KRAS and YAP1 Converge to Regulate EMT and Tumor Survival

Cancer cells that express oncogenic alleles of RAS typically require sustained expression of the mutant allele for survival, but the molecular basis of this oncogene dependency remains incompletely understood. To identify genes that can functionally substitute for oncogenic RAS, we systematically expressed 15,294 open reading frames in a human KRAS-dependent colon cancer cell line engineered to express an inducible KRAS-specific shRNA. We found 147 genes that promoted survival upon KRAS suppression. In particular, the transcriptional coactivator YAP1 rescued cell viability in KRAS-dependent cells upon suppression of KRAS and was required for KRAS-induced cell transformation. Acquired resistance to Kras suppression in a Kras-driven murine lung cancer model also involved increased YAP1 signaling. KRAS and YAP1 converge on the transcription factor FOS and activate a transcriptional program involved in regulating the epithelial-mesenchymal transition (EMT). Together, these findings implicate transcriptional regulation of EMT by YAP1 as a significant component of oncogenic RAS signaling.

[1]  Bert Gunter,et al.  Improved Statistical Methods for Hit Selection in High-Throughput Screening , 2003, Journal of biomolecular screening.

[2]  Kakajan Komurov,et al.  Core epithelial-to-mesenchymal transition interactome gene-expression signature is associated with claudin-low and metaplastic breast cancer subtypes , 2010, Proceedings of the National Academy of Sciences.

[3]  H. Kajiyama,et al.  Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells. , 2007, International journal of oncology.

[4]  Andrew D Sharrocks,et al.  Immediate-early gene activation by the MAPK pathways: what do and don't we know? , 2012, Biochemical Society transactions.

[5]  Xiang-Jiao Yang,et al.  YAP, TAZ, and Yorkie: a conserved family of signal-responsive transcriptional coregulators in animal development and human disease. , 2009, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[6]  Lior Pachter,et al.  Sequence Analysis , 2020, Definitions.

[7]  F. Gage,et al.  YAP regulates neural progenitor cell number via the TEA domain transcription factor. , 2008, Genes & development.

[8]  Robert A. Weinberg,et al.  Creation of human tumour cells with defined genetic elements , 1999, Nature.

[9]  H. Varmus,et al.  Induction and apoptotic regression of lung adenocarcinomas by regulation of a K-Ras transgene in the presence and absence of tumor suppressor genes. , 2001, Genes & development.

[10]  K. Guan,et al.  The Hippo-YAP pathway: new connections between regulation of organ size and cancer. , 2008, Current opinion in cell biology.

[11]  Martin Vingron,et al.  TransFind—predicting transcriptional regulators for gene sets , 2010, Nucleic Acids Res..

[12]  Sirio Dupont.  Role of YAP/TAZ in mechanotransduction , 2011 .

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

[14]  M. Sudol.  YAP1 oncogene and its eight isoforms , 2013, Oncogene.

[15]  K. Harvey,et al.  Control of Tissue Growth and Cell Transformation by the Salvador/Warts/Hippo Pathway , 2012, PloS one.

[16]  Thomas M Green,et al.  A public genome-scale lentiviral expression library of human ORFs , 2011, Nature Methods.

[17]  Shan Jiang,et al.  Yap1 Activation Enables Bypass of Oncogenic Kras Addiction in Pancreatic Cancer , 2014, Cell.

[18]  Jill P. Mesirov,et al.  β-Catenin-Driven Cancers Require a YAP1 Transcriptional Complex for Survival and Tumorigenesis , 2012, Cell.

[19]  G. Stamp,et al.  Binding of Ras to Phosphoinositide 3-Kinase p110α Is Required for Ras- Driven Tumorigenesis in Mice , 2007, Cell.

[20]  Martin Renqiang Min,et al.  An integrated encyclopedia of DNA elements in the human genome , 2012 .

[21]  A. Balmain,et al.  Interactions Between Wildtype and Mutant Ras Genes in Lung and Skin Carcinogenesis , 2012, Oncogene.

