Retinoic acid and TGF-ß signalling cooperate to overcome MYCN-induced retinoid resistance

David J. Duffy (Corresponding Author), Aleksandar Krstic, Melinda Halasz, Thomas Schwarzl, Anja Konietzny, Kristiina Iljin, Desmond G. Higgins, Walter Kolch

Research output: Contribution to journalArticleScientificpeer-review

9 Citations (Scopus)

Abstract

Background: Retinoid therapy is widely employed in clinical oncology to differentiate malignant cells into their more benign counterparts. However, certain high-risk cohorts, such as patients with MYCN-amplified neuroblastoma, are innately resistant to retinoid therapy. Therefore, we employed a precision medicine approach to globally profile the retinoid signalling response and to determine how an excess of cellular MYCN antagonises these signalling events to prevent differentiation and confer resistance.
Methods: We applied RNA sequencing (RNA-seq) and interaction proteomics coupled with network-based systems level analysis to identify targetable vulnerabilities of MYCN-mediated retinoid resistance. We altered MYCN expression levels in a MYCN-inducible neuroblastoma cell line to facilitate or block retinoic acid (RA)-mediated neuronal differentiation. The relevance of differentially expressed genes and transcriptional regulators for neuroblastoma outcome were then confirmed using existing patient microarray datasets. 
Results: We determined the signalling networks through which RA mediates neuroblastoma differentiation and the inhibitory perturbations to these networks upon MYCN overexpression. We revealed opposing regulation of RA and MYCN on a number of differentiation-relevant genes, including LMO4, CYP26A1, ASCL1, RET, FZD7 and DKK1. Furthermore, we revealed a broad network of transcriptional regulators involved in regulating retinoid responsiveness, such as Neurotrophin, PI3K, Wnt and MAPK, and epigenetic signalling. Of these regulators, we functionally confirmed that MYCN-driven inhibition of transforming growth factor beta (TGF-β) signalling is a vulnerable node of the MYCN network and that multiple levels of cross-talk exist between MYCN and TGF-β. Co-targeting of the retinoic acid and TGF-β pathways, through RA and kartogenin (KGN; a TGF-ß signalling activating small molecule) combination treatment, induced the loss of viability of MYCN-amplified retinoid-resistant neuroblastoma cells.
Conclusions: Our approach provides a powerful precision oncology tool for identifying the driving signalling networks for malignancies not primarily driven by somatic mutations, such as paediatric cancers. By applying global omics approaches to the signalling networks regulating neuroblastoma differentiation and stemness, we have determined the pathways involved in the MYCN-mediated retinoid resistance, with TGF-β signalling being a key regulator. These findings revealed a number of combination treatments likely to improve clinical response to retinoid therapy, including co-treatment with retinoids and KGN, which may prove valuable in the treatment of high-risk MYCN-amplified neuroblastoma.
Original languageEnglish
Article number15
Pages (from-to)15
JournalGenome Medicine
Volume9
Issue number1
DOIs
Publication statusPublished - 10 Feb 2017
MoE publication typeA1 Journal article-refereed

Fingerprint

Retinoids
Tretinoin
Transforming Growth Factor beta
Neuroblastoma
Therapeutics
RNA Sequence Analysis
Precision Medicine
Medical Oncology
Gene Regulatory Networks
Nerve Growth Factors
Regulator Genes
Phosphatidylinositol 3-Kinases
Epigenomics
Proteomics
Neoplasms
Pediatrics
Cell Line
Mutation

Keywords

  • neuroblastoma
  • kartogenin (KGN)
  • RepSox
  • MYC (c-MYC)
  • differentiation
  • mRNA sequencing (mRNA-seq)
  • ChIP sequencing (ChIP-seq)
  • transforming growth factor beta (TGF-ß) signalling
  • interaction proteomics
  • precision medicine
  • genome medicine
  • Wnt ß-catenin signalling
  • cancer
  • systems medicine
  • neuronal differentiation
  • differentiation therapy

