Chen Kai,Wu Zhixiang,Wu Yeming.Proteomic profiling-based clinical risk analysis and molecular subtypic identification in neuroblastoma[J].Journal of Clinical Pediatric Surgery,2026,(04):316-322.[doi:10.3760/cma.j.cn101785-20251228-00138]
基于蛋白质组学特征的神经母细胞瘤临床危险度分层与分子亚型研究
- Title:
- Proteomic profiling-based clinical risk analysis and molecular subtypic identification in neuroblastoma
- 摘要:
- 目的 应用高通量蛋白质组学技术解析神经母细胞瘤(neuroblastoma,NB)的分子特征,以完善临床风险分层,并探索潜在治疗靶点。方法 回顾性纳入2012年10月至2015年2月在上海交通大学医学院附属新华医院接受手术治疗的150例NB患儿肿瘤组织样本,基于定量蛋白质组学平台构建表达谱。通过聚类分析比较不同临床风险等级(高危、中危、低危)及原发部位(肾上腺、非肾上腺)的蛋白质表达差异;运用无监督聚类定义蛋白质组亚型,应用脂肪酸氧化抑制剂(Etomoxir等)开展体外功能实验,验证高危亚型的代谢依赖性。结果 蛋白质组表达谱与临床风险高度匹配:高危组样本显著富集氧化磷酸化、 MYC靶点、E2F靶点及DNA修复通路;而低危组则富集上皮-间质转化及分化相关蛋白。原发于肾上腺的肿瘤预后显著差于非肾上腺来源,前者富集MYC信号、氧化磷酸化及胚胎发育相关蛋白,呈去分化状态;后者则表现出明显的免疫激活(补体通路、体液免疫反应)特征。无监督聚类将样本分为4种分子亚型:基质型(PAM1)预后极佳,富集细胞外基质蛋白;剪切型(PAM2)以RNA加工相关蛋白激活为特征;免疫型(PAM3)虽有淋巴细胞浸润但存在免疫逃逸;代谢型(PAM4)预后最差,能量代谢极度活跃。体外实验显示,NB细胞对脂肪酸氧化抑制剂Etomoxir高度敏感,可显著抑制细胞存活率,提示PAM4亚型高度依赖线粒体能量代谢。结论 本研究构建了NB的蛋白质组学分子全景,提出了4种分子亚型的分型模型,不仅揭示了肾上腺来源肿瘤的胚胎发育特征,更发现了极高危亚型PAM4对脂肪酸氧化的代谢依赖性,为NB的精准分型及高危患者的线粒体代谢靶向治疗提供了理论与实验依据。
- Abstract:
- Objective Neuroblastoma (NB) is characterized by profound clinical heterogeneity.Current clinical indicators have failed to fully elucidate its complex biological nature.This study was intended to utilize in-depth proteomic analysis to resolve the molecular features of NB,refine risk stratifications and identify potential therapeutic targets.Methods A cohort of 150 NB children was enrolled for deep characterization using high-throughput quantitative proteomics.Molecular differences across different clinical risk levels (high/intermediate/low risk) and primary sites (adrenal vs non-adrenal) were examined via supervised clustering.Unsupervised clustering was applied for defining proteomic subtypes.And metabolic dependency of high-risk subtypes was validated through cellular experiments using such a metabolic inhibitor as Etomoxir.Results Proteomic expression profiling hinted at high consistency with clinical risk stratification.The high-risk group was significantly enriched in oxidative phosphorylation,MYC targets,E2F targets and DNA repair pathways while the low-risk group exhibited stronger features of epithelial-mesenchymal transition (EMT) and differentiation.Adrenal-derived tumors demonstrated a significantly worse prognosis as compared to non-adrenal-derived tumors.The former was enriched with MYC signaling,oxidative phosphorylation and embryonic development-related proteins,reflecting a dedifferentiated state; the latter was characterized by obvious immune activation (complement and humoral immunity).Unsupervised clustering categorized NB into four distinct subtypes.PAM1 (stromal-type): Excellent prognosis and enrichment of extracellular matrix proteins.PAM2 (splicing-type): Characterized by RNA processing activation.PAM3 (immune-type): High death risk due to immune escape despite lymphocyte infiltration.PAM4 (metabolic-type): The worst prognosis and extremely active energy metabolism.Experimental validation confirmed that NB cells were specifically sensitive to fatty acid oxidation (FAO) inhibitor Etomoxir,significantly suppressing cell viability,indicating a high reliance of PAM4 subtype on mitochondrial energy metabolism.Conclusions This study constructs a proteomic molecular landscape of neuroblastoma and proposes a precise four-subtype molecular model.Our findings reveal the embryonic features of adrenal-origin tumors and pinpoint the metabolic dependency of ultra-high-risk subtype (PAM4) on fatty acid oxidation.These results provide theoretical and experimental rationales for precision stratification and mitochondrial metabolism-targeted therapy for NB.
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备注/Memo
收稿日期:2025-12-28。
基金项目:国家自然科学基金(82472715)
通讯作者:吴晔明,Email:wuymsh@163.com