Src promotes castration-recurrent prostate cancer through androgen receptor-dependent canonical and non-canonical transcriptional signatures

Progression of prostate cancer (PC) to castration-recurrent growth (CRPC) remains dependent on sustained expression and transcriptional activity of the androgen receptor (AR). A major mechanism contributing to CRPC progression is through the direct phosphorylation and activation of AR by Src-family (SFK) and ACK1 tyrosine kinases. However, the AR-dependent transcriptional networks activated by Src during CRPC progression have not been elucidated. Here, we show that activated Src (Src527F) induces androgen-independent growth in human LNCaP cells, concomitant with its ability to induce proliferation/survival genes normally induced by dihydrotestosterone (DHT) in androgen-dependent LNCaP and VCaP cells. Src induces additional gene signatures unique to CRPC cell lines, LNCaP-C4-2 and CWR22Rv1, and to CRPC LuCaP35.1 xenografts. By comparing the Src-induced AR-cistrome and/or transcriptome in LNCaP to those in CRPC and LuCaP35.1 tumors, we identified an 11-gene Src-regulated CRPC signature consisting of AR-dependent, AR binding site (ARBS)-associated genes whose expression is altered by DHT in LNCaP[Src527F] but not in LNCaP cells. The differential expression of a subset (DPP4, BCAT1, CNTNAP4, CDH3) correlates with earlier PC metastasis onset and poorer survival, with the expression of BCAT1 required for Src-induced androgen-independent proliferation. Lastly, Src enhances AR binding to non-canonical ARBS enriched for FOXO1, TOP2B and ZNF217 binding motifs; cooperative AR/TOP2B binding to a non-canonical ARBS was both Src- and DHT-sensitive and correlated with increased levels of Src-induced phosphotyrosyl-TOP2B. These data suggest that CRPC progression is facilitated via Src-induced sensitization of AR to intracrine androgen levels, resulting in the engagement of canonical and non-canonical ARBS-dependent gene signatures.

[1]  Michael D. Wilson,et al.  Topoisomerase II beta interacts with cohesin and CTCF at topological domain borders , 2016, Genome Biology.

[2]  H. Lehrach,et al.  Transcriptional signature induced by a metastasis‐promoting c‐Src mutant in a human breast cell line , 2016, The FEBS journal.

[3]  Joshua M. Stuart,et al.  N-Myc Drives Neuroendocrine Prostate Cancer Initiated from Human Prostate Epithelial Cells. , 2016, Cancer cell.

[4]  A. Imai,et al.  Significance of preoperative butyrylcholinesterase as an independent predictor of biochemical recurrence-free survival in patients with prostate cancer treated with radical prostatectomy , 2016, International Journal of Clinical Oncology.

[5]  P. O’Donnell,et al.  Saracatinib as a metastasis inhibitor in metastatic castration‐resistant prostate cancer: A University of Chicago Phase 2 Consortium and DOD/PCF Prostate Cancer Clinical Trials Consortium Study , 2016, The Prostate.

[6]  R. Vessella,et al.  The biology and clinical implications of prostate cancer dormancy and metastasis , 2015, Journal of Molecular Medicine.

[7]  S. Cockell,et al.  Genome-wide ChIP-seq analysis of human TOP2B occupancy in MCF7 breast cancer epithelial cells , 2015, Biology Open.

[8]  H. Lincet,et al.  The dark side of ZNF217, a key regulator of tumorigenesis with powerful biomarker value , 2015, Oncotarget.

[9]  Henry W. Long,et al.  The androgen receptor cistrome is extensively reprogrammed in human prostate tumorigenesis , 2015, Nature Genetics.

[10]  M. Katsogiannou,et al.  The hallmarks of castration-resistant prostate cancers. , 2015, Cancer treatment reviews.

[11]  H. Klocker,et al.  Adaptive responses of androgen receptor signaling in castration-resistant prostate cancer , 2015, Oncotarget.

[12]  J. Schalken,et al.  Enzalutamide: targeting the androgen signalling pathway in metastatic castration‐resistant prostate cancer , 2015, BJU International.

[13]  Youri Hoogstrate,et al.  C/D-box snoRNA-derived RNA production is associated with malignant transformation and metastatic progression in prostate cancer , 2015, Oncotarget.

[14]  Lawrence D. True,et al.  Integrative Clinical Genomics of Advanced Prostate Cancer , 2015, Cell.

[15]  Vladimir A. Richter,et al.  Regulatory Role of Small Nucleolar RNAs in Human Diseases , 2015, BioMed research international.

