Constitutive IP₃ signaling underlies the sensitivity of B-cell cancers to the Bcl-2/IP₃ receptor disruptor BIRD-2.

ABSTRACT: Anti-apoptotic Bcl-2 proteins are upregulated in different cancers, including diffuse large B-cell lymphoma (DLBCL) and chronic lymphocytic leukemia (CLL), enabling survival by inhibiting pro-apoptotic Bcl-2-family members and inositol 1,4,5-trisphosphate (IP₃) receptor (IP₃R)-mediated Ca²⁺-signaling. A peptide tool (Bcl-2/IP₃R Disruptor-2; BIRD-2) was developed to abrogate the interaction of Bcl-2 with IP₃Rs by targeting Bcl-2's BH4 domain. BIRD-2 triggers cell death in primary CLL cells and in DLBCL cell lines. Particularly, DLBCL cells with high levels of IP₃R2 were sensitive to BIRD-2. Here, we report that BIRD-2-induced cell death in DLBCL cells does not only depend on high IP₃R2-expression levels, but also on constitutive IP₃ signaling, downstream of the tonically active B-cell receptor. The basal Ca²⁺ level in SU-DHL-4 DLBCL cells was significantly elevated due to the constitutive IP₃ production. This constitutive IP₃ signaling fulfilled a pro-survival role, since inhibition of phospholipase C (PLC) using U73122 (2.5?µM) caused cell death in SU-DHL-4 cells. Milder inhibition of IP₃ signaling using a lower U73122 concentration (1?µM) or expression of an IP₃ sponge suppressed both BIRD-2-induced Ca²⁺ elevation and apoptosis in SU-DHL-4 cells. Basal PLC/IP₃ signaling also fulfilled a pro-survival role in other DLBCL cell lines, including Karpas 422, RI-1 and SU-DHL-6 cells, whereas PLC inhibition protected these cells against BIRD-2-evoked apoptosis. Finally, U73122 treatment also suppressed BIRD-2-induced cell death in primary CLL, both in unsupported systems and in co-cultures with CD40L-expressing fibroblasts. Thus, constitutive IP₃ signaling in lymphoma and leukemia cells is not only important for cancer cell survival, but also represents a vulnerability, rendering cancer cells dependent on Bcl-2 to limit IP₃R activity. BIRD-2 seems to switch constitutive IP₃ signaling from pro-survival into pro-death, presenting a plausible therapeutic strategy.

SUBMITTER: Bittremieux M 

PROVIDER: S-EPMC6370760 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature