Related Experiment Video

Updated: Sep 10, 2025

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

Published on: August 11, 2017

Multiomics and Single-Cell Insights Reveal a Lysophosphatidic Acid-Mediated Resistant Mechanism to Third-generation

Ruyun Gao¹, Ning Lou², Sheng Yang¹

¹Department of Medical Oncology, Beijing Key Laboratory of Key Technologies for Early Clinical Trial Evaluation of Innovative Drugs for Major Diseases, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research

|August 25, 2025

Abstract

Purpose:

Third-generation EGFR tyrosine kinase inhibitors (TKI) have revolutionized the treatment of EGFR-mutant non-small cell lung cancer (NSCLC). However, acquired resistance remains a significant challenge. This study investigates the metabolic mechanisms driving third-generation EGFR-TKI resistance.

Experimental Design:

We conducted plasma metabolomics analysis on 216 longitudinal samples from 186 patients with NSCLC enrolled in the clinical trial of rezivertinib (NCT03386955). Additionally, multiomics profiling of rezivertinib-resistant cell lines, functional in vitro experiments, and single-cell RNA sequencing analyses of 215 patients with NSCLC were integrated to reveal underlying mechanisms.

Results:

Nonresponder patients exhibited elevated glycerophospholipids and dysregulated lysophospholipid (LPL) metabolism. Unsupervised clustering identified two patient subgroups, with cluster 1 (characterized by high LPL levels) associated with poorer survival (P = 0.022). A metabolite-based predictive model achieved robust performance [AUC: 0.7762 (training) and 0.7485 (test)]. Longitudinal analyses demonstrated LPLs and lysophosphatidic acid (LPA) accumulation during the resistance process. Integrated multiomics analyses highlighted epithelial-mesenchymal transition and glycerophospholipid reprogramming in rezivertinib-resistant cells. Functional assays confirmed that LPA promoted cell migration and invasion and attenuated the efficacy of third-generation EGFR-TKI, whereas disruption of the LPA-LPA receptor signaling axis reversed LPA-mediated resistance. Single-cell RNA sequencing identified an LPA-secreting malignant subset (cluster c4), characterized by enhanced epithelial-mesenchymal transition activation and extensive microenvironmental cross-talk through Wnt, TGF-β, and extracellular matrix signals.

Conclusions:

Our study highlights the pivotal role of LPA-mediated signaling and metabolic reprogramming in third-generation EGFR-TKI resistance. Targeting LPA production or its downstream pathways may offer novel therapeutic strategies to overcome resistance. This study provides critical metabolic insights for managing EGFR-mutant NSCLC.

Insights

Lysophosphatidic acid (LPA) metabolism drives resistance to third-generation EGFR tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Targeting LPA pathways may overcome this resistance, offering new therapeutic strategies for NSCLC patients.

Area of Science:

Oncology
Metabolomics
Molecular Biology

Background:

Third-generation EGFR TKIs have improved NSCLC treatment.
Acquired resistance to these TKIs remains a significant clinical challenge.
Understanding resistance mechanisms is crucial for developing effective therapies.

Purpose of the Study:

To investigate the metabolic mechanisms underlying resistance to third-generation EGFR TKIs.
To identify key metabolic pathways and biomarkers associated with treatment resistance.

Main Methods:

Plasma metabolomics on 216 longitudinal samples from 186 NSCLC patients.
Multi-omics profiling of resistant cell lines.
Functional in vitro assays and single-cell RNA sequencing (scRNA-seq).

Main Results:

Elevated glycerophospholipids and dysregulated lysophospholipid (LPL) metabolism were observed in non-responders.
High LPL levels correlated with poorer survival.
LPA accumulation and epithelial-mesenchymal transition (EMT) were identified in resistant cells.
LPA promoted cell migration, invasion, and TKI resistance; targeting LPA-LPAR reversed resistance.
scRNA-seq revealed an LPA-secreting malignant subset with activated EMT and microenvironmental crosstalk.

Conclusions:

LPA-mediated signaling and metabolic reprogramming are pivotal in third-generation EGFR-TKI resistance.
Targeting LPA production or its signaling pathways presents a novel therapeutic strategy.
These findings offer critical metabolic insights for managing EGFR-mutant NSCLC resistance.

Read full analysis

More Related Videos

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

Published on: June 26, 2019

Patient-Derived Tumor Explants As a "Live" Preclinical Platform for Predicting Drug Resistance in Patients

Patient-Derived Tumor Explants As a "Live" Preclinical Platform for Predicting Drug Resistance in Patients

Published on: February 7, 2021

Related Concept Videos

Treatment Resistant Cancers

Treatment Resistant Cancers

Cancer is the second leading cause of death in the United States. A cancer cell is genetically unstable and hence can mutate faster. They can also modify their microenvironment and escape immune surveillance. The difficulties in treating cancer are further compounded by the emergence of rapid resistance to anticancer drugs. The most common ways to attain resistance in cancer cells include alteration in drug transport and metabolism, modification of drug target, elevated DNA damage response, or...

Targeted Cancer Therapies

Targeted Cancer Therapies

The targeted cancer therapies, also known as “molecular targeted therapies,” take advantage of the molecular and genetic differences between the cancer cells and the normal cells. It needs a thorough understanding of the cancer cells to develop drugs that can target specific molecular aspects that drive the growth, progression, and spread of cancer cells without affecting the growth and survival of other normal cells in the body.
There are several types of targeted therapies against...

Mitogens and the Cell Cycle

Mitogens and the Cell Cycle

Mitogens and their receptors play a crucial role in controlling the progression of the cell cycle. However, the loss of mitogenic control over cell division leads to tumor formation. Therefore, mitogens and mitogen receptors play an important role in cancer research. For instance, the epidermal growth factor (EGF) - a type of mitogen and its transmembrane receptor (EGFR), decides the fate of the cell's proliferation. When EGF binds to EGFR, a member of the ErbB family of tyrosine kinase...

Articles linked to this work by shared authors, journal, and citation graph.

Safety, efficacy, and pharmacokinetics of eratrectinib (VC004) in solid tumors with NTRK fusions: phase 1 results from a phase 1/2 study.

Experimental hematology & oncology·2026

Serplulimab Plus Chemotherapy, with or without HLX04, versus Chemotherapy as First-Line Treatment for Nonsquamous NSCLC: Final Survival Analysis of the Phase III ASTRUM-002 Study.

Cancer communications (London, England)·2026

Rezivertinib in EGFR-Mutated Non-Small Cell Lung Cancer Patients with Central Nervous System Metastasis: Central Nervous System Efficacy from the Phase III REZOR Study.

Cancer communications (London, England)·2026

Digital spatial profiling uncovers transcriptomic features of distinct plasma cell-like phenotypes in diffuse large B-cell lymphoma.

Blood advances·2026

DSG2+ Cancer Stem Cells Co-Located With FAP+ Myofibroblasts in the Tumor Boundary That Determines the Efficacy of Immunotherapy in Non-Small Cell Lung Cancer.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)·2026

Efficacy and Safety of Afatinib Plus Bevacizumab as First-Line Treatment for Advanced NSCLC Patients With Epidermal Growth Factor Receptor (EGFR) Mutations: A Multicenter, Phase II Trial.

Thoracic cancer·2025

See all related articles