MUC1 and Other Sialoglycoconjugates Inhibit Adenovirus-mediated Gene Transfer to Epithelial Cells

Abstract

Recombinant adenoviruses are currently being evaluated as gene transfer vectors for the treatment of airway diseases. Recent evidence indicates that gene transfer to differentiated airway epithelial cells is inefficient. We hypothesized that apical membrane glycoconjugates, such as the transmembrane mucin MUC1, reduce the efficiency of adenovirus-mediated gene transfer. To address this, studies were performed in primary bronchial epithelial and Madin Darby canine kidney (MDCK) cells transduced to express human MUC1. Colocalization of MUC1 and an adenoviral lacZ transgene in the bronchial epithelial cells revealed that at several multiplicities of infection, the percentage of cells expressing lacZ was five-fold less in MUC1-expressing cells. Moreover, lacZ expression was three- to eight-fold lower in MUC1-expressing than in control MDCK cells, demonstrating that MUC1 interferes with gene transfer and is not merely a phenotypic marker of a cell that is refractory to adenovirus infection. Neuraminidase pretreatment of cells to remove sialic acid residues prior to viral adsorption increased the efficiency of gene transfer two- to five-fold in human airway and MDCK cells, and in a xenograft model of human airway. This effect was also observed in cultured cells that do not express MUC1, suggesting that other sialylated glycoconjugates impact on the efficiency of gene transfer. An inhibitory effect of negatively charged glycoconjugates on adenovirus binding was further supported by the finding that adsorption of adenovirus with a polycation significantly increased gene transfer efficiency. These data demonstrate for the first time that sialoglycoconjugates on epithelial cells reduce the efficiency of adenovirus-mediated gene transfer.

Use of protamine to augment adenovirus-mediated cancer gene therapy

“…This phenomenon was not observed in any of our in vitro studies where vector and protamine could be added simultaneously with good effect. Arcasoy et al 35 reported similar observations when using various polycations in vitro and in vivo. In contrast, Kaplan et al 36 demonstrated that ex vivo preparations of Ad/polycation complexes were effective in augmenting gene transfer of LacZ in mouse lung in vivo.…”

“…31 In addition to these two receptors, the interaction of charges on the surface of the cell membrane appears to play a significant role in cell binding and entry of adenovirus. Arcasoy et al 35 have shown that Ad-mediated transducibility of epithelial cells is increased when the negatively charged sialic acid residue are removed from the epithelial cell membranes. The fiber receptor mediated pathway is not involved in the increased gene transfer observed with the use of polycations.…”

“…particles. [31][32][33][34] In one study, 35 the removal of negatively charged sialic acid residues on cultured polarized epithelial cells was shown to significantly improve the efficiency of adenovirus-mediated gene transfer. The same investigators also demonstrated that gene transfer could be augmented by addition of polycations such as polybrene, poly-l-lysine, DEAE-dextran and protamine.…”

Effects of bronchopulmonary inflammation induced by Pseudomonas aeruginosa on adenovirus-mediated gene transfer to airway epithelial cells in mice

“…42 This may be relevant because related molecules such as MUC1 and other sialylated glycoconjugates have been shown to decrease adenovirus-mediated gene transfer, possibly by interfering with viral attachment or internalization. 43 In the future, better understanding of the mechanisms by which this inflammation interferes with adenovirusmediated gene transfer may lead to more specific approaches to enhancing gene transfer in CF patients with inflamed airways.…”

The cell surface mucin MUC1 limits the severity of influenza A virus infection

“…As many microbial pathogens are known to trigger shedding of the MUC1 extracellular domain (ED) after binding, [24][25][26][27] we tested whether this phenomenon could also be caused by IAV. Supernatants were collected from PR8-infected CHO-K1 control and MUC1 þ cell cultures and the amount of MUC1-ED present in the supernatant was determined by ELISA.…”