Functional Evaluation of Computationally Designed IL-10 in IL-10 KO Mice

Area of Science:

• Biotechnology
• Immunology
• Gene Therapy

Background:

• Interleukin-10 (IL-10) has therapeutic promise for inflammatory diseases but also exhibits pro-inflammatory functions.
• Developing safer IL-10 variants is crucial for effective therapeutic applications.

Purpose of the Study:

• To design and evaluate IL-10 mutants with reduced pro-inflammatory activity for potential gene therapy applications.
• To assess the efficacy and safety of IL-10 mutant gene therapy in a mouse model of inflammatory bowel disease (IBD).

Main Methods:

• Structure-based computational design was used to create three IL-10 mutants.
• Mutant and wild-type IL-10 activity was tested in cell lines.
• Recombinant adeno-associated virus serotype 8 (rAAV8) vectors were used for gene therapy in IL-10 knockout mice.

Main Results:

• IL-10 mutants displayed significantly lower activity in IL-10-responsive cell lines compared to wild-type IL-10.
• Gene therapy with rAAV8 vectors resulted in high transgene expression in mice.
• IL-10 gene therapy improved body weight, reduced colon injury, and prevented IBD development.
• IL-10 mutant gene therapy showed reduced stimulation of CD8 T and NK cells compared to wild-type IL-10 therapy.

Conclusions:

• Engineered IL-10 mutants offer comparable protective effects to wild-type IL-10 in an IBD mouse model.
• These mutants demonstrate potential for reduced pro-inflammatory function, indicating therapeutic promise.
• Further safety and efficacy studies are warranted for IL-10 mutant gene therapy in various disease models.