[22]  Jiandie D. Lin,et al.  TEAD mediates YAP-dependent gene induction and growth control. , 2008, Genes & development.

[23]  Jennifer S. Yu,et al.  Inflammation and Hras signaling control epithelial-mesenchymal transition during skin tumor progression. , 2013, Genes & development.

[24]  Nir Hacohen,et al.  Genome-scale loss-of-function screening with a lentiviral RNAi library , 2006, Nature Methods.

[25]  H. Varmus,et al.  Regulation of transgenes in three-dimensional cultures of primary mouse mammary cells demonstrates oncogene dependence and identifies cells that survive deinduction. , 2009, Genes & development.

[26]  M. Baccarini,et al.  Raf-1 addiction in Ras-induced skin carcinogenesis. , 2009, Cancer cell.

[27]  M. Karin.  The regulation of AP-1 activity by mitogen-activated protein kinases. , 1996, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[28]  G. Boccaccio,et al.  Mammalian Smaug Is a Translational Repressor That Forms Cytoplasmic Foci Similar to Stress Granules* , 2005, Journal of Biological Chemistry.

[29]  K. Lim,et al.  Reduction in the requirement of oncogenic Ras signaling to activation of PI3K/AKT pathway during tumor maintenance. , 2005, Cancer cell.

[30]  G. Z. Cheng,et al.  Twist transcriptionally up-regulates AKT2 in breast cancer cells leading to increased migration, invasion, and resistance to paclitaxel. , 2007, Cancer research.

[31]  J. Inazawa,et al.  YAP is a candidate oncogene for esophageal squamous cell carcinoma. , 2011, Carcinogenesis.

[32]  J. Minna,et al.  Restoring E-cadherin expression increases sensitivity to epidermal growth factor receptor inhibitors in lung cancer cell lines. , 2006, Cancer research.

[33]  Jianmin Zhang,et al.  Transforming properties of YAP, a candidate oncogene on the chromosome 11q22 amplicon , 2006, Proceedings of the National Academy of Sciences.

[34]  M. DePamphilis,et al.  TEAD/TEF transcription factors utilize the activation domain of YAP65, a Src/Yes-associated protein localized in the cytoplasm. , 2001, Genes & development.

[35]  H. Varmus,et al.  Oncogene cooperation in tumor maintenance and tumor recurrence in mouse mammary tumors induced by Myc and mutant Kras , 2008, Proceedings of the National Academy of Sciences.

[36]  Sreenath V. Sharma,et al.  Oncogene addiction: setting the stage for molecularly targeted cancer therapy. , 2007, Genes & development.

[37]  B. Schaffhauser,et al.  Epithelial Mesenchymal Transition by C-Fos Estrogen Receptor Activation Involves Nuclear Translocation of β-Catenin and Upregulation of β-Catenin/Lymphoid Enhancer Binding Factor-1 Transcriptional Activity , 2000, The Journal of cell biology.

[38]  R. DePinho,et al.  Endogenous oncogenic K-ras(G12D) stimulates proliferation and widespread neoplastic and developmental defects. , 2004, Cancer cell.

[39]  D. Pan,et al.  The hippo signaling pathway in development and cancer. , 2010, Developmental cell.

[40]  K. Lau,et al.  Non-redundancy within the RAS oncogene family: Insights into mutational disparities in cancer , 2009, Molecules and cells.

[41]  M. Sudol,et al.  Yes-associated protein (YAP65) is a proline-rich phosphoprotein that binds to the SH3 domain of the Yes proto-oncogene product. , 1994, Oncogene.

[42]  Fusca.  Product , 1972, The Veterinary record.

[43]  M. Sudol,et al.  Nuclear localization and pro‐apoptotic signaling of YAP2 require intact PDZ‐binding motif , 2009, Genes to cells : devoted to molecular & cellular mechanisms.

