Flagship Biosciences, Westminster, CO, USA.
Athenaeum Pathology Consulting, Sudbury, MA, USA.
Sci Rep. 2021 May 10;11(1):9920. doi: 10.1038/s41598-021-89060-3.
Dickkopf-1 (DKK1) is a secreted modulator of Wnt signaling that is frequently overexpressed in tumors and associated with poor clinical outcomes. DKN-01 is a humanized monoclonal therapeutic antibody that binds DKK1 with high affinity and has demonstrated clinical activity in gastric/gastroesophageal junction (G/GEJ) patients with elevated tumoral expression of DKK1. Here we report on the validation of a DKK1 RNAscope chromogenic in situ hybridization assay to assess DKK1 expression in G/GEJ tumor tissue. To reduce pathologist time, potential pathologist variability from manual scoring and support pathologist decision making, a digital image analysis algorithm that identifies tumor cells and quantifies the DKK1 signal was developed. Following CLIA guidelines the DKK1 RNAscope chromogenic in situ hybridization assay and digital image analysis algorithm were successfully validated for sensitivity, specificity, accuracy, and precision. The DKK1 RNAscope assay in conjunction with the digital image analysis solution is acceptable for prospective screening of G/GEJ adenocarcinoma patients. The work described here will further advance the companion diagnostic development of our DKK1 RNAscope assay and could generally be used as a guide for the validation of RNAscope assays with digital image quantification.
Dickkopf-1（DKK1）是一种分泌型 Wnt 信号调节剂，在肿瘤中经常过表达，并与不良的临床结局相关。DKN-01 是一种人源化的单克隆治疗性抗体，与 DKK1 具有高亲和力，并且在肿瘤中 DKK1 表达升高的胃/胃食管交界处（G/GEJ）患者中显示出临床活性。在这里，我们报告了一种 DKK1 RNAscope 显色原位杂交检测方法，用于评估 G/GEJ 肿瘤组织中的 DKK1 表达。为了减少病理学家的时间，减少手动评分的潜在病理学家变异性，并支持病理学家的决策，开发了一种数字图像分析算法，该算法可以识别肿瘤细胞并量化 DKK1 信号。根据 CLIA 指南，成功验证了 DKK1 RNAscope 显色原位杂交检测方法和数字图像分析算法的灵敏度、特异性、准确性和精密度。DKK1 RNAscope 检测与数字图像分析解决方案相结合，可用于 G/GEJ 腺癌患者的前瞻性筛查。这里描述的工作将进一步推进我们的 DKK1 RNAscope 检测的伴随诊断开发，并且通常可以用作具有数字图像定量的 RNAscope 检测验证的指南。
