Phosphatase PP2A enhances MCL-1 protein half-life in multiple myeloma cells.
Cell Line, Tumor
Gene Expression Regulation, Neoplastic
Half-Life
Humans
JNK Mitogen-Activated Protein Kinases
/ metabolism
Lymphoma, Large B-Cell, Diffuse
/ enzymology
Multiple Myeloma
/ enzymology
Myeloid Cell Leukemia Sequence 1 Protein
/ genetics
Phosphorylation
Protein Phosphatase 2
/ genetics
Protein Processing, Post-Translational
Protein Stability
Proteolysis
Journal
Cell death & disease
ISSN: 2041-4889
Titre abrégé: Cell Death Dis
Pays: England
ID NLM: 101524092
Informations de publication
Date de publication:
03 03 2021
03 03 2021
Historique:
received:
18
05
2020
accepted:
14
12
2020
revised:
10
12
2020
entrez:
4
3
2021
pubmed:
5
3
2021
medline:
15
9
2021
Statut:
epublish
Résumé
Multiple myeloma (MM), a treatable but incurable malignancy, is characterized by the growth of clonal plasma cells in protective niches in the bone marrow. MM cells depend on expression of BCL-2 family proteins, in particular MCL-1, for survival. The regulation of MCL-1 is complex and cell type-dependent. Unraveling the exact mechanism by which MCL-1 is overexpressed in MM may provide new therapeutic strategies for inhibition in malignant cells, preferably limiting side effects in healthy cells. In this study, we reveal that one cause of overexpression could be stabilization of the MCL-1 protein. We demonstrate this in a subset of MM and diffuse large B cell lymphoma (DLBCL) cell lines and MM patient samples. We applied a phosphatase siRNA screen to identify phosphatases responsible for MCL-1 stabilization in MM, and revealed PP2A as the MCL-1 stabilizing phosphatase. Using the PP2A inhibitor okadaic acid, we validated that PP2A dephosphorylates MCL-1 at Ser159 and/or Thr163, and thereby stabilizes MCL-1 in MM cells with long MCL-1 half-life, but not in DLBCL cells. Combined kinase and phosphatase inhibition experiments suggest that the MCL-1 half-life in MM is regulated by the counteracting functions of JNK and PP2A. These findings increase the understanding of the mechanisms by which MCL-1 is post-translationally regulated, which may provide novel strategies to inhibit MCL-1 in MM cells.
Identifiants
pubmed: 33658484
doi: 10.1038/s41419-020-03351-7
pii: 10.1038/s41419-020-03351-7
pmc: PMC7930201
doi:
Substances chimiques
MCL1 protein, human
0
Myeloid Cell Leukemia Sequence 1 Protein
0
JNK Mitogen-Activated Protein Kinases
EC 2.7.11.24
Protein Phosphatase 2
EC 3.1.3.16
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
229Références
Adams, J. M. & Cory, S. The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26, 1324–1337 (2007).
pubmed: 17322918
pmcid: 2930981
doi: 10.1038/sj.onc.1210220
Slomp, A. & Peperzak, V. Role and regulation of pro-survival BCL-2 proteins in multiple myeloma. Front. Oncol. 8, 533 (2018).
pubmed: 30524962
pmcid: 6256118
doi: 10.3389/fonc.2018.00533
Zhang, B., Gojo, I. & Fenton, R. G. Myeloid cell factor-1 is a critical survival factor for multiple myeloma. Blood 99, 1885–1893 (2002).
pubmed: 11877256
doi: 10.1182/blood.V99.6.1885
Derenne, S. et al. Antisense strategy shows that Mcl-1 rather than Bcl-2 or Bcl-x L is an essential survival protein of human myeloma cells. Blood 100, 194–199 (2002).
pubmed: 12070027
doi: 10.1182/blood.V100.1.194
Tiedemann, R. E. et al. Identification of molecular vulnerabilities in human multiple myeloma cells by RNA interference lethality screening of the druggable genome. Cancer Res. 72, 757–768 (2012).
pubmed: 22147262
doi: 10.1158/0008-5472.CAN-11-2781
Wuillème-Toumi, S. et al. Mcl-1 is overexpressed in multiple myeloma and associated with relapse and shorter survival. Leukemia 19, 1248–1252 (2005).
pubmed: 15902294
doi: 10.1038/sj.leu.2403784
Kotschy, A. et al. The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models. Nature 538, 477–482 (2016).
pubmed: 27760111
doi: 10.1038/nature19830
Caenepeel, S. et al. AMG 176, a selective MCL1 inhibitor is effective in hematological cancer models alone and in combination with established therapies. Cancer Discov. 8, 1582–1597 (2018).
pubmed: 30254093
doi: 10.1158/2159-8290.CD-18-0387
Tron, A. E. et al. Discovery of Mcl-1-specific inhibitor AZD5991 and preclinical activity in multiple myeloma and acute myeloid leukemia. Nat. Commun. 9, 5341 (2018).
