Schmitz Daniel, Kazdal Daniel, Allgäuer Michael, Trunk Marcus, Vornhusen Sylke, Nahm Anna-Maria, Doll Matthias, Weingärtner Simon, Endris Volker, Penzel Roland, Kirchner Martina, Brandt Regine, Neumann Olaf, Sültmann Holger, Budczies Jan, Kienle Peter, Magdeburg Richard, Hetjens Svetlana, Schirmacher Peter, Bergmann Frank, Rudi Jochen, Stenzinger Albrecht, Volckmar Anna-Lena
Department of Gastroenterology, Oncology and Diabetology, Theresienkrankenhaus and St. Hedwigsklinik, University of Heidelberg, Heidelberg, Germany.
Institute of Pathology, University Hospital of Heidelberg, Heidelberg, Germany.
Genes Chromosomes Cancer. 2021 Jul;60(7):489-497. doi: 10.1002/gcc.22946. Epub 2021 Mar 16.
Pancreatic cysts or dilated pancreatic ducts are often found by cross-sectional imaging, but only mucinous lesions can become malignant. Therefore, distinction between mucinous and non-mucinous lesions is crucial for adequate patient management. We performed a prospective study including targeted next generation sequencing (NGS) of cell-free DNA in the diagnostic endoscopic ultrasound (EUS)-guided workup. Pancreatic cyst(s) or main duct fluid obtained by EUS-guided FNA was analysed by carcinoembryonic antigen (CEA), cytology and deep targeted NGS of 14 known gastrointestinal cancer genes (AKT1, BRAF, CTNNB1, EGFR, ERBB2, FBXW7, GNAS, KRAS, MAP2K1, NRAS, PIK3CA, SMAD4, TP53, APC) with a limit of detection down to variant allele frequency of 0.01%. Results were correlated to histopathology and clinical follow-up. One hundred and thirteen patients with pancreatic cyst(s) and/or a dilated pancreatic main duct (≥5 mm) were screened. Sixty-six patients had to be excluded, mainly due to inoperability or small cyst size (≤10 mm). Forty-seven patients were enrolled for further analysis. A final diagnosis was available in 27 cases including 8 negative controls. In 43/47 (91.5%) of patients a KRAS- and/or GNAS-mutation was diagnosed by NGS. 27.0% of the KRAS-mutated and 10.0% of the GNAS-mutated lesions harbored multiple mutations. KRAS/GNAS-testing by NGS, cytology, and CEA had a sensitivity and specificity of 94.7/100%, 38.1/100%, and 42.1/75.0%, respectively. KRAS/GNAS-testing was significantly superior to CEA (P = .0209) and cytology (P = .0016). In conclusion, KRAS/GNAS-testing by deep targeted NGS is a suitable method to distinguish mucinous from non-mucinous pancreatic lesions, suggesting its usage as a single diagnostic test. Results must be confirmed in a larger cohort.
胰腺囊肿或扩张的胰管常通过断层成像发现，但只有黏液性病变会发生恶变。因此，区分黏液性和非黏液性病变对于患者的恰当管理至关重要。我们进行了一项前瞻性研究，包括在诊断性内镜超声（EUS）引导检查中对游离DNA进行靶向二代测序（NGS）。通过EUS引导下细针穿刺（FNA）获取的胰腺囊肿或主胰管液，采用癌胚抗原（CEA）、细胞学检查以及对14个已知胃肠道癌基因（AKT1、BRAF、CTNNB1、EGFR、ERBB2、FBXW7、GNAS、KRAS、MAP2K1、NRAS、PIK3CA、SMAD4、TP53、APC）进行深度靶向NGS分析，检测限低至变异等位基因频率0.01%。结果与组织病理学和临床随访相关。筛选了113例有胰腺囊肿和/或扩张的主胰管（≥5毫米）的患者。66例患者因主要是无法手术或囊肿较小（≤10毫米）而被排除。47例患者纳入进一步分析。27例患者有最终诊断结果，包括8例阴性对照。在47例患者中的43例（91.5%）通过NGS检测到KRAS和/或GNAS突变。27.0%的KRAS突变病变和10.0%的GNAS突变病变存在多个突变。通过NGS、细胞学检查和CEA检测KRAS/GNAS的敏感性和特异性分别为94.7/100%、38.1/100%和42.1/75.0%。KRAS/GNAS检测显著优于CEA（P = 0.0209）和细胞学检查（P = 0.0016）。总之，通过深度靶向NGS检测KRAS/GNAS是区分胰腺黏液性和非黏液性病变的合适方法，表明可将其用作单一诊断测试。结果必须在更大的队列中得到证实。
