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ALK inhibitor resistance in ALK(F1174L)-driven neuroblastoma is associated with AXL activation and induction of EMT
D N Debruyne1, N Bhatnagar1, B Sharma1
1Department of Pediatric Hematology/Oncology, Dana-Farber Cancer Institute and Boston Children's Hospital, Boston, MA, USA.
Abstract:
The crizotinib-resistant ALK(F1174L) mutation arises de novo in neuroblastoma (NB) and is acquired in ALK translocation-driven cancers, lending impetus to the development of novel anaplastic lymphoma kinase (ALK) inhibitors with different modes of action. The diaminopyrimidine TAE684 and its derivative ceritinib (LDK378), which are structurally distinct from crizotinib, are active against NB cells expressing ALK(F1174L). Here we demonstrate acquired resistance to TAE684 and LDK378 in ALK(F1174L)-driven human NB cells that is linked to overexpression and activation of the AXL tyrosine kinase and epithelial-to-mesenchymal transition (EMT). AXL phosphorylation conferred TAE684 resistance to NB cells through upregulated extracellular signal-regulated kinase (ERK) signaling. Inhibition of AXL partly rescued TAE684 resistance, resensitizing these cells to this compound. AXL activation in resistant cells was mediated through increased expression of the active form of its ligand, GAS6, that also served to stabilize the AXL protein. Although ectopic expression of AXL and TWIST2 individually in TAE684-sensitive parental cells led to the elevated expression of mesenchymal markers and invasive capacity, only AXL overexpression induced resistance to TAE684 as well. TAE684-resistant cells showed greater sensitivity to HSP90 inhibition than did their parental counterparts, with downregulation of AXL and AXL-mediated ERK signaling. Our studies indicate that aberrant AXL signaling and development of an EMT phenotype underlie resistance of ALK(F1174L)-driven NB cells to TAE684 and its derivatives. We suggest that the combination of ALK and AXL or HSP90 inhibitors be considered to delay the emergence of such resistance.
Insights
Resistance to anaplastic lymphoma kinase (ALK) inhibitors in neuroblastoma involves AXL tyrosine kinase activation and epithelial-to-mesenchymal transition (EMT). Combining ALK inhibitors with AXL or HSP90 inhibitors may overcome this resistance.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- The anaplastic lymphoma kinase (ALK) F1174L mutation is a key driver in neuroblastoma (NB) and other cancers, leading to resistance against ALK inhibitors like crizotinib.
- Novel ALK inhibitors, including TAE684 and ceritinib, show activity against ALK(F1174L)-mutated NB cells, but acquired resistance remains a challenge.
Purpose of the Study:
- To investigate the mechanisms of acquired resistance to TAE684 and ceritinib in ALK(F1174L)-driven human neuroblastoma cells.
- To explore the role of AXL tyrosine kinase and epithelial-to-mesenchymal transition (EMT) in mediating this resistance.
- To evaluate potential therapeutic strategies combining ALK inhibitors with AXL or HSP90 inhibitors.
Main Methods:
- Generation and characterization of TAE684-resistant ALK(F1174L)-mutated neuroblastoma cell lines.
- Analysis of AXL tyrosine kinase, GAS6, TWIST2, and extracellular signal-regulated kinase (ERK) signaling pathways.
- Assessment of epithelial-to-mesenchymal transition (EMT) markers and invasive capacity.
- Evaluation of drug sensitivity to AXL and HSP90 inhibitors in resistant cells.
Main Results:
- Acquired resistance to TAE684 and ceritinib was linked to AXL tyrosine kinase overexpression and activation, alongside EMT.
- AXL phosphorylation and subsequent extracellular signal-regulated kinase (ERK) pathway activation conferred resistance.
- GAS6 ligand upregulation stabilized AXL protein, mediating resistance.
- Ectopic AXL expression induced resistance and EMT, while TWIST2 overexpression promoted EMT but not resistance.
- Resistant cells exhibited increased sensitivity to HSP90 inhibition, leading to AXL downregulation.
Conclusions:
- Aberrant AXL signaling and EMT are critical mechanisms underlying resistance to ALK inhibitors in ALK(F1174L)-driven neuroblastoma.
- Targeting AXL or HSP90 in combination with ALK inhibitors presents a promising strategy to overcome or delay resistance.
- Further investigation into combined therapeutic approaches is warranted for effective neuroblastoma treatment.
