Validation of the systematic scoring of immunohistochemically stained tumour tissue microarrays using <i>QuPath</i> digital image analysis

Loughrey, Maurice B.; Bankhead, Peter; Coleman, Helen; Hagan, Ryan S; Craig, Stephanie G.; McCorry, Amy M B; Gray, Ronan T.; McQuaid, Stephen; Dunne, Philip D.; Hamilton, Peter; James, Jacqueline; Salto-Tellez, Manuel

doi:10.1111/his.13516

Cited by 68 publications

(63 citation statements)

References 23 publications

(69 reference statements)

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“…DIA of all DAB PD-L1 SP263 IHC stained cases was performed using the open source DIA program QuPath v0.1.2, developed at Queen's University Belfast [5,[17][18][19][20]. All IHC slides were scanned at 40× on an Aperio AT2 digital scanner (Leica Biosystems, Vista, CA, USA).…”

Section: Pd-l1 Ihc Image Analysismentioning

confidence: 99%

“…All IHC slides were scanned at 40× on an Aperio AT2 digital scanner (Leica Biosystems, Vista, CA, USA). A robust workflow and rigorous quality control steps were taken to remove unsuitable areas for analysis (e.g., necrosis, tissue folds, normal structures, and non-specific staining), this was confirmed by a second reviewer with frequent consultation, as described [5,[17][18][19][20]. Briefly, digital annotations were made, within which cell detection was conducted using default parameters within QuPath.…”

Section: Pd-l1 Ihc Image Analysismentioning

confidence: 99%

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Improving the Diagnostic Accuracy of the PD-L1 Test with Image Analysis and Multiplex Hybridization

Humphries

Bingham

Sidi

et al. 2020

Cancers

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Targeting of the programmed cell death protein (PD-1)/programmed death-ligand 1 (PD-L1) axis with checkpoint inhibitors has changed clinical practice in non-small cell lung cancer (NSCLC). However, clinical assessment remains complex and ambiguous. We aim to assess whether digital image analysis (DIA) and multiplex immunofluorescence can improve the accuracy of PD-L1 diagnostic testing. A clinical cohort of routine NSCLC patients reflex tested for PD-L1 (SP263) immunohistochemistry (IHC), was assessed using DIA. Samples of varying assessment difficulty were assessed by multiplex immunofluorescence. Sensitivity, specificity, and concordance was evaluated between manual diagnostic evaluation and DIA for chromogenic and multiplex IHC. PD-L1 expression by DIA showed significant concordance (R² = 0.8248) to manual assessment. Sensitivity and specificity was 86.8% and 91.4%, respectively. Evaluation of DIA scores revealed 96.8% concordance to manual assessment. Multiplexing enabled PD-L1+/CD68+ macrophages to be readily identified within PD-L1+/cytokeratin+ or PD-L1-/cytokeratin+ tumor nests. Assessment of multiplex vs. chromogenic IHC had a sensitivity and specificity of 97.8% and 91.8%, respectively. Deployment of DIA for PD-L1 diagnostic assessment is an accurate process of case triage. Multiplex immunofluorescence provided higher confidence in PD-L1 assessment and could be offered for challenging cases by centers with appropriate expertise and specialist equipment.

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Section: Pd-l1 Ihc Image Analysismentioning

confidence: 99%

Section: Pd-l1 Ihc Image Analysismentioning

confidence: 99%

Improving the Diagnostic Accuracy of the PD-L1 Test with Image Analysis and Multiplex Hybridization

Humphries

Bingham

Sidi

et al. 2020

Cancers

Self Cite

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“…Median PD-L1 was 32 (interquartile range: 29-50) for metastatic and 24 (19)(20)(21)(22)(23)(24)(25)(26)(27) for primary cell lines (Wilcoxon rank sum test p = 0.02), and the highest value was in a metastatic cell line. Expression of MET was similar in primary and metastatic cell lines, although the interquartile range was higher for metastatic (17-41) than primary (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29) tumor cells (Wilcoxon rank sum test p = 0.44), and the highest value of MET was in a metastatic cell line (Table S1).…”

