BACKGROUND There is a substantial unmet clinical need for an accurate and effective blood biomarker for neuroendocrine neoplasms (NEN). We therefore evaluated, under real-world conditions in an ENETS Center of Excellence (CoE), the clinical utility of the NETest as a liquid biopsy and compared its utility with chromogranin A (CgA) measurement. METHODS The cohorts were: gastroenteropancreatic NEN (GEP-NEN; n = 253), bronchopulmonary NEN (BPNEN; n = 64), thymic NEN (n = 1), colon cancer (n = 37), non-small-cell lung cancer (NSCLC; n = 63), benign lung disease (n = 59), and controls (n = 86). In the GEPNEN group, 164 (65%) had image-positive disease (IPD, n = 135) or were image-negative but resection-margin/biopsy-positive (n = 29), and were graded as G1 (n = 106), G2 (n = 49), G3 (n = 7), or no data (n = 2). The remainder (n = 71) had no evidence of disease (NED). In the BPNEN group, 43/64 (67%) had IPD. Histology revealed typical carcinoids (TC, n = 14), atypical carcinoids (AC, n = 14), small-cell lung cancer (SCLC, n = 11), and large-cell neuroendocrine carcinoma (LCNEC, n = 4). Disease status (stable or progressive) was evaluated according to RECIST v1.1. Blood sampling involved NETest (n = 563) and NETest/CgA analysis matched samples (n = 178). NETest was performed by PCR (on a scale of 0-100), with a score ≥20 reflecting a disease-positive status and >40 reflecting progressive disease. CgA positivity was determined by ELISA. Samples were deidentified and measurements blinded. The Kruskal-Wallis, Mann-Whitney U, and McNemar tests, and the area under the curve (AUC) of the receiver-operating characteristics (ROC) were used in the statistical analysis. RESULTS In the GEPNEN group, NETest was significantly higher (34.4 ± 1.8, p < 0.0001) in disease-positive patients than in patients with NED (10.5 ± 1, p < 0.0001), colon cancer patients (18 ± 4, p < 0.0004), and controls (7 ± 0.5, p < 0.0001). Sensitivity for detecting disease compared to controls was 89% and specificity was 94%. NETest levels were increased in G2 vs. G1 (39 ± 3 vs. 32 ± 2, p = 0.02) and correlated with stage (localized: 26 ± 2 vs. regional/distant: 40 ± 3, p = 0.0002) and progression (55 ± 5 vs. 34 ± 2 in stable disease, p = 0.0005). In the BPNEN group, diagnostic sensitivity was 100% and levels were significantly higher in patients with bronchopulmonary carcinoids (BPC; 30 ± 1.3) who had IPD than in controls (7 ± 0.5, p < 0.0001), patients with NED (24.1 ± 1.3, p < 0.005), and NSCLC patients (17 ± 3, p = 0.0001). NETest levels were higher in patients with poorly differentiated BPNEN (LCNEC + SCLC; 59 ± 7) than in those with BPC (30 ± 1.3, p = 0.0005) or progressive disease (57.8 ± 7), compared to those with stable disease (29.4 ± 1, p < 0.0001). The AUC for differentiating disease from controls was 0.87 in the GEPNEN group and 0.99 in BPC patients (p < 0.0001). Matched CgA analysis was performed in 178 patients. In the GEPNEN group (n = 135), NETest was significantly more accurate for detecting disease (99%) than CgA positivity (53%; McNemar test χ2 = 87, p < 0.0001). In the BPNEN group (n = 43), NETest was significantly more accurate for disease detection (100%) than CgA positivity (26%; McNemar's test χ2 = 30, p < 0.0001). CONCLUSIONS The NETest is an accurate diagnostic for GEPNEN and BPNEN. It exhibits tumor biology correlation with grading, staging, and progression. CgA as a biomarker is significantly less accurate than NETest. The NETest has substantial clinical utility that can facilitate patient management.

