Inhibition of phosphoinositide-3 kinases / ameliorates pulmonary granuloma by rescuing Treg function in a sarcoidosis model

Sarcoidosis is a multisystem inflammatory disease characterized by the development of Th1/Th17/regulatory T cells (Tregs)-related non-caseating granulomas. Phosphoinositide-3 kinases delta/gamma (PI3K delta/gamma) play an important role in the maintenance of effective immunity, especially for Tregs homeostasis and stability. In the present study, superoxide dismutase A (SodA) stimulation was used to establish the sarcoidosis mouse model. The second immune stimulus was accompanied by CAL-101 (PI3K delta inhibitor) or AS-605240 (PI3K delta/gamma inhibitor) treatment. To detect the effect of the PI3K delta/gamma inhibitor on the morphology of pulmonary granuloma and the activation of the PI3K signaling pathway, hematoxylin and eosin staining and immunofluorescence and western blotting was used, respectively. Fluorescence-activated cell sorting analysis and reverse transcription-quantitative PCR were adopted to detect the effect of the PI3K delta/gamma inhibitor on the SodA-induced sarcoidosis mouse model in respect to immune cell disorder and the function of Treg cells, with CD4(+)CD25(-) T cells and CD4(+)CD25(+) T cells sorted by magnetic cell sorting. The results demonstrated that the inhibition of PI3K delta/gamma by transtracheal CAL-101/AS-605240 administration facilitated pulmonary granuloma formation. These therapeutic effects were associated with certain mechanisms, including suppressing the aberrantly activated PI3K/Akt signaling in both pulmonary granuloma and Tregs, particularly rescuing the suppressive function of Tregs. Notably, CAL-101 was more effective in immune modulation compared with AS-605240 and could overcome the aberrantly activated Akt in the lung and Tregs. These results suggest that PI3K/Akt signaling, especially the PI3K delta subunit, can play a key role in optimal Tregs-mediated protection against pulmonary sarcoidosis. Therefore, transtracheal usage of PI3K delta/gamma inhibitors is an attractive therapy that may be developed into a new immune-therapeutic principle for sarcoidosis in the future.