背景:虽然不常见,甲状腺髓样癌(MTC)占甲状腺癌死亡的很大比例。最近的研究已经验证了两级国际甲状腺髓样癌分级系统(IMTCGS)来预测临床结果。5%的Ki67增殖指数(Ki67PI)截止值将低等级MTC与高等级MTC分开。在这项研究中,我们将数字图像分析(DIA)与手动计数(MC)进行了比较,以确定MTC队列中的Ki67PI,探索遇到的挑战。
方法:两位病理学家对来自85个MTC的可用载玻片进行了审查。每个病例的Ki67PI通过免疫组织化学记录,用Aperio®载玻片扫描仪以40倍放大倍率扫描,并使用QuPath®DIA平台进行量化。同样的热点被截图,彩色印刷,盲目计算。对于每种情况,计数超过500个MTC细胞。每个MTC使用IMTCGS标准进行分级。
结果:在我们的MTC队列中(n=85),在IMTGS中,84.7%和15.3%为低等级和高等级。在整个队列中,QuPath®DIA表现良好(R2=0.9891),但与MC相比似乎欠佳。QuPath®在高级别病例(R2=0.99)中的表现优于低级别病例(R2=0.7071)。总的来说,用MC或DIA测定的Ki67PI不影响IMTGS等级。遇到的DIA挑战包括优化小区检测、重叠的原子核,和组织伪影。遇到的MC挑战包括背景染色,与正常元素的形态重叠,计算时间。
结论:我们的研究强调了DIA在定量MTC的Ki67PI中的实用性,并且可以作为与有丝分裂活动和坏死的其他标准一起进行分级的辅助手段。
BACKGROUND: Although uncommon, medullary thyroid carcinoma (MTC) accounts for a significant proportion of thyroid cancer deaths. Recent studies have validated the two-tier International Medullary Thyroid Carcinoma Grading System (IMTCGS) to predict clinical outcomes. A 5% Ki67 proliferative index (Ki67PI) cut-off separates low-grade from high-grade MTC. In this study, we compared digital image analysis (DIA) to manual counting (MC) for determining the Ki67PI in a MTC cohort, and explored the challenges encountered.
METHODS: Available slides from 85 MTCs were reviewed by two pathologists. The Ki67PI was documented by immunohistochemistry for each case, scanned with the Aperio® slide scanner at 40× magnification, and quantified using the QuPath® DIA platform. The same hotspots were screenshot, printed in color, and blindly counted. For each case, over 500 MTC cells were counted. Each MTC was graded using IMTCGS criteria.
RESULTS: In our MTC cohort (n = 85), 84.7 and 15.3% were low- and high-grade with the IMTCGS. In the entire cohort, QuPath® DIA performed well (R2 = 0.9891) but appeared to undercall compared to MC. QuPath® performed better in high-grade cases (R2 = 0.99) compared to low-grade cases (R2 = 0.7071). Overall, Ki67PI determined with either MC or DIA did not affect IMTCGS grade. Encountered DIA challenges include optimizing cell detection, overlapping nuclei, and tissue artifacts. Encountered MC challenges include background staining, morphologic overlap with normal elements, and counting time.
CONCLUSIONS: Our study highlights the utility of DIA in quantifying Ki67PI for MTC and can serve as an adjunct for grading in conjunction with the other criteria of mitotic activity and necrosis.