MRGPRX2 facilitates IgE-mediated systemic anaphylaxis in a newly established knock-in mouse model.

BACKGROUND

In addition to FcεRI, a subtype of human mast cells (MCs) expresses Mas-related G protein-coupled receptor X2 (MRGPRX2; mouse counterpart MrgprB2). Although MrgprB2 contributes to IgE-mediated passive systemic anaphylaxis (PSA) in vivo, an MRGPRX2 inhibitor, compound 9 (C9), does not block MrgprB2- or IgE-mediated MC degranulation in vitro.

OBJECTIVE

Our aim was to generate mice expressing human MRGPRX2 to study receptor function in vitro and PSA in vivo.

METHODS

The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing approach was utilized to replace endogenous MrgprB2 with human MRGPRX2 in mice (MRGPRX2-KI mice). MRGPRX2 expression in the skin, gingiva, trachea, and colon were evaluated by using an anti-human MRGPRX2 antibody. Peritoneal MCs (PMCs) cultured from wild-type, MRGPRX2-KI, and MrgprB2 mice were used to study agonists-induced degranulation. The effects of selective MRGPRX2 inhibitors (C9 and compound 9-6 [C9-6]) on substance P- or IgE-mediated MC degranulation in vitro and IgE-mediated PSA in vivo were tested.

RESULTS

MRGPRX2-expressing MCs were present in tissues of MRGPRX2-KI mice. Most of the agonists tested induced greater degranulation at lower concentrations in PMCs from MRGPRX2-KI mice than in cells from wild-type mice. Furthermore, C9 and C9-6 inhibited degranulation in MRGPRX2-KI PMCs in response to substance P. In contrast, they had no effect on IgE-mediated degranulation in vitro but did inhibit PSA in MRGPRX2-KI mice in vivo.

CONCLUSIONS

MRGPRX2-KI mice provide a readily available source of primary MCs for signaling studies. Furthermore, transactivation of MRGPRX2 contributes to IgE-mediated PSA, suggesting that MRGPRX2-KI mice could be utilized as a preclinical model for testing novel therapeutics targeting MRGPRX2 and its cross talk with FcεRI.