IGF2 mRNA binding protein 3 (IMP3) promotes glioma cell migration by enhancing the translation of RELA/p65

The diffusely infiltrative nature of glioblastoma (GBM) makes them highly recurrent. IGF2 mRNA-binding protein 3 (IMP3), a GBM upregulated RNA binding protein, promotes glioma cell migration. An integrative bioinformatics analysis identified p65 (RELA), a subunit of NF-κB heterodimer as a target and an important mediator of IMP3 promoted glioma cell migration. IMP3 increased p65 protein levels without any change in p65 transcript levels, but promoted its polysome association. RIP-PCR demonstrated the binding of IMP3 to p65 transcript. UV crosslinking experiments with in vitro transcribed RNA confirmed the specific and direct binding of IMP3 to sites on p65 3′UTR. Further, IMP3 induced luciferase activity from p65 3′UTR reporter carrying wild type sites but not mutated sites. Exogenous overexpression of p65 from a 3′UTR-less construct rescued the reduced migration of glioma cells in IMP3 silenced condition. In addition, IMP3 silencing inhibited glioma stem-like cell maintenance and migration. The exogenous overexpression of 3′UTR-less p65 significantly alleviated the inhibition of neurosphere formation observed in IMP3 silenced glioma stem-like cells. Further, we show that IMP3 is transcriptionally activated by NF-κB pathway indicating the presence of a positive feedback loop between IMP3 and p65. This study establishes p65 as a novel target of IMP3 in increasing glioma cell migration and underscores the significance of IMP3-p65 feedback loop for therapeutic targeting in GBM.

[1]  Kulandaivelu Mahalingam,et al.  Elucidation of the genetic and epigenetic landscape alterations in RNA binding proteins in glioblastoma , 2016, Oncotarget.

[2]  T. Cooper,et al.  Roles for RNA-binding proteins in development and disease , 2016, Brain Research.

[3]  Jun Ma,et al.  MiR-608 inhibits the migration and invasion of glioma stem cells by targeting macrophage migration inhibitory factor. , 2016, Oncology reports.

[4]  Sol Katzman,et al.  RNA-binding protein IGF2BP3 targeting of oncogenic transcripts promotes hematopoietic progenitor proliferation. , 2016, The Journal of clinical investigation.

[5]  H. Goel,et al.  IMP3 promotes stem-like properties in triple-negative breast cancer by regulating SLUG , 2016, Oncogene.

[6]  E. Schwarz,et al.  Increased insulin mRNA binding protein-3 expression correlates with vascular enhancement of renal cell carcinoma by intravenous contrast-CT and is associated with bone metastasis , 2015, Journal of bone oncology.

[7]  S. Kesari,et al.  Effect of the JAK2/STAT3 inhibitor SAR317461 on human glioblastoma tumorspheres , 2015, Journal of Translational Medicine.

[8]  Jian Zheng,et al.  CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186 , 2015, Oncotarget.

[9]  N. Chow,et al.  Overexpression of the RNA-binding proteins Lin28B and IGF2BP3 (IMP3) is associated with chemoresistance and poor disease outcome in ovarian cancer , 2015, British Journal of Cancer.

[10]  Ming Zhou,et al.  Enhanced IMP3 Expression Activates NF-кB Pathway and Promotes Renal Cell Carcinoma Progression , 2015, PloS one.

[11]  N. Agarwal,et al.  Survival Outcomes and Tumor IMP3 Expression in Patients with Sarcomatoid Metastatic Renal Cell Carcinoma , 2015, Journal of oncology.

[12]  S. Gerstberger,et al.  A census of human RNA-binding proteins , 2014, Nature Reviews Genetics.

[13]  K. Taniuchi,et al.  IGF2BP3-mediated translation in cell protrusions promotes cell invasiveness and metastasis of pancreatic cancer , 2014, Oncotarget.

[14]  B. Kazeminezhad,et al.  Usefulness of insulin-like growth factor II mRNA-binding protein 3 (IMP3) as a new marker for the diagnosis of esophageal adenocarcinoma in challenging cases. , 2014, The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology.

