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Cellular and Molecular Bioengineering

Cellular and Molecular Bioengineering - Poster Session A

Poster H16 - A Specific GSK-3 Inhibitor Attenuates Interferon-gamma Induced CXCL10 Expression by Keratinocytes

Thursday, October 24, 2024
10:00 AM - 11:00 AM EST
Location: Exhibit Hall E, F & G

Introduction:  Human melanocytes reside within the skin and are responsible for its hue. A skin disease termed vitiligo causes the loss of skin pigmentation due to the destruction of melanocytes. Over 70 million people worldwide are afflicted with this autoimmune disorder, yet only one drug, with a limited response rate, is available. A key step in the pathogenesis of vitiligo is interferon-gamma (IFN-γ) induction of CXCL10 expression by keratinocyte skin cells. We have been investigating the use of a novel organic compound, termed COB-187, as a possible therapeutic for immune-based diseases. In several models of inflammation, COB-187 has been found to inhibit induced CXCL10 expression. Thus, we sought to probe the hypothesis that COB-187 attenuates IFN-γ induction of CXCL10 protein expression by cultured keratinocytes.

Materials and

Methods:  Keratinocytes from ATCC (PCS-200-011) were cultured and treated with IFN-γ (InvivoGen) and COB-187 or DMSO carrier control for 6 hours. Subsequently, the supernatants were collected and analyzed for CXCL10 protein levels via an ELISA assay (ThermoFisher, KAC2361). From the data, the IC50, which is the concentration of the compound needed to decrease the CXCL10 signal by 50%, was determined through regression. An MTS assay, which measures metabolic activity, was used to determine the effect on viability.

Results, Conclusions, and Discussions: 

Results: We found that (i) IFN-γ induces CXCL10 expression by a keratinocyte cell line and (ii) COB-187 dramatically attenuates IFN-γ induced expression by keratinocytes. Human keratinocytes were treated with media alone or 10 ng/ml IFN-γ, in the presence of DMSO (carrier control), or COB-187 (1, 10, or 20 μM). After 6 hours of incubation, the supernatants were harvested and analyzed for the presence of CXCL10 via ELISA. COB-187 significantly inhibited IFN-γ induced CXCL10 protein expression with an IC50 of around 1 μM. An MTS assay was conducted to give insight into cell viability after treatments. These results indicated that COB-187 had little to no effect on cell viability.

Conclusion and

Discussion: While appearing simply cosmetic, vitiligo is an autoimmune disease impacting skin pigmentation and those with the disease can experience additional complications including hearing problems, other immune disorders, and emotional distress. Currently, there is only one FDA-approved drug for vitiligo, ruxolitinib, with a limited response and high cost. Ruxolitinib is a Janas kinase (JAK) inhibitor. The results presented here suggest that COB-187 could be the starting point for a novel therapeutic approach for vitiligo. Specifically, COB-187 does not appear to be a JAK inhibitor as it did not inhibit any JAK kinases in a previous kinase screen (Noori et al., Am J Phys, 2019). Thus, COB-187 appears to provide an opportunity to treat vitiligo via a unique mechanism. Given that COB-187 has been shown to be a highly potent and specific GSK-3 inhibitor (Noori et al., Am J Phys, 2019), the findings suggest that inhibition of GSK-3 could be a target for vitiligo. From a broader perspective, these studies suggest that COB-187 could have therapeutic potential for other diseases that involve aberrant CXCL10 expression (e.g. cancer, pathological inflammation).

Acknowledgements (Optional):  This work was funded by the National Institutes of Health (R15GM110602), the National Science Foundation (CDS&E Grant 1953311).