Thayer School of Engineering, Dartmouth College, Hanover, NH, 03755, USA.
Department of Pathology and Laboratory Medicine, Dartmouth Hitchcock Medical Center, Lebanon, NH, 03766, USA.
Mol Imaging Biol. 2023 Oct;25(5):911-922. doi: 10.1007/s11307-022-01771-9. Epub 2023 Jun 23.
Reliable and rapid identification of tumor in the margins of breast specimens during breast-conserving surgery to reduce repeat surgery rates is an active area of investigation. Dual-stain difference imaging (DDSI) is one of many approaches under evaluation for this application. This technique aims to topically apply fluorescent stain pairs (one targeted to a receptor-of-interest and the other a spectrally distinct isotype), image both stains, and compute a normalized difference image between the two channels. Prior evaluation and optimization in a variety of preclinical models produced encouraging diagnostic performance. Herein, we report on a pilot clinical study which evaluated HER2-targeted DDSI on 11 human breast specimens.
Gross sections from 11 freshly excised mastectomy specimens were processed using a HER2-receptor-targeted DDSI protocol shortly after resection. After staining with the dual-probe protocol, specimens were imaged on a fluorescence scanner, followed by tissue fixation for hematoxylin and eosin and anti-HER2 immunohistochemical staining. Receiver operator characteristic curves and area under the curve (AUC) analysis were used to assess diagnostic performance of the resulting images. Performance values were also compared to expression level determined from IHC staining.
Eight of the 11 specimens presented with distinguishable invasive ductal carcinoma and/or were not affected by an imaging artifact. In these specimens, the DDSI technique provided an AUC = 0.90 ± 0.07 for tumor-to-adipose tissue and 0.81 ± 0.15 for tumor-to-glandular tissue, which was significantly higher than AUC values recovered from images of the targeted probe alone. DDSI values and diagnostic performance did not correlate with HER2 expression level, and tumors with low HER2 expression often produced high AUC, suggesting that even the low expression levels were enough to help distinguish tumor.
The results from this preliminary study of rapid receptor-specific staining in human specimens were consistent with prior preclinical results and demonstrated promising diagnostic potential.
在保乳手术中可靠且快速地识别肿瘤边缘，以降低重复手术率，这是目前研究的热点。双染差异成像（DDSI）是用于此应用的众多方法之一。该技术旨在局部应用荧光染色对（一种针对感兴趣的受体，另一种是光谱上不同的同型），同时对两种染色进行成像，并计算两个通道之间的归一化差异图像。在各种临床前模型中进行的早期评估和优化产生了令人鼓舞的诊断性能。在此，我们报告了一项针对 11 个人类乳腺标本的 HER2 靶向 DDSI 的初步临床研究。
在切除后不久，使用 HER2 受体靶向 DDSI 方案对 11 个新鲜切除的乳房标本的大体切片进行处理。使用双探针方案染色后，将标本在荧光扫描仪上进行成像，然后对组织进行固定，用于苏木精和伊红染色和抗 HER2 免疫组织化学染色。使用接收者操作特性曲线和曲线下面积（AUC）分析来评估所得图像的诊断性能。性能值还与免疫组织化学染色确定的表达水平进行了比较。
在 11 个标本中，有 8 个标本呈现出可区分的浸润性导管癌，或不受成像伪影的影响。在这些标本中，DDSI 技术为肿瘤与脂肪组织的 AUC 值为 0.90 ± 0.07，肿瘤与腺组织的 AUC 值为 0.81 ± 0.15，明显高于单独靶向探针图像的 AUC 值。DDSI 值和诊断性能与 HER2 表达水平无关，并且 HER2 表达水平低的肿瘤通常产生高 AUC，这表明即使是低表达水平也足以帮助区分肿瘤。
这项针对人类标本中快速受体特异性染色的初步研究结果与之前的临床前结果一致，并显示出有希望的诊断潜力。
