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Published on: May 14, 2012
Non-polarized secretion of mouse interferon-beta from gene-transferred human intestinal Caco-2 cells
K Kawabata1, M Kondo, Y Watanabe
1Department of Drug Delivery Research, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Purpose:
The intestinal epithelium is considered to be a feasible target for somatic gene therapy. To this end, Caco-2 cells derived from human colon carcinoma were transfected with a mouse interferon-beta (IFN-beta) expression vector and several stable sublines were established; this hetero-specific cytokine allows unexpected cellular effects to be avoided. Using the highest mouse IFN-beta-producing sublines, the mode of IFN secretion was examined.
Methods:
The secretion polarity of mouse IFN-beta in its gene-transduced Caco-2 sublines was studied in a bicameral culture system in which the chambers were separated by microporous filters.
Results:
Mouse IFN-beta was secreted to the same extent from both apical and basolateral surfaces of the transduced cells regardless of cell aging.
Conclusions:
These results suggest that in the intestinal epithelium exogenous gene products such as IFNs can be delivered to both the luminal and blood sides in vivo. Thus, the intestinal epithelium may be suitable for systemic and local delivery of therapeutic proteins by gene transfer.
Insights
Somatic gene therapy using the intestinal epithelium is promising. Mouse interferon-beta (IFN-beta) was secreted equally from both sides of engineered Caco-2 cells, suggesting dual delivery potential.
Area of Science:
- Cell Biology
- Molecular Biology
- Gastroenterology
Background:
- The intestinal epithelium presents a viable target for somatic gene therapy.
- Engineered Caco-2 cells, derived from human colon carcinoma, were utilized to study gene expression.
- Mouse interferon-beta (IFN-beta) was chosen as a hetero-specific cytokine to mitigate unintended cellular effects.
Purpose of the Study:
- To investigate the secretion polarity of mouse IFN-beta in gene-transduced Caco-2 cell sublines.
- To assess the feasibility of using the intestinal epithelium for therapeutic protein delivery via gene transfer.
Main Methods:
- Stable Caco-2 sublines expressing mouse IFN-beta were established.
- A bicameral culture system with microporous filters was employed to study secretion polarity.
- The highest IFN-beta-producing sublines were selected for detailed analysis.
Main Results:
- Mouse IFN-beta demonstrated secretion from both apical and basolateral surfaces of the transduced Caco-2 cells.
- The secretion polarity remained consistent irrespective of cell aging.
- This indicates a non-polarized secretion pattern for the exogenous gene product.
Conclusions:
- Exogenous gene products, like IFNs, can be delivered to both luminal and blood sides of the intestinal epithelium in vivo.
- The intestinal epithelium is a suitable candidate for the systemic and local delivery of therapeutic proteins through gene transfer.
- This approach holds potential for novel therapeutic strategies in gastrointestinal diseases.
