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Updated: Jul 1, 2025
Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
Published on: September 19, 2018
Efficacy of tyrosine kinase inhibitors examined by a combination of Raman micro-spectroscopy and a deep wavelet
Irina Schuler1,2, Martin Schuler1,2, Tatjana Frick1,2
1Center for Protein Diagnostics, Ruhr-University Bochum, Bochum, Germany. Samir.Elmashtoly@leibniz-ipht.de.
Abstract:
HER2 is a crucial therapeutic target in breast cancer, and the survival rate of breast cancer patients has increased because of this receptor's inhibition. However, tumors have shown resistance to this therapeutic strategy due to oncogenic mutations that decrease the binding of several HER2-targeted drugs, including lapatinib, and confer resistance to this drug. Neratinib can overcome this drug resistance and effectively inhibit HER2 signaling and tumor growth. In the present study, we examined the efficacy of lapatinib and neratinib using breast cancer cells by Raman microscopy combined with a deep wavelet scattering-based multivariate analysis framework. This approach discriminated between control cells and drug-treated cells with high accuracy, compared to classical principal component analysis. Both lapatinib and neratinib induced changes in the cellular biochemical composition. Furthermore, the Raman results were compared with the results of several in vitro assays. For instance, drug-treated cells exhibited (i) inhibition of ERK and AKT phosphorylation, (ii) inhibition of cellular proliferation, (iii) cell-cycle arrest, and (iv) apoptosis as indicated by western blotting, real-time cell analysis (RTCA), cell-cycle analysis, and apoptosis assays. Thus, the observed Raman spectral changes are attributed to cell-cycle arrest and apoptosis. The results also indicated that neratinib is more potent than lapatinib. Moreover, the uptake and distribution of lapatinib in cells were visualized through its label-free marker bands in the fingerprint region using Raman spectral imaging. These results show the prospects of Raman microscopy in drug evaluation and presumably in drug discovery.
Insights
Neratinib overcomes HER2 drug resistance more effectively than lapatinib in breast cancer cells. Raman microscopy combined with advanced analysis visualized drug effects and cellular changes, aiding drug evaluation.
Area of Science:
- Biophysics
- Cell Biology
- Cancer Research
Background:
- HER2 is a key therapeutic target in breast cancer, but resistance to drugs like lapatinib limits efficacy.
- Neratinib demonstrates potential to overcome this resistance and inhibit HER2 signaling.
- Novel methods are needed to evaluate drug efficacy and understand resistance mechanisms.
Purpose of the Study:
- To investigate the efficacy of lapatinib and neratinib in breast cancer cells.
- To utilize Raman microscopy and deep wavelet scattering analysis for drug effect assessment.
- To compare the potency of neratinib and lapatinib and visualize drug uptake.
Main Methods:
- Breast cancer cells were treated with lapatinib and neratinib.
- Raman microscopy combined with deep wavelet scattering multivariate analysis was employed.
- Results were validated using western blotting, RTCA, cell-cycle, and apoptosis assays.
Main Results:
- The Raman-based approach accurately discriminated between control and drug-treated cells.
- Both drugs induced significant changes in cellular biochemical composition, linked to cell-cycle arrest and apoptosis.
- Neratinib exhibited greater potency than lapatinib in inhibiting HER2 signaling and tumor growth.
- Raman spectral imaging visualized lapatinib uptake and distribution within cells.
Conclusions:
- Raman microscopy offers a powerful label-free tool for evaluating drug efficacy in cancer cells.
- Neratinib shows promise as a more potent therapeutic agent against HER2-resistant breast cancer.
- The study highlights the potential of Raman spectroscopy in drug discovery and development.