Section: Expression Of Met and Pd-l1 In Melanoma Cell Linesmentioning

confidence: 98%

Correlation of MET and PD-L1 Expression in Malignant Melanoma

Song

Desar

Pengo

et al. 2020

Cancers

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The proto-oncogene MET, the hepatocyte growth factor (HGF) receptor, is a transmembrane receptor tyrosine kinase (RTK) with a prominent role in tumor metastasis and resistance to anti-cancer therapies. Melanoma demonstrates relatively frequent MET aberrations, including MET gene amplification. Concurrently, programmed death-ligand 1 (PD-L1), with its ability to evade anti-tumor immune responses, has emerged as a prominent therapeutic target in melanoma and other malignancies and its expression is used as a predictive biomarker of response to immunotherapy. We performed immunohistochemistry analysis of MET and PD-L1 in 18 human melanoma cell lines derived from both primary and metastatic lesions, and in a human melanoma tissue microarray containing one hundreds melanocytic lesions, including primary cutaneous melanomas, primary mucosal melanomas, metastatic melanomas and benign melanocytic nevi as controls. After color deconvolution, each core was segmented to isolate staining and calculate the percentage of positive cells. Overall, MET expression was higher in tumors with increased PD-L1 expression. Moreover, a robust correlation between MET and PD-L1 expression was found in samples from metastatic melanoma and not in primary cutaneous or mucosal melanoma. These data suggest that relative expression levels of these proteins in combination is a marker of advanced disease and testing for expression of these markers should be considered in patients with melanoma.

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“…ImageJ and MATLAB), scriptable data mining, and rapid processing and export of spatial, morphological and intensity‐based data. There is also good interobserver reproducibility of bioimage data using QuPath in the TMA setting [100].…”

Section: Brightfield‐based Mihc/ifmentioning

confidence: 99%

“…The technology is, however, limited by its ability to perform digital scoring of biomarkers with more complex patterns of staining, such as mismatch repair proteins or immune checkpoint inhibitors like PD‐L1. This arises from the nature of staining multiple cell populations (tumor and immune) and the distinction between tumor and immune cell staining being difficult due to a patchy pattern of staining [100]. Several studies have illustrated the potential of QuPath in TMA biomarker scoring [99, 101, 102], and it is also currently actively utilized by researchers to analyze mIHC/IF samples in the study of cancer immunotherapy [103].…”

Section: Brightfield‐based Mihc/ifmentioning

confidence: 99%

Overview of multiplex immunohistochemistry/immunofluorescence techniques in the era of cancer immunotherapy

Tan

Nerurkar

Cai

et al. 2020

Cancer Communications

407

318

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Conventional immunohistochemistry (IHC) is a widely used diagnostic technique in tissue pathology. However, this technique is associated with a number of limitations, including high inter-observer variability and the capacity to label only one marker per tissue section. This review details various highly multiplexed techniques that have emerged to circumvent these constraints, allowing simultaneous detection of multiple markers on a single tissue section and the comprehensive study of cell composition, cellular functional and cell-cell interactions. Among these techniques, multiplex Immunohistochemistry/Immunofluorescence (mIHC/IF) has emerged to be particularly promising. mIHC/IF provides high-throughput multiplex staining and standardized quantitative analysis for highly reproducible, efficient and cost-effective tissue studies. This technique has immediate potential for translational research and clinical practice, particularly in the era of cancer immunotherapy.

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Validation of the systematic scoring of immunohistochemically stained tumour tissue microarrays using QuPath digital image analysis

Abstract: Scoring of immunohistochemically stained tumour TMAs using QuPath is functional and reproducible, even among users of limited experience of digital pathology images, and more accurate than manual scoring.

Cited by 68 publications

References 23 publications

Improving the Diagnostic Accuracy of the PD-L1 Test with Image Analysis and Multiplex Hybridization

Improving the Diagnostic Accuracy of the PD-L1 Test with Image Analysis and Multiplex Hybridization

Correlation of MET and PD-L1 Expression in Malignant Melanoma

Overview of multiplex immunohistochemistry/immunofluorescence techniques in the era of cancer immunotherapy

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