Validation of a DKK1 RNAscope Chromogenic in Situ Hybridization Assay for Gastric and Gastroesophageal Junction Adenocarcinoma Tumors

Overview

Journal Sci Rep

Specialty Science

Date 2021 May 11

PMID 33972574

Citations 9

Authors

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

Affiliations

Soon will be listed here.

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.

Citing Articles

DKN-01 in Combination With Tislelizumab and Chemotherapy as First-Line Therapy in Advanced Gastric or Gastroesophageal Junction Adenocarcinoma: DisTinGuish.

Klempner S, Sonbol M, Wainberg Z, Uronis H, Chiu V, Scott A J Clin Oncol. 2024; 43(3):339-349.

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Characterization of cell states in biliary tract cancers identifies mechanisms of therapeutic resistance in a phase II trial of DKN-01/nivolumab.

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Visualization of Individual RNA Molecules by Proximity Ligation-Based Chromogenic In Situ Hybridization Assay.

Xia X, Jiang M, Lin C, Ke R Methods Mol Biol. 2024; 2822:143-156.

PMID: 38907917 DOI: 10.1007/978-1-0716-3918-4_11.

Correlation of TRPA1 RNAscope and Agonist Responses.

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