Virginia Research Day 2025

Medical Student Research Biomedical

20 The Effect Of Novel P1 Adensoine Receptor Modulators on Systemic Lupus Erythematosus (SLE) Serum Inflammatory Cytokines in NZB/W Mice

Shawn Dziepak, MS 1 ; Hilary Montano 2 ; Emma Peohler 1 ; Tyler McCormick 1 ; Chris Reilly, PhD 1,2 Corresponding author: sdziepak@vt.vcom.edu

1 Edward Via College of Osteopathic Medicine - Virginia Campus 2 Virginia-Maryland College of Veterinary Medicine, Blacksburg, Virginia USA

However, A2AR agonist FC-values were higher than or similar to the PBS treatment group in cytokines IL-6, IL-17A, and IL-27. Comparing all treatment groups, both dexamethasone and the A2BR antagonist produced FC-values than the A2AR agonist in 11/12 cytokines, the exception being IL1-beta. When compared to dexamethasone, the A2BR antagonist produced lower or similar FC-values in every inflammatory cytokines. The A2BR antagonist was shown to yield the greatest efficacy in immune cytokine reduction, supported by the lowest fold change values in 8/12 cytokines in the NZB/W SLE mouse model. While the A2AR agonist did not reduce immune cytokine expression as effectively as the A2BR antagonist or dexamethasone, it still reduced cytokine production in most of the analytes. Based on this data, both novel compounds demonstrated a reduction in immune cytokines and should be further investigated as promising agents to reduce SLE mediated inflammatory markers. cytokine except for IL1-beta and IL-27. Conclusion: All treatment conditions demonstrated a reduction in the majority of

Background: Systemic Lupus Erythematosus (SLE) is a devastating chronic systemic auto inflammatory disease that is associated with significant morbidity and mortality. The specific etiology of SLE is not known, however, it is the resultant loss of immunotolerance and dysregulated widespread inflammation. Corticosteroids and other immunosuppressants are the mainstay of treatment aimed at reducing the inflammatory burden of SLE but neither specifically targets autoimmunity nor aims to restore immunotolerance. P1 purinergic receptors are anti-inflammatory G-protein coupled receptors involved in extracellular adenosine metabolism and are current biological targets of interest in SLE. P1 receptors, specifically A2AR and A2BR, are promising targets for autoimmunity suppression due to their inherent immunoregulatory functions and dysregulated expression in SLE. Our experimental aim was to compare the respective effects of dexamethasone, an A2AR agonist, and a A2BR antagonist on immune cytokine profiles in a SLE mouse model.

Methods: NZB/W SLE model mice were assigned to four experimental groups: PBS (experimental control), Dexamethasone (treatment control), an A2AR agonist (ADO-6001), or an A2BR antagonist (ADO-5024). All mice underwent treatment for 13 weeks while serum was collected weekly. A panel of 12 inflammatory cytokines, BAFF, CCL2, CXCL-10, IFN-gamma, IL1-beta, IL-2, IL-4, IL-6, IL-10, IL-17A, IL-27, and TNF-alpha were analyzed and compared before and after treatment using a commercially available custom mouse cytokine kit with the LUMINEX FlexMap3D multiplex instrument. The average net mean florescent intensity (MFI) was measured for each analyte and compared within and between experimental groups. Fold change (FC) values were calculated from net mean MFI to analyze intragroup and intergroup analyte changes. Results: All analytes passed kit specific quality metrics and produced linear regression curves with R-values of >0.99. dexamethasone and the A2BR antagonist demonstrated lower FC-values based on mean net MFI values compared to PBS in all but one cytokine, IL1-beta. The A2AR agonist demonstrated lower FC-values in 9/12 cytokines compared to PBS.

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2025 Research Recognition Day