Lung cancer is the deadliest malignancy in the United States. Non–small cell lung cancer (NSCLC) accounts for 85% of cases and is frequently driven by activating mutations in the gene encoding the KRAS GTPase (e.g., KRASG12D). Our previous work demonstrated that Argonaute 2 (AGO2)—a component of the RNA-induced silencing complex (RISC)—physically interacts with RAS and promotes its downstream signaling. We therefore hypothesized that AGO2 could promote KRASG12D-dependent NSCLC in vivo. To test the hypothesis, we evaluated the impact of Ago2 knockout in the KPC (LSL-Kras ^G12D/+;p53^f/f;Cre) mouse model of NSCLC. In KPC mice, intratracheal delivery of adenoviral Cre drives lung-specific expression of a stop-floxed KRASG12D allele and biallelic ablation of p53. Simultaneous biallelic ablation of floxed Ago2 inhibited KPC lung nodule growth while reducing proliferative index and improving pathological grade. We next applied the KP^HetC model, in which the Clara cell–specific CCSP-driven Cre activates KRASG12D and ablates a single p53 allele. In these mice, Ago2 ablation also reduced tumor size and grade. In both models, Ago2 knockout inhibited ERK phosphorylation (pERK) in tumor cells, indicating impaired KRAS signaling. RNA sequencing (RNA-seq) of KPC nodules and nodule-derived organoids demonstrated impaired canonical KRAS signaling with Ago2 ablation. Strikingly, accumulation of pERK in KPC organoids depended on physical interaction of AGO2 and KRAS. Taken together, our data demonstrate a pathogenic role for AGO2 in KRAS-dependent NSCLC. Given the prevalence of this malignancy and current difficulties in therapeutically targeting KRAS signaling, our work may have future translational relevance.

肺癌是美国最致命的恶性肿瘤。非小细胞肺癌 (NSCLC) 占病例的 85%，通常是由编码 KRAS GTP 酶（例如KRASG12D）的基因中的激活突变引起的。我们之前的工作表明，Argonaute 2 (AGO2)——RNA 诱导沉默复合物 (RISC) 的一个组成部分——与 RAS 发生物理相互作用并促进其下游信号传导。因此，我们假设 AGO2 可以在体内促进KRASG12D依赖性 NSCLC。为了检验这一假设，我们评估了Ago2敲除对NSCLC KPC ( LSL-Kras ^G12D/+ ;p53 ^f/f ;Cre ) 小鼠模型的影响。在 KPC 小鼠中，气管内递送腺病毒 Cre 可驱动 stop-floxed KRASG12D等位基因的肺部特异性表达和p53的双等位基因消融。同时双等位基因消融 floxed Ago2可抑制 KPC 肺结节生长，同时降低增殖指数并改善病理分级。接下来我们应用了KP ^Het C模型，其中 Clara 细胞特异性CCSP驱动的 Cre 激活KRASG12D并消除单个p53等位基因。在这些小鼠中，Ago2消融还减小了肿瘤的大小和级别。在这两种模型中，Ago2敲除都会抑制肿瘤细胞中的 ERK 磷酸化 (pERK)，表明 KRAS 信号传导受损。KPC结节和结节衍生类器官的 RNA 测序 (RNA-seq)表明， Ago2消融导致经典 KRAS 信号传导受损。引人注目的是，KPC 类器官中 pERK 的积累取决于 AGO2 和 KRAS 的物理相互作用。综上所述，我们的数据证明了 AGO2 在 KRAS 依赖性 NSCLC 中的致病作用。考虑到这种恶性肿瘤的流行以及目前针对 KRAS 信号传导治疗的困难，我们的工作可能具有未来的转化相关性。