Characterization of a Steroid Receptor Coactivator Small Molecule Stimulator that Overstimulates Cancer Cells and Leads to Cell Stress and Death.

By integrating growth pathways on which cancer cells rely, steroid receptor coactivators (SRC-1, SRC-2, and SRC-3) represent emerging targets in cancer therapeutics. High-throughput screening for SRC small molecule inhibitors (SMIs) uncovered MCB-613 as a potent SRC small molecule "stimulator" (SMS). We demonstrate that MCB-613 can super-stimulate SRCs' transcriptional activity. Further investigation revealed that MCB-613 increases SRCs' interactions with other coactivators and markedly induces ER stress coupled to the generation of reactive oxygen species (ROS). Because cancer cells overexpress SRCs and rely on them for growth, we show that we can exploit MCB-613 to selectively induce excessive stress in cancer cells. This suggests that over-stimulating the SRC oncogenic program can be an effective strategy to kill cancer cells.

[1]  Thomas J. Raub,et al.  Determination of the Affinity of Drugs toward Serum Albumin by Measurement of the Quenching of the Intrinsic Tryptophan Fluorescence of the Protein , 1999, The Journal of pharmacy and pharmacology.

[2]  Jun Wang,et al.  Role of ABL family kinases in cancer: from leukaemia to solid tumours , 2013, Nature Reviews Cancer.

[3]  P. Meltzer,et al.  In breast cancer, amplification of the steroid receptor coactivator gene AIB1 is correlated with estrogen and progesterone receptor positivity. , 1998, Clinical cancer research : an official journal of the American Association for Cancer Research.

[4]  G. Gores,et al.  Proteasome inhibition-induces endoplasmic reticulum dysfunction and cell death of human cholangiocarcinoma cells. , 2007, World journal of gastroenterology.

[5]  L. Landmesser,et al.  Ultrastructural differences during embryonic cell death in normal and peripherally deprived ciliary ganglia , 1976, The Journal of cell biology.

[6]  B. O’Malley,et al.  Normal and cancer-related functions of the p160 steroid receptor co-activator (SRC) family , 2009, Nature Reviews Cancer.

[7]  Q. Jin,et al.  Dual Activators of Protein Kinase R (PKR) and Protein Kinase R‐Like Kinase (PERK) Identify Common and Divergent Catalytic Targets , 2013, Chembiochem : a European journal of chemical biology.

[8]  D. Bredesen,et al.  An alternative, nonapoptotic form of programmed cell death. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[9]  L. Penn,et al.  Reflecting on 25 years with MYC , 2008, Nature Reviews Cancer.

[10]  Y. Zeng,et al.  Correlation of AIB1 overexpression with advanced clinical stage of human colorectal carcinoma. , 2005, Human pathology.

[11]  Yusuke Nakamura,et al.  Amplification and over‐expression of the AIB1 nuclear receptor co‐activator gene in primary gastric cancers , 2000, International journal of cancer.

[12]  G. Semenza,et al.  Inhibition of lactate dehydrogenase A induces oxidative stress and inhibits tumor progression , 2010, Proceedings of the National Academy of Sciences.

[13]  C. Sander,et al.  Integrative genomic profiling of human prostate cancer. , 2010, Cancer cell.

[14]  Eun Hee Kim,et al.  Superoxide anion and proteasomal dysfunction contribute to curcumin-induced paraptosis of malignant breast cancer cells. , 2010, Free radical biology & medicine.

[15]  A. Maitra,et al.  Overexpression of the Nuclear Receptor Coactivator AIB1 (SRC-3) during Progression of Pancreatic Adenocarcinoma , 2004, Clinical Cancer Research.

[16]  X. Guan,et al.  Prognostic significance of c‐myc and AIB1 amplification in hepatocellular carcinoma , 2002, Cancer.

[17]  W. Sellers,et al.  High tumor incidence and activation of the PI3K/AKT pathway in transgenic mice define AIB1 as an oncogene. , 2004, Cancer cell.

[18]  D. Aswad,et al.  Regulation of transcription by a protein methyltransferase. , 1999, Science.

[19]  Ross A. Hamilton,et al.  Proteomic analysis of coregulators bound to ERα on DNA and nucleosomes reveals coregulator dynamics. , 2013, Molecular cell.