[16]  D. Mankoff,et al.  Castration-Resistant Prostate Cancer Bone Metastasis Response Measured by 18F-Fluoride PET After Treatment with Dasatinib and Correlation with Progression-Free Survival: Results from American College of Radiology Imaging Network 6687 , 2015, The Journal of Nuclear Medicine.

[17]  F. Claessens,et al.  Emerging mechanisms of enzalutamide resistance in prostate cancer , 2014, Nature Reviews Urology.

[18]  B. Győrffy,et al.  A functional interplay between ZNF217 and Estrogen Receptor alpha exists in luminal breast cancers , 2014, Molecular oncology.

[19]  Zhiquan Hu,et al.  CAMK2N1 inhibits prostate cancer progression through androgen receptor-dependent signaling , 2014, Oncotarget.

[20]  W. Isaacs,et al.  AR-V7 and resistance to enzalutamide and abiraterone in prostate cancer. , 2014, The New England journal of medicine.

[21]  S. Yadav,et al.  Recent insights into NF‐κB signalling pathways and the link between inflammation and prostate cancer , 2014, BJU international.

[22]  B. Foster,et al.  Differential Requirement for Src Family Tyrosine Kinases in the Initiation, Progression, and Metastasis of Prostate Cancer , 2014, Molecular Cancer Research.

[23]  Bryan M. Gillard,et al.  A Genome-Wide RNAi Screen Identifies FOXO4 as a Metastasis-Suppressor through Counteracting PI3K/AKT Signal Pathway in Prostate Cancer , 2014, PloS one.

[24]  P. Farnham,et al.  Global analysis of ZNF217 chromatin occupancy in the breast cancer cell genome reveals an association with ERalpha , 2014, BMC Genomics.

[25]  I. Gelman.  Androgen Receptor Activation in Castration-Recurrent Prostate Cancer: The Role of Src-Family and Ack1 Tyrosine Kinases , 2014, International journal of biological sciences.

[26]  D. Tindall,et al.  Modulation of Androgen Receptor by FOXA1 and FOXO1 Factors in Prostate Cancer , 2014, International journal of biological sciences.

[27]  Zhiquan Hu,et al.  The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer , 2014, Oncotarget.

[28]  David C. Smith,et al.  A randomized phase II study of cediranib alone versus cediranib in combination with dasatinib in docetaxel resistant, castration resistant prostate cancer patients , 2014, Investigational New Drugs.

[29]  O. Sartor,et al.  Castration-resistant prostate cancer: adaptive responses in the androgen axis. , 2014, Cancer treatment reviews.

[30]  T. Meehan,et al.  An atlas of active enhancers across human cell types and tissues , 2014, Nature.

[31]  F. Claessens,et al.  Comparative Genomic and Transcriptomic Analyses of LNCaP and C4-2B Prostate Cancer Cell Lines , 2014, PloS one.

[32]  P. Corn,et al.  Src signaling pathways in prostate cancer , 2014, Cancer and Metastasis Reviews.

[33]  Chih-Hsin Tang,et al.  Prostate cancer-derived CCN3 induces M2 macrophage infiltration and contributes to angiogenesis in prostate cancer microenvironment , 2014, Oncotarget.

[34]  T. Kuzel,et al.  CCN3/NOV gene expression in human prostate cancer is directly suppressed by the androgen receptor , 2014, Oncogene.

[35]  D. Zheng,et al.  Glucocorticoid Receptor Confers Resistance to Antiandrogens by Bypassing Androgen Receptor Blockade , 2013, Cell.

[36]  Joaquim Bellmunt,et al.  Docetaxel and dasatinib or placebo in men with metastatic castration-resistant prostate cancer (READY): a randomised, double-blind phase 3 trial. , 2013, The Lancet. Oncology.

[37]  K. Pienta,et al.  Metastatic castration-resistant prostate cancer reveals intrapatient similarity and interpatient heterogeneity of therapeutic kinase targets , 2013, Proceedings of the National Academy of Sciences.

[38]  Bryan M. Gillard,et al.  Src controls castration recurrence of CWR22 prostate cancer xenografts , 2013, Cancer medicine.

[39]  N. Sharifi.  Mechanisms of androgen receptor activation in castration-resistant prostate cancer. , 2013, Endocrinology.

[40]  Gang Shao,et al.  A clinically relevant androgen receptor mutation confers resistance to second-generation antiandrogens enzalutamide and ARN-509. , 2013, Cancer discovery.

