Transient cytochalasin-D treatment induces apically administered rAAV2 across tight junctions for transduction of enterocytes.
Enteropathogens are known to disrupt apical actin filaments and/or tight-junction barriers of intestinal epithelial cells to promote infection. In this study, we show that a controlled, cytochalasin-D (Cyto-D)-mediated disruption of actin filaments and tight junctions enhanced the apical delivery of the gene-therapy vector recombinant adeno-associated virus serotype 2 (rAAV2). This increase in transduction efficiency can be attributed to the enhanced delivery of rAAV2 across the Cyto-D disrupted tight junctions, allowing basolateral entry of rAAV2. Previously, we have shown that MG101 and doxorubicin are capable of overcoming proteasome-mediated transduction barriers of rAAV2 in enterocytes. In this study, when Cyto-D was combined with MG101 and doxorubicin in apical delivery of rAAV2 to transduce the differentiated Caco-2 enterocytes, a synergistic >2300-fold increase in transgene expression was achieved. We conclude that Cyto-D is capable of permeating the polarized enterocytes for rAAV2 transduction, which may potentially be a useful device to facilitate intestinal gene transfer via the gut lumen.
Fu
Ya-Yuan
YY
Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan, ROC.
Sibley
Eric
E
Division of Pediatric Gastroenterology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Tang
Shiue-Cheng
SC
Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan, ROC.
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000821:virology; D018938:Caco-2 Cells; D015638:Cytochalasin D / Q000378:metabolism / Q000494:pharmacology*; D000229:Dependovirus / Q000235:genetics / Q000502:physiology*; D020895:Enterocytes / Q000187:drug effects / Q000821:virology; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D011995:Recombination, Genetic; D019108:Tight Junctions / Q000187:drug effects / Q000821:virology; D014161:Transduction, Genetic
D015638:Cytochalasin D