Proteasome modulating agents induce rAAV2-mediated transgene expression in human intestinal epithelial cells

Tang, Shiue–Cheng; Sambanis, Athanassios; Sibley, Eric

doi:10.1016/j.bbrc.2005.03.245

Cited by 18 publications

(15 citation statements)

References 41 publications

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“…The results of this project demonstrate that colonic epithelial cells are susceptible to in vitro and ex vivo transduction with AAV pseudotypes 2/1, 2/1 and 2/5. Similar to previous studies [32], HT29 and T84 cells were poorly transduced by AAV. However, non-polarized Caco2 cells provide a good intestinal cell type for AAV2/1, 2/2 and 2/5 transduction.…”

Section: Discussionsupporting

confidence: 88%

Gene Delivery to Intestinal Epithelial Cells In vitro and In vivo with Recombinant Adeno-Associated Virus Types 1, 2 and 5

et al. 2007

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Intestinal disorders such as inflammatory bowel disease (IBD) result in chronic illness requiring lifelong therapy. Our aim was to evaluate the efficacy of recombinant adeno-associated virus (AAV) vector-mediated gene delivery to intestinal epithelial cells in vitro and in vivo. Human colon epithelial cell lines and colon biopsies were transduced using AAV pseudotypes 2/1, 2/2, and 2/5 encoding green fluorescence protein (GFP). Mice were administered the same vectors through oral, enema, intraperitoneal (IP) injection and superior mesenteric artery (SMA) injection routes. Tropism and efficiency were determined by microscopy, flow cytometry, immunohistochemistry and PCR. Caco2 cells were more permissive to AAV transduction. Human colon epithelial cells in organ culture were more effectively transduced by AAV2/2. SMA injection provided the most effective means of vector gene transfer to small intestine and colonic epithelial cells in vivo. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript applicable for the investigation of IBD pathogenesis and novel therapeutic options, but also revealed the need for further studies to identify more efficient pseudotypes.

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Section: Discussionsupporting

confidence: 88%

Gene Delivery to Intestinal Epithelial Cells In vitro and In vivo with Recombinant Adeno-Associated Virus Types 1, 2 and 5

et al. 2007

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“…1C), carrying the expression cassette of EGFP only, were used to associate with PEI for polyplex preparation. These two plasmids were constructed as previously described [26,27]. In addition to these two plasmids, a third plasmid, pCMV-lacZ (Fig.…”

Section: Cell Culture and Plasmid Dna Preparationmentioning

confidence: 99%

Electrostatic immobilization of DNA polyplexes on small intestinal submucosa for tissue substrate-mediated transfection

2008

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“…Previously, we have revealed proteasome-mediated barriers to rAAV2 transduction in intestinal epithelial cells (Tang et al, 2005). Delivery of rAAV2 vectors to polarized human Caco-2 enterocytes in transwells resulted in preferential transduction at the basolateral surface following treatment that could be enhanced by treatment with the proteasomemodulating agents MG101 and doxorubicin.…”

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confidence: 95%

“…Previously, we have shown that the proteasome-modulating agents MG101 and doxorubicin are capable of overcoming proteasome-mediated transduction barriers to rAAV2 in enterocytes (Tang et al, 2005). Cyto-D (1 mg ml…”

mentioning

confidence: 99%

“…Transduction barriers have been identified at different stages of rAAV2-host cell interaction; these stages include internalization, endocytosis, ubiquitination-proteasome pathway, uncoating and second-strand DNA synthesis (Tang et al, 2005;Duan et al, 2000;Thomas et al, 2004;Ferrari et al, 1996). Our results suggest that apical rAAV2 transduction of the polarized intestinal epithelium is limited in part by microvilli and tight junctions.…”

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confidence: 99%

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Transient cytochalasin-D treatment induces apically administered rAAV2 across tight junctions for transduction of enterocytes

Sibley

Tang

2008

Journal of General Virology

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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.

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Proteasome modulating agents induce rAAV2-mediated transgene expression in human intestinal epithelial cells

Cited by 18 publications

References 41 publications

Gene Delivery to Intestinal Epithelial Cells In vitro and In vivo with Recombinant Adeno-Associated Virus Types 1, 2 and 5

Gene Delivery to Intestinal Epithelial Cells In vitro and In vivo with Recombinant Adeno-Associated Virus Types 1, 2 and 5

Electrostatic immobilization of DNA polyplexes on small intestinal submucosa for tissue substrate-mediated transfection

Transient cytochalasin-D treatment induces apically administered rAAV2 across tight junctions for transduction of enterocytes

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