Oral Presentation Advances in Neuroblastoma Research Congress 2016

MYCN amplified neuroblastomas require TEAD4 to orchestrate transcriptional programs, exposing a therapeutic vulnerability (#56)

Gonzalo Lopez 1 2 , Presha Rajbhandari 1 3 , Jiyang Yu 1 , Ruth Rodriguez-Barrueco 4 , Archana Iyer 1 , Mariano J Alvarez 1 , Jo L Harenza 2 , Derek Oldridge 2 5 , Daniel Martinez 2 , Mark Yarmarkovich 5 , Jo Vandesompele 6 7 , Pieter Mestdagh 6 7 , Jose M Silva 4 , Antonio Iavarone 8 , Anna Lasorella 8 , John M Maris 2 5 , Andrea Califano 1 3 8
  1. Department of Systems Biology, Columbia University, New York, NY, USA
  2. Division of Oncology and Center for Childhood Cancer Research, Childrens Hospital Of Philadelphia, Philadelphia, PA, USA
  3. Department of Biological Sciences, Columbia University, New York, NY, USA
  4. Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, NY, USA
  5. Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, NY, USA
  6. Center for Medical Genetics, Ghent University, Ghent, Belgium
  7. Cancer Research Institute , Ghent University, Ghent, Belgium
  8. Institute for Cancer Genetics, Columbia University, New York, NY, USA

BACKGROUND: High-risk neuroblastomas (NBL) display substantial clinical and molecular heterogeneity. Despite progress understanding NBL genetics, little progress has been made in procuring personalized treatments. Here we sought to determine effector molecular mechanisms downstream of genetic alterations sustaining different tumor subtypes1.

METHODS: We first defined molecular subtypes from the TARGET and NRC Consortium gene expression datasets by consensus clustering and identified specific master regulators (MRs) of transcriptional programs using VIPER/ssMARINA algorithm2 available as a bioconductor R package. We performed in vitro and in vivo validation of MRs by RNAi screening followed by experimental and computational assays to elucidate the interdependencies between top MRs and their biological role. Finally, we studied tissue microarrays (TMA) and applied multivariate Cox regression to understand the clinical relevance of our findings.

RESULTS: We identified MRs from three distinct molecular subtypes of high-risk NBL, conserved across cohorts. siRNA and shRNA screens identified a TEAD4-MYCN positive feed forward loop as a key NBL state maintenance of MYCN dysregulated tumors. RNA-seq upon MYCN and TEAD4 knockdown showed that MYCN controls cell growth and represses differentiation and that TEAD4 is essential for G1 to S transition and DNA repair. Specifically, TEAD4 transactivates AURKA, CDKs, Cyclin D, E2Fs, DNA replication factors, checkpoint kinases and ubiquitin ligases. Consistently, TEAD4 inhibition induced critical loss of NBL cell viability, both in vitro and in vivo. TMA staining shows increased TEAD4 protein levels in high risk tumors (P = 0.021); expression/activity is an independent predictor of survival in multivariable analysis (P = 0.0084 corrected for age, stage and MYCN status).

CONCLUSION: TEAD4 plays a critical role in maintaining the high proliferative state of undifferentiated NBLs with sustained growth promoted by MYCN dysregulation. The critical effect of disrupting TEAD4-MYCN module on cell viability and its clinical relevance brings forward TEAD4 as an excellent candidate target for therapeutic intervention.

  1. Chen, J. C. et al. Identification of Causal Genetic Drivers of Human Disease through Systems-Level Analysis of Regulatory Networks. Cell 159, 402-414, doi:10.1016/j.cell.2014.09.021 (2014).
  2. Aytes, A. et al. Cross-species regulatory network analysis identifies a synergistic interaction between FOXM1 and CENPF that drives prostate cancer malignancy. Cancer cell 25, 638-651, doi:10.1016/j.ccr.2014.03.017 (2014).