Table_2_NOTCH1 mutation associates with impaired immune response and decreased relapse-free survival in patients with resected T1-2N0 laryngeal cancer.docx

BackgroundPatients with early-stage laryngeal cancer, even stage T1-2N0, are at considerable risk of recurrence and death. The genetic and immunologic characteristics of recurrent laryngeal cancer remain unclear.
MethodsA total of 52 T1-2N0 laryngeal cancer patients were enrolled. Of these, 42 tissue samples were performed by targeted DNA sequencing, and 21 cases were performed by NanoString immuno-oncology targeted RNA sequencing to identify the distinct molecular bases and immunologic features associated with relapse in patients with early laryngeal cancer, respectively.
ResultsTo the best to our knowledge, we present for the first time an overview of the genomic mutation spectrum of early-stage laryngeal cancers. A total of 469 genomic alterations were detected in 211 distinct cancer-relevant genes, and the genes found to be mutated in more than five patients (>10%) included tumor protein p53 (TP53, 78.5%), FAT atypical cadherin 1 (FAT1, 26%), LDL receptor related protein 1B (LRP1B, 19%), cyclin dependent kinase inhibitor 2A (CDKN2A, 17%), tet methylcytosine dioxygenase 2 (TET2, 17%), notch receptor 1 (NOTCH1, 12%) and neuregulin 1 (NRG1, 12%). Recurrent laryngeal cancer demonstrated a higher tumor mutation burden (TMB), as well as higher LRP1B mutation and NOTCH1 mutation rates. Univariate and multivariate analyses revealed that high TMB (TMB-H) and NOTCH1 mutation are independent genetic factors that are significantly associated with shorter relapse-free survival (RFS). Simultaneously, the results of the transcriptome analysis presented recurrent tumors with NOTCH1 mutation displayed upregulation of the cell cycle pathway, along with decreased B cells score, T cells score, immune signature score and tumor-infiltrating lymphocytes (TILs) score. The Cancer Genome Atlas (TCGA)-laryngeal cancer dataset also revealed weakened immune response and impaired adhesion functions in NOTCH1-mutant patients.
ConclusionsGenomic instability and impaired immune response are key features of the immunosurveillance escape and recurrence of early laryngeal cancer after surgery. These findings revealed immunophenotypic attenuation in recurrent tumors and provided valuable information for improving the management of these high-risk patients. Due to the small number of patients in this study, these differences need to be further validated in a larger cohort.

背景 即使是T1-2N0期的早期喉癌患者，仍面临较高的复发与死亡风险。目前复发性喉癌的遗传与免疫特征仍不明确。
方法 本研究共纳入52例T1-2N0期喉癌患者。其中，42份组织样本接受了靶向DNA测序，21例样本接受了NanoString肿瘤免疫靶向RNA测序，分别用以明确早期喉癌患者复发相关的独特分子基础与免疫特征。
结果 据我们所知，本研究首次系统呈现了早期喉癌的基因组突变谱特征。研究在211个明确与癌症相关的基因中，共检测到469个基因组变异。在超过5例患者（占比>10%）中发生突变的基因包括：肿瘤蛋白p53（TP53，78.5%）、FAT非典型钙黏蛋白1（FAT1，26%）、低密度脂蛋白受体相关蛋白1B（LRP1B，19%）、细胞周期蛋白依赖性激酶抑制剂2A（CDKN2A，17%）、四甲基胞嘧啶双加氧酶2（TET2，17%）、Notch受体1（NOTCH1，12%）以及神经调节蛋白1（NRG1，12%）。复发性喉癌的肿瘤突变负荷（Tumor Mutation Burden, TMB）更高，同时LRP1B与NOTCH1的突变率也更高。单因素与多因素分析显示，高肿瘤突变负荷（TMB-H）与NOTCH1突变是独立的遗传危险因素，与更短的无复发生存期（Relapse-Free Survival, RFS）显著相关。同时，转录组分析结果显示，携带NOTCH1突变的复发性肿瘤中，细胞周期通路出现上调，而B细胞评分、T细胞评分、免疫特征评分以及肿瘤浸润淋巴细胞（Tumor-Infiltrating Lymphocytes, TILs）评分均有所下降。癌症基因组图谱（The Cancer Genome Atlas, TCGA）-喉癌数据集的分析结果也证实，携带NOTCH1突变的患者存在免疫应答减弱与黏附功能受损的情况。
结论 基因组不稳定性与免疫应答受损是早期喉癌术后免疫监视逃逸与复发的关键特征。本研究揭示了复发性肿瘤的免疫表型衰减特征，为优化此类高危患者的临床管理提供了有价值的参考信息。由于本研究的患者样本量较小，上述差异仍需在更大规模的队列研究中进一步验证。