PO.BCS01.03 · 生物信息与计算
剖析EGFR突变型非小细胞肺癌新辅助奥希替尼治疗后耐药性发展的机制
Dissecting the mechanisms of drug resistance development post neoadjuvant osimertinib treatment in EGFR-mutant non-small lung cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：尽管EGFR突变型非小细胞肺癌（NSCLC）接受奥希替尼治疗，药物耐受性残留细胞（DTPs）仍通过可逆性适应而存活。早期干预可能是延迟或预防奥希替尼获得性耐药出现的有效策略。在本研究中，我们通过单细胞RNA测序表征了在新辅助奥希替尼治疗后存活的DTPs，并识别出处于获得耐药性之前早期过渡阶段的DTPs。
方法：新辅助奥希替尼每日给药，进行两个28天周期，随后行手术切除，并进行为期3年的辅助奥希替尼治疗。我们对17例携带EGFR突变（E19del，N=7；L858R，N=10）的NSCLC患者的配对治疗前和治疗后样本进行了单细胞RNA测序。分析基于Python Scanpy软件包进行。
结果：肿瘤上皮细胞被划分为11个亚簇。基于模块、特征和调控子活性的增加，我们对其进行了亚簇划分和相应注释。高DTP特征包括Multi Adaptive（多重适应）、EMT、Immune Evasion（免疫逃逸）、ERS/UPR、Cycling（周期）、AT1和Hyper Ciliated（高度纤毛化）簇。AT2、Chromatin Accessible（染色质可及）、Intermediate（中间）和Detox Ciliated（解毒纤毛化）簇表现出较弱的DTP特征。新辅助奥希替尼治疗后，Multi Adaptive、ERS/UPR和EMT簇的比例显著增加。Multi Adaptive簇具有最高的DTP特征，而Immune Evasion簇则表达已知招募免疫抑制细胞的基因。ERS/UPR簇显示XBP1调控子活性以及内质网应激和未折叠蛋白反应特征的增加。Cycling簇的特征是E2F调控子活性增加，伴随增殖基因上调。AT1簇具有强烈的Wnt信号传导和TEAD4调控子活性增加。Hyper Ciliated簇显示纤毛发生基因表达增加。轨迹分析显示，普遍表达肺泡标志物的EMT、Immune Evasion、Cycling和AT1簇起源于AT2，然后在Intermediate簇分化为三个谱系。基因表达与拟时序的关联分析显示，I型干扰素和TGFbeta通路在所有谱系中均显著增强。
结论：我们的分析揭示了EGFR突变型NSCLC中DTPs群体的多样性和异质性，以及新辅助奥希替尼治疗后DTPs的演化轨迹。靶向这些DTPs特征可能有望应用于开发新型治疗策略，以克服对奥希替尼的耐药性。
查看英文原文 English abstract
Introduction: Despite treatment with osimertinib in EGFR -mutant non-small cell lung cancer (NSCLC), drug-tolerant persisters (DTPs) survive through reversible adaptations. Early intervention may be an effective strategy to delay or prevent the emergence of acquired resistance to osimertinib. In this study, we characterized DTPs that survived neoadjuvant osimertinib treatment via single-cell RNA sequencing, and identified DTPs at an early transition stage before they acquire resistance.
Methods: Neoadjuvant osimertinib was administered daily for two 28-day cycles followed by surgical resection and adjuvant osimertinib for 3 years. We performed single-cell RNA sequencing on matched pre- and post- treatment samples from 17 patients with NSCLC harboring EGFR mutations (E19del, N=7; L858R, N=10). The analysis was performed based on the Python Scanpy package.
Results: Tumor epithelial cells were divided into 11 subclusters. Based on the increase in modules, signatures, and regulon activity, we subclustered and annotated them accordingly. High DTP features included the Multi Adaptive, EMT, Immune Evasion, ERS/UPR, Cycling, AT1, and Hyper Ciliated clusters. The AT2, Chromatin Accessible, Intermediate, and Detox Ciliated clusters exhibited weak DTP features. After neoadjuvant osimertinib, the proportion of Multi Adaptive, ERS/UPR, and EMT clusters were significantly increased. The Multi Adaptive cluster had the highest DTP signature whereas the Immune Evasion cluster expressed genes known for recruiting immunosuppressive cells. The ERS/UPR cluster showed increase in XBP1 regulon activity as well as endoplasmic reticulum stress and unfolded protein response signature. The Cycling cluster was characterized by increased activity of E2F regulons with upregulated proliferation genes. The AT1 cluster had strong Wnt signaling and increased TEAD4 regulon activity. The Hyper Ciliated cluster showed increased expression of ciliogenesis genes. Trajectory analysis revealed that EMT, Immune Evasion, Cycling, and AT1 clusters which commonly express alveolar markers, originated from AT2 and then diverge into three lineages at the Intermediate cluster. Association analysis of gene expression with pseudotime revealed that type I interferon and TGFbeta pathways were significantly augmented in all lineages.
Conclusion: Our analysis shows the diversity and heterogeneity of the DTPs population and the evolutionary trajectories of DTPs after neoadjuvant osimertinib treatment in EGFR -mutant NSCLC. Targeting these DTPs features may potentially be applied for development of novel therapeutic strategies to overcome resistance to osimertinib.
利益披露 Disclosure
S. Choi, None..
J. Lee, None..
H. Jee, None..
H. Shim, None..
B. Park, None..
C. Lee, None..
M. Hong, None..
B. Cho, None..
S. Lim, None.