Fukuda Koji, Nanjo Shigeki, Takeuchi Shinji, Chakrabarti Turja, Brown Tyiesha, Dev Sahadevan Sharon Wesley, Arai Sachiko, Sato Shigeki, Kotani Hiroshi, Nishiyama Akihiro, Sakaguchi Hiroyuki, Ohtsubo Koushiro, Taniguchi Hiroaki, Blakely Collin M, Bivona Trever G, Yano Seiji
Division of Innovative Cancer Control Research, Cancer Research Institute, Kanazawa University, Kanazawa, Japan; Nano Life Science Institute, Kanazawa University, Kanazawa, Japan.
Department of Respiratory Medicine, Faculty of Medicine, Institute of Medical, Pharmaceutical, and Health Sciences, Kanazawa University, Kanazawa, Japan.
J Thorac Oncol. 2025 Jun 13. doi: 10.1016/j.jtho.2025.06.007.
Osimertinib, a third-generation EGFR tyrosine kinase inhibitor (EGFR-TKI), is commonly used as a first-line treatment for EGFR-mutant NSCLC. Nevertheless, despite its efficacy, resistance remains a major clinical challenge with unknown underlying mechanisms. This study aimed to investigate the mechanisms driving osimertinib resistance and identify therapeutic strategies.
Using a mouse model of leptomeningeal carcinomatosis, we induced osimertinib resistance and performed next-generation sequencing to characterize resistance-associated mutations. We also analyzed clinical samples to correlate ARID1A status with progression-free survival and overall survival in patients receiving osimertinib.
Mutations in the AT-rich interacting domain-containing protein 1A (ARID1A) gene were the most prevalent in resistant cells. Functional assays revealed that ARID1A knockout in parental cells and wild-type ARID1A gene expression in resistant cells were critical in conferring osimertinib resistance. A Clustered Regularly Interspaced Short Palindromic Repeats-Cas9 knockout screen identified WEE1 kinase as a potent enhancer of apoptosis in ARID1A-mutant osimertinib-resistant cells. Mechanistically, ARID1A-mutant cells exhibited reduced expression of genes involved in cell cycle regulation and DNA repair, rendering them particularly sensitive to WEE1 inhibition. In the leptomeningeal carcinomatosis mouse model, the combined inhibition of EGFR and WEE1 significantly suppressed tumor growth. Clinically, patients with ARID1A mutations treated with osimertinib had significantly shorter median progression-free survival (6.25 versus 18.0 months, p = 0.0036) and overall survival (17.0 versus 34.0 months, p = 0.024) than did those with wild-type ARID1A.
These findings suggest that ARID1A mutations are critical biomarkers for osimertinib resistance and highlight WEE1 inhibition as a promising therapeutic approach for ARID1A-mutant osimertinib-resistant NSCLC.
奥希替尼是一种第三代表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI），常用于表皮生长因子受体（EGFR）突变的非小细胞肺癌（NSCLC）的一线治疗。然而，尽管其疗效显著，但耐药性仍然是一个主要的临床挑战，其潜在机制尚不清楚。本研究旨在探究导致奥希替尼耐药的机制，并确定治疗策略。
利用软脑膜癌病小鼠模型，诱导奥希替尼耐药，并进行二代测序以鉴定与耐药相关的突变。我们还分析了临床样本，以将富含AT区相互作用蛋白1A（ARID1A）的状态与接受奥希替尼治疗的患者的无进展生存期和总生存期相关联。
富含AT区相互作用蛋白1A（ARID1A）基因的突变在耐药细胞中最为常见。功能分析表明，亲本细胞中的ARID1A基因敲除和耐药细胞中的野生型ARID1A基因表达对于赋予奥希替尼耐药性至关重要。一项成簇规律间隔短回文重复序列-Cas9基因敲除筛选确定WEE1激酶是ARID1A突变的奥希替尼耐药细胞中凋亡的有效增强剂。从机制上讲，ARID1A突变细胞中参与细胞周期调控和DNA修复的基因表达降低，使其对WEE1抑制特别敏感。在软脑膜癌病小鼠模型中，EGFR和WEE1的联合抑制显著抑制了肿瘤生长。临床上，接受奥希替尼治疗的ARID1A突变患者的中位无进展生存期（6.25个月对18.0个月，p = 0.0036）和总生存期（17.0个月对34.0个月，p = 0.024）明显短于野生型ARID1A患者。
这些发现表明，ARID1A突变是奥希替尼耐药的关键生物标志物，并突出了WEE1抑制作为ARID1A突变的奥希替尼耐药NSCLC的一种有前景的治疗方法。
