Majumder Shounak, Halfdanarson Thorvardur R, Berger Calise K, Foote Patrick H, Cao Xiaoming, McGlinch Maria C, Gysbers Brianna J, de La Fuente Jaime, Robran Mariah J, Doering Karen A, Burger Kelli N, Bamlet William E, Oberg Ann L, Mahoney Douglas W, Graham Rondell P, Taylor William R, Petersen Gloria M, Kisiel John B
Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota.
Division of Medical Oncology, Mayo Clinic, Rochester, Minnesota.
Gastro Hep Adv. 2022 Apr 2;1(3):409-416. doi: 10.1016/j.gastha.2022.01.006. eCollection 2022.
Methylated DNA markers (MDMs) accurately identify several different cancer types, but there are limited data for pancreatic neuroendocrine tumors (pNETs). We aimed to identify MDM candidates in tissue that differentiate pNETs from normal pancreas.
wUsing DNA from frozen normal pancreas (13) and pNET (51) tissues, we performed reduced representation bisulfite sequencing for MDM discovery. Validation in independent formalin fixed paraffin embedded tissues used pNET cases (67; solid = 50, cystic = 17), normal pancreas (24), and buffy coat (36) controls. Primary pNET MDM distributions were compared with lung (36), small bowel (36) NETs, and metastatic pNET (25) tissues. The discrimination accuracy was summarized as the area under the receiver operator characteristic curve (AUC) with corresponding 95% confidence intervals (CIs). Fisher's linear discriminant analysis was performed to estimate a linear discriminate score (LDS) differentiating normal from pNET tissue and applied to all patient groups; discrimination accuracy of the LDS was summarized as the bootstrap cross-validated AUC.
Median AUC for distinguishing normal pancreas from pNET tissue was 0.91 (interquartile range: 0.80-0.93). The cross-validated AUC for the LDS discriminating normal pancreatic tissue from primary and metastatic pNETs was 0.957 (95% CI 0.858-1.0, < .0001) and 0.963 (95% CI 0.865-1.0, < .0001), respectively. The LDS for the MDM panel was significantly higher for primary pNET, metastatic pNET, lung NET, and small bowel NET, each compared with normal pancreas tissue ( < .0001). There was no statistical difference between primary pNET and metastatic pNET ( = .1947).
In independent tissue validation, MDMs accurately discriminate pNETs from normal pancreas. These results provide scientific rationale for exploration of these tissue MDMs in a plasma-based assay for clinical application.
甲基化DNA标志物（MDMs）能够准确识别多种不同类型的癌症，但关于胰腺神经内分泌肿瘤（pNETs）的数据有限。我们旨在识别能将pNETs与正常胰腺区分开来的组织中的MDM候选物。
我们使用来自冷冻的正常胰腺（13例）和pNET（51例）组织的DNA，进行简化代表性亚硫酸氢盐测序以发现MDMs。在独立的福尔马林固定石蜡包埋组织中进行验证，使用pNET病例（67例；实性=50例，囊性=17例）、正常胰腺（24例）和血沉棕黄层（36例）作为对照。将原发性pNET的MDM分布与肺（36例）、小肠（36例）NETs以及转移性pNET（25例）组织进行比较。辨别准确性以受试者操作特征曲线（AUC）下的面积及相应的95%置信区间（CIs）进行总结。进行Fisher线性判别分析以估计区分正常组织与pNET组织的线性判别分数（LDS），并应用于所有患者组；LDS的辨别准确性以自助法交叉验证的AUC进行总结。
区分正常胰腺与pNET组织的AUC中位数为0.91（四分位间距：0.80 - 0.93）。区分正常胰腺组织与原发性和转移性pNETs的LDS的交叉验证AUC分别为0.957（95%CI 0.858 - 1.0，P <.0001）和0.963（95%CI 0.865 - 1.0，P <.0001）。与正常胰腺组织相比，原发性pNET、转移性pNET、肺NET和小肠NET的MDM面板的LDS均显著更高（P <.0001）。原发性pNET与转移性pNET之间无统计学差异（P =.1947）。
在独立的组织验证中，MDMs能准确区分pNETs与正常胰腺。这些结果为在基于血浆的检测中探索这些组织MDMs用于临床应用提供了科学依据。