[41]  Joshua M. Korn,et al.  An F876L mutation in androgen receptor confers genetic and phenotypic resistance to MDV3100 (enzalutamide). , 2013, Cancer discovery.

[42]  W. Ye,et al.  A phase II trial of dasatinib in patients with metastatic castration-resistant prostate cancer treated previously with chemotherapy , 2013, Anti-cancer drugs.

[43]  E. Campo,et al.  Functional analysis of sucrase-isomaltase mutations from chronic lymphocytic leukemia patients. , 2013, Human molecular genetics.

[44]  Michael V. Fiandalo,et al.  The role of intracrine androgen metabolism, androgen receptor and apoptosis in the survival and recurrence of prostate cancer during androgen deprivation therapy. , 2013, Current drug targets.

[45]  M. Ittmann,et al.  ERK and AKT Signaling Drive MED1 Overexpression in Prostate Cancer in Association with Elevated Proliferation and Tumorigenicity , 2013, Molecular Cancer Research.

[46]  P. Paliwal,et al.  Long-term use of dasatinib in patients with metastatic castration-resistant prostate cancer after receiving the combination of dasatinib and docetaxel , 2013, Cancer management and research.

[47]  E. Kremmer,et al.  Identification of ZNF217, hnRNP‐K, VEGF‐A and IPO7 as targets for microRNAs that are downregulated in prostate carcinoma , 2013, International journal of cancer.

[48]  K. Silverstein,et al.  Androgen receptor splice variants mediate enzalutamide resistance in castration-resistant prostate cancer cell lines. , 2013, Cancer research.

[49]  I. Mills,et al.  The androgen receptor induces a distinct transcriptional program in castration-resistant prostate cancer in man. , 2013, Cancer cell.

[50]  Steve Y. Cho,et al.  A phase 2 study of KX2-391, an oral inhibitor of Src kinase and tubulin polymerization, in men with bone-metastatic castration-resistant prostate cancer , 2013, Cancer Chemotherapy and Pharmacology.

[51]  J. Mohler,et al.  Biology of castration-recurrent prostate cancer. , 2012, The Urologic clinics of North America.

[52]  Kurt Miller,et al.  Increased survival with enzalutamide in prostate cancer after chemotherapy. , 2012, The New England journal of medicine.

[53]  V. Morsch,et al.  Cholinesterase activities and biochemical determinations in patients with prostate cancer: influence of Gleason score, treatment and bone metastasis. , 2012, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[54]  D. Coppola,et al.  Ack1-mediated Androgen Receptor Phosphorylation Modulates Radiation Resistance in Castration-resistant Prostate Cancer* , 2012, The Journal of Biological Chemistry.

[55]  Benjamin J. Raphael,et al.  The Mutational Landscape of Lethal Castrate Resistant Prostate Cancer , 2012, Nature.

[56]  Jun Liu,et al.  Cooperation between Polycomb and androgen receptor during oncogenic transformation. , 2012, Genome research.

[57]  T. Graeber,et al.  Oncogene-specific activation of tyrosine kinase networks during prostate cancer progression , 2012, Proceedings of the National Academy of Sciences.

[58]  Wei Li,et al.  Definition of a FoxA1 Cistrome that is crucial for G1 to S-phase cell-cycle transit in castration-resistant prostate cancer. , 2011, Cancer research.

[59]  P. Nelson,et al.  Androgen receptor gene expression in prostate cancer is directly suppressed by the androgen receptor through recruitment of lysine-specific demethylase 1. , 2011, Cancer cell.

[60]  D. Tindall,et al.  Androgen receptor rediscovered: the new biology and targeting the androgen receptor therapeutically. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[61]  I. Mills,et al.  The androgen receptor fuels prostate cancer by regulating central metabolism and biosynthesis , 2011, The EMBO journal.

[62]  M. Gross,et al.  Once-daily dasatinib: expansion of phase II study evaluating safety and efficacy of dasatinib in patients with metastatic castration-resistant prostate cancer. , 2011, Urology.

[63]  D. Tindall,et al.  Minireview: Alternative activation pathways for the androgen receptor in prostate cancer. , 2011, Molecular endocrinology.

[64]  Allen D. Delaney,et al.  LNCaP Atlas: Gene expression associated with in vivo progression to castration-recurrent prostate cancer , 2010, BMC Medical Genomics.

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

[66]  Martin J. Aryee,et al.  Androgen-induced TOP2B mediated double strand breaks and prostate cancer gene rearrangements , 2010, Nature Genetics.

