CREPT promotes LUAD progression by enhancing the CDK9 and RNAPII assembly to promote ERK-driven gene transcription.

IF 13.3 1区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
Theranostics Pub Date : 2025-07-25 eCollection Date: 2025-01-01 DOI:10.7150/thno.115572
Mengdi Li, Yuting Lin, Jiayu Wang, He Yang, Danhui Ma, Ye Tian, Yi Wang, Liu Yang, Umar Farooq, Yinyin Wang, Fangli Ren, Jian Sheng, Guoqing Zhang, Liang Chen, Jun Li, Xiangnan Li, Zhijie Chang
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引用次数: 0

Abstract

Background: 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. Methods: 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). Results: CREPT overexpression correlated with poor LUAD patient survival and enhanced tumorigenesis in EGFR or KRAS mutant LUAD cells. CREPT deletion impaired LUAD initiation and progression in the CC10-rtTA;TetO-KRASG12D 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-KRASG12D 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. Conclusion: 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.

悄悄通过增强CDK9和RNAPII的组装来促进erk驱动的基因转录,从而促进LUAD的进展。
背景:尽管EGFR和kras靶向治疗肺腺癌(LUAD)取得了进展,但对于对当前治疗无反应或耐药的患者,需要新的靶点。本研究证实了悄悄在LUAD进展中调控erk下游基因转录的关键作用。方法:采用人LUAD组织、EGFR/KRAS突变LUAD细胞系和小鼠模型,研究悄悄表达和功能。Micro-CT监测肿瘤进展。采用腺相关病毒(AAV)介导的悄悄清除作为治疗策略。RNA测序和荧光素酶报告分析鉴定了差异表达基因(DEGs)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占用。结果:在EGFR或KRAS突变的LUAD细胞中,悄悄过表达与较差的LUAD患者生存和增强的肿瘤发生相关。悄悄缺失损害了CC10-rtTA中LUAD的启动和进展;TetO-KRASG12D小鼠模型。从机制上讲,在ERK激活后,爬行细胞促进了CDK9与RNA聚合酶II (RNAPII)的组装,增强了KRAS-ERK-Elk-1信号下游恶性肿瘤相关基因的转录。蹑手蹑脚缺失和突变体R106A和S134A破坏了CREPT- rnapii的相互作用,降低了CDK9在Elk-1下游基因启动子上的占用及其表达。CC10-rtTA的目标都有所增加;TetO-KRASG12D和异种移植小鼠模型导致肿瘤生长停止。此外,在人源化小鼠模型中,aav介导的悄悄沉默抑制肿瘤进展,并显示出与派姆单抗的协同作用潜力。结论:我们的研究结果强调了悄悄作为LUAD进展的关键调节因子,并提示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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Theranostics
Theranostics MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
25.40
自引率
1.60%
发文量
433
审稿时长
1 months
期刊介绍: Theranostics serves as a pivotal platform for the exchange of clinical and scientific insights within the diagnostic and therapeutic molecular and nanomedicine community, along with allied professions engaged in integrating molecular imaging and therapy. As a multidisciplinary journal, Theranostics showcases innovative research articles spanning fields such as in vitro diagnostics and prognostics, in vivo molecular imaging, molecular therapeutics, image-guided therapy, biosensor technology, nanobiosensors, bioelectronics, system biology, translational medicine, point-of-care applications, and personalized medicine. Encouraging a broad spectrum of biomedical research with potential theranostic applications, the journal rigorously peer-reviews primary research, alongside publishing reviews, news, and commentary that aim to bridge the gap between the laboratory, clinic, and biotechnology industries.
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