Increased Transduction of Human Intestinal Epithelial Cells by Adenoviral Vectors in Inflammatory Bowel Disease

Schmiedlin-Ren, Phyllissa; Kesisoglou, Filippos; Mapili, J.; Sabek, Sayed E; Barnett, Jeffrey L.; Chey, William D.; Roessler, Blake J.

doi:10.1097/01.mib.0000158535.54428.a5

Cited by 7 publications

(6 citation statements)

References 39 publications

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“…Ad5 and Ad5SHORT Transduction Studies in Differentiated and Undifferentiated Caco-2 Cells. As previously reported, , transduction of Caco-2 cells by adenoviral vectors was low. Application of the Ad5SHORT vectors led to further significant decrease in transduction efficiency.…”

Section: Resultssupporting

confidence: 65%

Chimeric Ad5 Vectors Expressing the Short Fiber of Ad41 Show Reduced Affinity for Human Intestinal Epithelium

et al. 2005

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Altering adenovirus tropism has attracted increased attention in recent years to improve gene delivery. We constructed a recombinant Ad5 vector carrying the non-CAR (coxsackievirus and adenovirus receptor) binding short fiber of enterotropic Ad41 (Ad5SHORT) and tested its transduction efficiency on enterocytes. Ad5SHORT was engineered, in high titers similar to the parent vector, by homologous recombination in Escherichia coli BJ5183 (recBC sbcBC) and propagated on C7 cells. Western blotting confirmed the presence of Ad41 short fiber on Ad5SHORT while lack of CAR-binding was evident by the low transduction of CHO-CAR cells. Transduction efficiency of enterocytes, the natural target tissue for the fiber-"donor" virus Ad41, was tested in human intestinal biopsy cultures and in Caco-2 cells, including ulcerative colitis tissue and mucosal wound healing models. Ad5SHORT exhibited up to 23-fold lower transduction levels compared to Ad5 in human intestinal biopsy cultures and up to 13-fold in the in vitro systems. The differences with the in vitro systems were more pronounced when less differentiated cells were used. These studies highlight the potential for using this chimeric Ad5/Ad41 vector as a scaffold for the development of retargeted adenoviral vectors. Finally, our results suggest that the short fiber does not appear to be mediating, at least by itself, the increased enterocyte affinity of Ad41.

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Section: Resultssupporting

confidence: 65%

Chimeric Ad5 Vectors Expressing the Short Fiber of Ad41 Show Reduced Affinity for Human Intestinal Epithelium

et al. 2005

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“…In the present study, LV-transduced cells in human explant samples were found primarily in the subepithelial region of the LP whereas in other colorectal explant studies using adenoviral vectors [42] transduction was commonly observed in the basolateral region. However, it is difficult to compare our data with other published studies as the cell tropism may differ between vectors.…”

Section: Discussionmentioning

confidence: 50%

Effective in vivo and ex vivogene transfer to intestinal mucosa by VSV-G-pseudotyped lentiviral vectors

et al. 2010

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BackgroundGene transfer to the gastrointestinal (GI) mucosa is a therapeutic strategy which could prove particularly advantageous for treatment of various hereditary and acquired intestinal disorders, including inflammatory bowel disease (IBD), GI infections, and cancer.MethodsWe evaluated vesicular stomatitis virus glycoprotein envelope (VSV-G)-pseudotyped lentiviral vectors (LV) for efficacy of gene transfer to both murine rectosigmoid colon in vivo and human colon explants ex vivo. LV encoding beta-galactosidase (LV-β-Gal) or firefly-luciferase (LV-fLuc) reporter genes were administered by intrarectal instillation in mice, or applied topically for ex vivo transduction of human colorectal explant tissues from normal individuals. Macroscopic and histological evaluations were performed to assess any tissue damage or inflammation. Transduction efficiency and systemic biodistribution were evaluated by real-time quantitative PCR. LV-fLuc expression was evaluated by ex vivo bioluminescence imaging. LV-β-Gal expression and identity of transduced cell types were examined by histochemical and immunofluorescence staining.ResultsImaging studies showed positive fLuc signals in murine distal colon; β-Gal-positive cells were found in both murine and human intestinal tissue. In the murine model, β-Gal-positive epithelial and lamina propria cells were found to express cytokeratin, CD45, and CD4. LV-transduced β-Gal-positive cells were also seen in human colorectal explants, consisting mainly of CD45, CD4, and CD11c-positive cells confined to the LP.ConclusionsWe have demonstrated the feasibility of LV-mediated gene transfer into colonic mucosa. We also identified differential patterns of mucosal gene transfer dependent on whether murine or human tissue was used. Within the limitations of the study, the LV did not appear to induce mucosal damage and were not distributed beyond the distal colon.