[44]  M. Karin,et al.  c-Fos transcriptional activity stimulated by H-Ras-activated protein kinase distinct from JNK and ERK , 1994, Nature.

[45]  K. Irvine,et al.  Regulation of Hippo signaling by EGFR-MAPK signaling through Ajuba family proteins. , 2013, Developmental cell.

[46]  M. Wigler,et al.  Identification and Validation of Oncogenes in Liver Cancer Using an Integrative Oncogenomic Approach , 2006, Cell.

[47]  G. Suske.  The Sp-family of transcription factors. , 1999, Gene.

[48]  Christof Fellmann,et al.  Toolkit for evaluating genes required for proliferation and survival using tetracycline-regulated RNAi , 2011, Nature Biotechnology.

[49]  R. Hynes,et al.  The Hippo pathway target, YAP, promotes metastasis through its TEAD-interaction domain , 2012, Proceedings of the National Academy of Sciences.

[50]  Cole Trapnell,et al.  Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. , 2010, Nature biotechnology.

[51]  R. Jaenisch,et al.  YAP1 Increases Organ Size and Expands Undifferentiated Progenitor Cells , 2007, Current Biology.

[52]  Hammad Qureshi.  Contributions , 1974, Livre Blanc de la Recherche en Mécanique.

[53]  G. Stamp,et al.  RalGDS is required for tumor formation in a model of skin carcinogenesis. , 2005, Cancer cell.

[54]  M. Yaniv,et al.  Transformation by ras modifies AP1 composition and activity , 1997, Oncogene.

[55]  H. Sasaki,et al.  Mammalian Tead proteins regulate cell proliferation and contact inhibition as transcriptional mediators of Hippo signaling , 2008, Development.

[56]  Anne E Carpenter,et al.  A Lentiviral RNAi Library for Human and Mouse Genes Applied to an Arrayed Viral High-Content Screen , 2006, Cell.

[57]  Patricia Greninger,et al.  A gene expression signature associated with "K-Ras addiction" reveals regulators of EMT and tumor cell survival. , 2009, Cancer cell.

[58]  Li Li,et al.  Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control. , 2007, Genes & development.

[59]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[60]  D. Feldser,et al.  Response and resistance to NF-κB inhibitors in mouse models of lung adenocarcinoma. , 2011, Cancer discovery.

[61]  Ronald C. Petersen,et al.  Conversion , 2006, Neurology.

[62]  D. Bar-Sagi,et al.  RAS oncogenes: weaving a tumorigenic web , 2011, Nature Reviews Cancer.

[63]  W. W. Nichols,et al.  Antisense-fos RNA causes partial reversion of the transformed phenotypes induced by the c-Ha-ras oncogene , 1990, Molecular and cellular biology.

[64]  Wolfgang Henrich,et al.  The prognostic significance of epithelial-mesenchymal transition in breast cancer. , 2002, Anticancer research.

[65]  Nathanael Gray,et al.  Factors underlying sensitivity of cancers to small-molecule kinase inhibitors , 2009, Nature Reviews Drug Discovery.

[66]  Tom H. Pringle,et al.  The human genome browser at UCSC. , 2002, Genome research.

[67]  L. Staudt,et al.  Critical role for transcriptional repressor Snail2 in transformation by oncogenic RAS in colorectal carcinoma cells , 2010, Oncogene.

[68]  Jianmin Zhang,et al.  YAP-dependent induction of amphiregulin identifies a non-cell-autonomous component of the Hippo pathway , 2009, Nature Cell Biology.

[69]  H. Schwarz,et al.  Activation of an inducible c-FosER fusion protein causes loss of epithelial polarity and triggers epithelial-fibroblastoid cell conversion , 1992, Cell.

[70]  B. Spiegelman,et al.  c-fos is required for malignant progression of skin tumors , 1995, Cell.

[71]  T. Jacks,et al.  Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase , 2009, Nature Protocols.

[72]  Data production leads,et al.  An integrated encyclopedia of DNA elements in the human genome , 2012 .