title: Identification and pharmacological inactivation of the MYCN gene network as a therapeutic strategy for neuroblastic tumor cells.
abstract: The MYC family of transcription factors consists of three well characterized members, c-MYC, L-MYC, and MYCN, deregulated in the majority of human cancers. In neuronal tumors such as neuroblastoma, MYCN is frequently activated by gene amplification, and reducing its expression by RNA interference has been shown to promote growth arrest and apoptosis of tumor cells. From a clinical perspective, RNA interference is not yet a viable option, and small molecule inhibitors of transcription factors are difficult to develop. We therefore planned to identify, at the global level, the genes interacting functionally with MYCN required to promote fitness of tumor cells facing oncogenic stress. To find genes whose inactivation is synthetically lethal to MYCN, we implemented a genome-wide approach in which we carried out a drop-out shRNA screen using a whole genome library that was delivered into isogenic neuroblastoma cell lines expressing or not expressing MYCN. After the screen, we selected for in-depth analysis four shRNAs targeting AHCY, BLM, PKMYT1, and CKS1B. These genes were chosen because they are directly regulated by MYC proteins, associated with poor prognosis of neuroblastoma patients, and inhibited by small molecule compounds. Mechanistically, we found that BLM and PKMYT1 are required to limit oncogenic stress and promote stabilization of the MYCN protein. Cocktails of small molecule inhibitors of CKS1B, AHCY, BLM, and PKMYT1 profoundly affected the growth of all neuroblastoma cell lines but selectively caused death of MYCN-amplified cells. Our findings suggest that drugging the MYCN network is a promising avenue for the treatment of high risk, neuroblastic cancers.
authors.affiliation: From the Brunel Institute of Cancer Genetics and Pharmacogenomics, Brunel University London, London UB8 3PH, United Kingdom and the Institute of Child Health, University College London, London WC1N 1EH, United Kingdom.
authors.forename: Olesya
authors.initials: O
authors.lastname: Chayka
authors.affiliation: the Institute of Child Health, University College London, London WC1N 1EH, United Kingdom.
authors.forename: Cosimo Walter
authors.initials: CW
authors.lastname: D'Acunto
authors.affiliation: the Institute of Child Health, University College London, London WC1N 1EH, United Kingdom.
authors.forename: Odette
authors.initials: O
authors.lastname: Middleton
authors.affiliation: From the Brunel Institute of Cancer Genetics and Pharmacogenomics, Brunel University London, London UB8 3PH, United Kingdom and.
authors.forename: Maryam
authors.initials: M
authors.lastname: Arab
authors.affiliation: From the Brunel Institute of Cancer Genetics and Pharmacogenomics, Brunel University London, London UB8 3PH, United Kingdom and the Institute of Child Health, University College London, London WC1N 1EH, United Kingdom arturo.sala@brunel.ac.uk.
authors.forename: Arturo
authors.initials: A
authors.lastname: Sala
mesh_terms: D017209:Apoptosis; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002469:Cell Separation; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D016259:Genes, myc; D055106:Genome-Wide Association Study; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008565:Membrane Proteins / Q000378:metabolism; D000071447:N-Myc Proto-Oncogene Protein; D009447:Neuroblastoma / Q000188:drug therapy / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D015513:Oncogene Proteins / Q000378:metabolism*; D011379:Prognosis; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D053484:RecQ Helicases / Q000378:metabolism
chemical_list: C459801:MYCN protein, human; D008565:Membrane Proteins; D000071447:N-Myc Proto-Oncogene Protein; D009687:Nuclear Proteins; D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D011505:Protein-Tyrosine Kinases; C490679:PKMYT1 protein, human; D017346:Protein Serine-Threonine Kinases; C097151:Bloom syndrome protein; D053484:RecQ Helicases
keywords: Anticancer Drug; Cancer Biology; Cancer Therapy; MYCN; Myc (c-Myc); Neuroblastoma; Synthetic Lethality; shRNA Screen