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

Area of Science:

• Molecular Biology
• Gastroenterology
• Gene Therapy

Background:

• Intestinal gene transfer is a promising therapeutic strategy for various diseases.
• Recombinant adeno-associated virus serotype 2 (rAAV2) vectors show limited transduction efficiency in intestinal epithelial cells.
• Proteasome inhibitors have previously enhanced rAAV2 transduction in airway cells.

Purpose of the Study:

• To investigate if proteasome inhibition can enhance rAAV2 transduction in intestinal epithelial cells.
• To evaluate the synergistic effect of MG101 and Doxorubicin on rAAV2-mediated transgene expression.

Main Methods:

• Utilized undifferentiated and differentiated Caco-2 cell lines, along with HT-29, T84, and HCT-116 cells.
• Treated cells with proteasome-modulating agents MG101 (N-acetyl-L-leucyl-L-leucyl-L-norleucine) and Doxorubicin.
• Measured rAAV2-mediated luciferase transgene expression to assess transduction efficiency.

Main Results:

• Combined treatment with MG101 and Doxorubicin synergistically increased rAAV2 luciferase expression by over 400-fold in undifferentiated Caco-2 cells.
• In differentiated Caco-2 cells, treatment enhanced transduction from the basolateral surface.
• Enhanced rAAV2 transduction was observed across multiple human intestinal epithelial cell lines (Caco-2, HT-29, T84, HCT-116).

Conclusions:

• MG101 and Doxorubicin exert generic effects that enhance rAAV2 transduction in intestinal epithelial cells.
• Proteasome-modulating agents can significantly improve the efficiency of intestinal gene transfer.
• This approach holds potential for developing more effective intestinal gene therapy protocols.