Site-specific gene delivery in vivo through engineered Sendai viral envelopes

Abstract

ABSTRACT

A Histidine Switch in Hemagglutinin-Neuraminidase Triggers Paramyxovirus-Cell Membrane Fusion

“…Although F proteins of some of the simian virus 5 strains and mutant F protein of NDV induce cell fusion independently (39), coexpression of homologous HN proteins (type specific) accentuates the fusion potential of the virus. Similarly, our previous studies established that Sendai viral envelope devoid of HN protein (F-virosomes [FV]) can fuse with liver cells in culture and in whole animals, but cografting of its HN protein in the same envelope significantly enhances the fusion activity (3,4,34). These findings indicate that a homotypic F-HN interaction is essential for efficient membrane fusion (41).…”

“…These results demonstrate that peptide sequences of HN proteins containing His 247 and His 245 (SH or PH) act as a trigger for F-mediated cell-cell fusion in the two virus species selected. We had established previously that the F glycoprotein of SeV could specifically and strongly bind to a sugar receptor, ASGPR, on the surface of human hepatoblastoma cells (HepG2) in culture and in whole animal leading to complete fusion of viral envelopes devoid of HN proteins (FV) with liver cell plasma membrane (3,30,34,45). However, since the rate and extent of such fusion were dramatically reduced, a critical role of HN in triggering F protein for a superfusion condition was hypothesized (45).…”

“…While developing the FV-based novel liver-specific drug/ gene delivery vehicle (4,30,34) exploiting the high affinity of F-protein to asialoglycoprotein receptors (ASGPR) on the hepatocyte surface, we observed significant reduction in membrane fusion activity in the absence of its native attachment protein, HN. The fusion efficiency of FV increased on cografting a histidyl residue of a cationic amphiphile (L H ) in the virosome membrane (L H F-virosome) (45).…”

Sustained Activation of Mitogen-Activated Protein Kinases and Activator Protein 1 by the Hepatitis B Virus X Protein in Mouse Hepatocytes In Vivo

“…The F-virosomes containing HBx DNA were prepared following our standardized protocol (53). In brief, 75 g of each plasmid was incubated with a detergent-solubilized fraction of Sendai virus containing its envelope devoid of hemagglutinin-neuraminidase protein.…”

“…Exploiting the membrane-fusogenic ability of Sendai virus F glycoprotein and the high affinity of the terminal ␤-galactosecontaining ligand (present on F glycoprotein) for the asialoglycoprotein receptor on the hepatocyte surface, we have developed an efficient virus-based delivery system (F-virosomes) (55a) to transfer foreign genes to mouse hepatocytes in vivo (6,7,53). Using this system, we have now introduced the HBx gene into the whole animal and examined the effect of hepatocyte-expressed HBx protein on signaling cascades.…”