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Updated: Jan 11, 2026
Profiling Sensitivity to Targeted Therapies in EGFR-Mutant NSCLC Patient-Derived Organoids
Published on: November 22, 2021
Single-cell RNA profiling reveals an immunosuppressive microenvironment in EGFR double-mutant non-small cell lung
Jiao Yang1, Jun-Wei Su1, Hong-Rui Li2,3,4
1Guangdong Lung Cancer Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, China.
Background:
Non-small cell lung cancer (NSCLC) patients with double-driver gene mutations are reported with poorer survival outcomes and reduced therapy responses compared to single-mutant (SM) patients. While substantial progress has been made in treating cancers with single-driver gene alterations, the therapeutic implications and tumor microenvironment of double-mutant (DM) NSCLC remain largely unexplored because of its rarity and poor prognosis. This study aims to delineate the landscape of the immune microenvironment within DM NSCLC.
Methods:
We employed single-cell RNA sequencing (scRNA-seq) to generate a comprehensive transcriptomic atlas from 25 EGFR mutant NSCLC samples. To assess immune microenvironment changes over time, time-series scRNA-seq and multiplex immunofluorescence analyses were performed in two DM patients. In vitro, EGFR-mutant cell lines were constructed to assess potential therapeutic responses for future clinical applications.
Results:
The scRNA-seq platform scFocuSCOPE accurately identified and characterized rare, mutation-bearing cancer cells at the single-cell level. DM cancer cells exhibited a strong tendency toward angiogenesis, suggesting an invasive phenotype. DM patients had a more suppressed immune microenvironment, with fewer dysfunctional T lymphocytes. The observation of fewer immune cells and high programmed death ligand-1 (PD-L1) expression in DM cases, probably related to immune evasion and poorer prognosis. In one DM patient, PD-L1 expression remained unchanged after targeted therapy but decreased after immunotherapy. In vitro, EGFR/ERBB2 DM cells showed greater sensitivity to dual-targeted therapies than to single-agent treatments.
Conclusions:
scFocuSCOPE precisely delineated tumor heterogeneity and immune suppression in EGFR-DM NSCLC. The complex immune landscape of EGFR-DM tumors offers valuable insights for future mechanistic studies and personalized therapies.
Insights
Double-mutant non-small cell lung cancer (NSCLC) exhibits a suppressed immune microenvironment and invasive characteristics. This study reveals insights into immune evasion and potential dual-targeted therapies for double-mutant NSCLC patients.
Area of Science:
- Oncology
- Immunology
- Genomics
Background:
- Double-mutant (DM) non-small cell lung cancer (NSCLC) presents poorer prognoses and reduced treatment efficacy compared to single-mutant (SM) NSCLC.
- The tumor microenvironment and therapeutic strategies for DM NSCLC remain understudied due to rarity and poor outcomes.
Purpose of the Study:
- To characterize the immune microenvironment of DM NSCLC.
- To explore potential therapeutic avenues for DM NSCLC.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) was utilized to analyze 25 EGFR-mutant NSCLC samples.
- Time-series scRNA-seq and multiplex immunofluorescence were performed on DM patients.
- In vitro EGFR-mutant cell lines were established for therapeutic response assessment.
Main Results:
- scFocuSCOPE platform identified rare, mutation-bearing cancer cells.
- DM cancer cells displayed increased angiogenesis and an invasive phenotype.
- DM NSCLC showed a suppressed immune microenvironment with fewer T lymphocytes and high PD-L1 expression, suggesting immune evasion.
Conclusions:
- scFocuSCOPE accurately identified tumor heterogeneity and immune suppression in EGFR-DM NSCLC.
- The findings provide insights into the complex immune landscape of DM NSCLC, informing future research and personalized treatment strategies.