Cite this

Duffy, D. J., Krstic, A., Halasz, M., Schwarzl, T., Konietzny, A., Iljin, K., ... Kolch, W. (2017). Retinoic acid and TGF-ß signalling cooperate to overcome MYCN-induced retinoid resistance. Genome Medicine, 9(1), 15. [15]. https://doi.org/10.1186/s13073-017-0407-3
Duffy, David J. ; Krstic, Aleksandar ; Halasz, Melinda ; Schwarzl, Thomas ; Konietzny, Anja ; Iljin, Kristiina ; Higgins, Desmond G. ; Kolch, Walter. / Retinoic acid and TGF-ß signalling cooperate to overcome MYCN-induced retinoid resistance. In: Genome Medicine. 2017 ; Vol. 9, No. 1. pp. 15.
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title = "Retinoic acid and TGF-{\ss} signalling cooperate to overcome MYCN-induced retinoid resistance",
abstract = "Background: Retinoid therapy is widely employed in clinical oncology to differentiate malignant cells into their more benign counterparts. However, certain high-risk cohorts, such as patients with MYCN-amplified neuroblastoma, are innately resistant to retinoid therapy. Therefore, we employed a precision medicine approach to globally profile the retinoid signalling response and to determine how an excess of cellular MYCN antagonises these signalling events to prevent differentiation and confer resistance.Methods: We applied RNA sequencing (RNA-seq) and interaction proteomics coupled with network-based systems level analysis to identify targetable vulnerabilities of MYCN-mediated retinoid resistance. We altered MYCN expression levels in a MYCN-inducible neuroblastoma cell line to facilitate or block retinoic acid (RA)-mediated neuronal differentiation. The relevance of differentially expressed genes and transcriptional regulators for neuroblastoma outcome were then confirmed using existing patient microarray datasets. Results: We determined the signalling networks through which RA mediates neuroblastoma differentiation and the inhibitory perturbations to these networks upon MYCN overexpression. We revealed opposing regulation of RA and MYCN on a number of differentiation-relevant genes, including LMO4, CYP26A1, ASCL1, RET, FZD7 and DKK1. Furthermore, we revealed a broad network of transcriptional regulators involved in regulating retinoid responsiveness, such as Neurotrophin, PI3K, Wnt and MAPK, and epigenetic signalling. Of these regulators, we functionally confirmed that MYCN-driven inhibition of transforming growth factor beta (TGF-β) signalling is a vulnerable node of the MYCN network and that multiple levels of cross-talk exist between MYCN and TGF-β. Co-targeting of the retinoic acid and TGF-β pathways, through RA and kartogenin (KGN; a TGF-{\ss} signalling activating small molecule) combination treatment, induced the loss of viability of MYCN-amplified retinoid-resistant neuroblastoma cells.Conclusions: Our approach provides a powerful precision oncology tool for identifying the driving signalling networks for malignancies not primarily driven by somatic mutations, such as paediatric cancers. By applying global omics approaches to the signalling networks regulating neuroblastoma differentiation and stemness, we have determined the pathways involved in the MYCN-mediated retinoid resistance, with TGF-β signalling being a key regulator. These findings revealed a number of combination treatments likely to improve clinical response to retinoid therapy, including co-treatment with retinoids and KGN, which may prove valuable in the treatment of high-risk MYCN-amplified neuroblastoma.",
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Duffy, DJ, Krstic, A, Halasz, M, Schwarzl, T, Konietzny, A, Iljin, K, Higgins, DG & Kolch, W 2017, 'Retinoic acid and TGF-ß signalling cooperate to overcome MYCN-induced retinoid resistance', Genome Medicine, vol. 9, no. 1, 15, pp. 15. https://doi.org/10.1186/s13073-017-0407-3

Retinoic acid and TGF-ß signalling cooperate to overcome MYCN-induced retinoid resistance. / Duffy, David J. (Corresponding Author); Krstic, Aleksandar; Halasz, Melinda; Schwarzl, Thomas; Konietzny, Anja; Iljin, Kristiina; Higgins, Desmond G.; Kolch, Walter.

In: Genome Medicine, Vol. 9, No. 1, 15, 10.02.2017, p. 15.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Retinoic acid and TGF-ß signalling cooperate to overcome MYCN-induced retinoid resistance

AU - Duffy, David J.

AU - Krstic, Aleksandar

AU - Halasz, Melinda

AU - Schwarzl, Thomas

AU - Konietzny, Anja

AU - Iljin, Kristiina

AU - Higgins, Desmond G.

