Mcl-1 Degradation Is Required for Targeted Therapeutics to Eradicate Colon Cancer Cells.

The Bcl-2 family protein Mcl-1 is often degraded in cancer cells subjected to effective therapeutic treatment, and defective Mcl-1 degradation has been associated with intrinsic and acquired drug resistance. However, a causal relationship between Mcl-1 degradation and anticancer drug responses has not been directly established, especially in solid tumor cells where Mcl-1 inhibition alone is insufficient to trigger cell death. In this study, we present evidence that Mcl-1 participates directly in determining effective therapeutic responses in colon cancer cells. In this setting, Mcl-1 degradation was induced by a variety of multikinase inhibitor drugs, where it relied upon GSK3β phosphorylation and FBW7-dependent ubiquitination. Specific blockade by genetic knock-in (KI) abolished apoptotic responses and conferred resistance to kinase inhibitors. Mcl-1-KI also suppressed the antiangiogenic and anti-hypoxic effects of kinase inhibitors in the tumor microenvironment. Interestingly, these same inhibitors also induced the BH3-only Bcl-2 family protein PUMA, which is required for apoptosis. Degradation-resistant Mcl-1 bound and sequestered PUMA from other prosurvival proteins to maintain cell survival, which was abolished by small-molecule Mcl-1 inhibitors. Our findings establish a pivotal role for Mcl-1 degradation in the response of colon cancer cells to targeted therapeutics, and they provide a useful rational platform to develop Mcl-1-targeting agents that can overcome drug resistance. Cancer Res; 77(9); 2512-21. ©2017 AACR.

[1]  A. Strasser,et al.  The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models , 2016, Nature.

[2]  Y. Fernández-Marrero,et al.  Survival control of malignant lymphocytes by anti-apoptotic MCL-1 , 2016, Leukemia.

[3]  Jian Yu,et al.  FBW7 mutations mediate resistance of colorectal cancer to targeted therapies by blocking Mcl-1 degradation , 2016, Oncogene.

[4]  A. Venditti,et al.  Venetoclax: Bcl-2 inhibition for the treatment of chronic lymphocytic leukemia. , 2016, Drugs of today.

[5]  A. Letai,et al.  Mitochondria-Judges and Executioners of Cell Death Sentences. , 2016, Molecular cell.

[6]  E. Olejniczak,et al.  Discovery of 2-Indole-acylsulfonamide Myeloid Cell Leukemia 1 (Mcl-1) Inhibitors Using Fragment-Based Methods. , 2016, Journal of medicinal chemistry.

[7]  K. Ravichandran,et al.  Do not let death do us part: ‘find-me’ signals in communication between dying cells and the phagocytes , 2016, Cell Death and Differentiation.

[8]  Jian Yu,et al.  Abstract 667: Regorafenib inhibits colorectal tumor growth through PUMA-mediated apoptosis , 2015 .

[9]  Scott A. Erickson,et al.  Structure-guided design of a series of MCL-1 inhibitors with high affinity and selectivity. , 2015, Journal of medicinal chemistry.

[10]  S. Desagher,et al.  Mcl-1 Ubiquitination: Unique Regulation of an Essential Survival Protein , 2014, Cells.

[11]  T. Cierpicki,et al.  3-Substituted-N-(4-Hydroxynaphthalen-1-yl)arylsulfonamides as a Novel Class of Selective Mcl-1 Inhibitors: Structure-Based Design, Synthesis, SAR, and Biological Evaluation , 2014, Journal of medicinal chemistry.

[12]  Richard W. Kriwacki,et al.  Many players in BCL-2 family affairs. , 2014, Trends in biochemical sciences.

[13]  K. Bencardino,et al.  Regorafenib in metastatic colorectal cancer , 2014, Expert review of anticancer therapy.

[14]  Tao Zhang,et al.  A Novel Small-Molecule Inhibitor of Mcl-1 Blocks Pancreatic Cancer Growth In Vitro and In Vivo , 2013, Molecular Cancer Therapeutics.

[15]  C. Billard.  BH3 Mimetics: Status of the Field and New Developments , 2013, Molecular Cancer Therapeutics.

[16]  Yeul-Hong Kim,et al.  Regorafenib for metastatic colorectal cancer , 2013, The Lancet.

[17]  G. Brix,et al.  Regorafenib Inhibits Growth, Angiogenesis, and Metastasis in a Highly Aggressive, Orthotopic Colon Cancer Model , 2013, Molecular Cancer Therapeutics.

[18]  Jian Yu,et al.  Targeting Bax interaction sites reveals that only homo-oligomerization sites are essential for its activation , 2013, Cell Death and Differentiation.

[19]  D. Sargent,et al.  Regorafenib monotherapy for previously treated metastatic colorectal cancer (CORRECT): an international, multicentre, randomised, placebo-controlled, phase 3 trial , 2013, The Lancet.

[20]  Brian A. Chauder,et al.  Discovery of potent myeloid cell leukemia 1 (Mcl-1) inhibitors using fragment-based methods and structure-based design. , 2013, Journal of medicinal chemistry.

[21]  J. Opferman,et al.  A competitive stapled peptide screen identifies a selective small molecule that overcomes MCL-1-dependent leukemia cell survival. , 2012, Chemistry & biology.

