Extracellular and ER-stored Ca2+ contribute to BIRD-2-induced cell death in diffuse large B-cell lymphoma cells. Issue 1 (December 2018)

• Record Type: Journal Article
• Authors: Bittremieux, Mart; La Rovere, Rita; Schuermans, Marleen; Luyten, Tomas; Mikoshiba, Katsuhiko; Vangheluwe, Peter; Parys, Jan; Bultynck, Geert
• Is Part Of: Cell death discovery. Volume 4:Issue 1(2018)
• Page Start: 1
• Page End: 16
• Publication Date: 2018-12
• Subjects: Cell death -- Periodicals; 571.936
• DOI: 10.1038/s41420-018-0118-6
• Languages: English
• ISSNs: 2058-7716

Abstract

Abstract The anti-apoptotic protein Bcl-2 is upregulated in several cancers, including diffuse large B-cell lymphoma (DLBCL) and chronic lymphocytic leukemia (CLL). In a subset of these cancer cells, Bcl-2 blocks Ca2+ -mediated apoptosis by suppressing the function of inositol 1, 4, 5-trisphosphate (IP3 ) receptors (IP3 Rs) located at the endoplasmic reticulum (ER). A peptide tool, called Bcl-2/IP3 receptor disruptor-2 (BIRD-2), was developed to disrupt Bcl-2/IP3 R complexes, triggering pro-apoptotic Ca2+ signals and killing Bcl-2-dependent cancer cells. In DLBCL cells, BIRD-2 sensitivity depended on the expression level of IP3 R2 channels and constitutive IP3 signaling downstream of the B-cell receptor. However, other cellular pathways probably also contribute to BIRD-2-provoked cell death. Here, we examined whether BIRD-2-induced apoptosis depended on extracellular Ca2+ and more particularly on store-operated Ca2+ entry (SOCE), a Ca2+ -influx pathway activated upon ER-store depletion. Excitingly, DPB162-AE, a SOCE inhibitor, suppressed BIRD-2-induced cell death in DLBCL cells. However, DPB162-AE not only inhibits SOCE but also depletes the ER Ca2+ store. Treatment of the cells with YM-58483 and GSK-7975A, two selective SOCE inhibitors, did not protect against BIRD-2-induced apoptosis. Similar data were obtained by knocking down STIM1 using small interfering RNA. Yet, extracellular Ca2+ contributed to BIRD-2 sensitivity in DLBCL, since the extracellular Ca2+ buffer ethylene glycol tetraacetic acid (EGTA) blunted BIRD-2-triggered apoptosis. The protective effects observed with DPB162-AE are likely due to ER Ca2+ -store depletion, since a similar protective effect could be obtained using the sarco/endoplasmic reticulum Ca2+ -ATPase inhibitor thapsigargin. Thus, both the ER Ca2+ -store content and extracellular Ca2+, but not SOCE, are critical factors underlying BIRD-2-provoked cell death.