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Updated: May 10, 2026
Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
Published on: September 19, 2018
Changes in signaling pathways induced by vandetanib in a human medullary thyroid carcinoma model, as analyzed by
Sophie Broutin1, Frédéric Commo, Leanne De Koning
1Institut Gustave-Roussy, CNRS UMR8200, Villejuif, France ; broutin@igr.fr.
Background:
Medullary thyroid carcinoma (MTC) is a rare tumor that is due to activating mutations in the proto-oncogene RET. Vandetanib, a tyrosine-kinase inhibitor, has been recently approved to treat adult patients with metastatic MTC. The aim of this study was to investigate changes in signaling pathways induced by vandetanib treatment in preclinical MTC models, using the reverse-phase protein array method (RPPA).
Methods:
The human TT cell line was used to assess in vitro and in vivo activity of vandetanib. Protein extracts from TT cells or TT xenografted mice, treated by increasing concentrations of vandetanib for different periods of time, were probed with a set of 12 antibodies representing major signaling pathways, using RPPA. Results were validated using two distinct protein detection methods, western-immunoblotting and immunohistochemistry.
Results:
Vandetanib displays antiproliferative and antiangiogenic activities and inhibits RET auto-phosphorylation. MAPK and AKT pathways were the two major signaling pathways inhibited by vandetanib. Interestingly, phosphorylated levels of NFκB-p65 were significantly increased by vandetanib. Comparable results were obtained in both the in vitro and in vivo approaches as well as for the protein detection methods, although some discrepancies were observed between RPPA and western-immunoblotting.
Conclusions:
Results confirmed the reliability and the utility of RPPA for screening global changes induced in signaling pathways by kinase inhibitors. MAPK and AKT were identified as the main pathways involved in vandetanib response in MTC models. Our results also suggest alternative routes for controlling the disease and provide a rationale for the development of therapeutic combinations based on the comprehensive identification of molecular events induced by inhibitors.
Insights
Vandetanib treatment for medullary thyroid carcinoma (MTC) inhibits MAPK and AKT pathways. This study used reverse-phase protein array (RPPA) to reveal these and other signaling changes in preclinical MTC models.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Medullary thyroid carcinoma (MTC) is a rare cancer driven by RET proto-oncogene mutations.
- Vandetanib, a tyrosine-kinase inhibitor, is approved for metastatic MTC in adults.
- Investigating vandetanib's effects on signaling pathways in MTC models is crucial.
Purpose of the Study:
- To explore vandetanib-induced signaling pathway alterations in preclinical MTC models.
- To assess the utility of reverse-phase protein array (RPPA) for this investigation.
- To identify key pathways modulated by vandetanib in MTC.
Main Methods:
- Utilized the human TT cell line for in vitro and in vivo studies.
- Employed RPPA with 12 antibodies targeting major signaling pathways.
- Validated RPPA findings using western-immunoblotting and immunohistochemistry.
- Treated cells and TT xenografted mice with varying vandetanib concentrations and durations.
Main Results:
- Vandetanib demonstrated antiproliferative and antiangiogenic effects.
- Inhibition of RET auto-phosphorylation, MAPK, and AKT pathways by vandetanib.
- Observed a significant increase in phosphorylated NFκB-p65 levels.
- Consistent results across in vitro/in vivo models and detection methods, with minor RPPA/western-blotting discrepancies.
Conclusions:
- RPPA is a reliable method for screening kinase inhibitor-induced pathway changes.
- MAPK and AKT pathways are central to vandetanib's response in MTC.
- Findings support exploring alternative therapeutic strategies and combination therapies for MTC.
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