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Updated: May 5, 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
11 Genetic Stability and Oncogenesis Research Unit, National Center for Scientific Research (CNRS UMR 8200), Gustave Roussy and University of Paris-Sud , Villejuif, France .
Background:
Medullary thyroid carcinoma (MTC) is a rare tumor that is caused by 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 autophosphorylation. The 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. However, some discrepancies were observed between RPPA and Western immunoblotting, possibly due to lack of specificity of the primary antibodies used.
Conclusions:
Overall, our results confirmed the interest 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 effectively treats medullary thyroid carcinoma (MTC) by inhibiting RET signaling, impacting MAPK and AKT pathways. This study highlights RPPA
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.
- Understanding vandetanib's molecular effects is crucial for optimizing treatment.
Purpose of the Study:
- To investigate signaling pathway alterations induced by vandetanib in preclinical MTC models.
- To evaluate the utility of Reverse-Phase Protein Array (RPPA) for this analysis.
Main Methods:
- Utilized the human TT cell line for in vitro and in vivo studies.
- Applied RPPA with 12 antibodies targeting major signaling pathways.
- Validated results using Western immunoblotting and immunohistochemistry.
Main Results:
- Vandetanib demonstrated antiproliferative and antiangiogenic effects, inhibiting RET autophosphorylation.
- The MAPK and AKT pathways were significantly inhibited by vandetanib.
- Increased phosphorylated NFκB-p65 levels were observed, suggesting complex pathway modulation.
Conclusions:
- RPPA is effective for screening kinase inhibitor-induced pathway changes in MTC.
- MAPK and AKT pathways are key targets of vandetanib in MTC.
- Findings support exploring combination therapies for MTC based on identified molecular events.
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