Li Mengdi, Lin Yuting, Wang Jiayu, Yang He, Ma Danhui, Tian Ye, Wang Yi, Yang Liu, Farooq Umar, Wang Yinyin, Ren Fangli, Sheng Jian, Zhang Guoqing, Chen Liang, Li Jun, Li Xiangnan, Chang Zhijie
State Key Laboratory of Membrane Biology, School of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing, 100084, China.
Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing, 102218, China.
Theranostics. 2025 Jul 25;15(16):8337-8359. doi: 10.7150/thno.115572. eCollection 2025.
Despite advancements in EGFR- and KRAS-targeted therapies for lung adenocarcinoma (LUAD), novel targets are needed for patients unresponsive or resistant to current treatments. This study demonstrates the critical role of CREPT in modulating ERK-downstream gene transcription in LUAD progression. CREPT expression and function were investigated using human LUAD tissues, EGFR/KRAS mutant LUAD cell lines, and mouse models. Micro-CT was used to monitor tumor progression. Adeno-associated virus (AAV)-mediated CREPT depletion was employed as a therapeutic strategy. RNA sequencing and luciferase reporter assays identified differentially expressed genes (DEGs) and affected signaling pathways. Protein interactions and CDK9 occupancy were assessed using multiplex immunofluorescence, immunoprecipitation, and chromatin immunoprecipitation (ChIP). CREPT overexpression correlated with poor LUAD patient survival and enhanced tumorigenesis in EGFR or KRAS mutant LUAD cells. deletion impaired LUAD initiation and progression in the CC10-rtTA;TetO- mouse model. Mechanistically, CREPT promoted CDK9 assembly with RNA polymerase II (RNAPII) following ERK activation, enhancing transcription of malignancy-related genes downstream of KRAS-ERK-Elk-1 signaling. CREPT depletion and the mutants R106A and S134A disrupting CREPT-RNAPII interaction reduced CDK9 occupancy at Elk-1 downstream gene promoters and their expression. Targeting CREPT in both CC10-rtTA;TetO- and xenograft mouse models resulted in tumor growth arrest. Furthermore, in a humanized mouse model, AAV-mediated CREPT silencing inhibited tumor progression and showed synergistic potential with pembrolizumab. Our findings highlight CREPT as a pivotal regulator of LUAD progression and suggest it could be a potential therapeutic target for patients with EGFR or KRAS mutations insensitive or resistant to targeted therapies.
尽管针对肺腺癌（LUAD）的表皮生长因子受体（EGFR）和 Kirsten 大鼠肉瘤病毒癌基因（KRAS）靶向治疗取得了进展，但对于对当前治疗无反应或耐药的患者仍需要新的靶点。本研究证明了 CREPT 在调节 LUAD 进展中 ERK 下游基因转录方面的关键作用。使用人 LUAD 组织、EGFR/KRAS 突变型 LUAD 细胞系和小鼠模型研究了 CREPT 的表达和功能。采用微型计算机断层扫描（Micro-CT）监测肿瘤进展。采用腺相关病毒（AAV）介导的 CREPT 缺失作为治疗策略。通过 RNA 测序和荧光素酶报告基因检测确定差异表达基因（DEG）和受影响的信号通路。使用多重免疫荧光、免疫沉淀和染色质免疫沉淀（ChIP）评估蛋白质相互作用和 CDK9 占据情况。CREPT 过表达与 LUAD 患者生存率低相关，并增强了 EGFR 或 KRAS 突变型 LUAD 细胞的肿瘤发生。在 CC10-rtTA;TetO-小鼠模型中，CREPT 缺失损害了 LUAD 的起始和进展。从机制上讲，CREPT 在 ERK 激活后促进 CDK9 与 RNA 聚合酶 II（RNAPII）组装，增强 KRAS-ERK-Elk-1 信号下游恶性相关基因的转录。CREPT 缺失以及破坏 CREPT-RNAPII 相互作用的突变体 R106A 和 S134A 降低了 CDK9 在 Elk-1 下游基因启动子处的占据及其表达。在 CC10-rtTA;TetO-和异种移植小鼠模型中靶向 CREPT 导致肿瘤生长停滞。此外，在人源化小鼠模型中，AAV 介导的 CREPT 沉默抑制肿瘤进展，并显示出与帕博利珠单抗的协同潜力。我们的研究结果突出了 CREPT 作为 LUAD 进展的关键调节因子，并表明它可能是对靶向治疗不敏感或耐药的 EGFR 或 KRAS 突变患者的潜在治疗靶点。
