Live-cell FRET imaging-based efficacy prediction of EGFR-TKIs in EGFR L858R-mutant NSCLC.
- 1Department of Pain Management, the First Affiliated Hospital of Jinan University, Guangzhou, China.
- 2Department of Thoracic Surgery, The First Affiliated Hospital of Jinan University, Guangzhou, China.
- 3MOE Key Laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China.
Abstract
Objectives
Functional assays can assist individualised selection of treatment regimens by evaluating the ex vivo response of tumour cells to anticancer therapeutics. However, the two major strategies used to assess responses to targeted drugs, target-based profiling and phenotypic profiling, have inherent limitations. Target-based profiling does not directly capture downstream cell-fate outcomes, whereas phenotypic profiling provides limited mechanistic information on drug action.Methods
Live-cell fluorescence resonance energy transfer (FRET) microscopy was used to integrate the interaction between EGFRL858R and GRB2 with cellular phenotypic features, thereby predicting the efficacy of EGFR-tyrosine kinase inhibitors (TKIs) at the single-cell level.Results
The target-based drug score (T-score), derived from the interaction between EGFRL858R and GRB2 in living cells co-expressing CFP-EGFRL858R and YFP-GRB2, enabled evaluation of the on-target activity of EGFR-TKIs against EGFRL858R. The phenotype drug score (P-score), constructed from cellular phenotype features, characterised the phenotypic responses induced by EGFR-TKIs. The TP-score, integrating the T- and P-scores, accurately predicted the efficacy of EGFR-TKIs. In living H3255 cells, the TP-score of osimertinib (OSI) was higher than that of gefitinib (GEF). Notably, in H3255 cell-derived xenograft (CDX) mouse models, OSI showed superior in vivo antitumour efficacy compared with GEF. Furthermore, live-cell TP scoring revealed that primary tumour cells from a patient with NSCLC did not respond to OSI, a finding consistent with the absence of actionable EGFR mutations in this patient.Conclusions
This study demonstrates the clinical feasibility of predicting EGFR-TKI efficacy by integrating EGFRL858R-GRB2 interactions with cellular phenotypic features in single living cells.
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BibTeX
@article{li2026livecell,
title = {{Live-cell FRET imaging-based efficacy prediction of EGFR-TKIs in EGFR L858R-mutant NSCLC.}},
author = {Xin Li and Lingao Dai and Pingshan Yang and Shuanghe Li and Chuan Peng and Shiyao Li and Beini Sun and Songwang Cai and Xiaoping Wang and Tongsheng Chen},
journal = {Clinical and translational medicine},
year = {2026},
month = oct,
doi = {10.1002/ctm2.70878},
url = {https://europepmc.org/article/MED/42926325},
note = {CC BY},
}
RIS
TY - JOUR
TI - Live-cell FRET imaging-based efficacy prediction of EGFR-TKIs in EGFR L858R-mutant NSCLC.
AU - Xin Li
AU - Lingao Dai
AU - Pingshan Yang
AU - Shuanghe Li
AU - Chuan Peng
AU - Shiyao Li
AU - Beini Sun
AU - Songwang Cai
AU - Xiaoping Wang
AU - Tongsheng Chen
PY - 2026
DA - 2026/10/01
JO - Clinical and translational medicine
DO - 10.1002/ctm2.70878
UR - https://europepmc.org/article/MED/42926325
AB - Objectives Functional assays can assist individualised selection of treatment regimens by evaluating the ex vivo response of tumour cells to anticancer therapeutics. However, the two major strategies used to assess responses to targeted drugs, target-based profiling and phenotypic profiling, have inherent limitations. Target-based profiling does not directly capture downstream cell-fate outcomes, whereas phenotypic profiling provides limited mechanistic information on drug action.Methods Live-cell fluorescence resonance energy transfer (FRET) microscopy was used to integrate the interaction between EGFRL858R and GRB2 with cellular phenotypic features, thereby predicting the efficacy of EGFR-tyrosine kinase inhibitors (TKIs) at the single-cell level.Results The target-based drug score (T-score), derived from the interaction between EGFRL858R and GRB2 in living cells co-expressing CFP-EGFRL858R and YFP-GRB2, enabled evaluation of the on-target activity of EGFR-TKIs against EGFRL858R. The phenotype drug score (P-score), constructed from cellular phenotype features, characterised the phenotypic responses induced by EGFR-TKIs. The TP-score, integrating the T- and P-scores, accurately predicted the efficacy of EGFR-TKIs. In living H3255 cells, the TP-score of osimertinib (OSI) was higher than that of gefitinib (GEF). Notably, in H3255 cell-derived xenograft (CDX) mouse models, OSI showed superior in vivo antitumour efficacy compared with GEF. Furthermore, live-cell TP scoring revealed that primary tumour cells from a patient with NSCLC did not respond to OSI, a finding consistent with the absence of actionable EGFR mutations in this patient.Conclusions This study demonstrates the clinical feasibility of predicting EGFR-TKI efficacy by integrating EGFRL858R-GRB2 interactions with cellular phenotypic features in single living cells.
SN - 2001-1326
ER -