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“…To determine the biological effects of DRA, we also conducted in vivo experiments. Previous work has shown that normal mature intestinal epithelial cells are extremely resistant to transduction by adenovirus, whereas tissue from IBD patients is transduced significantly more readily than normal tissue [46]. Therefore, Ad-DRA was administered on days 1, 3, and 5 following the initial DSS treatment.…”

Section: Discussionmentioning

confidence: 99%

SLC26A3 (DRA) prevents TNF-alpha-induced barrier dysfunction and dextran sulfate sodium-induced acute colitis

Ding

Dongxiao

Mengke

et al. 2018

Laboratory Investigation

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SLC26A3 encodes a Cl/HCO ion transporter that is also known as downregulated in adenoma (DRA) and is involved in HCO/mucus formation. The role of DRA in the epithelial barrier has not been previously established. In this study, we investigated the in vivo and in vitro mechanisms of DRA in the colon epithelial barrier. Immunofluorescence (IF) and co-immunoprecipitation (co-IP) studies reveal that DRA binds directly to tight junction (TJ) proteins and affects the expression of TJ proteins in polarized Caco-2BBe cells. Similarly, DRA colocalizes with ZO-1 in the intestinal epithelium. Knockdown or overexpression of DRA leads to alterations in TJ proteins and epithelial permeability. In addition, TNF-α treatment downregulates DRA by activating NF-кB and subsequently affecting intestinal epithelial barrier integrity. Furthermore, overexpression of DRA partly reverses the TNF-α-induced damage by stabilizing TJ proteins. Neutralization of TNF-α in dextran sulfate sodium (DSS)-induced colitis mice demonstrates improved the outcomes, and the therapeutic effect of the TNF-α neutralizing mAb is mediated in part by the preservation of DRA expression. These data suggest that DRA may be one of the therapeutic targets of TNF-α. Moreover, DRA delivered by adenovirus vector significantly prevents the exacerbation of colitis and improves epithelial barrier function by promoting the recovery of TJ proteins in DSS-treated mice. In conclusion, DRA plays a role in protecting the epithelial barrier and may be a therapeutic target in gut homeostasis.

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Increased Transduction of Human Intestinal Epithelial Cells by Adenoviral Vectors in Inflammatory Bowel Disease

Abstract: We conclude that intestinal epithelial cells from IBD patients are more readily transfected by standard adenoviral vectors than are those from control patients. These results suggest that targeting genes to inflamed intestine through the luminal route may be possible.

Cited by 7 publications

References 39 publications

Chimeric Ad5 Vectors Expressing the Short Fiber of Ad41 Show Reduced Affinity for Human Intestinal Epithelium

Chimeric Ad5 Vectors Expressing the Short Fiber of Ad41 Show Reduced Affinity for Human Intestinal Epithelium

Effective in vivo and ex vivogene transfer to intestinal mucosa by VSV-G-pseudotyped lentiviral vectors

SLC26A3 (DRA) prevents TNF-alpha-induced barrier dysfunction and dextran sulfate sodium-induced acute colitis

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