Abstract 5573: Validation of Aspyre Clinical Test for Lung at low input levels: Simplified genomic profiling for NSCLC with minimal tissue requirements

Abstract Although many assays exist for detecting actionable variants in NSCLC, few of them are suitable for use in low quantity or quality specimens, which significantly contribute to biomarker testing failures. Aspyre Clinical Test for Lung (Tissue) is a robust simplified genomic profiling panel that tests for 114 biomarkers in NSCLC across 11 genes (ALK, BRAF, EGFR, ERBB2, KRAS, MET, RET, ROS1, NTRK1/2/3) and was able to rescue 96% of samples that failed NGS-based testing. Aspyre Clinical Test for Lung (Tissue) was initially validated for 20ng DNA and 6ng RNA input from FFPE lung tissue samples, though it was predicted to have robust performance at even lower inputs. In this study we tested the impact of using low input levels down to 5 ng DNA and 1.5 ng RNA on the sensitivity (LoD95), specificity, analytical accuracy and analytical precision of Aspyre Clinical Test for Lung (Tissue). The study included FFPE lung tissue samples from patients with NSCLC (> 10% tumor content), variant-negative FFPE tissue from tonsil tissue, and contrived samples with controlled variant allele fractions of mutant oligonucleotides in an FFPE-based background.The LoD95 for eleven different single nucleotide variants or insertions/deletions from the Aspyre Clinical Test for Lung panel were established at ≤ 3% VAF, and one SNV at 7.5% VAF. The sensitivity for seven gene fusions, which are detected by Aspyre through RNA, was ≤ 100 copies, and for MET exon 14 skipping at ≤ 200 copies. The specificity was 100% with no false positive results for any healthy tonsil tissue samples (30 samples in 60 replicates). The analytical accuracy study found no discordant variant calls between Aspyre and 1) 19 contrived samples at 2x LoD95 or 2) 29 NSCLC patient samples compared to both an orthogonal target-enrichment NGS and Aspyre at higher inputs. Results were replicable and precise across operators, reagent lots, runs, and real-time PCR instruments.This low-input study demonstrates the opportunity to deploy Aspyre Clinical Test for Lung (Tissue) for slide samples with few cells, as well as small biopsy specimens, ensuring reliable genomic profiling even in limited or challenging samples. Combined with a high success rate and a proven 2-day turnaround time, this expands access to precision medicine for NSCLC patients. Citation Format: Christina Xyrafaki, Katherine Knudsen, Magdalena Stolarek-Januszkiewicz, Rebecca Palmer, Sam Abujudeh, Ryan Evans, Eleanor Gray, Amanda Green, Robert Osborne, Barnaby Balmforth, Wendy Levin, Shari Brown. Validation of Aspyre Clinical Test for Lung at low input levels: Simplified genomic profiling for NSCLC with minimal tissue requirements [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5573.