Wu Jianzhuang, Xie Kexin, Zhang Yixuan, Zhang Weiyi, Cheng Rongjie, Zhang Yaliang, Xia Yugui, Liu Tongyan, Yin Rong, Qiu Yudong, Xu Tao, Li Rutian, Sun Qi, Yan Chao
Department of Pancreatic and Metabolic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China.
J Exp Clin Cancer Res. 2025 Mar 25;44(1):106. doi: 10.1186/s13046-025-03369-9.
KRAS is the most frequently mutated oncogene in human cancers, with KRAS being a prevalent driver mutation in 12-13% non-small cell lung cancer (NSCLC) cases. Despite breakthroughs in KRAS inhibitors such as sotorasib (AMG-510) and adagrasib (MRTX-849), clinical resistance remains a challenging issue, highlighting the need for deeper understanding of the molecular mechanisms underlying KRAS-driven oncogenic signaling in NSCLC. Previously, we identified RASON as a novel regulator of KRAS signaling in pancreatic cancer. Herein, we aim to explore the role of RASON in KRAS-driven NSCLC and its therapeutic potential.
Immunohistochemistry analysis of NSCLC patient cohorts was performed to demonstrate the correlation between RASON expression and NSCLC progression. Immunoblotting was performed to evaluate the effects of RASON on KRAS downstream signaling. In vitro and in vivo assays including cell proliferation, sphere formation, tumor implantation and genetic mouse models were performed to determine the oncogenic role of RASON. RNA-seq analysis was utilized to identify the key signaling pathway regulated by RASON. Immunofluorescence, immunoprecipitation, nuclear magnetic resonance and biochemistry assays were used to validate the interaction between KRAS and RASON. Phagocytosis assay and flow cytometry were conducted to explore the effects of RASON on the tumor immune microenvironment. Pharmacological inhibition in subcutaneous xenograft model was used to determine the therapeutical potential of RASON.
RASON is overexpressed in NSCLC with KRAS mutation and correlates with poor patient prognosis. Genetic knockout of RASON significantly reduced lung tumor burden in LSL-KRAS; Trp53 mice. In KRAS-mutant lung cancer cell lines, RASON overexpression enhanced, while CRISPR-mediated knockout suppressed, both in vitro proliferation and in vivo tumor growth. Mechanistically, RASON directly binds KRAS, stabilizes it in the GTP-bound hyperactive state and promotes downstream signaling. RASON knockout significantly reduced CD47 expression, enhancing macrophage-mediated phagocytosis and anti-tumor immunity. Therapeutically, antisense oligonucleotides targeting RASON not only exhibited tumor-suppressive effects, but also synergized with the KRAS inhibitor AMG-510 to significantly enhance anti-tumor efficacy.
This study reveals RASON as a key oncogenic regulator of KRAS signaling, driving lung tumorigenesis and progression, and identifies RASON as a promising therapeutic target for KRAS mutant non-small cell lung cancer.
KRAS是人类癌症中最常发生突变的致癌基因，在12% - 13%的非小细胞肺癌（NSCLC）病例中，KRAS是一种常见的驱动突变。尽管在KRAS抑制剂如索托拉西布（AMG - 510）和阿达格拉西布（MRTX - 849）方面取得了突破，但临床耐药性仍然是一个具有挑战性的问题，这凸显了深入了解NSCLC中KRAS驱动的致癌信号传导分子机制的必要性。此前，我们在胰腺癌中鉴定出RASON是KRAS信号传导的一种新型调节因子。在此，我们旨在探讨RASON在KRAS驱动的NSCLC中的作用及其治疗潜力。
对NSCLC患者队列进行免疫组织化学分析，以证明RASON表达与NSCLC进展之间的相关性。进行免疫印迹以评估RASON对KRAS下游信号传导的影响。进行包括细胞增殖、球体形成、肿瘤植入和基因小鼠模型在内的体外和体内实验，以确定RASON的致癌作用。利用RNA测序分析来鉴定由RASON调节的关键信号通路。使用免疫荧光、免疫沉淀、核磁共振和生化分析来验证KRAS与RASON之间的相互作用。进行吞噬作用分析和流式细胞术以探讨RASON对肿瘤免疫微环境的影响。在皮下异种移植模型中进行药理抑制，以确定RASON的治疗潜力。
RASON在具有KRAS突变的NSCLC中过表达，并且与患者预后不良相关。RASON基因敲除显著降低了LSL - KRAS；Trp53小鼠的肺肿瘤负荷。在KRAS突变的肺癌细胞系中，RASON过表达增强了体外增殖和体内肿瘤生长，而CRISPR介导的敲除则抑制了这些过程。从机制上讲，RASON直接结合KRAS，将其稳定在GTP结合的高活性状态并促进下游信号传导。RASON敲除显著降低了CD47表达，增强了巨噬细胞介导的吞噬作用和抗肿瘤免疫力。在治疗方面，靶向RASON的反义寡核苷酸不仅表现出肿瘤抑制作用，还与KRAS抑制剂AMG - 510协同作用，显著增强了抗肿瘤疗效。
本研究揭示RASON是KRAS信号传导的关键致癌调节因子，驱动肺肿瘤发生和进展，并确定RASON是KRAS突变型非小细胞肺癌的一个有前景的治疗靶点。
