RESUMO
BACKGROUND: Mas-related G-protein coupled receptor X2 (MRGPRX2) is a promiscuous receptor on mast cells that mediates IgE-independent degranulation and has been implicated in multiple mast cell-mediated disorders, including chronic urticaria, atopic dermatitis, and pain disorders. Although it is a promising therapeutic target, few potent, selective, small molecule antagonists have been identified, and functional effects of human MRGPRX2 inhibition have not been evaluated in vivo. OBJECTIVE: We identified and characterized novel, potent, and selective orally active small molecule MRGPRX2 antagonists for potential treatment of mast cell-mediated disease. METHODS: Antagonists were identified using multiple functional assays in cell lines overexpressing human MRGPRX2, LAD2 mast cells, human peripheral stem cell-derived mast cells, and isolated skin mast cells. Skin mast cell degranulation was evaluated in Mrgprb2em(-/-) knockout (KO) and Mrgprb2em(MRGPRX2) transgenic human MRGPRX2 knock-in (KI) mice by assessment of agonist-induced skin vascular permeability. Ex vivo skin mast cell degranulation and associated histamine release was evaluated by microdialysis of human skin tissue samples. RESULTS: MRGPRX2 antagonists potently inhibited agonist-induced MRGPRX2 activation and mast cell degranulation in all mast cell types tested, in an IgE-independent manner. Orally administered MRGPRX2 antagonists also inhibited agonist-induced degranulation and resulting vascular permeability in MRGPRX2 KI mice. In addition, antagonist treatment dose dependently inhibited agonist-induced degranulation in ex vivo human skin. CONCLUSION: MRGPRX2 small molecule antagonists potently inhibited agonist-induced mast cell degranulation in vitro and in vivo as well as ex vivo in human skin, supporting potential therapeutic utility as a novel treatment for multiple human diseases involving clinically relevant mast cell activation.
RESUMO
Lauritzen Canal, a portion of San Francisco Bay near Richmond, California, USA, was heavily contaminated with dichlorodiphenyltrichloroethane (DDT) and dieldrin as a result of releases from a pesticide-formulating firm. In 1996 and 1997, 82,000 m3 of contaminated sediment was removed from the canal by dredging. This study evaluated the success of the dredging based largely on body burdens of DDT and its metabolites (sigmaDDT) in resident biota, with some data on sediment- and water-contaminant levels and sediment toxicity testing. Sediment disturbance during dredging introduced a pulse of sigmaDDT into the Lauritzen Canal ecosystem, and body burdens of fish and invertebrates increased 2- to 76-fold, depending on the species. Approximately 1 1/2 years after remediation, 11 of 14 indicators showed contamination comparable with or worse than the contamination that existed prior to dredging. Monitoring of mussels up to four years postdredging suggests some modest improvement, although the sigmaDDT body burden of canal mussels remained far above the norm for San Francisco Bay. The elevated sigmaDDT body burdens in biota that persisted for years after remediation reflect recent exposure and are not merely a result of slow metabolic elimination of the sigmaDDT pulse associated with dredging. Sediment sigmaDDT concentrations were low immediately after dredging, but within months, the canal bottom became covered with a veneer of fine sediment as contaminated as that that had been removed. The source of this material has not been conclusively established, but we suspect it came from slumping and erosion from the flanks of the canal beneath docks and around pilings where dredging was not done. In retrospect, either capping in place or more thorough dredging may have been more successful in reducing pesticide exposure of the biota, although there were difficulties associated with both alternatives.