[20]  E. Jeffes,et al.  Living T9 glioma cells expressing membrane macrophage colony-stimulating factor produce immediate tumor destruction by polymorphonuclear leukocytes and macrophages via a "paraptosis"-induced pathway that promotes systemic immunity against intracranial T9 gliomas. , 2002, Blood.

[21]  S. Schreiber,et al.  Synthesis, cellular evaluation, and mechanism of action of piperlongumine analogs , 2012, Proceedings of the National Academy of Sciences.

[22]  A. Wellstein,et al.  The nuclear receptor coactivator amplified in breast cancer-1 is required for Neu (ErbB2/HER2) activation, signaling, and mammary tumorigenesis in mice. , 2008, Cancer research.

[23]  D. Neal,et al.  Expression of RAC 3, a steroid hormone receptor co-activator in prostate cancer , 2001, British Journal of Cancer.

[24]  Brian York,et al.  Steroid Receptor Coactivator (SRC) Family: Masters of Systems Biology* , 2010, The Journal of Biological Chemistry.

[25]  M. Venkatachalam,et al.  A Novel Role for MAP1 LC3 in Non-Autophagic Cytoplasmic Vacuolation Death of Cancer Cells , 2009, Oncogene.

[26]  M. Ittmann,et al.  Steroid receptor coactivator-3/AIB1 promotes cell migration and invasiveness through focal adhesion turnover and matrix metalloproteinase expression. , 2008, Cancer Research.

[27]  S. Schreiber,et al.  Discovery of Small-Molecule Enhancers of Reactive Oxygen Species That are Nontoxic or Cause Genotype-Selective Cell Death , 2013, ACS chemical biology.

[28]  R. Karlsson,et al.  Surface plasmon resonance detection and multispot sensing for direct monitoring of interactions involving low-molecular-weight analytes and for determination of low affinities. , 1995, Analytical biochemistry.

[29]  D. Felsher,et al.  Reversible tumorigenesis by MYC in hematopoietic lineages. , 1999, Molecular cell.

[30]  N. Heisterkamp,et al.  Tyrosine 207 in CRKL is the BCR/ABL phosphorylation site , 1997, Oncogene.

[31]  M. Southey,et al.  Overexpression of the steroid receptor coactivator AIB1 in breast cancer correlates with the absence of estrogen and progesterone receptors and positivity for p53 and HER2/neu. , 2001, Cancer research.

[32]  E. Manivannan,et al.  Advances in chalcones with anticancer activities. , 2014, Recent patents on anti-cancer drug discovery.

[33]  U. Hofmann,et al.  Oncogenic Stress Induced by Acute Hyper-Activation of Bcr-Abl Leads to Cell Death upon Induction of Excessive Aerobic Glycolysis , 2011, PloS one.

[34]  Sivanesan Dakshanamurthy,et al.  Tyrosine Phosphorylation of the Nuclear Receptor Coactivator AIB1/SRC-3 Is Enhanced by Abl Kinase and Is Required for Its Activity in Cancer Cells , 2008, Molecular and Cellular Biology.

[35]  E. McDermott,et al.  Inverse relationship between ER-β and SRC-1 predicts outcome in endocrine-resistant breast cancer , 2004, British Journal of Cancer.

[36]  Xiao-Ming Yin,et al.  Absence of Bax switched MG132-induced apoptosis to non-apoptotic cell death that could be suppressed by transcriptional or translational inhibition , 2007, Apoptosis.

[37]  Hideaki Kurioka,et al.  Elevated expression levels of NCOA3, TOP1, and TFAP2C in breast tumors as predictors of poor prognosis , 2003, Cancer.

[38]  K. Tew,et al.  Redox metabolism and malignancy. , 2010, Current opinion in pharmacology.

[39]  S. Gruber,et al.  Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway , 2006, Nature Cell Biology.

[40]  E. McDermott,et al.  Expression of SRC-1, AIB1, and PEA3 in HER2 mediated endocrine resistant breast cancer; a predictive role for SRC-1 , 2004, Journal of Clinical Pathology.

[41]  Jianming Xu,et al.  The steroid receptor coactivator-1 regulates twist expression and promotes breast cancer metastasis. , 2009, Cancer research.

[42]  Yun Lu,et al.  A morphologically conserved nonapoptotic program promotes linker cell death in Caenorhabditis elegans. , 2007, Developmental cell.