AU - Kolch, Walter

PY - 2017/2/10

Y1 - 2017/2/10

N2 - Background: Retinoid therapy is widely employed in clinical oncology to differentiate malignant cells into their more benign counterparts. However, certain high-risk cohorts, such as patients with MYCN-amplified neuroblastoma, are innately resistant to retinoid therapy. Therefore, we employed a precision medicine approach to globally profile the retinoid signalling response and to determine how an excess of cellular MYCN antagonises these signalling events to prevent differentiation and confer resistance.Methods: We applied RNA sequencing (RNA-seq) and interaction proteomics coupled with network-based systems level analysis to identify targetable vulnerabilities of MYCN-mediated retinoid resistance. We altered MYCN expression levels in a MYCN-inducible neuroblastoma cell line to facilitate or block retinoic acid (RA)-mediated neuronal differentiation. The relevance of differentially expressed genes and transcriptional regulators for neuroblastoma outcome were then confirmed using existing patient microarray datasets. Results: We determined the signalling networks through which RA mediates neuroblastoma differentiation and the inhibitory perturbations to these networks upon MYCN overexpression. We revealed opposing regulation of RA and MYCN on a number of differentiation-relevant genes, including LMO4, CYP26A1, ASCL1, RET, FZD7 and DKK1. Furthermore, we revealed a broad network of transcriptional regulators involved in regulating retinoid responsiveness, such as Neurotrophin, PI3K, Wnt and MAPK, and epigenetic signalling. Of these regulators, we functionally confirmed that MYCN-driven inhibition of transforming growth factor beta (TGF-β) signalling is a vulnerable node of the MYCN network and that multiple levels of cross-talk exist between MYCN and TGF-β. Co-targeting of the retinoic acid and TGF-β pathways, through RA and kartogenin (KGN; a TGF-ß signalling activating small molecule) combination treatment, induced the loss of viability of MYCN-amplified retinoid-resistant neuroblastoma cells.Conclusions: Our approach provides a powerful precision oncology tool for identifying the driving signalling networks for malignancies not primarily driven by somatic mutations, such as paediatric cancers. By applying global omics approaches to the signalling networks regulating neuroblastoma differentiation and stemness, we have determined the pathways involved in the MYCN-mediated retinoid resistance, with TGF-β signalling being a key regulator. These findings revealed a number of combination treatments likely to improve clinical response to retinoid therapy, including co-treatment with retinoids and KGN, which may prove valuable in the treatment of high-risk MYCN-amplified neuroblastoma.

AB - Background: Retinoid therapy is widely employed in clinical oncology to differentiate malignant cells into their more benign counterparts. However, certain high-risk cohorts, such as patients with MYCN-amplified neuroblastoma, are innately resistant to retinoid therapy. Therefore, we employed a precision medicine approach to globally profile the retinoid signalling response and to determine how an excess of cellular MYCN antagonises these signalling events to prevent differentiation and confer resistance.Methods: We applied RNA sequencing (RNA-seq) and interaction proteomics coupled with network-based systems level analysis to identify targetable vulnerabilities of MYCN-mediated retinoid resistance. We altered MYCN expression levels in a MYCN-inducible neuroblastoma cell line to facilitate or block retinoic acid (RA)-mediated neuronal differentiation. The relevance of differentially expressed genes and transcriptional regulators for neuroblastoma outcome were then confirmed using existing patient microarray datasets. Results: We determined the signalling networks through which RA mediates neuroblastoma differentiation and the inhibitory perturbations to these networks upon MYCN overexpression. We revealed opposing regulation of RA and MYCN on a number of differentiation-relevant genes, including LMO4, CYP26A1, ASCL1, RET, FZD7 and DKK1. Furthermore, we revealed a broad network of transcriptional regulators involved in regulating retinoid responsiveness, such as Neurotrophin, PI3K, Wnt and MAPK, and epigenetic signalling. Of these regulators, we functionally confirmed that MYCN-driven inhibition of transforming growth factor beta (TGF-β) signalling is a vulnerable node of the MYCN network and that multiple levels of cross-talk exist between MYCN and TGF-β. Co-targeting of the retinoic acid and TGF-β pathways, through RA and kartogenin (KGN; a TGF-ß signalling activating small molecule) combination treatment, induced the loss of viability of MYCN-amplified retinoid-resistant neuroblastoma cells.Conclusions: Our approach provides a powerful precision oncology tool for identifying the driving signalling networks for malignancies not primarily driven by somatic mutations, such as paediatric cancers. By applying global omics approaches to the signalling networks regulating neuroblastoma differentiation and stemness, we have determined the pathways involved in the MYCN-mediated retinoid resistance, with TGF-β signalling being a key regulator. These findings revealed a number of combination treatments likely to improve clinical response to retinoid therapy, including co-treatment with retinoids and KGN, which may prove valuable in the treatment of high-risk MYCN-amplified neuroblastoma.

KW - neuroblastoma

KW - kartogenin (KGN)

KW - RepSox

KW - MYC (c-MYC)

KW - differentiation

KW - mRNA sequencing (mRNA-seq)

KW - ChIP sequencing (ChIP-seq)

KW - transforming growth factor beta (TGF-ß) signalling

KW - interaction proteomics

KW - precision medicine

KW - genome medicine

KW - Wnt ß-catenin signalling

KW - cancer

KW - systems medicine

KW - neuronal differentiation

KW - differentiation therapy

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