Accounting for intensity variation in image analysis of large-scale multiplexed clinical trial datasets
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Authors
Frei, A. L.McGuigan, A.
Sinha, R. R.
Glaire, M. A.
Jabbar, F.
Gneo, L.
Tomasevic, T.
Harkin, A.
Iveson, T. J.
Saunders, Mark P
Oein, K.
Maka, N.
Pezella, F.
Campo, L.
Hay, J.
Edwards, J.
Sansom, O. J.
Kelly, C.
Tomlinson, I.
Kildal, W.
Kerr, R. S.
Kerr, D. J.
Danielsen, H. E.
Domingo, E.
Church, D. N.
Koelzer, V. H.
Affiliation
Department of Pathology and Molecular Pathology, University Hospital Zurich, University of Zurich, Zurich, SwitzerlandIssue Date
2023
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Multiplex immunofluorescence (mIF) imaging can provide comprehensive quantitative and spatial information for multiple immune markers for tumour immunoprofiling. However, application at scale to clinical trial samples sourced from multiple institutions is challenging due to pre-analytical heterogeneity. This study reports an analytical approach to the largest multi-parameter immunoprofiling study of clinical trial samples to date. We analysed 12,592 tissue microarray (TMA) spots from 3,545 colorectal cancers sourced from more than 240 institutions in two clinical trials (QUASAR 2 and SCOT) stained for CD4, CD8, CD20, CD68, FoxP3, pan-cytokeratin, and DAPI by mIF. TMA slides were multi-spectrally imaged and analysed by cell-based and pixel-based marker analysis. We developed an adaptive thresholding method to account for inter- and intra-slide intensity variation in TMA analysis. Applying this method effectively ameliorated inter- and intra-slide intensity variation improving the image analysis results compared with methods using a single global threshold. Correlation of CD8 data derived by our mIF analysis approach with single-plex chromogenic immunohistochemistry CD8 data derived from subsequent sections indicates the validity of our method (Spearman's rank correlation coefficients ρ between 0.63 and 0.66, p ≪ 0.01) as compared with the current gold standard analysis approach. Evaluation of correlation between cell-based and pixel-based analysis results confirms equivalency (ρ > 0.8, p ≪ 0.01, except for CD20 in the epithelial region) of both analytical approaches. These data suggest that our adaptive thresholding approach can enable analysis of mIF-stained clinical trial TMA datasets by digital pathology at scale for precision immunoprofiling.Citation
Frei AL, McGuigan A, Sinha RR, Glaire MA, Jabbar F, Gneo L, et al. Accounting for intensity variation in image analysis of large-scale multiplexed clinical trial datasets. The journal of pathology Clinical research. 2023 Sep 11. PubMed PMID: 37697694. Epub 2023/09/12. eng.Journal
Journal of Pathol Clinical ResearchDOI
10.1002/cjp2.342PubMed ID
37697694Additional Links
https://doi.org/10.1002/cjp2.342Type
ArticleLanguage
enae974a485f413a2113503eed53cd6c53
10.1002/cjp2.342