pubmed: 30559424
pmcid: 6297231
doi: 10.1038/s41467-018-07551-w
Peperzak, V. et al. Mcl-1 is essential for the survival of plasma cells. Nat. Immunol. 14, 290–297 (2013).
pubmed: 23377201
pmcid: 4041127
doi: 10.1038/ni.2527
Vikström, I. et al. Mcl-1 is essential for germinal center formation and B cell memory. Science 330, 1095–1100 (2010).
pubmed: 20929728
pmcid: 2991396
doi: 10.1126/science.1191793
Vikström, I. et al. MCL-1 is required throughout B-cell development and its loss sensitizes specific B-cell subsets to inhibition of BCL-2 or BCL-XL. Cell Death Dis. 7, e2345 (2016).
pubmed: 27560714
pmcid: 5108322
doi: 10.1038/cddis.2016.237
Opferman, J. T. et al. Obligate role of anti-apoptotic MCL-1 in the survival of hematopoietic stem cells. Science 307, 1101–1105 (2005).
pubmed: 15718471
doi: 10.1126/science.1106114
Thomas, R. L. et al. Loss of MCL-1 leads to impaired autophagy and rapid development of heart failure. Genes Dev. 27, 1365–1377 (2013).
pubmed: 23788623
pmcid: 3701192
doi: 10.1101/gad.215871.113
Malone, C. D. et al. Mcl-1 regulates the survival of adult neural precursor cells. Mol. Cell. Neurosci. 49, 439–447 (2012).
pubmed: 22357134
doi: 10.1016/j.mcn.2012.02.003
Kozopas, K. M., Yang, T., Buchan, H. L., Zhou, P. & Craig, R. W. MCL1, a gene expressed in programmed myeloid cell differentiation, has sequence similarity to BCL2. Proc. Natl Acad. Sci. USA 90, 3516–3520 (1993).
pubmed: 7682708
doi: 10.1073/pnas.90.8.3516
pmcid: 46331
Yang, T., Kozopas, K. M. & Craig, R. W. The intracellular distribution and pattern of expression of Mcl-1 overlap with, but are not identical to, those of Bcl-2. J. Cell Biol. 128, 1173–1184 (1995).
pubmed: 7896880
doi: 10.1083/jcb.128.6.1173
Nijhawan, D. et al. Elimination of Mcl-1 is required for the initiation of apoptosis following ultraviolet irradiation. Genes Dev. 17, 1475–1486 (2003).
pubmed: 12783855
pmcid: 196078
doi: 10.1101/gad.1093903
Maurer, U., Charvet, C., Wagman, A. S., Dejardin, E. & Green, D. R. Glycogen synthase kinase-3 regulates mitochondrial outer membrane permeabilization and apoptosis by destabilization of MCL-1. Mol. Cell 21, 749–760 (2006).
pubmed: 16543145
doi: 10.1016/j.molcel.2006.02.009
Thomas, L. W., Lam, C. & Edwards, S. W. Mcl-1; the molecular regulation of protein function. FEBS Lett. 584, 2981–2989 (2010).
pubmed: 20540941
doi: 10.1016/j.febslet.2010.05.061
Mojsa, B., Lassot, I. & Desagher, S. Mcl-1 ubiquitination: unique regulation of an essential survival protein. Cells 3, 418–437 (2014).
pubmed: 24814761
pmcid: 4092850
doi: 10.3390/cells3020418
Ding, Q. et al. Down-regulation of myeloid cell leukemia-1 through inhibiting Erk/Pin 1 pathway by sorafenib facilitates chemosensitization in breast cancer. Cancer Res. 68, 6109–6118 (2008).
pubmed: 18676833
pmcid: 2676572
doi: 10.1158/0008-5472.CAN-08-0579
Inoshita, S. et al. Phosphorylation and inactivation of myeloid cell leukemia 1 by JNK in response to oxidative stress. J. Biol. Chem. 277, 43730–43734 (2002).
pubmed: 12223490
doi: 10.1074/jbc.M207951200
Kodama, Y. et al. Antiapoptotic effect of c-Jun N-terminal Kinase-1 through Mcl-1 stabilisation in TNF-induced hepatocyte apoptosis. Gastroenterology 136, 1423–1434 (2009).
pubmed: 19249395
doi: 10.1053/j.gastro.2008.12.064
Ding, Q. et al. Degradation of Mcl-1 by b-TrCP mediates glycogen synthase kinase 3-induced tumor suppression and chemosensitization. Mol. Cell. Biol. 27, 4006–4017 (2007).
