Journal Article

Tumour‐derived substrate‐adherent cells promote neuroblastoma survival through secreted trophic factors

Or, Penelope M.‐Y., Ho, Wayne L.H., Chan, Andrew M., Wang, Yubing, Li, Lisha, Li, Jing, Liu, Tian, Wai Wong, Chi

2021

Overview

Neuroblastoma (NB) is the most common extracranial solid tumour in children. NB is highly heterogeneous and is comprised of a mixture of neuroblastic cancer cells and stromal cells. We previously reported that N‐type cells (neuroblastic cells) and S‐type cells (substrate‐adherent cells) in the SK‐N‐SH cell line shared almost identical genetic backgrounds. Sublines of N‐ and S‐type cells were isolated from an early passage (P35) of SK‐N‐SH. Sequencing analysis revealed that all sublines harboured the anaplastic lymphoma kinase (ALK) F1174L mutation, indicating that they were tumour derived. Surprisingly, over 74% resembled S‐type cells. In coculture experiments, S‐type cells protected N‐type cells from apoptosis induced by the oncogenic ALK inhibitor TAE684. Western blotting analyses showed that ALK, protein kinase A (AKT) and STAT3 signalling were stimulated in the cocultures. Furthermore, the conditioned medium from S‐type cells activated these downstream signalling molecules in the N‐type cells. The activation of STAT3 in the N‐type cells was ALK‐independent, while AKT was regulated by the ALK activation status. To identify the responsible soluble factors, we used a combination of transcriptomic and proteomic analysis and found that plasminogen activator inhibitor 1, secreted protein acidic and cysteine rich, periostin and galectin‐1 were potential mediators of STAT3 signalling. The addition of recombinant proteins to the tumour cells treated with the ALK inhibitor partially enhanced cell viability. Overall, the tumour‐derived S‐type cells prevented apoptosis in the N‐type cells via ALK‐independent STAT3 activation triggered by secreted factors. The inhibition of these factors in combination with ALK inhibition could provide a new direction for targeted therapies to treat high‐risk NB. Neuroblastoma S‐type cells promote survival of N‐type tumor cells through cell‐to‐cell contact, which is associated with upregulation of p‐STAT3 in N‐type cells. Independently, S‐type cells secrete trophic factors that promote the survival of N‐type cells. Trophic factors could act either by stimulating p‐STAT3, or by upregulating p‐AKT and p‐ERK1/2 through an p‐ALK‐dependent pathway. The inhibition of these factors in combination with ALK inhibition could provide a new strategy for treatment of high‐risk neuroblastoma.

• Publisher: John Wiley & Sons, Inc,John Wiley and Sons Inc,Wiley
• Subject: Adherent cells / AKT protein / ALK / Analysis / Anaplastic Lymphoma Kinase - antagonists & inhibitors / Anaplastic Lymphoma Kinase - genetics / Anaplastic Lymphoma Kinase - metabolism / Antibodies / Apoptosis / Bone marrow / Cell activation / Cell Adhesion / Cell Proliferation / Cell Survival / Cell viability / Coculture Techniques / Crizotinib / Culture Media, Conditioned / Gene expression / Genomes / Humans / Kinases / Ligands / Lymphoma / Lymphomas / Mass spectrometry / Mass Spectrometry - methods / Metastasis / Mutation / Neuroblastoma / Neuroblastoma - enzymology / Neuroblastoma - metabolism / Neuroblastoma - pathology / Osteonectin / Plasminogen activator inhibitors / Protein kinase A / Protein Kinase Inhibitors - pharmacology / Protein kinases / Protein-tyrosine kinase / Proteins / Proteomics / Pyrimidines - pharmacology / Recombinant proteins / Scientific equipment and supplies industry / Scientific imaging / Sequence analysis / Sequence Analysis, RNA - methods / Software / Solid tumors / STAT3 / Stat3 protein / STAT3 Transcription Factor - metabolism / Stromal cells / substrate‐adherent cells / Transcriptome / Trophic factors / Tumors / tumour microenvironment / Western blotting