Zhu Hongtao, Cheng Lidong, Liu Dan, Ma Xiaoyu, Chen Zhiye, Fan Heng, Li Ran, Zhang Yang, Mi Hailong, Li Jun, Zhang Suojun, Yu Xingjiang, Shu Kai
Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Neurosurgery, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Neuro Oncol. 2025 Mar 7;27(3):695-710. doi: 10.1093/neuonc/noae224.
Glioma stem cells (GSCs) are the root cause of tumorigenesis, recurrence, and therapeutic resistance in glioblastoma (GBM), the most prevalent and lethal type of primary adult brain malignancy. The exploitation of novel methods targeting GSCs is crucial for the treatment of GBM. In this study, we investigate the function of the novel ROR1-GRB2-c-Fos axis in GSCs maintenance and GBM progression.
The expression characteristics of ROR1 in GBM and GSCs were assessed by bioinformatic analysis, patient specimens, and patient-derived GSCs. Lentivirus-mediated gene knockdown and overexpression were conducted to evaluate the effect of ROR1 on GSCs proliferation and self-renewal both in vitro and in vivo. The downstream signaling of ROR1 in GSCs maintenance was unbiasedly determined by RNA-seq and validated both in vitro and in vivo. Finally, rescue assays were performed to further validate the function of the ROR1-GRB2-c-Fos axis in GSCs maintenance and GBM progression.
ROR1 is upregulated in GBM and preferentially expressed in GSCs. Disruption of ROR1 markedly impairs GSC proliferation and self-renewal, and inhibits GBM growth in vivo. Moreover, ROR1 stabilizes GRB2 by directly binding and reducing its lysosomal degradation, and ROR1 knockdown significantly inhibits GRB2/ERK/c-Fos signaling in GSCs. Importantly, ectopic expression of c-Fos counteracts the effects caused by ROR1 silencing both in vitro and in vivo.
ROR1 plays essential roles in GSCs maintenance through binding to GRB2 and activation of ERK/c-Fos signaling, which highlights the therapeutic potential of targeting the ROR1-GRB2-c-Fos axis.
胶质瘤干细胞（GSCs）是胶质母细胞瘤（GBM）发生、复发和治疗耐药的根本原因，GBM是成人原发性脑恶性肿瘤中最常见且致命的类型。开发针对GSCs的新方法对GBM的治疗至关重要。在本研究中，我们探究了新型ROR1 - GRB2 - c - Fos轴在GSCs维持和GBM进展中的作用。
通过生物信息学分析、患者标本和患者来源的GSCs评估ROR1在GBM和GSCs中的表达特征。进行慢病毒介导的基因敲低和过表达，以评估ROR1对GSCs体外和体内增殖及自我更新的影响。通过RNA测序无偏地确定ROR1在GSCs维持中的下游信号，并在体外和体内进行验证。最后，进行挽救实验以进一步验证ROR1 - GRB二 - c - Fos轴在GSCs维持和GBM进展中的作用。
ROR1在GBM中上调并在GSCs中优先表达。ROR1的破坏显著损害GSC增殖和自我更新，并在体内抑制GBM生长。此外，ROR1通过直接结合并减少其溶酶体降解来稳定GRB2，ROR1敲低显著抑制GSCs中的GRB2/ERK/c - Fos信号。重要的是，c - Fos的异位表达在体外和体内均抵消了ROR1沉默所引起的影响。
ROR1通过与GRB2结合并激活ERK/c - Fos信号在GSCs维持中起重要作用，这突出了靶向ROR1 - GRB2 - c - Fos轴的治疗潜力。
