Activation of the Unfolded Protein Response Contributes toward the Antitumor Activity of Vorinostat

Abstract

Histone deacetylase (HDAC) inhibitors represent an emerging class of anticancer agents progressing through clinical trials. Although their primary target is thought to involve acetylation of core histones, several nonhistone substrates have been identified, including heat shock protein (HSP) 90, which may contribute towards their antitumor activity. Glucose-regulated protein 78 (GRP78) is a member of the HSP family of molecular chaperones and plays a central role in regulating the unfolded protein response (UPR). Emerging data suggest that GRP78 is critical in cellular adaptation and survival associated with oncogenesis and may serve as a cancer-specific therapeutic target. On the basis of shared homology with HSP family proteins, we sought to determine whether GRP78 could serve as a molecular target of the HDAC inhibitor vorinostat. Vorinostat treatment led to GRP78 acetylation, dissociation, and subsequent activation of its client protein double-stranded RNA-activated protein-like endoplasmic reticulum kinase (PERK). Investigations in a panel of cancer cell lines identified that UPR activation after vorinostat exposure is specific to certain lines. Mass spectrometry performed on immunoprecipitated GRP78 identified lysine-585 as a specific vorinostat-induced acetylation site of GRP78. Downstream activation of the UPR was confirmed, including eukaryotic initiating factor 2alpha phosphorylation and increase in ATF4 and C/EBP homologous protein expression. To determine the biologic relevance of UPR activation after vorinostat, RNA interference of PERK was performed, demonstrating significantly decreased sensitivity to vorinostat-induced cytotoxicity. Collectively, these findings indicate that GRP78 is a biologic target of vorinostat, and activation of the UPR through PERK phosphorylation contributes toward its antitumor activity.

Identification and Characterization of a Novel Human Methyltransferase Modulating Hsp70 Protein Function through Lysine Methylation

“…4, E and F). Interestingly, previous studies have reported that peptides covering Lys-585 in human HSPA5 (corresponding to Lys-586 in mouse HSPA5) are acetylated (24,25). Notably, trimethylation and acetylation give very similar changes in molecular mass, ϩ42.0106 Da, and ϩ42.0470 Da, respectively, and cannot be distinguished by lowresolution MS.…”

Autophagy potentiates the anti-cancer effects of the histone deacetylase inhibitors in hepatocellular carcinoma

“…Upon ER stress, PERK, IRE1 and ATF6 are released from GRP78 and become activated. 51 A recent study indicates that SAHA induces acetylation of GRP78, leading to dissociation and activation of PERK, 52 providing a direct regulation of ER stress by HDAC inhibition.…”

Histone Deacetylases Promote ER Stress Induced Epithelial Mesenchymal Transition in Human Lung Epithelial Cells

“…In certain tumor cells, HDACs localized in ER affect the occurrence of ER stress [21-23]. In this study, we provided the first evidence that tunicamycin and bleomycin induced ER stress in alveolar epithelial cells via the upregulation of HDAC2/6, which mediate the formation of EMT.…”