[15]  Y. Zhang,et al.  Role of IGF2BP3 in trophoblast cell invasion and migration , 2014, Cell Death and Disease.

[16]  Hui Zhou,et al.  starBase v2.0: decoding miRNA-ceRNA, miRNA-ncRNA and protein–RNA interaction networks from large-scale CLIP-Seq data , 2013, Nucleic Acids Res..

[17]  D. Friedmann-Morvinski.  Glioblastoma heterogeneity and cancer cell plasticity. , 2014, Critical reviews in oncogenesis.

[18]  K. Somasundaram,et al.  A DNA methylation prognostic signature of glioblastoma: identification of NPTX2-PTEN-NF-κB nexus. , 2013, Cancer research.

[19]  M. Gujrati,et al.  Essential role of cooperative NF-κB and Stat3 recruitment to ICAM-1 intronic consensus elements in the regulation of radiation-induced invasion and migration in glioma , 2013, Oncogene.

[20]  C. Ménard,et al.  Glioblastoma management in the temozolomide era: have we improved outcome? , 2013, Journal of Neuro-Oncology.

[21]  O. Myklebost,et al.  Comparison of glioma stem cells to neural stem cells from the adult human brain identifies dysregulated Wnt- signaling and a fingerprint associated with clinical outcome. , 2013, Experimental cell research.

[22]  L. Pfeffer,et al.  Constitutive Activation of Signal Transducer and Activator of Transcription 3 (STAT3) and Nuclear Factor κB Signaling in Glioblastoma Cancer Stem Cells Regulates the Notch Pathway* , 2013, The Journal of Biological Chemistry.

[23]  S. Dooley,et al.  Reciprocal regulation by TLR4 and TGF-β in tumor-initiating stem-like cells. , 2013, The Journal of clinical investigation.

[24]  G. Stark,et al.  PHF20 regulates NF-κB signalling by disrupting recruitment of PP2A to p65 , 2013, Nature Communications.

[25]  Simon Kasif,et al.  An aberrant transcription factor network essential for Wnt signaling and stem cell maintenance in glioblastoma. , 2013, Cell reports.

[26]  Angélica Figueroa,et al.  Posttranscriptional regulation by RNA-binding proteins during epithelial-to-mesenchymal transition , 2013, Cellular and Molecular Life Sciences.

[27]  Jun Wang,et al.  Multiple Functions of the RNA-Binding Protein HuR in Cancer Progression, Treatment Responses and Prognosis , 2013, International journal of molecular sciences.

[28]  Raymond Moran,et al.  PhysBinder: improving the prediction of transcription factor binding sites by flexible inclusion of biophysical properties , 2013, Nucleic Acids Res..

[29]  A. Mercurio,et al.  IMP3 Protein Promotes Chemoresistance in Breast Cancer Cells by Regulating Breast Cancer Resistance Protein (ABCG2) Expression* , 2013, The Journal of Biological Chemistry.

[30]  G. Pelicci,et al.  Cancer stem cell contribution to glioblastoma invasiveness , 2013, Stem Cell Research & Therapy.

[31]  N. Onishi,et al.  Up-Regulation of Imp3 Confers In Vivo Tumorigenicity on Murine Osteosarcoma Cells , 2012, PLoS ONE.

[32]  Saumitra Das,et al.  Targeting Ribosome assembly on the HCV RNA using a small RNA molecule , 2012, RNA biology.

[33]  Norman E. Davey,et al.  Insights into RNA Biology from an Atlas of Mammalian mRNA-Binding Proteins , 2012, Cell.

[34]  Hui Liu,et al.  AnimalTFDB: a comprehensive animal transcription factor database , 2011, Nucleic Acids Res..

[35]  Tzong-Shiue Yu,et al.  A restricted cell population propagates glioblastoma growth after chemotherapy , 2012 .

[36]  G. Brewer,et al.  RNA-binding Protein Insulin-like Growth Factor mRNA-binding Protein 3 (IMP-3) Promotes Cell Survival via Insulin-like Growth Factor II Signaling after Ionizing Radiation* , 2011, The Journal of Biological Chemistry.

