NFE2L2预测ESCC放化疗敏感性及调控机制

放化疗是中晚期食管鳞癌（ESCC）的主要治疗手段，但不同患者放化疗敏感性差异巨大，探索该差异的机制，寻找有效的预测指标具有重要临床意义。NFE2L2在ESCC中突变频率较高，其下游通路可调控肿瘤的放化疗敏感性，但具体分子机制有待阐释。前期研究提示：NFE2L2突变状态与治疗过程的筛选相关，突变患者预后较差。生信分析发现：NFE2L2的热点突变和功能性异常转录本均可导致其下游活化，激活非经典WNT通路。为进一步研究NFE2L2调控放化疗敏感性的机制，利用基因编辑技术获得热点突变与第2外显子跳跃的细胞克隆；并通筛选ESCC治疗抵抗细胞系，在分子层面上深入探讨NFE2L2轴的调控方式，探索其与WNT通路的关系。在细胞、动物及临床样本水平验证NFE2L2轴与放化疗抵抗的关系，利用血液ctDNA检测NFE2L2突变载荷的动态变化，预测ESCC患者的放化疗敏感性与预后，为个体化治疗提供依据。

Chemoradiotherapy (CRT) is the main treatment method for advanced esophageal squamous cell carcinoma (ESCC). However, the biological heterogeneity leads to diverse treatment outcomes, which highlights the clinical significance of providing insights into intrinsic reasons and effective response predictors. NFE2L2 frequently mutates in ESCC and plays a notable role in theraputic resistance, while the underlying mechanisms need further exploration. Our previous work has suggested that NFE2L2 mutation, which indicates poor prognosis, may attribute to positive selection during CRT. Systematic bioinformatic analysis found that the hot-spot mutation and functional abnormality transcripts of NFE2L2 could up-regulate its downstream molecules, including genes involed in non-canonical WNT pathway.. To investigate the role of NFE2L2-axis in CRT sensitivity and its regulation on WNT pathway, several ESCC cell lines are established for mechanism study. CRISPR/Cas9 technique is applied to obtain genome-edited cell clones bearing NFE2L2 mutations or second-exon-skipped transcripts, while chemotherapeutic agents and radiation are used to select CRT-resistant cell clones. The validation works are performed on multiple levels as cells, mice and clinical samples. Moreover, by dynamically detecting NFE2L2 mutation in ctDNA, the therapeutic effect and tumor recurrence could be predicted from its temporal mutation load, which is a promsing biomarker for individualized ESCC treatment.