Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors
Abstract 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/gastro...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-89060-3 |
| _version_ | 1830145080192663552 |
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| author | Charles Caldwell James B. Rottman Will Paces Elizabeth Bueche Sofia Reitsma Joseph Gibb Vitria Adisetiyo Michael S. Haas Heidi Heath Walter Newman Jason Baum Roberto Gianani Michael H. Kagey |
| author_facet | Charles Caldwell James B. Rottman Will Paces Elizabeth Bueche Sofia Reitsma Joseph Gibb Vitria Adisetiyo Michael S. Haas Heidi Heath Walter Newman Jason Baum Roberto Gianani Michael H. Kagey |
| author_sort | Charles Caldwell |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-17T10:18:05Z |
| format | Article |
| id | doaj.art-386bfb9934384f32ac9a8bd2228acb37 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-17T10:18:05Z |
| publishDate | 2021-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-386bfb9934384f32ac9a8bd2228acb372022-12-21T21:52:52ZengNature PortfolioScientific Reports2045-23222021-05-0111111210.1038/s41598-021-89060-3Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumorsCharles Caldwell0James B. Rottman1Will Paces2Elizabeth Bueche3Sofia Reitsma4Joseph Gibb5Vitria Adisetiyo6Michael S. Haas7Heidi Heath8Walter Newman9Jason Baum10Roberto Gianani11Michael H. Kagey12Flagship BiosciencesAthenaeum Pathology ConsultingFlagship BiosciencesFlagship BiosciencesFlagship BiosciencesFlagship BiosciencesFlagship BiosciencesLeap Therapeutics Inc.Leap Therapeutics Inc.Leap Therapeutics Inc.Leap Therapeutics Inc.Flagship BiosciencesLeap Therapeutics Inc.Abstract 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.https://doi.org/10.1038/s41598-021-89060-3 |
| spellingShingle | Charles Caldwell James B. Rottman Will Paces Elizabeth Bueche Sofia Reitsma Joseph Gibb Vitria Adisetiyo Michael S. Haas Heidi Heath Walter Newman Jason Baum Roberto Gianani Michael H. Kagey Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors Scientific Reports |
| title | Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors |
| title_full | Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors |
| title_fullStr | Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors |
| title_full_unstemmed | Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors |
| title_short | Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors |
| title_sort | validation of a dkk1 rnascope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors |
| url | https://doi.org/10.1038/s41598-021-89060-3 |
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