Aligning digital CD8⁺ scoring and targeted next‐generation sequencing with programmed death ligand 1 expression: a pragmatic approach in early‐stage squamous cell lung carcinoma

Abstract

Aims To study programmed death ligand 1 (PD‐L1) expression, tumour‐infiltrating T lymphocytes (TILs) and the molecular context in patients with early‐stage squamous cell lung carcinomas (SCCs). Methods and results The study included samples from 40 patients (discovery cohort) and 29 patients (validation cohort) diagnosed with early‐stage SCC. PD‐L1 immunohistochemistry (IHC) was performed with three commercially available clones (E1L3N, SP263 and SP142). CD8+ TILs were scored with a digital algorithm. All tumours were analysed with targeted next‐generation sequencing (NGS). Additionally, TP53 mutations were investigated with direct sequencing. In both cohorts, we observed a significant association between CD8+ TILs density and high PD‐L1 IHC expression in tumour cells (TCs). Furthermore, high SP142 PD‐L1 expression in immune cells (ICs) was also associated significantly with CD8+ TILs density. Therefore, CD8+ TILs density discriminated between patients with high versus low PD‐L1 IHC expression with excellent sensitivity and specificity. Interestingly, the highest percentages of PD‐L1‐positive TCs with the three antibodies were found in samples with cyclin‐dependent kinase 6 (CDK6) amplification, with high amplification of proto‐oncogene C‐Myc (CMYC) or with cyclin D1–PI3 kinase subunit alpha (CCND1–PIK3CA) co‐amplification. High SP142 PD‐L1 IHC expression in ICs showed a non‐significant correlation with TP53 mutations. Conversely, most cases with fibroblast growth factor receptor 1 (FGFR1) amplification were negative for all PD‐L1 clones. Conclusions Our preliminary results support the use of digital CD8+ TILs scoring and targeted NGS alongside PD‐L1 expression. The approach presented herein could help define patients with SCCs candidates to immune checkpoints inhibitors.

Prognostic impact of CD8 and programmed death-ligand 1 expression in patients with resectable non-small cell lung cancer

“…While some studies used the proportion of CD8-positive cells, 13,23,24 others including this study determined the cut-off value of CD8 by its absolute number. 22,42 Statistical parameters used to identify the cut-off value were also various, including mean, median, quartile, and previously reported values. 13,[22][23][24]42,43 Subsequent analyses will be necessary to establish definitive cut-off value for CD8 positivity.…”

“…22,42 Statistical parameters used to identify the cut-off value were also various, including mean, median, quartile, and previously reported values. 13,[22][23][24]42,43 Subsequent analyses will be necessary to establish definitive cut-off value for CD8 positivity. Fifth, PD-L1 expression was determined using particular PD-L1 detection antibodies and IHC.…”

ESMO recommendations on the standard methods to detect NTRK fusions in daily practice and clinical research

“…In addition to the AMP technology, other NGS platforms can offer the possibility to test for NTRK fusions. These include the GeneTrails Solid Tumor Fusion Gene Panel (Knight Diagnostic Laboratories), designed to detect fusions involving 20 target genes including NTRK1, NTRK2, NTRK3 [57]; the Universal Fusion/Expression Profile (Neogenomics), an assay capable of detecting different classes of genomic abnormalities such as fusion transcripts and transcriptomic gene expression levels in 1385 genes (NTRK1, NTRK2, NTRK3 included); and the Oncomine assays (ThermoFisher Scientific), which cover fusion variants including NTRK1, NTRK2 and NTRK3 [58].…”

Updated guidelines for predictive biomarker testing in advanced non-small-cell lung cancer: a National Consensus of the Spanish Society of Pathology and the Spanish Society of Medical Oncology

“…For this reason, KRAS testing is currently not indicated as an individual test but it is appropriate that the study of the KRAS gene is included in extended panels [21,22]. With regards to other potential biomarkers predictive of an immune response, the microsatellite instability and the immune microenvironment study should be highlighted, from the viewpoint of RNA expression and tissue determination of multiple immune cells [17,58,86,87].…”