pubmed: 17387146
doi: 10.1128/MCB.00620-06
Zhong, Q., Gao, W., Du, F. & Wang, X. Mule/ARF-BP1, a BH3-only E3 ubiquitin ligase, catalyzes the polyubiquitination of Mcl-1 and regulates apoptosis. Cell 121, 1085–1095 (2005).
pubmed: 15989957
doi: 10.1016/j.cell.2005.06.009
Inuzuka, H. et al. SCFFBW7 regulates cellular apoptosis by targeting MCL1 for ubiquitylation and destruction. Nature 471, 104–109 (2011).
pubmed: 21368833
pmcid: 3076007
doi: 10.1038/nature09732
Harley, M. E., Allan, L. A., Sanderson, H. S. & Clarke, P. R. Phosphorylation of Mcl-1 by CDK–cyclin B1 initiates its Cdc20-dependent destruction during mitotic arrest. EMBO J. 29, 2407–2420 (2010).
pubmed: 20526282
pmcid: 2910263
doi: 10.1038/emboj.2010.112
Magiera, M. M. et al. Trim17-mediated ubiquitination and degradation of Mcl-1 initiate apoptosis in neurons. Cell Death Differ. 20, 281–292 (2013).
pubmed: 22976837
doi: 10.1038/cdd.2012.124
Schwickart, M. et al. Deubiquitinase USP9X stabilizes MCL1 and promotes tumour cell survival. Nature 463, 103–107 (2010).
pubmed: 20023629
doi: 10.1038/nature08646
Wang, B. et al. Role of Ku70 in deubiquitination of Mcl-1 and suppression of apoptosis. Cell Death Differ. 21, 1160–1169 (2014).
pubmed: 24769731
pmcid: 4207484
doi: 10.1038/cdd.2014.42
Zhang, S. et al. Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors. Nat. Commun. 9, 1–12 (2018).
Khalil, A. M., Cambier, J. C. & Shlomchik, M. J. B cell receptor signal transduction in the GC is short-circuited by high phosphatase activity. Science 336, 1178–1182 (2012).
pubmed: 22555432
pmcid: 3777391
doi: 10.1126/science.1213368
Peperzak, V., Slinger, E., Ter Burg, J. & Eldering, E. Functional disparities among BCL-2 members in tonsillar and leukemic B-cell subsets assessed by BH3-mimetic profiling. Cell Death Differ. 24, 111–119 (2017).
pubmed: 27689871
doi: 10.1038/cdd.2016.105
Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25, 402–408 (2001).
pubmed: 11846609
doi: 10.1006/meth.2001.1262
Hornbeck, P. V. et al. PhosphoSitePlus, 2014: mutations, PTMs and recalibrations. Nucleic Acids Res. 43, D512–D520 (2015).
pubmed: 25514926
doi: 10.1093/nar/gku1267
Kong, M., Ditsworth, D., Lindsten, T. & Thompson, C. B. α4 is an essential regulator of PP2A phosphatase activity. Mol. Cell 36, 51–60 (2009).
pubmed: 19818709
pmcid: 2761955
doi: 10.1016/j.molcel.2009.09.025
Virshup, D. M. & Shenolikar, S. From promiscuity to precision: protein phosphatases get a makeover. Mol. Cell 33, 537–545 (2009).
doi: 10.1016/j.molcel.2009.02.015
pubmed: 19285938
Huang, T. et al. CDK7 inhibitor THZ1 inhibits MCL1 synthesis and drives cholangiocarcinoma apoptosis in combination with BCL2/BCL-XL inhibitor ABT-263. Cell Death Dis. 10, 602 (2019).
pubmed: 31399555
pmcid: 6688996
doi: 10.1038/s41419-019-1831-7
Hanamura, I. et al. Frequent gain of chromosome band 1q21 in plasma-cell dyscrasias detected by fluorescence in situ hybridization: incidence increases from MGUS to relapsed myeloma and is related to prognosis and disease progression following tandem stem-cell transplantatio. Blood 108, 1724–1733 (2006).
pubmed: 16705089
pmcid: 1895503
doi: 10.1182/blood-2006-03-009910
Gupta, V. A. et al. Bone marrow microenvironment–derived signals induce Mcl-1 dependence in multiple myeloma. Blood 129, 1969–1979 (2017).
pubmed: 28151428
pmcid: 5383873
doi: 10.1182/blood-2016-10-745059
Slomp, A. et al. Multiple myeloma with amplification of chromosome 1q is highly sensitive to MCL-1 targeting. Blood Adv. 3, 4202–4214 (2019).
pubmed: 31856269
pmcid: 6929383
doi: 10.1182/bloodadvances.2019000702
Cui, J. & Placzek, W. J. Post-transcriptional regulation of anti-apoptotic BCL2 family members. Int. J. Mol. Sci. 19, 308 (2018).
