TRAIL Triggers CRAC-Dependent Calcium Influx and Apoptosis through the Recruitment of Autophagy Proteins to Death-Inducing Signaling Complex

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively kills various cancer cell types, but also leads to the activation of signaling pathways that favor resistance to cell death. Here, we investigated the as yet unknown roles of calcium signaling and autophagy regulatory proteins during TRAIL-induced cell death in leukemia cells. Taking advantage of the Gene Expression Profiling Interactive Analysis (GEPIA) project, we first found that leukemia patients present a unique TRAIL receptor gene expression pattern that may reflect their resistance to TRAIL. The exposure of NB4 acute promyelocytic leukemia cells to TRAIL induces intracellular Ca2+ influx through a calcium release-activated channel (CRAC)-dependent mechanism, leading to an anti-apoptotic response. Mechanistically, we showed that upon TRAIL treatment, two autophagy proteins, ATG7 and p62/SQSTM1, are recruited to the death-inducing signaling complex (DISC) and are essential for TRAIL-induced Ca2+ influx and cell death. Importantly, the treatment of NB4 cells with all-trans retinoic acid (ATRA) led to the upregulation of p62/SQSTM1 and caspase-8 and, when added prior to TRAIL stimulation, significantly enhanced DISC formation and the apoptosis induced by TRAIL. In addition to uncovering new pleiotropic roles for autophagy proteins in controlling the calcium response and apoptosis triggered by TRAIL, our results point to novel therapeutic strategies for sensitizing leukemia cells to TRAIL.

Digital spatial profiling of human parathyroid tumors reveals cellular and molecular alterations linked to vitamin D deficiency

“…Notably, expression of GABBR1 ( gamma aminobutyric acid type B receptor 1 ), which our group has recently shown to oppose CaSR-mediated calcium sensing in the parathyroid ( 34 ), is significantly elevated in Def-Ts. Additional genes found to be significantly upregulated in these tumors act in signaling pathways controlling apoptosis (TNFSF10, tumor necrosis superfamily member 10 ) ( 35 , 36 ), cellular stress response (SGK1, serum glucocorticoid regulated kinase 1 ) ( 37 ), inositol-derived second messenger production (IPMK, inositol polyphosphate multikinase ) ( 38 ), and energetic metabolism (AMPD3, adenosine monophosphate deaminase 3 ) ( 39 ). Despite the higher preoperative PTH observed in vitamin D-deficient PHPT patients, PTH transcript abundance was not significantly different between tissue sources ( P = 0.2862 by one-way ANOVA) or when comparing chief to oxyphil cells ( P = 0.7284 by unpaired t -test).…”

TNF-Related Apoptosis-Inducing Ligand: Non-Apoptotic Signalling

“…Interestingly, both TRAIL- and FasL-induced pro-metastatic pathways are associated with an early increase in intracellular Ca 2+ and tyrosine kinase signalling [ 114 , 415 , 416 ]. The use of isogenic stable cancer cells deficient for either DR4 or DR5 [ 114 ], demonstrated that TRAIL-induced pro-metastatic signalling was solely triggered by DR5 and correlated with a rapid Ca 2+ flux [ 114 , 417 , 418 ]. Furthermore, early increased cytosolic Ca 2+ was shown to be activated upon TRAIL exposure in both Jurkat and NB4 leukemia cells, protecting the latter from apoptosis [ 418 ].…”

“…The use of isogenic stable cancer cells deficient for either DR4 or DR5 [ 114 ], demonstrated that TRAIL-induced pro-metastatic signalling was solely triggered by DR5 and correlated with a rapid Ca 2+ flux [ 114 , 417 , 418 ]. Furthermore, early increased cytosolic Ca 2+ was shown to be activated upon TRAIL exposure in both Jurkat and NB4 leukemia cells, protecting the latter from apoptosis [ 418 ]. It was found in these cells that recruitment of both p62 and ATG7 to complex I was required for calcium influx induced by TRAIL [ 418 ].…”

“…Furthermore, early increased cytosolic Ca 2+ was shown to be activated upon TRAIL exposure in both Jurkat and NB4 leukemia cells, protecting the latter from apoptosis [ 418 ]. It was found in these cells that recruitment of both p62 and ATG7 to complex I was required for calcium influx induced by TRAIL [ 418 ].…”

“…Like TRAIL, FasL also induces an increase in cytosolic Ca 2+ , associated with cell-motility and metastasis [ 386 , 415 , 419 , 420 , 421 ]. Intracellular increase in Ca 2+ is generally induced by PLCγ1 and IP3R activation, due to ER Ca 2+ release [ 422 ], but may also be triggered, as demonstrated in leukemia cells, after ORAI1 activation and CRAC channels opening [ 418 ]. Autophagy Related 7 (ATG7) [ 423 ] and Sequestosome 1 (p62/SQSTM1) [ 424 ], are two autophagic proteins related to ORAI1 and CRAC channels, whose recruitment to DR5 induce the release of Ca 2+ from the ER [ 418 ].…”