Predicting Response to Programmed Cell Death Protein-1 or Programmed Death-Ligand 1 Blockade in NSCLC—Is Multiplex Immunohistochemistry or Immunofluorescence the Answer?

Although immune checkpoint inhibitors (ICIs) have provided a welcome paradigm shift in treating NSCLC with remarkable survival benefits some patients, their success has been tempered by low response rates and difficulty identifying accurate predictive biomarkers. Effective biomarkers to help identify responders, minimize unnecessary cost toxicity nonresponders, when use different combination treatments would be of great clinical benefit. A variety investigated spanning from expression single protein, such as programmed death-ligand 1 (PD-L1), global “immune signatures” estimates tumor antigenicity using various assays assess mutational burden (TMB).1Camidge D.R. Doebele R.C. Kerr K.M. Comparing contrasting for immunotherapy targeted therapy NSCLC.Nat Rev Clin Oncol. 2019; 16: 341-355Crossref PubMed Scopus (166) Google Scholar, 2Lantuejoul S. Sound-Tsao M. Cooper W.A. et al.PD-L1 testing lung cancer 2019: perspective the IASLC pathology committee.J Thorac 2020; 15: 499-519Abstract Full Text PDF (108) 3Sholl L.M. Hirsch F.R. Hwang D. al.The promises challenges mutation an biomarker: International Association Study Lung Cancer Pathology 1409-1424Abstract (56) Scholar Nevertheless, given complex dynamic interplay vast array inhibitory stimulatory signals within microenvironment, it is not surprising that solitary parameter, PD-L1 or TMB, insufficient reliably responders.1Camidge Gene profiling approaches (e.g., NanoString) evaluate multiple mRNA levels simultaneously providing more assessment elements specimen, but date, they disappointing biomarkers.4Lu Stein J.E. Rimm D.L. al.Comparison biomarker modalities predicting PD-1/PD-L1 blockade: systematic review meta-analysis.JAMA 5: 1195-1204Crossref (191) Could nuanced approach proteins together spatial multiplex immunohistochemistry (mIHC) immunofluorescence (IF) capture complexity improve stratification patients ICIs? mIHC IF can density cell subsets, markers, localization section formalin-fixed, paraffin-embedded (FFPE) tissue enabling comprehensive microenvironment.5Hofman P. Badoual C. Henderson F. al.Multiplexed molecular cancer—just about ready prime-time?.Cancers (Basel). 11: 283Crossref (51) 6Tan W.C.C. Nerurkar S.N. Cai H.Y. al.Overview immunohistochemistry/immunofluorescence techniques era immunotherapy.Cancer Commun (Lond). 40: 135-153Crossref (68) 7Taube J.M. Akturk G. Angelo Society Immunotherapy statement on best practices (IHC) staining validation.J Immunother Cancer. 8e000155Crossref (44) Interestingly, recent meta-analysis anti–programmed death protein-1 (PD-1) therapies across types, including NSCLC, found were superior PD-L1, gene profiling.4Lu These included CD8, PD-1, TILs dormant T subsets. In this issue Journal Thoracic Oncology, proof-of-principle study Yeong al.8Yeong J, Suteja L, Simoni Y, al. Intratumoral CD39+CD8+ cells predict blockade [e-pub ahead print]. J https://doi.org/10.1016/j.jtho.2021.04.016, accessed July 6, 2021.Google reveals (mIF) antigen-specific cytotoxic subsets sections FFPE associated higher objective ICIs. Their built data previous discovery work multiplexed mass cytometry deep profiling, which reported subpopulation tumor-specific characterized coexpression CD39 CD89Simoni Y. Becht E. Fehlings al.Bystander CD8+ are abundant phenotypically distinct human tumour infiltrates.Nature. 2018; 557: 575-579Crossref (467) hypothesized subset could outcome. immunomodulatory molecule elicited microenvironment marker functionally exhausted cells.10Canale F.P. Ramello M.C. Núñez N. al.CD39 defines exhaustion tumor-infiltrating cells.Cancer Res. 78: 115-128Crossref (130) poised target tumors if function restored, making them ideal targeting ICIs PD-1 pathway central regulator exhaustion.10Canale Of three used clinically applicable assay, only CD39+ identified mIF correlated significantly data, although association was perfect (r = 0.624). The assay then reveal ratio had among 35 who received axis second advanced line treatment. No other clinicopathological age, smoking status, EGFR CD39, ICI treatment response, cohort small heterogeneous mutations matched Previous studies revealed rather than combinations markers specific instances incorporating effective at ICIs, CD3, CD4, granzyme B.11Gettinger Choi J. Mani al.A TIL phenotype non-small carcinomas sensitive blockers.Nat Commun. 