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. 2025 May 20;6(5):102121.
doi: 10.1016/j.xcrm.2025.102121. Epub 2025 May 6.

Genetic biomarker study of sunvozertinib for clinical prognosis and prediction in NSCLC with EGFR exon 20 insertion mutation

Affiliations

Genetic biomarker study of sunvozertinib for clinical prognosis and prediction in NSCLC with EGFR exon 20 insertion mutation

Yan Xu et al. Cell Rep Med. .

Abstract

This is a report of biomarker analysis for sunvozertinib, a leading epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) targeting EGFR exon 20 insertion mutation (exon20ins) non-small cell lung cancer (NSCLC). There is a positive correlation between positive EGFR exon20ins in plasma circulating tumor DNA (ctDNA) and advanced disease. Shorter progression-free survival and lower objective response rate (45.8% vs. 68.0%) were observed in patients with positive EGFR exon20ins compared to those with negative status. Droplet digital PCR analysis showed that the EGFR exon20ins allele in ctDNA decreased over time in 85.7% of patients, with the earliest clearance occurred after 1 week of sunvozertinib treatment. Acquired EGFR C797S is identified as a potential on-target resistance mutation to sunvozertinib. Finally, efforts are undertaken to investigate therapeutic approaches that aim to overcome the putative acquired resistance to sunvozertinib.

Keywords: EGFR exon20ins; NSCLC; biomarker; sunvozertinib.

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Conflict of interest statement

Declaration of interests Y.X. has received partial research funding from AstraZeneca outside the submitted work. J.C.-H.Y. reports institutional fees from Amgen for advisory works; grants, personal fees, and institutional fee from AstraZeneca for advisory works; institutional fee from Bayer for advisory works; institutional fees from Boehringer Ingelheim for advisory works; institutional fees from Bristol Myers Squibb for advisory works; institutional fee from Daiichi Sankyo for advisory works; institutional fee from Eli Lilly for advisory works; institutional fee from Merck KGaA, Darmstadt, Germany, for advisory works; institutional fee from Merck Sharp & Dohme for advisory works; institutional fee from Novartis for advisory works; institutional fee from Pfizer for advisory works; grants and institutional fee from Roche/Genentech for advisory works; institutional fee and travel fee from Takeda Oncology for advisory works; institutional fee from Yuhan Pharmaceuticals for advisory works; institutional fee from Janssen Pharmaceuticals for advisory works; institutional fee from Gilead Sciences Inc., for advisory works; institutional fee from GSK for advisory works; personal fee from BeiGene for advisory works; institutional fee from Regeneron Pharmaceutical for advisory works; institutional fee from ArriVent for advisory works; institutional fee from AnHeart Therapeutics for advisory works; and travel fee from Dizal Pharmaceuticals to major conference. D.P. reports consulting, advisory role, or lectures: AstraZeneca, AbbVie, Bristol Myers Squibb, Boehringer Ingelheim, Celgene, Daiichi Sankyo, Eli Lilly, Merck, Novartis, Janssen, Pfizer, Roche, Pierre Fabre, Takeda, ArriVent, Mirati, Seagen, and GSK; clinical trial research as a principal investigator or co-investigator (institutional financial