Huang Haijin, Feng Yanping, Xu Yuhui, Liu Jianping, Peng Wei, Zeng Linshan, Zeng Yong, Liu Jinping, He Xiao, Liu Haijin
Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China.
Jiangxi Provincial Clinical Research Center for Vascular Anomalies, The First Affiliated Hospital of GanNan Medical University, Ganzhou, Jiangxi, China.
Pediatr Res. 2025 May 15. doi: 10.1038/s41390-025-04074-1.
Intrinsic resistance reduces the effectiveness of many anticancer therapies. Anlotinib, a small-molecule multi-targeted tyrosine kinase inhibitor, has shown potential in treating hepatoblastoma. This study investigates the role of γ-aminobutyric acid (GABA) in anlotinib resistance using in vivo and in vitro models.
HuH-6 hepatoblastoma cells were implanted into nude mice to assess the effects of anlotinib on tumor growth. Neurotransmitter-targeted metabolomics was performed to analyze neurotransmitter metabolism in xenograft tumor tissues. In vitro, HuH-6 and HepG2 cells were treated with anlotinib to evaluate changes in GABA synthesis, degradation, and associated protein expression.
Anlotinib significantly inhibited HuH-6 tumor growth but was less effective than cisplatin. Neurotransmitter-targeted metabolomics showed tumors treated with anlotinib exhibited increased GABA levels. Anlotinib treatment also upregulated the protein expression of GAD1, a key enzyme in GABA synthesis. In vitro, anlotinib treatment enhanced GABA release and GAD1 expression in hepatoblastoma cells. Exogenous GABA stimulation promoted cell proliferation in vitro and tumor growth in vivo. Notably, GAD1 knockdown enhanced anlotinib's inhibitory effects on hepatoblastoma in vitro and in vivo.
Anlotinib induces intrinsic resistance in hepatoblastoma by upregulating GAD1 and increasing GABA accumulation. Targeting GAD1 may enhance anlotinib's therapeutic efficacy and help overcome resistance.
Anlotinib upregulates GAD1 to enhance GABA synthesis, driving intrinsic resistance in hepatoblastoma by activating tumor-promoting GABA signaling in the tumor microenvironment. First identification of the GAD1/GABA axis as a critical mediator of anlotinib resistance, expanding understanding of neurotransmitter-driven drug tolerance in pediatric cancers. GAD1 knockdown synergizes with anlotinib to overcome resistance, establishing a combinatorial strategy to enhance antitumor efficacy in preclinical models. Proposes targeting GABA metabolism to optimize anlotinib-based therapies, addressing unmet needs in refractory hepatoblastoma treatment.
内在抗性降低了许多抗癌疗法的有效性。安罗替尼是一种小分子多靶点酪氨酸激酶抑制剂，已显示出治疗肝母细胞瘤的潜力。本研究使用体内和体外模型研究γ-氨基丁酸（GABA）在安罗替尼抗性中的作用。
将HuH-6肝母细胞瘤细胞植入裸鼠体内，以评估安罗替尼对肿瘤生长的影响。进行靶向神经递质的代谢组学分析，以分析异种移植肿瘤组织中的神经递质代谢。在体外，用安罗替尼处理HuH-6和HepG2细胞，以评估GABA合成、降解及相关蛋白表达的变化。
安罗替尼显著抑制HuH-6肿瘤生长，但效果不如顺铂。靶向神经递质的代谢组学显示，接受安罗替尼治疗的肿瘤GABA水平升高。安罗替尼治疗还上调了GABA合成中的关键酶GAD1的蛋白表达。在体外，安罗替尼治疗增强了肝母细胞瘤细胞中GABA的释放和GAD1的表达。外源性GABA刺激促进体外细胞增殖和体内肿瘤生长。值得注意的是，敲低GAD1增强了安罗替尼在体外和体内对肝母细胞瘤的抑制作用。
安罗替尼通过上调GAD1和增加GABA积累诱导肝母细胞瘤的内在抗性。靶向GAD1可能增强安罗替尼的治疗效果并有助于克服抗性。
安罗替尼上调GAD1以增强GABA合成，通过激活肿瘤微环境中促进肿瘤的GABA信号传导驱动肝母细胞瘤的内在抗性。首次确定GAD1/GABA轴是安罗替尼抗性的关键介质，扩展了对儿童癌症中神经递质驱动的药物耐受性的理解。敲低GAD1与安罗替尼协同克服抗性，在临床前模型中建立了增强抗肿瘤疗效的联合策略。提出靶向GABA代谢以优化基于安罗替尼的疗法，满足难治性肝母细胞瘤治疗中未满足的需求。