9: 3196Crossref (80) 12Thommen D.S. Koelzer V.H. Herzig transcriptionally PD-1+ pool potential non-small-cell treated blockade.Nat Med. 24: 994-1004Crossref (400) 13Mazzaschi Madeddu Falco A. al.Low lymphocytes confers immune-privileged prognostic value.Clin 407-419Crossref (123) 14Wu S.P. Liao R.Q. Tu al.Stromal PD-L1-positive regulatory PD-1-positive CD8-positive define immunotherapy.J 13: 521-532Abstract (66) 15Althammer Tan T.H. Spitzmüller al.Automated image analysis biopsies anti-PD-L1 therapy.J 7: 121Crossref (40) 16Herbst R.S. Soria J.C. Kowanetz al.Predictive correlates antibody MPDL3280A patients.Nature. 2014; 515: 563-567Crossref (3287) Now, protein enable assessed same subcellular location, cellular constituents locations stromal intratumoral.5Hofman There approaches, each own advantages disadvantages brightfield fluorescent direct indirect methods antigen detection, latter signal amplification achieve greater sensitivity. If antibodies host species used, inactivation binding required between cycle immunolabelling acquisition, chemical heat-based approaches.5Hofman Scholar,6Tan Chromogenic advantage being analyzable conventional microscope, evaluation difficult unless validated software available. Vectra platform (Akoya Biosciences, Menlo Park, CA) likely most often technology, number limited, ranging six nine, possibly 12 future. Conversely, up 30 40 one highly consisting metal-based DNA barcoding–based IF. includes DSP GeoMx Spatial profiler (NanoString Technologies, Inc., Seattle, WA) allows high-plex quantification 84 1800 18,000 targets new transcriptome atlas whole panels, respectively. general, significant requiring minimal (the IHC assay), unlike genetic-based TMB generally require 10 times much tissue. Multiplex chromogenic well suited digital rapid resolution extraction large amounts data. By contrast, IHC, inexpensive, typically scored subjective semiquantitative manner pathologists limited ability multiplexing must compartments types. offers many benefits, technology without challenges. labor intensive slower specialized expensive equipment, platforms, storage widely available laboratories. accuracy reproducibility laboratories established, careful time-consuming optimization validation ensure performs assay.7Taube This particularly important owing false positives completely deactivated cycle. Assessment few regions interest, mIF, slide faster requires less acquisition may representative assessment.5Hofman addition, preanalytical variables affect assays, inadequate fixation prolonged cold ischemic time, just relevant assays. Furthermore, similar continuous variables, threshold distinguish “positive” “negative” needs determined marker. Different clones, protocols, cutoffs, all need standardized value. Considering community faced standardizing even IF, far face hurdles reach standards. Lower plex better applications, whereas research discovery. Even chance applicability potentially achieved specialized, costly microscopes. aside, promising decisions immunotherapy. Far done, however, convince scientific clinic. Much larger ideally wider homogeneous patient populations protocols those no targetable genetic alterations first-line ICI, prospective trial. Direct comparison established where appropriate, mIHC, also compare performance determine truly NSCLC. provides opportunity hope moving toward personalized medicine, regulation nature categorical unlikely ever match mutation-based therapies. there room incremental gains maximize benefit risk patients. right concept attempting translate costly, experimental approach, clear we yet. Whether value over outweighing disadvantages, into remains open question. Wendy Cooper: Conceptualization, Writing—original draft. Sylvie Lantuejoul, Mari Mino-Kenudson: Writing—review editing. Cells Predict Response Programmed Cell Death Protein-1 Ligand-1 Blockade Patients With NSCLCJournal OncologyVol. 16Issue 8PreviewProgrammed (PD-L1) currently metastatic Despite stratification, responses vary. Thus, search novel improved prediction ongoing. Previously, time-of-flight (CyTOF), our group demonstrated represent antigen-specific, treatment-naive Full-Text