Inhibition of PIM1 blocks the autophagic flux to sensitize glioblastoma cells to ABT-737-induced apoptosis

• Published in: Biochimica et biophysica acta. Molecular cell research Vol. 1866; no. 2; pp. 175 - 189
• Main Authors: Remy, J., Linder, B., Weirauch, U., Konovalova, J., Marschalek, R., Aigner, A., Kögel, D.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2019
• Subjects: Apoptosis; Autophagy; BH3-mimetics; Brain tumors; PIM1; GSK3B; zVAD; CASP8 inhibitor; ACTB; CASP9; CHX; MAP1LC3B; CASP8; CASP3; CASP4; PIM1; TBP; IM; BH3-mimetics; EL; AZD; Ctrl; DDIT3; AKT1S1; BCL2-like 1; VDAC1; CASP9 inhibitor; GAPDH; SQSTM1; Brain tumors; XIAP; FCCP; Baf A1; L; Autophagy; ABT; BCL2L11; TMRM; KD; PMAIP1; GB; MCL1; HSPA5; KO; AKT; CQ; MTOR; ptf-LC3; TIC; SGI; CYCS; PI; BCL2L2; Apoptosis
• ISSN: 0167-4889; 1879-2596
• DOI: 10.1016/j.bbamcr.2018.10.017

Overcoming apoptosis resistance is one major issue in glioblastoma (GB) therapies. Accumulating evidence indicates that resistance to apoptosis in GB is mediated via upregulation of pro-survival BCL2-family members. The synthetic BH3-mimetic ABT-737 effectively targets BCL2, BCL2 like 1 and BCL2 like 2 but still barely affects cell survival which is presumably due to its inability to inhibit myeloid cell leukemia 1 (MCL1). The constitutively active serine/threonine kinase proviral integration site for moloney murine leukemia virus 1 (PIM1) was recently found to be overexpressed in GB patient samples and to maintain cell survival in these tumors. For different GB cell lines, Western Blot, mitochondrial fractionation, fluorescence microscopy, effector caspase assays, flow cytometry, and an adult organotypic brain slice transplantation model were used to investigate the putative PIM1/MCL1 signaling axis regarding potential synergistic effects with ABT-737. We demonstrate that combination of the PIM1 inhibitor SGI-1776 or the pan-PIM kinase inhibitor AZD1208 with ABT-737 strongly sensitizes GB cells to apoptosis. Unexpectedly, this effect was found to be MCL1-independent, but could be partially blocked by caspase 8 (CASP8) inhibition. Remarkably, the analysis of autophagy markers in combination with the observation of massive accumulation and hampered degradation of autophagosomes suggests a completely novel function of PIM1 as a late stage autophagy regulator, maintaining the autophagic flux at the level of autophagosome/lysosome fusion. Our data indicate that PIM1 inhibition and ABT-737 synergistically induce apoptosis in an MCL1-independent but CASP8-dependent manner in GB. They also identify PIM1 as a suitable target for overcoming apoptosis resistance in GB. •PIM1 inhibition sensitizes glioblastoma cells to ABT-737-induced apoptosis.•Synergistic apoptosis occurs in an MCL1-independent, but caspase 8-dependent manner.•Inhibition of PIM1 triggers accumulation and hampered degradation of autophagosomes.•Autophagosomes serve as a platform for caspase 8 recruitment and activation.•Our results indicate a novel function of PIM1 as a late stage autophagy regulator.