pmcid: 5796252
doi: 10.3390/ijms19010308
Senichkin, V. V., Streletskaia, A. Y., Gorbunova, A. S., Zhivotovsky, B. & Kopeina, G. S. Saga of Mcl-1: regulation from transcription to degradation. Cell Death Differ. 27, 405–419 (2020).
pubmed: 31907390
pmcid: 7206148
doi: 10.1038/s41418-019-0486-3
Morel, C., Carlson, S. M., White, F. M. & Davis, R. J. Mcl-1 integrates the opposing actions of signaling pathways that mediate survival and apoptosis. Mol. Cell. Biol. 29, 3845–3852 (2009).
pubmed: 19433446
pmcid: 2704749
doi: 10.1128/MCB.00279-09
Nifoussi, S. K. et al. Inhibition of protein phosphatase 2A (PP2A) prevents Mcl-1 protein dephosphorylation at the Thr-163/Ser-159 phosphodegron, dramatically reducing expression in Mcl-1-amplified lymphoma cells. J. Biol. Chem. 289, 21950–21959 (2014).
pubmed: 24939844
pmcid: 4139212
doi: 10.1074/jbc.M114.587873
Manning, G., Whyte, D. B., Martinez, R., Hunter, T. & Sudarsanam, S. The protein kinase complement of the human genome. Science 298, 1912–1934 (2002).
pubmed: 12471243
doi: 10.1126/science.1075762
Arena, S., Benvenuti, S. & Bardelli, A. Genetic analysis of the kinome and phosphatome in cancer. Cell. Mol. Life Sci. 62, 2092–2099 (2005).
pubmed: 16132230
doi: 10.1007/s00018-005-5205-1
Sacco, F., Perfetto, L., Castagnoli, L. & Cesareni, G. The human phosphatase interactome: an intricate family portrait. FEBS Lett. 586, 2732–2739 (2012).
pubmed: 22626554
pmcid: 3437441
doi: 10.1016/j.febslet.2012.05.008
Rowland, M. A., Harrison, B. & Deeds, E. J. Phosphatase specificity and pathway insulation in signaling networks. Biophys. J. 108, 986–996 (2015).
pubmed: 25692603
pmcid: 4336360
doi: 10.1016/j.bpj.2014.12.011
Eleftheriadou, O. et al. Expression and regulation of type 2A protein phosphatases and alpha4 signalling in cardiac health and hypertrophy. Basic Res. Cardiol. 112, 37 (2017).
pubmed: 28526910
pmcid: 5438423
doi: 10.1007/s00395-017-0625-2
Kong, M. et al. The PP2A-associated protein α4 is an essential inhibitor of apoptosis. Science 306, 695–698 (2004).
pubmed: 15499020
doi: 10.1126/science.1100537
Seshacharyulu, P., Pandey, P., Datta, K. & Batra, S. K. Phosphatase: PP2A structural importance, regulation and its aberrant expression in cancer. Cancer Lett. 335, 9–18 (2013).
pubmed: 23454242
pmcid: 3665613
doi: 10.1016/j.canlet.2013.02.036
O’Connor, C. M. et al. Inactivation of PP2A by a recurrent mutation drives resistance to MEK inhibitors. Oncogene 39, 703–717 (2020).
doi: 10.1038/s41388-019-1012-2
pubmed: 31541192
Sangodkar, J. et al. All roads lead to PP2A: exploiting the therapeutic potential of this phosphatase. FEBS J. 283, 1004–1024 (2016).
pubmed: 26507691
doi: 10.1111/febs.13573
Eichhorn, P. J. A., Creyghton, M. P. & Bernards, R. Protein phosphatase 2A regulatory subunits and cancer. Biochim. Biophys. Acta 1795, 1–15 (2009).
pubmed: 18588945
Hong, C. S. et al. LB100, a small molecule inhibitor of PP2A with potent chemo- and radio-sensitizing potential. Cancer Biol. Ther. 16, 821–833 (2015).
pubmed: 25897893
pmcid: 4623051
doi: 10.1080/15384047.2015.1040961
De Munter, S., Köhn, M. & Bollen, M. Challenges and opportunities in the development of protein phosphatase-directed therapeutics. ACS Chem. Biol. 8, 36–45 (2013).
pubmed: 23214403
doi: 10.1021/cb300597g
Chung, V. et al. Safety, tolerability, and preliminary activity of LB-100, an inhibitor of protein phosphatase 2A, in patients with relapsed solid tumors: an open-label, dose escalation, first-in-human, phase I trial. Clin. Cancer Res. 23, 3277–3284 (2017).
pubmed: 28039265
doi: 10.1158/1078-0432.CCR-16-2299
Lai, D. et al. PP2A inhibition sensitizes cancer stem cells to ABL tyrosine kinase inhibitors in BCR-ABFL+ human leukemia. Sci. Transl. Med. 10, eaan8735 (2018).
pubmed: 29437150
doi: 10.1126/scitranslmed.aan8735