Validation of a rapid HLA-DQA105 pharmacogenomics assay to identify at-risk resistance to anti-tumor necrosis factor therapy among patients with inflammatory bowel disease

• Published in: American journal of clinical pathology 2023-08, Vol.160 (2), p.194-199
• Main Authors: DelBaugh, Regina M, Cook, Leanne J, Siegel, Corey A, Tsongalis, Gregory J, Khan, Wahab A
• Format: Article
• Language: English
• Subjects: Adalimumab; Antagonists; Complications and side effects; Development and progression; DNA probes; DQA1 protein; Genetic aspects; Genome-wide association studies; Genomics; Genotypes; Genotyping; Histocompatibility antigen HLA; Histocompatibility antigens; HLA histocompatibility antigens; Immunogenicity; Inflammatory bowel disease; Inflammatory bowel diseases; Intestine; Laboratories; Necrosis; Pharmacogenetics; Pharmacogenomics; Scientific equipment and supplies industry; Tumor necrosis factor; Tumor necrosis factor-TNF; Tumors
• ISSN: 0002-9173; 1943-7722
• DOI: 10.1093/ajcp/aqad036

The HLA-DQA1*05 variant (rs2097432) is associated with increased risk of immunogenicity to tumor necrosis factor antagonists, with subsequent resistance to therapy in patients with inflammatory bowel disease. Identification of these patients would optimize personalized therapeutic selection. Genomic DNA was extracted from 80 deidentified samples in an unselected patient population with an unknown rs2097432 genotype. Split sample analysis was performed using a reference laboratory. Primer probes for a TaqMan quantitative polymerase chain reaction (qPCR) assay (Thermo Fisher Scientific) were custom designed. Synthesized genomic-block fragments were used as controls. All qPCR reactions were performed using a TaqMan GTXpress Master Mix (Thermo Fisher Scientific) on the Applied Biosystems 7500 system under fast cycling conditions. Of 80 samples, 50% were wild-type reference genotypes, 22.5% were heterozygous, and 27.5% were homozygous variant calls, comparable to population data. Split analysis samples between 2 independent laboratories were 100% concordant. The detection limit tested across genomic-block controls processed in duplicate was reproducible on sample input from 10 ng titrated down to 1.25 ng across 2 independent runs. Further, analytical specificity assessed with previous wild-type reference and homozygous variant DNA spiked into genomic-block controls produced appropriate heterozygous genotypes. Here we present validation of a lab-developed test for a rapid HLA-DQA1*05 (rs2097432) pharmacogenomics assay targeting a hotspot identified by genome-wide association studies. Targeted genotyping employed here will allow for expeditious personalized therapeutic selection.