GSK682753A is a selective and highly potent inverse agonist of the Epstein-Barr virus-induced receptor 2 (EBI2) with an IC50 of 53.6nM[1]. GSK682753A is commonly used in studies on EBI2/GPR183-related immune cell migration, B cell function, and inflammatory signaling[1]. GSK682753A has poor plasma stability in vivo[2].
In vitro, HEK293 cells expressing EBI2 were serum-starved overnight and treated with different concentrations of GSK682753A for 10min. GSK682753A inhibited constitutive EBI2-mediated ERK phosphorylation, blocked downstream MAPK/ERK signaling[1]. CHO cells stably expressing human EBI2 were treated with GSK682753A at 100nM, 1μM and 10μM in the presence of 1nM 7α,25-OHC for 30min. GSK682753A blocked 7α,25-OHC-induced EBI2/GPR183-mediated G protein activation in the [35S]GTPγS binding assay[3]. Mouse splenic B cells isolated from WT mice or B cell-specific human EBI2-overexpressing transgenic mice were incubated with GSK682753A (10μM; 4h) in a Transwell migration assay. GSK682753A inhibited the migration of both WT and EBI2-overexpressing B cells, reducing migration to approximately 50% of the basal migration level of WT B cells[3]. PMA-differentiated THP-1 macrophages activated with LPS (100ng/mL) and IFN-γ (10ng/mL) were treated with GSK682753A (10μM; 8h). GSK682753A reduced IL-6 and TNF-α production in this inflammatory macrophage model[4].
References:
[1]. POTENT, I. O. A., & AGONIST, E. I. (2011). Ligand Modulation of the Epstein-Barr Virus-induced Seven-transmembrane Receptor EBI2. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 286(33), 29292-29302.
[2]. Braden, K., Giancotti, L. A., Chen, Z., DeLeon, C., Latzo, N., Boehm, T., ... & Salvemini, D. (2020). GPR183-oxysterol axis in spinal cord contributes to neuropathic pain. The Journal of Pharmacology and Experimental Therapeutics, 375(2), 367-375.
[3]. Benned-Jensen, T., Madsen, C. M., Arfelt, K. N., Smethurst, C., Blanchard, A., Jepras, R., & Rosenkilde, M. M. (2013). Small molecule antagonism of oxysterol-induced Epstein–Barr virus induced gene 2 (EBI2) activation. FEBS Open Bio, 3, 156-160.
[4]. Bartlett, S., Gemiarto, A. T., Ngo, M. D., Sajiir, H., Hailu, S., Sinha, R., ... & Ronacher, K. (2020). GPR183 regulates interferons, autophagy, and bacterial growth during Mycobacterium tuberculosis infection and is associated with TB disease severity. Frontiers in immunology, 11, 601534.