary murine glia following exposure to vesicular stomatitis virus

“…Viral stocks were prepared by infecting monolayer cultures of baby hamster kidney (BHK-21) cells (ATCC, CCL-10) with VSV (Indiana strain) or by infecting African green monkey kidney cells (Vero; ATCC, CCL-81) with Sendai virus (SeV; Fushimi strain) with an enhanced green fluorescent protein (eGFP) gene just downstream of the P gene (described in Bitzer et al , 2003) at a low multiplicity of infection (MOI) of 0.05 plaque-forming units (PFU) per cell and incubated for 48 h at 34°C in SFM4Mega Vir protein-free medium (Hyclone). The cell-free medium containing released viruses was collected and viral titers were quantified using a standard plaque assay of serial dilutions of VSV or SeV on BHK-21 or Vero cells, respectively, at 34°C.…”

Cytopathogenesis of Sendai Virus in Well-Differentiated Primary Pediatric Bronchial Epithelial Cells

“…SeV is virulent in rodents, but despite extensive antigenic and genetic similarity to human parainfluenza virus type 1 (hPIV1), it is not known to cause disease in humans (33). Interest in rSeV as a vector is exemplified by the fact that (i) its genome can easily be manipulated to stably express heterologous genes (9), (ii) it does not undergo homologous recombination, (iii) cell transduction is independent of the cell cycle, (iv) there is no DNA phase during replication and hence no possibility of cell transformation, and (v) its cell or tissue tropisms and replication competency may be modulated by reverse genetics and appropriate rescue systems (5,8).…”

Gene Therapy for Peripheral Arterial Disease and Nursing Implications: Clinical Experience on the use of Sendai Viral Vector

“…The SeV genome cannot integrate into the host genome, thereby reducing the risk for insertional mutagenesis [32,33] A new gene transfer vector based on a non-transmissible recombinant SeVvector carrying the FGF-2 gene (SeV/dF-hFGF2) was investigated in a first-in-man gene therapy clinical trial for CLI patients with no other option, and was completed in March 2011 [34]. A total of 12 patients, one limb per patient, were treated with SeV/ dF-hFGF2 (product name DVC1-0101) in a four-doseescalation design.…”