Digital image analysis of multiplex fluorescence IHC in colorectal cancer recognizes the prognostic value of CDX2 and its negative correlation with SOX2

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Digital image analysis of multiplex fluorescence IHC in colorectal cancer recognizes the prognostic value of CDX2 and its negative correlation with SOX2

Tuesday, 26.11.2019

Flourescence-based multiplex immunohistochemistry (mIHC) combined with multispectral imaging and digital image analysis (DIA) is a quantitative high-resolution method for determination of protein expression in tissue. We applied this method for five biomarkers (CDX2, SOX2, SOX9, E-cadherin, and β-catenin) using tissue microarrays of a Norwegian unselected series of primary colorectal cancer. The data were compared with previously obtained chromogenic IHC data of the same tissue cores, visually assessed by the Allred method. We found comparable results between the methods, although confirmed that DIA offered improved resolution to differentiate cases with high and low protein expression. However, we experienced inherent challenges with digital image analysis of membrane staining, which was better assessed visually. DIA and mIHC enabled quantitative analysis of biomarker coexpression on the same tissue section at the single-cell level, revealing a strong negative correlation between the differentiation markers CDX2 and SOX2. Both methods confirmed known prognostic associations for CDX2, but DIA improved data visualization and detection of clinicopathological and biological associations. In summary, mIHC combined with DIA is an efficient and reliable method to evaluate protein expression in tissue, here shown to recapitulate and improve detection of known clinicopathological and survival associations for the emerging biomarker CDX2, and is therefore a candidate approach to standardize CDX2 detection in pathology laboratories.

 

Authors and Affiliations:

Lopes N1,2,3,4, Bergsland CH1,4,5, Bjørnslett M1,4, Pellinen T4,6, Svindland A4,5, Nesbakken A4,5,7, Almeida R2,8,9, Lothe RA1,4,5, David L2,3,8, Bruun J10,11.

1 Department of Molecular Oncology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.

2 i3S-Institute for Research and Innovation in Health, University of Porto, Porto, Portugal.

3 IPATIMUP-Institute of Molecular Pathology and Immunology, University of Porto, Porto, Portugal.

4 K.G. Jebsen Colorectal Cancer Research Centre, Division of Cancer Medicine, Oslo University Hospital, Oslo, Norway.

5 Institute for Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway.

6 Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Helsinki, Finland.

7 Department of Gastrointestinal Surgery, Oslo University Hospital, Oslo, Norway.

8 Faculty of Medicine, University of Porto, Porto, Portugal.

9 Department of Biology, Faculty of Sciences, University of Porto, Porto, Portugal.

10 Department of Molecular Oncology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.

11 K.G. Jebsen Colorectal Cancer Research Centre, Division of Cancer Medicine, Oslo University Hospital, Oslo, Norway

 

Abstract:

Flourescence-based multiplex immunohistochemistry (mIHC) combined with multispectral imaging and digital image analysis (DIA) is a quantitative high-resolution method for determination of protein expression in tissue. We applied this method for five biomarkers (CDX2, SOX2, SOX9, E-cadherin, and β-catenin) using tissue microarrays of a Norwegian unselected series of primary colorectal cancer. The data were compared with previously obtained chromogenic IHC data of the same tissue cores, visually assessed by the Allred method. We found comparable results between the methods, although confirmed that DIA offered improved resolution to differentiate cases with high and low protein expression. However, we experienced inherent challenges with digital image analysis of membrane staining, which was better assessed visually. DIA and mIHC enabled quantitative analysis of biomarker coexpression on the same tissue section at the single-cell level, revealing a strong negative correlation between the differentiation markers CDX2 and SOX2. Both methods confirmed known prognostic associations for CDX2, but DIA improved data visualization and detection of clinicopathological and biological associations. In summary, mIHC combined with DIA is an efficient and reliable method to evaluate protein expression in tissue, here shown to recapitulate and improve detection of known clinicopathological and survival associations for the emerging biomarker CDX2, and is therefore a candidate approach to standardize CDX2 detection in pathology laboratories.

 

JournalLaboratory Investigation

 

LinKhttps://www.nature.com/articles/s41374-019-0336-4