[22]  Jian Yu,et al.  The Multi-Targeted Kinase Inhibitor Sunitinib Induces Apoptosis in Colon Cancer Cells via PUMA , 2012, PloS one.

[23]  Jian Yu,et al.  Inhibiting oncogenic signaling by sorafenib activates PUMA via GSK3β and NF-κB to suppress tumor cell growth , 2011, Oncogene.

[24]  P. Fisher,et al.  An Optically Pure Apogossypolone Derivative as Potent Pan-Active Inhibitor of Anti-Apoptotic Bcl-2 Family Proteins , 2011, Front. Oncol..

[25]  D. Zopf,et al.  Regorafenib (BAY 73‐4506): A new oral multikinase inhibitor of angiogenic, stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity , 2011, International journal of cancer.

[26]  D. Hanahan,et al.  Hallmarks of Cancer: The Next Generation , 2011, Cell.

[27]  Adam R. Johnson,et al.  Sensitivity to antitubulin chemotherapeutics is regulated by MCL1 and FBW7 , 2011, Nature.

[28]  Jian Yu,et al.  PUMA Induction by FoxO3a Mediates the Anticancer Activities of the Broad-Range Kinase Inhibitor UCN-01 , 2010, Molecular Cancer Therapeutics.

[29]  S. Edwards,et al.  Mcl‐1; the molecular regulation of protein function , 2010, FEBS letters.

[30]  Derek Y. Chiang,et al.  The landscape of somatic copy-number alteration across human cancers , 2010, Nature.

[31]  F. Bazan,et al.  Deubiquitinase USP9X stabilizes MCL1 and promotes tumour cell survival , 2010, Nature.

[32]  J. Llovet,et al.  Preclinical overview of sorafenib, a multikinase inhibitor that targets both Raf and VEGF and PDGF receptor tyrosine kinase signaling , 2008, Molecular Cancer Therapeutics.

[33]  Erinna F. Lee,et al.  Structure of the BH3 domains from the p53-inducible BH3-only proteins Noxa and Puma in complex with Mcl-1. , 2008, Journal of molecular biology.

[34]  Laurent Bélec,et al.  Small molecule obatoclax (GX15-070) antagonizes MCL-1 and overcomes MCL-1-mediated resistance to apoptosis , 2007, Proceedings of the National Academy of Sciences.

[35]  C. Tse,et al.  Influence of Bcl-2 family members on the cellular response of small-cell lung cancer cell lines to ABT-737. , 2007, Cancer research.

[36]  Apurva A Desai,et al.  Sorafenib in advanced clear-cell renal-cell carcinoma. , 2007, The New England journal of medicine.

[37]  John Calvin Reed,et al.  Mechanisms of apoptosis sensitivity and resistance to the BH3 mimetic ABT-737 in acute myeloid leukemia. , 2006, Cancer cell.

[38]  C. Scott,et al.  The BH3 mimetic ABT-737 targets selective Bcl-2 proteins and efficiently induces apoptosis via Bak/Bax if Mcl-1 is neutralized. , 2006, Cancer cell.

[39]  U. Landegren,et al.  Direct observation of individual endogenous protein complexes in situ by proximity ligation , 2006, Nature Methods.

[40]  H. Abaan,et al.  Structure-based design of potent small-molecule inhibitors of anti-apoptotic Bcl-2 proteins. , 2006, Journal of medicinal chemistry.

[41]  Peng Wang,et al.  PUMA Dissociates Bax and Bcl-XL to Induce Apoptosis in Colon Cancer Cells* , 2006, Journal of Biological Chemistry.

[42]  Jian Yu,et al.  PUMA Sensitizes Lung Cancer Cells to Chemotherapeutic Agents and Irradiation , 2006, Clinical Cancer Research.

[43]  M. Hung,et al.  Degradation of Mcl-1 by β-TrCP Mediates Glycogen Synthase Kinase 3-Induced Tumor Suppression and Chemosensitization , 2006, Molecular and Cellular Biology.

[44]  D. Green,et al.  Glycogen synthase kinase-3 regulates mitochondrial outer membrane permeabilization and apoptosis by destabilization of MCL-1. , 2006, Molecular cell.

[45]  Xiaodong Wang,et al.  Mule/ARF-BP1, a BH3-Only E3 Ubiquitin Ligase, Catalyzes the Polyubiquitination of Mcl-1 and Regulates Apoptosis , 2005, Cell.

[46]  S. Korsmeyer,et al.  Obligate Role of Anti-Apoptotic MCL-1 in the Survival of Hematopoietic Stem Cells , 2005, Science.

[47]  K. Kinzler,et al.  PUMA mediates the apoptotic response to p53 in colorectal cancer cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[48]  J. Opferman,et al.  Delving deeper: MCL-1's contributions to normal and cancer biology. , 2013, Trends in cell biology.

[49]  Yonghong Xiao,et al.  SCFFBW 7 regulates cellular apoptosis by targeting MCL 1 for ubiquitylation and destruction , 2011 .

[50]  A. Strasser,et al.  The BCL-2 protein family: opposing activities that mediate cell death , 2008, Nature Reviews Molecular Cell Biology.