[67]  S. Rabbani,et al.  SKI-606 (Bosutinib) Blocks Prostate Cancer Invasion, Growth, and Metastasis In vitro and In vivo through Regulation of Genes Involved in Cancer Growth and Skeletal Metastasis , 2010, Molecular Cancer Therapeutics.

[68]  Bin Fang,et al.  Ack1 Mediated AKT/PKB Tyrosine 176 Phosphorylation Regulates Its Activation , 2010, PloS one.

[69]  Daniel Gioeli,et al.  Dasatinib inhibits site-specific tyrosine phosphorylation of androgen receptor by Ack1 and Src kinases , 2010, Oncogene.

[70]  W. Schultze‐Seemann,et al.  Comparison of Gene Expression in LNCaP Prostate Cancer Cells after Treatment with Bicalutamide or 5-Alpha-Reductase Inhibitors , 2010, Urologia Internationalis.

[71]  R. Vessella,et al.  Novel biomarkers for prostate cancer including noncoding transcripts. , 2009, The American journal of pathology.

[72]  Steven J. M. Jones,et al.  Identification of novel androgen-responsive genes by sequencing of LongSAGE libraries , 2009, BMC Genomics.

[73]  V. Brunton,et al.  Src Family Kinase Activity Is Up-Regulated in Hormone-Refractory Prostate Cancer , 2009, Clinical Cancer Research.

[74]  Gurkamal Chatta,et al.  A phase II trial of the Src-kinase inhibitor AZD0530 in patients with advanced castration-resistant prostate cancer: a California Cancer Consortium study , 2009, Anti-cancer drugs.

[75]  Clifford A. Meyer,et al.  Model-based Analysis of ChIP-Seq (MACS) , 2008, Genome Biology.

[76]  H. Mukhtar,et al.  Src kinase potentiates androgen receptor transactivation function and invasion of androgen-independent prostate cancer C4-2 cells , 2008, Oncogene.

[77]  D. Tindall,et al.  Androgen receptor (AR) coregulators: a diversity of functions converging on and regulating the AR transcriptional complex. , 2007, Endocrine reviews.

[78]  K. Pienta,et al.  A hierarchical network of transcription factors governs androgen receptor-dependent prostate cancer growth. , 2007, Molecular cell.

[79]  A. Mortazavi,et al.  Genome-Wide Mapping of in Vivo Protein-DNA Interactions , 2007, Science.

[80]  J. Mohler,et al.  Activated Cdc42-associated kinase Ack1 promotes prostate cancer progression via androgen receptor tyrosine phosphorylation , 2007, Proceedings of the National Academy of Sciences.

[81]  H. Kung,et al.  Nonreceptor tyrosine kinases in prostate cancer. , 2007, Neoplasia.

[82]  Zhiyong Guo,et al.  Regulation of androgen receptor activity by tyrosine phosphorylation. , 2006, Cancer cell.

[83]  Peter A. Jones,et al.  Locus-Wide Chromatin Remodeling and Enhanced Androgen Receptor-Mediated Transcription in Recurrent Prostate Tumor Cells , 2006, Molecular and Cellular Biology.

[84]  D. Tindall,et al.  Ligand-independent Androgen Receptor Activity Is Activation Function-2-independent and Resistant to Antiandrogens in Androgen Refractory Prostate Cancer Cells* , 2006, Journal of Biological Chemistry.

[85]  R. Vessella,et al.  FOXO1A is a candidate for the 13q14 tumor suppressor gene inhibiting androgen receptor signaling in prostate cancer. , 2006, Cancer research.

[86]  H. Kung,et al.  Activity of the Src-kinase inhibitor AZD0530 in androgen-independent prostate cancer (AIPC): Pre-clinical rationale for a phase II trial. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[87]  S. Tooze,et al.  Synaptotagmin IV is necessary for the maturation of secretory granules in PC12 cells , 2006, The Journal of cell biology.

[88]  S. Powers,et al.  Metastatic properties and genomic amplification of the tyrosine kinase gene ACK1. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[89]  Daniel J. Blankenberg,et al.  Galaxy: a platform for interactive large-scale genome analysis. , 2005, Genome research.

[90]  K. Tomer,et al.  Testosterone and Dihydrotestosterone Tissue Levels in Recurrent Prostate Cancer , 2005, Clinical Cancer Research.

[91]  T. Hubbard,et al.  NestedMICA: sensitive inference of over-represented motifs in nucleic acid sequence , 2005, Nucleic acids research.

