Nuclear factor-κB activation by transforming growth factor-β1 drives tumour microenvironment-mediated drug resistance in neuroblastoma

Br J Cancer. 2024 Jul;131(1):90-100. doi: 10.1038/s41416-024-02686-8. Epub 2024 May 28.

Abstract

Background: Intrinsic and extrinsic factors in the tumour microenvironment (TME) contribute to therapeutic resistance. Here we demonstrate that transforming growth factor (TGF)-β1 produced in the TME increased drug resistance of neuroblastoma (NB) cells.

Methods: Human NB cell lines were tested in vitro for their sensitivity to Doxorubicin (DOX) and Etoposide (ETOP) in the presence of tumour-associated macrophages (TAM) and mesenchymal stromal cells/cancer-associated fibroblasts (MSC/CAF). These experiments were validated in xenotransplanted and primary tumour samples.

Results: Drug resistance was associated with an increased expression of efflux transporter and anti-apoptotic proteins. Upregulation was dependent on activation of nuclear factor (NF)-κB by TGF-β-activated kinase (TAK1) and SMAD2. Resistance was reversed upon pharmacologic and genetic inhibitions of NF-κB, and TAK1/SMAD2. Interleukin-6, leukaemia inhibitory factor and oncostatin M were upregulated by this TGF-β/TAK1/NF-κB/SMAD2 signalling pathway contributing to drug resistance via an autocrine loop activating STAT3. An analysis of xenotransplanted NB tumours revealed an increased presence of phospho (p)-NF-κB in tumours co-injected with MSC/CAF and TAM, and these tumours failed to respond to Etoposide but responded if treated with a TGF-βR1/ALK5 inhibitor. Nuclear p-NF-κB was increased in patient-derived tumours rich in TME cells.

Conclusions: The data provides a novel insight into a targetable mechanism of environment-mediated drug resistance.

MeSH terms

  • Animals
  • Cancer-Associated Fibroblasts / drug effects
  • Cancer-Associated Fibroblasts / metabolism
  • Cancer-Associated Fibroblasts / pathology
  • Cell Line, Tumor
  • Doxorubicin* / pharmacology
  • Drug Resistance, Neoplasm*
  • Etoposide / pharmacology
  • Humans
  • MAP Kinase Kinase Kinases / metabolism
  • Mice
  • NF-kappa B* / metabolism
  • Neuroblastoma* / drug therapy
  • Neuroblastoma* / genetics
  • Neuroblastoma* / metabolism
  • Neuroblastoma* / pathology
  • Signal Transduction / drug effects
  • Smad2 Protein / metabolism
  • Transforming Growth Factor beta1* / metabolism
  • Tumor Microenvironment* / drug effects
  • Xenograft Model Antitumor Assays

Substances

  • NF-kappa B
  • Transforming Growth Factor beta1
  • MAP kinase kinase kinase 7
  • Doxorubicin
  • Etoposide
  • MAP Kinase Kinase Kinases
  • Smad2 Protein
  • TGFB1 protein, human
  • SMAD2 protein, human