interests): AstraZeneca, Bristol Myers Squibb, Boehringer Ingelheim, Eli Lilly, Merck, Novartis, Pfizer, Roche, Medimmune, Sanofi-Aventis, Taiho Pharma, Novocure, Daiichi Sankyo, AbbVie, Janssen, Pierre Fabre, Takeda, ArriVent, Mirati, and Seagen; and travel, accommodations, and expenses: AstraZeneca, Roche, Novartis, and Pfizer. B.P.F. receives research support from Bristol Myers Squibb Foundation, Guardant Health, Bayer, Merck/MSD, Foundation Medicine, Illumina, Regeneron, AstraZeneca, Merus, Gilead, Catalyst, and OncoHost. E.F. reports receipt of personal honoraria for advisory board participation from AbbVie, Amgen, AstraZeneca, Bayer, Boehringer Ingelheim, BMS, Daiichi Sankyo, F. Hoffmann-La Roche, Genmab, Gilead, GSK, ITeos Therapeutics, Janssen, Johnson & Johnson, MSD, Novartis, Pierre Fabre, Pfizer, Regeneron, Turning Point, and Daiichi Sankyo; personal speaker honoraria from Amgen, AstraZeneca, BMS, Daiichi Sankyo, Eli Lilly, F. Hoffmann-La Roche, Johnson & Johnson, Genentech, Gilead, Janssen, Medical Trends, Medscape, Merck Serono, MSD, Novartis, PeerVoice, Pierre Fabre, Pfizer, Regeneron, and Touch Oncology; Board of Director role: Grifols; and financial support for meeting attendance and/or travel from AstraZeneca, Janssen, and Roche. E.F. is a principal investigator in trials (institutional financial support for clinical trials) sponsored by AstraZeneca, AbbVie, Amgen, Bayer, BeiGene, Boehringer Ingelheim, BMS, Daiichi Sankyo, Exelixis, F. Hoffmann-La Roche, Genentech, GSK, Janssen, MSD, Merck KGAA, Mirati, Novartis, Nuvalent, Pfizer, and Takeda. J.B.-B. reports personal fees from advisory boards (MSD and Roche); educational lectures from BMS, AstraZeneca, Pfizer, Takeda, Regeneron, Amgen, Merck, and Sanofi, outside the submitted work; and has received support for attending meetings and/or travel from Takeda, MSD, and Roche. D.R.C.: Ad hoc consulting Dizal. L.B. has received consulting fees from Pfizer, AnHeart, AstraZeneca, Regeneron, Genentech, Janssen, Novocure, Bayer, Daichi, BMS, Sanofi, Gilead, Teligene, BI, BioAtla, and Neuvogen. M.N. is on the advisory board for AstraZeneca, Daiichi Sankyo, Takeda, Novartis, EMD Serono, Janssen, Pfizer, Eli Lilly and Company, Bayer, Regeneron, BMS, and Genentech; a consultant for Caris Life Sciences (virtual tumor board); a speaker for Blueprint Medicines, Janssen, Mirati, and Takeda; reports travel support from AnHeart Therapeutics; and reports stock/stock options from MBrace Therapeutics. X.Z. and L.Z. are employees of Dizal Pharmaceutical and hold stock in Dizal Pharmaceutical. P.A.J.’s institution has received research funding from AstraZeneca, Daiichi Sankyo, PUMA, Eli Lilly, Boehringer Ingelheim, Revolution Medicines, and Takeda Oncology. P.A.J. reports consulting fees from AstraZeneca, Boehringer Ingelheim, Pfizer, Roche/Genentech, Chugai Pharmaceuticals, Eli Lilly pharmaceuticals, SFJ Pharmaceuticals, Voronoi, Daiichi Sankyo, Biocartis, Novartis, Takeda Oncology, Mirati Therapeutics, Transcenta, Silicon Therapeutics, Syndax, Nuvalent, Bayer, Esai, Allorion Therapeutics, Accutar Biotech, AbbVie, Monte Rosa, Scorpion Therapeutics, Merus, Frontier Medicines, Hongyun Biotechnology, Duality, Blueprint Medicines, and Dizal Pharmaceuticals; stock in Gatekeeper Pharmaceuticals; and a patent for EGFR mutations issued, licensed, and with royalties paid by LabCorp.