[92]  C. Sette,et al.  Expression of a truncated form of the c-Kit tyrosine kinase receptor and activation of Src kinase in human prostatic cancer. , 2004, The American journal of pathology.

[93]  Roberto Bianco,et al.  Involvement of growth factor receptors of the epidermal growth factor receptor family in prostate cancer development and progression to androgen independence. , 2003, Clinical prostate cancer.

[94]  R. Vessella,et al.  LuCaP 35: A new model of prostate cancer progression to androgen independence , 2003, The Prostate.

[95]  G. Jenster,et al.  AKT-Independent Protection of Prostate Cancer Cells from Apoptosis Mediated through Complex Formation between the Androgen Receptor and FKHR , 2003, Molecular and Cellular Biology.

[96]  G. McLean,et al.  Src-induced de-regulation of E-cadherin in colon cancer cells requires integrin signalling , 2002, Nature Cell Biology.

[97]  F. S. French,et al.  Androgen receptor stabilization in recurrent prostate cancer is associated with hypersensitivity to low androgen. , 2001, Cancer research.

[98]  K. Pienta,et al.  VCaP, a cell-based model system of human prostate cancer. , 2001, In vivo.

[99]  L. Hood,et al.  Prostate-localized and androgen-regulated expression of the membrane-bound serine protease TMPRSS2. , 1999, Cancer research.

[100]  W. Zhu,et al.  Identification of two novel cis-elements in the promoter of the prostate-specific antigen gene that are required to enhance androgen receptor-mediated transactivation. , 1997, Nucleic acids research.

[101]  D. Kassel,et al.  Correlation of the phosphorylation states of pp60c-src with tyrosine kinase activity: the intramolecular pY530-SH2 complex retains significant activity if Y419 is phosphorylated. , 1996, Biochemistry.

[102]  H. Hanafusa,et al.  Transcriptional down regulation of the nov proto-oncogene in fibroblasts transformed by p60v-src , 1996, Molecular and cellular biology.

[103]  G. Prins,et al.  Regulation of proliferation and production of prostate-specific antigen in androgen-sensitive prostatic cancer cells, LNCaP, by dihydrotestosterone. , 1995, Endocrinology.

[104]  M. Gleave,et al.  Derivation of androgen‐independent human LNCaP prostatic cancer cell sublines: Role of bone stromal cells , 1994, International journal of cancer.

[105]  M. Gleave,et al.  Autocrine regulation of prostate-specific antigen gene expression in a human prostatic cancer (LNCaP) subline. , 1993, Cancer research.

[106]  P. Troncoso,et al.  Molecular characterization of enzalutamide-treated bone metastatic castration-resistant prostate cancer. , 2015, European urology.

[107]  A. Burdova,et al.  Relation of ETS transcription factor family member ERG, androgen receptor and topoisomerase 2β expression to TMPRSS2-ERG fusion status in prostate cancer. , 2014, Neoplasma.

[108]  A. Burdova,et al.  Relation of ETS transcription factor family member ERG, androgen receptor and topoisomerase 2β expression to TMPRSS2-ERG fusion status in prostate cancer. , 2013, Neoplasma.

[109]  P. Corn,et al.  Small-molecule protein tyrosine kinase inhibitors for the treatment of metastatic prostate cancer. , 2012, Future medicinal chemistry.

[110]  Timothy L Bailey,et al.  BIOINFORMATICS ORIGINAL PAPER , 2022 .

[111]  I. Gelman.  Src-family tyrosine kinases as therapeutic targets in advanced cancer. , 2011, Frontiers in bioscience.

[112]  D. Tindall,et al.  Androgen regulation of gene expression. , 2010, Advances in cancer research.

[113]  Clifford A. Meyer,et al.  Supplemental Material References Open Access , 2008 .

[114]  J. Mohler.  Castration-recurrent prostate cancer is not androgen-independent. , 2008, Advances in experimental medicine and biology.

[115]  Joon-ha Ok,et al.  Clinical implications of neuroendocrine differentiation in prostate cancer , 2007, Prostate Cancer and Prostatic Diseases.

[116]  B. Knudsen,et al.  Prostate cancer and the met hepatocyte growth factor receptor. , 2004, Advances in cancer research.

[117]  Charles Elkan,et al.  The Value of Prior Knowledge in Discovering Motifs with MEME , 1995, ISMB.

[118]  F. S. French,et al.  Autologous down-regulation of androgen receptor messenger ribonucleic acid. , 1990, Molecular endocrinology.