TY - JOUR
T1 - Efavirenz and efavirenz-like compounds activate human, murine, and macaque hepatic IRE1a-XBP1
AU - Heck, Carley J.S.
AU - Hamlin, Allyson N.
AU - Bumpus, Namandjé N.
N1 - Funding Information:
This work was supported by the National Institutes of Health [Grant R01 GM103853 awarded to N.N.B. and Grant T32 GM008763 awarded to the Department of Pharmacology and Molecular Sciences at the Johns Hopkins School of Medicine], and a National Science Foundation Graduate Research Fellowship [DGE-1232825 awarded to C.J.S.H.]. https://doi.org/10.1124/mol.118.113647. s This article has supplemental material available at molpharm. aspetjournals.org.
Publisher Copyright:
Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.
PY - 2019/2
Y1 - 2019/2
N2 - Efavirenz (EFV), a widely used antiretroviral drug, is associated with idiosyncratic hepatotoxicity and dyslipidemia. Here we demonstrate that EFV stimulates the activation in primary hepatocytes of key cell stress regulators: inositol-requiring 1a (IRE1a) and X-box binding protein 1 (XBP1). Following EFV exposure, XBP1 splicing (indicating activation) was increased 35.7-fold in primary human hepatocytes. In parallel, XBP1 splicing and IRE1a phosphorylation (p-IRE1a, active IRE1a) were elevated 36.4-fold and 4.9-fold, respectively, in primary mouse hepatocytes. Of note, with EFV treatment, 47.2% of mouse hepatocytes were apoptotic; which was decreased to 23.9% in the presence of STF 083010, an inhibitor of XBP1 splicing. Experiments performed using pregnane X receptor (PXR)-null mouse hepatocytes revealed that EFV-mediated XBP1 splicing and hepatocyte death were not dependent on PXR, which is a nuclear receptor transcription factor that plays a crucial role in the cellular response to xenobiotics. Interestingly, incubation with the primary metabolite of EFV, 8-hydroxyefavirenz (8-OHEFV), only resulted in 10.3- and 2.9-fold increased XBP1 splicing in human and mouse hepatocytes and no change in levels of p-IRE1a in mouse hepatocytes. To further probe the structure-activity relationship of IRE1a-XBP1 activation by EFV, 16 EFV analogs were employed. Of these, an analog in which the EFV alkyne is replaced with an alkene and an analog in which the oxazinone oxygen is replaced by a carbon stimulated XBP1 splicing in human, mouse, and macaque hepatocytes. These data demonstrate that EFV and compounds sharing the EFV scaffold can activate IRE1a-XBP1 across human, mouse, and macaque species.
AB - Efavirenz (EFV), a widely used antiretroviral drug, is associated with idiosyncratic hepatotoxicity and dyslipidemia. Here we demonstrate that EFV stimulates the activation in primary hepatocytes of key cell stress regulators: inositol-requiring 1a (IRE1a) and X-box binding protein 1 (XBP1). Following EFV exposure, XBP1 splicing (indicating activation) was increased 35.7-fold in primary human hepatocytes. In parallel, XBP1 splicing and IRE1a phosphorylation (p-IRE1a, active IRE1a) were elevated 36.4-fold and 4.9-fold, respectively, in primary mouse hepatocytes. Of note, with EFV treatment, 47.2% of mouse hepatocytes were apoptotic; which was decreased to 23.9% in the presence of STF 083010, an inhibitor of XBP1 splicing. Experiments performed using pregnane X receptor (PXR)-null mouse hepatocytes revealed that EFV-mediated XBP1 splicing and hepatocyte death were not dependent on PXR, which is a nuclear receptor transcription factor that plays a crucial role in the cellular response to xenobiotics. Interestingly, incubation with the primary metabolite of EFV, 8-hydroxyefavirenz (8-OHEFV), only resulted in 10.3- and 2.9-fold increased XBP1 splicing in human and mouse hepatocytes and no change in levels of p-IRE1a in mouse hepatocytes. To further probe the structure-activity relationship of IRE1a-XBP1 activation by EFV, 16 EFV analogs were employed. Of these, an analog in which the EFV alkyne is replaced with an alkene and an analog in which the oxazinone oxygen is replaced by a carbon stimulated XBP1 splicing in human, mouse, and macaque hepatocytes. These data demonstrate that EFV and compounds sharing the EFV scaffold can activate IRE1a-XBP1 across human, mouse, and macaque species.
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U2 - 10.1124/mol.118.113647
DO - 10.1124/mol.118.113647
M3 - Article
C2 - 30442673
AN - SCOPUS:85059907440
SN - 0026-895X
VL - 95
SP - 185
EP - 195
JO - Molecular Pharmacology
JF - Molecular Pharmacology
IS - 2
ER -