Depleting interferon regulatory factor‐1(IRF‐1) with CRISPR/Cas9 attenuates inducible oxidative metabolism without affecting RA‐induced differentiation in HL‐60 human AML cells

Abstract

Abstract The known collaboration between all‐transretinoic acid and interferon motivates this study of the dependence of RA‐induced leukemic cell differentiation on interferon regulatory factor‐1 (IRF‐1), a transcription factor that is the main mediator of interferon effects. In the HL‐60 acute myeloid leukemia (AML) model that represents a rare RA‐responsive subtype of AML, IRF‐1 is not expressed until RA induces its prominent expression, and ectopic IRF‐1 expression enhances RA‐induced differentiation, motivating interest in how IRF‐1 is putatively needed for RA response. Accordingly, we created CRISPR/Cas9‐mediated IRF‐1 knockout HL‐60 cells. Contrary to expectation, loss of IRF‐1 did not diminish RA‐induced cellular signaling that propels differentiation, and RA‐induced cell differentiation markers, including CD38 and CD11b expression and G1/G0cell cycle arrest, were unaffected. However, elimination of IRF‐1 inhibited RA‐induced p47phox expression and inducible oxidative metabolism detected by reactive oxygen species (ROS), suggesting IRF‐1 is essential for mature granulocytic inducible oxidative metabolism. In the case of 1,25‐Dihydroxyvitamin D3‐induced differentiation to monocytes, IRF‐1 loss did not affect D3‐induced expression of CD38, CD11b, and CD14, and G1/0 arrest; but inhibited ROS production. Our data suggest that IRF‐1 is inessential for differentiation but upregulates p47phox expression for mature‐cell ROS production.

Keywords

• differentiation
• HL‐60 cells
• inducible oxidative metabolism
• interferon regulatory factor‐1(IRF‐1)
• retinoic acid