Hu Ya'nan, Liu Jinming, Tu Jian, Yang Min, He Qisheng, Li Fei, Xu Xiaojing, Ji Zhongqing, Xu Jianwei, Zhong Wentao, Yan Mengwen, Yang Ying, Zhang Huanxiang
Department of Cell Biology, MOE Key Laboratory of Geriatric Diseases and Immunology, Suzhou Medical College of Soochow University, Suzhou, China.
Department of Pathology, The Second Affiliated Hospital of Soochow University, Suzhou, China.
Cell Prolif. 2025 Aug;58(8):e70013. doi: 10.1111/cpr.70013. Epub 2025 Mar 11.
Exploring effective, prompt and universally applicable approaches for inducing the differentiation of glioblastoma (GBM) into terminally differentiated cells, such as astrocytes or neurons that cease cell division, is pivotal for the success of GBM differentiation therapy. In this study, a neuronal-specific promoter-reporter system was employed to screen small molecules that promote neural differentiation. The cocktail YFSS, consisting of Y27632, Forskolin, SB431542 and SP600125, which selectively targets the ROCK, cAMP, TGF-β and JNK signalling pathways, respectively, was found to effectively trigger differentiation in human GBM cells. This process yielded neuron-like cells within 7 days, inhibited GBM cell proliferation and reduced malignancy traits, such as stemness, migratory and invasive capabilities. Transcriptome sequencing revealed the pathways altered by YFSS, shedding light on its dual role in halting cell proliferation and initiating neuronal differentiation. A notable increase in CEND1 expression, a key molecule in cell cycle and neuronal differentiation regulation, was observed during differentiation. However, CEND1 alone could not replicate YFSS's high conversion efficiency and its depletion reduced the differentiation and restored proliferation of the GBM cells. In vivo, prolonged and localised YFSS application significantly curtailed tumour growth and extended survival in patient-derived xenograft mice models. In summary, our findings reveal that the small-molecule cocktail YFSS is an effective means for inducing neuronal differentiation in GBM cells, representing a novel and promising pathway for the advancement of GBM treatment.
探索有效、迅速且普遍适用的方法，促使胶质母细胞瘤（GBM）分化为终末分化细胞，如停止细胞分裂的星形胶质细胞或神经元，对于GBM分化治疗的成功至关重要。在本研究中，采用神经元特异性启动子-报告系统筛选促进神经分化的小分子。发现由Y27632、福斯高林、SB431542和SP600125组成的鸡尾酒YFSS能有效触发人GBM细胞的分化，该鸡尾酒分别选择性靶向ROCK、cAMP、TGF-β和JNK信号通路。这一过程在7天内产生神经元样细胞，抑制GBM细胞增殖并降低恶性特征，如干性、迁移和侵袭能力。转录组测序揭示了YFSS改变的信号通路，阐明了其在阻止细胞增殖和启动神经元分化中的双重作用。在分化过程中观察到细胞周期和神经元分化调节的关键分子CEND1表达显著增加。然而，单独的CEND1无法复制YFSS的高转化效率，其缺失会降低GBM细胞的分化并恢复增殖。在体内，在患者来源的异种移植小鼠模型中，长期局部应用YFSS显著抑制肿瘤生长并延长生存期。总之，我们的研究结果表明，小分子鸡尾酒YFSS是诱导GBM细胞神经元分化的有效手段，代表了GBM治疗进展的一条新的有前景的途径。