[37]  G. Mor,et al.  Inhibition of Aurora-A kinase induces cell cycle arrest in epithelial ovarian cancer stem cells by affecting NFĸB pathway , 2011, Cell cycle.

[38]  K. Thennarasu,et al.  Insulin Growth Factor-2 Binding Protein 3 (IGF2BP3) Is a Glioblastoma-specific Marker That Activates Phosphatidylinositol 3-Kinase/Mitogen-activated Protein Kinase (PI3K/MAPK) Pathways by Modulating IGF-2* , 2011, The Journal of Biological Chemistry.

[39]  Nathan D Price,et al.  A CD133-related gene expression signature identifies an aggressive glioblastoma subtype with excessive mutations , 2011, Proceedings of the National Academy of Sciences.

[40]  S. Senju,et al.  Peptides derived from human insulin‐like growth factor‐II mRNA binding protein 3 can induce human leukocyte antigen‐A2‐restricted cytotoxic T lymphocytes reactive to cancer cells , 2011, Cancer science.

[41]  J. Mosser,et al.  DNA methylation in glioblastoma: impact on gene expression and clinical outcome , 2010, BMC Genomics.

[42]  Antony Hsieh,et al.  IGFBP2 promotes glioma tumor stem cell expansion and survival. , 2010, Biochemical and biophysical research communications.

[43]  Scott B. Dewell,et al.  Transcriptome-wide Identification of RNA-Binding Protein and MicroRNA Target Sites by PAR-CLIP , 2010, Cell.

[44]  Roger E. McLendon,et al.  Targeting A20 Decreases Glioma Stem Cell Survival and Tumor Growth , 2010, PLoS biology.

[45]  S. Almo,et al.  ZBP1 recognition of beta-actin zipcode induces RNA looping. , 2010, Genes & development.

[46]  R. McLendon,et al.  Targeting Interleukin 6 Signaling Suppresses Glioma Stem Cell Survival and Tumor Growth , 2009, Stem cells.

[47]  D. Huntsman,et al.  IGF2BP3 (IMP3) expression is a marker of unfavorable prognosis in ovarian carcinoma of clear cell subtype , 2009, Modern Pathology.

[48]  Hui Wang,et al.  c-Myc Is Required for Maintenance of Glioma Cancer Stem Cells , 2008, PloS one.

[49]  T. Glisovic,et al.  RNA‐binding proteins and post‐transcriptional gene regulation , 2008, FEBS letters.

[50]  R. Britto,et al.  Novel Glioblastoma Markers with Diagnostic and Prognostic Value Identified through Transcriptome Analysis , 2008, Clinical Cancer Research.

[51]  Yusuke Nakamura,et al.  Identification of human leukocyte antigen‐A24‐restricted epitope peptides derived from gene products upregulated in lung and esophageal cancers as novel targets for immunotherapy , 2007, Cancer science.

[52]  Alexander Brawanski,et al.  CD133(+) and CD133(-) glioblastoma-derived cancer stem cells show differential growth characteristics and molecular profiles. , 2007, Cancer research.

[53]  T. Gilmore,et al.  Inhibitors of NF-kappaB signaling: 785 and counting. , 2006, Oncogene.

[54]  A. Baldwin,et al.  Nuclear factor-kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. , 2006, Oncogene.

[55]  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.

[56]  Xavier Matias-Guiu,et al.  NF-kB in development and progression of human cancer , 2005, Virchows Archiv.

[57]  Martin J. van den Bent,et al.  Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. , 2005, The New England journal of medicine.

[58]  P. Barker,et al.  TRAF6‐dependent NF‐kB transcriptional activity during mouse development , 2004, Developmental dynamics : an official publication of the American Association of Anatomists.

[59]  Jan Christiansen,et al.  Sequential dimerization of human zipcode-binding protein IMP1 on RNA: a cooperative mechanism providing RNP stability. , 2004, Nucleic acids research.

[60]  Cynthia Hawkins,et al.  Identification of a cancer stem cell in human brain tumors. , 2003, Cancer research.

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

[62]  A. Aggarwal,et al.  Structure of NF-kappaB p50/p65 heterodimer bound to the PRDII DNA element from the interferon-beta promoter. , 2002, Structure.