[43]  J. Qin,et al.  Selective phosphorylations of the SRC-3/AIB1 coactivator integrate genomic reponses to multiple cellular signaling pathways. , 2004, Molecular cell.

[44]  S. Yokoyama,et al.  Oncogenic Ras triggers cell suicide through the activation of a caspase-independent cell death program in human cancer cells , 1999, Oncogene.

[45]  B. O’Malley,et al.  Nuclear receptor coregulators: judges, juries, and executioners of cellular regulation. , 2007, Molecular cell.

[46]  P. Griffin,et al.  Bufalin is a potent small-molecule inhibitor of the steroid receptor coactivators SRC-3 and SRC-1. , 2014, Cancer research.

[47]  M. Beckmann,et al.  Gene Amplification and Expression of the Steroid Receptor Coactivator SRC3 (AIB1) in Sporadic Breast and Endometrial Carcinomas , 2001, Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme.

[48]  R. Weichselbaum,et al.  Activation of the Cytoplasmic c-Abl Tyrosine Kinase by Reactive Oxygen Species* , 2000, The Journal of Biological Chemistry.

[49]  L. Chin,et al.  Essential role for oncogenic Ras in tumour maintenance , 1999, Nature.

[50]  B. Rowan,et al.  Expression of estrogen receptor coregulators in normal and malignant human endometrium. , 2004, Gynecologic oncology.

[51]  P. Ray,et al.  Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. , 2012, Cellular signalling.

[52]  Eiji Kinoshita,et al.  Recognition of phosphate monoester dianion by an alkoxide-bridged dinuclear zinc(II) complex. , 2004, Dalton transactions.

[53]  E. Kubista,et al.  Expression of Sex Steroid Receptors and their Co-Factors in Normal and Malignant Breast Tissue: AIB1 is a Carcinoma-Specific Co-Activator , 2003, Breast Cancer Research and Treatment.

[54]  B. O’Malley,et al.  SRC-3 Coactivator Functional Lifetime Is Regulated by a Phospho-Dependent Ubiquitin Time Clock , 2007, Cell.

[55]  L. Neckers,et al.  Endoplasmic Reticulum Vacuolization and Valosin-Containing Protein Relocalization Result from Simultaneous Hsp90 Inhibition by Geldanamycin and Proteasome Inhibition by Velcade , 2006, Molecular Cancer Research.

[56]  P. Clarke,et al.  Developmental cell death: morphological diversity and multiple mechanisms , 2004, Anatomy and Embryology.

[57]  J. Pollack,et al.  Steroid receptor coactivator-3 expression in lung cancer and its role in the regulation of cancer cell survival and proliferation. , 2010, Cancer research.

[58]  Myles A Brown,et al.  Targeting the AIB1 oncogene through mammalian target of rapamycin inhibition in the mammary gland. , 2006, Cancer research.

[59]  B. O’Malley,et al.  Small molecule inhibition of the steroid receptor coactivators, SRC-3 and SRC-1. , 2011, Molecular endocrinology.

[60]  R. Evans,et al.  Nuclear Receptor Coactivator ACTR Is a Novel Histone Acetyltransferase and Forms a Multimeric Activation Complex with P/CAF and CBP/p300 , 1997, Cell.

[61]  D. Tenen,et al.  Reversibility of acute B-cell leukaemia induced by BCR–ABL1 , 2000, Nature Genetics.

[62]  R. Deberardinis,et al.  Inhibition of cancer cell proliferation by PPARγ is mediated by a metabolic switch that increases reactive oxygen species levels. , 2014, Cell metabolism.

[63]  R. Kaufman,et al.  The impact of the unfolded protein response on human disease , 2012, The Journal of cell biology.

[64]  P. Meltzer,et al.  AIB1, a steroid receptor coactivator amplified in breast and ovarian cancer. , 1997, Science.

[65]  C. Powers,et al.  Ribozyme Targeting Demonstrates That the Nuclear Receptor Coactivator AIB1 Is a Rate-limiting Factor for Estrogen-dependent Growth of Human MCF-7 Breast Cancer Cells* , 2001, The Journal of Biological Chemistry.

[66]  P. Griffin,et al.  Identification of Verrucarin A as a Potent and Selective Steroid Receptor Coactivator-3 Small Molecule Inhibitor , 2014, PloS one.