Figures

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Graphical abstract
Figure 1
Figure 1
Correlations between EGFR exon20ins in baseline plasma ctDNA and the number of metastatic sites, brain metastasis, and tumor response with sunvozertinib (A) Metastatic sites/lesions of patients with negative or positive EGFR exon20ins in ctDNA. (B) Sum of tumor diameters of patients with negative or positive EGFR exon20ins in ctDNA. (C) EGFR exon20ins abundance in patients with low number (<3) versus high number (≥3) of metastatic sites/lesions. (D) EGFR exon20ins abundance in patients with or without baseline brain metastasis. (E) ORRs in patients with negative or positive EGFR exon20ins in ctDNA. (F) Kaplan-Meier analysis of PFS according to EGFR exon20ins status in ctDNA. EGFR exon20ins status (positive or negative) was defined by NGS. EGFR exon20ins status was unknown due to too low DNA quantity or not achieving sufficient unique sequencing depths as required by NGS. (G) EGFR exon20ins abundance in patients with different tumor responses. One patient non-evaluable for tumor response was not included in the analysis. (H) ORRs in patients with low or high abundance of EGFR exon20ins in ctDNA. (I) Kaplan-Meier analysis of PFS according to EGFR exon20ins abundance in ctDNA. The Mann-Whitney test was used for the analysis in (A), (B), (C), (D), and (G). Fisher’s exact test was used for the analysis in (E) and (H). Significance was established when the p value was less than 0.05. All tests were two-sided. No further pairwise comparison was performed in (G) given the overall test was not significant. The heavy lines in (A) and (B) represent the median numbers of metastatic sites/lesions and median sums of the longest diameters of the target lesions in each group, respectively. The heavy lines in (C), (D), and (G) represent the median values of EGFR exon20ins abundance in each group. In (E)–(I), efficacy data at 300 mg were used for the analysis. In (F) and (I), median PFS was estimated by Kaplan-Meier method with 95% CIs. In (H) and (I), the median value of EGFR exon20ins abundance (3.7%) was used as a cutoff value to divide patients into two groups: low abundance (<3.7%) and high abundance (≥3.7%). CI, confidence interval; ctDNA, circulation tumor DNA; EGFR, epidermal growth factor receptor; exon20ins, exon 20 insertion mutation; mos, months; mPFS, median progression-free survival; NE, not evaluable; NGS, next-generation sequencing; ORR, objective response rate; PD, progressive disease; PFS, progression-free survival; PR, partial response; SD, stable disease.
Figure 2
Figure 2
Dynamic changes of EGFR exon20ins DNA copy number in plasma ctDNA with sunvozertinib treatment (A) Correlation between mutant allele frequency (MAF) of EGFR exon20ins in baseline plasma ctDNA samples tested by next-generation sequencing (NGS) and droplet digital PCR (ddPCR). Among the 34 patients with available MAF of EGFR exon20ins tested by ddPCR, seven patients did not have MAF of EGFR exon20ins tested by NGS due to too low DNA quantity or not achieving sufficient unique sequencing depths as required by NGS. Thus, MAF of EGFR exon20ins in baseline plasma ctDNA of 27 patients tested by NGS and ddPCR was used for the analysis. Each dot represents the MAF of EGFR exon20ins tested by NGS and ddPCR. Pearson’s correlation coefficient was applied for the analysis. (B) Dynamic changes of EGFR exon20ins DNA copy number with sunvozertinib treatment at 300 mg, and association with tumor response. ddPCR, droplet digital PCR; EGFR, epidermal growth factor receptor; exon20ins, exon 20 insertion mutation; MAF, mutant allele frequency; NGS, next-generation sequencing; PD, progressive disease; PR, partial response; SD, stable disease.
Figure 3
Figure 3
Potential genetic resistance mechanism of sunvozertinib (A) Genetic characteristics potentially related to resistance to sunvozertinib by next-generation sequencing. (B–D) In the index cases (subjects #001, 007, and 023), EGFR C797S mutation was identified at disease progression or time point around disease progression confirmed by image scans. Overlapping reads spanning EGFR exon20ins location and C797 contained both exon20ins and C797S mutations, indicating that the two mutations occurred in cis on the same allele. (E–G) Longitudinal monitoring of EGFR exon20ins and C797S mutation during the treatment by using droplet digital PCR. (H) Anti-proliferative effect of sunvozertinib in Ba/F3 cells expressing EGFR exon20ins SVD or SVD-C797S double-mutant protein. (I) Inhibition of pEGFR pathway with sunvozertinib in Ba/F3 cells expressing EGFR exon20ins SVD-C797S double-mutant protein. (J) Anti-proliferation activity of sunvozertinib or BDTX-1535 on KLN205 cells engineered with EGFR exon20ins SVD-C797S double mutations. BL, baseline; EGFR, epidermal growth factor receptor; exon20ins, exon 20 insertion mutation; PD, progressive disease; PR, partial response; SD, stable disease; SVD, D770_N771insSVD.
Figure 4
Figure 4
Antitumor activity of a JAK inhibitor golidocitinib in combination with chemotherapy in a xenograft model expressing EGFR exon20ins SVD-C797S (A) Tumor growth inhibition of a xenograft model expressing EGFR with exon20ins SVD and C797S by different treatments. Error bars represent the standard error of mean of the individual means. Two-way ANOVA analysis was used to compare treatment groups to vehicle control group. ∗∗p < 0.01; ∗∗∗p < 0.0005; ∗∗∗∗p < 0.0001. (B) pSTAT3 signals and (C) EGFR expression in tumor tissues post treatment. The tumor tissues from each treatment were collected at 2 h post the last dose of treatment (three mice per group). Error bars represent the standard deviation of individual means. Statistical analysis was performed using one-way ANOVA with Dunnett test. ∗∗p < 0.01; ns, not significant. Bid, twice daily; biw, twice weekly; EGFR, epidermal growth factor receptor; exon20ins, exon 20 insertion mutation; STAT3, signal transducer and activator of transcription 3; SVD, D770_N771insSVD; i.p., intraperitoneally; p.o., orally.

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