200P - CXCR1+ neutrophil infiltration orchestrates response to third-generation EGFR-TKI in EGFR mutant NSCLC

Background

Although third-generation EGFR-TKI is standard of care for patients with EGFR-mutant NSCLC, little is known about the predictors of response or resistance.

Methods

Here, we integrated single-cell RNA (scRNA) sequencing, bulk RNA sequencing, multiplexed immunofluorescence and flow cytometry data from pretreatment and post-resistant tumor samples of EGFR-mutant NSCLC patients received third-generation EGFR-TKIs.

Results

We show that resistant samples had a markedly enriched CXCR1⁺ neutrophils infiltration (P<0.01) than pretreatment samples, which were distinguished from other subtypes of neutrophils and displayed immunsupressive characteristics. Spatial analysis showed that increased CXCR1⁺ neutrophils predominantly infiltrated into the tumor core in resistant samples and the average distance of neutrophils to tumor cells markedly reduced from 33 to 19 μm. Deep analysis of scRNA and bulk RNA sequencing data revealed the increased interactions between CXCR1⁺ neutrophils and tumor cells and activated TNF-α/NF-κB signaling pathway in tumor cells of resistant samples. In vitro and in vivo experiments validated that CXCR1⁺ neutrophils resulted in resistance to third-generation EGFR-TKI via activating TNF-α/NF-κB signaling pathway in tumor cells. Importantly, patients with low pretreatment CXCR1⁺ neutrophil infiltration abundance had a dramatically longer progression-free survival (11.8 vs. 7.5 months; P=0.019) and overall survival (33.0 vs. 23.5 months; P=0.029) than those with high infiltration abundance.

Conclusions

In summary, the current findings suggest that CXCR1⁺ neutrophil infiltration in pretreatment tumor tissues was associated with the efficacy of third-generation EGFR-TKI and increased CXCR1⁺ neutrophil infiltration in posttreatment tumor tissues played a role in mediating the resistance to third-generation EGFR-TKI in EGFR-mutant NSCLC patients, underscoring the clinical significance of CXCR1⁺ neutrophils within the tumor microenvironment. Our findings may aid in identifying patients more likely to benefit from third-generation EGFR-TKI treatment.

Legal entity responsible for the study

The authors.

Funding

This study was supported in part by grants from the National Natural Science Foundation of China (No. 82102859, 82272875 and 12126605).

Disclosure

All authors have declared no conflicts of interest.