RhoA orchestrates glycolysis for TH2 cell differentiation and allergic airway inflammation.

Background Mitochondrial metabolism is known to be important for T-cell activation. However, its involvement in effector T-cell differentiation has just begun to gain attention. Importantly, how metabolic pathways are integrated with T-cell activation and effector cell differentiation and function remains largely unknown. Objective We sought to test our hypothesis that RhoA GTPase orchestrates glycolysis for T H 2 cell differentiation and T H 2-mediated allergic airway inflammation. Methods Conditional RhoA-deficient mice were generated by crossing RhoA flox/flox mice with CD2-Cre transgenic mice. Effects of RhoA on T H 2 differentiation were evaluated based on in vitro T H 2-polarized culture conditions and in vivo in ovalbumin-induced allergic airway inflammation. Cytokine levels were measured by using intracellular staining and ELISA. T-cell metabolism was measured by using the Seahorse XF24 Analyzer and flow cytometry. Results Disruption of RhoA inhibited T-cell activation and T H 2 differentiation in vitro and prevented the development of allergic airway inflammation in vivo , with no effect on T H 1 cells. RhoA deficiency in activated T cells led to multiple defects in metabolic pathways, such as glycolysis and oxidative phosphorylation. Importantly, RhoA couples glycolysis to T H 2 cell differentiation and allergic airway inflammation through regulating IL-4 receptor mRNA expression and T H 2-specific signaling events. Finally, inhibition of Rho-associated protein kinase, an immediate downstream effector of RhoA, blocked T H 2 differentiation and allergic airway inflammation. Conclusion RhoA is a key component of the signaling cascades leading to T H 2 differentiation and allergic airway inflammation at least in part through control of T-cell metabolism and the Rho-associated protein kinase pathway.