中文翻译：

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NETest 作为胃肠胰腺和支气管肺神经内分泌肿瘤液体活检的前瞻性评估：ENETS 卓越中心体验。

背景对于神经内分泌肿瘤(NEN)的准确且有效的血液生物标志物存在大量未满足的临床需求。因此，我们在 ENETS 卓越中心 (CoE) 的真实条件下评估了 NETest 作为液体活检的临床效用，并将其效用与嗜铬粒蛋白 A (CgA) 测量进行了比较。方法 队列为：胃肠胰腺 NEN（GEP-NEN；n = 253）、支气管肺 NEN（BPNEN；n = 64）、胸腺 NEN（n = 1）、结肠癌（n = 37）、非小细胞肺癌（NSCLC；n = 63）、良性肺部疾病（n = 59）和对照（n = 86）。在 GEPNEN 组中，164 名 (65%) 患有影像阳性疾病（IPD，n = 135）或影像阴性但切除边缘/活检阳性（n = 29），并被分级为 G1（n = 106） )、G2 (n = 49)、G3 (n = 7) 或无数据 (n = 2)。其余（n = 71）没有疾病迹象（NED）。在 BPNEN 组中，43/64 (67%) 患有 IPD。组织学显示典型类癌（TC，n = 14）、非典型类癌（AC，n = 14）、小细胞肺癌（SCLC，n = 11）和大细胞神经内分泌癌（LCNEC，n = 4）。根据 RECIST v1.1 评估疾病状态（稳定或进展）。血液采样涉及 NETest (n = 563) 和 NETest/CgA 分析匹配样本 (n = 178)。NETest 是通过 PCR 进行的（0-100 分），得分≥20 表示疾病阳性，>40 表示疾病进展。通过ELISA确定CgA阳性。样品被去识别化并且测量被盲化。Kruskal-Wallis、Mann-Whitney U 和 McNemar 检验，和受试者工作特征（ROC）的曲线下面积（AUC）用于统计分析。结果 在 GEPNEN 组中，疾病阳性患者的 NETest (34.4 ± 1.8, p < 0.0001) 显着高于 NED 患者 (10.5 ± 1, p < 0.0001)、结肠癌患者 (18 ± 4, p < 0.0004) ) 和对照 (7 ± 0.5, p < 0.0001)。与对照相比，检测疾病的敏感性为 89%，特异性为 94%。G2 与 G1 的 NETest 水平增加（39 ± 3 与 32 ± 2，p = 0.02）并与阶段（局部：26 ± 2 与区域/远距离：40 ± 3，p = 0.0002）和进展（ 55 ± 5 vs. 34 ± 2 病情稳定，p = 0.0005）。在 BPNEN 组中，诊断敏感性为 100%，支气管肺类癌（BPC；30±1. 3) 与对照组 (7 ± 0.5, p < 0.0001)、NED 患者 (24.1 ± 1.3, p < 0.005) 和 NSCLC 患者 (17 ± 3, p = 0.0001) 相比，患有 IPD 的患者。与疾病稳定的患者 (29.4) 相比，低分化 BPNEN (LCNEC + SCLC; 59 ± 7) 患者的 NETest 水平高于 BPC (30 ± 1.3, p = 0.0005) 或疾病进展 (57.8 ± 7) 患者± 1, p < 0.0001)。区分疾病与对照的 AUC 在 GEPNEN 组中为 0.87，在 BPC 患者中为 0.99 (p < 0.0001)。对 178 名患者进行了匹配的 CgA 分析。在 GEPNEN 组 (n = 135) 中，NETest 检测疾病 (99%) 的准确度明显高于 CgA 阳性 (53%；McNemar 检验 χ2 = 87，p < 0.0001)。在 BPNEN 组 (n = 43) 中，NETest 检测疾病 (100%) 的准确度明显高于 CgA 阳性 (26%; McNemar' s 检验 χ2 = 30，p < 0.0001)。结论 NETest 可准确诊断 GEPNEN 和 BPNEN。它表现出与分级、分期和进展的肿瘤生物学相关性。CgA 作为生物标志物的准确性明显低于 NETest。NETest 具有重要的临床实用性，可以促进患者管理。