Spatial transcriptomic analysis of tumour with high and low CAIX expression in TNBC tissue samples using GeoMx™ RNA assay
Suad A.K. Shamis1,2, Francesca Savioli1, Aula Ammar2, Sara S.F. Al-Badran2, Phimmada Hatthakarnkul2, Holly Leslie2, Elizabeth E.A. Mallon3, Nigel B. Jamieson2, Donald C. McMillan1 and Joanne Edwards2
1Academic Unit of Surgery, School of Medicine, University of Glasgow, Royal Infirmary, Alexandria Parade, 2Unit of Molecular Pathology, School of Cancer Sciences, University of Glasgow, Wolfson Wohl Cancer Research Centre, Garscube Estate and 3Department of Pathology, Queen Elizabeth University Hospital, Glasgow, United Kingdom
Corresponding Author: Suad A.K. Shamis, Academic Unit of Surgery, School of Medicine, University of Glasgow, Royal Infirmary, Alexandria Parade, G31 2ER, Glasgow, UK. e-mail: S.Shamis.1@research.gla.ac.uk
Summary. Purpose. Prognostic significance and gene signatures associated with carbonic anhydrase IX (CAIX) was investigated in triple negative breast cancer (TNBC) patients.
Methods. Immunohistochemistry (IHC) for CAIX was performed in tissue microarrays (TMAs) of 136 TNBC patients. In a subset of 52 patients Digital Spatial Profiler (DSP) was performed in tumour (pan-cytokeratin+) and stroma (pan-cytokeratin-). Differentially expressed genes (DEGs) with P<0.05 and log2 fold change (FC)>(±0.25 and ±0.3, for tumour and stromal compartment, respectively) were identified. Four genes were validated at the protein level.
Result. Cytoplasmic CAIX expression was independently associated with poor recurrence free survival in TNBC patients [hazard ratio (HR)=6.59, 95% confidence interval (CI): 1.47-29.58, P=0.014]. DEG analysis identified 4 up-regulated genes (CD68, HIF1A, pan-melanocyte, and VSIR) in the tumour region and 9 down-regulated genes in the stromal region (CD86, CD3E, MS4A1, BCL2, CCL5, NKG7, PTPRC, CD27, and FAS) when low versus high CAIX expression was explored. Employing IHC, high CD68 and HIF-1α was associated with poorer prognosis and high BCL2 and CD3 was associated with good prognosis.
Conclusions. DSP technology identified DEGs in TNBC. Selected genes validated by IHC showed involvement of CD3 and BCL2 expression within stroma and HIF-1α, and CD68 expression within tumour. However, further functional analysis is warranted. Histol Histopathol 39, 177-200 (2024)
Key words: Breast cancer, CAIX, Hypoxia gene expression, Nanostring nCounter techonology transcriptomics, HIF-1α, Apoptosis, BCL2, Infiltrating macrophages, CD68, Lymphocytes, CD3, Stromal microenvironment
DOI: 10.14670/HH-18-655
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©The Author(s) 2024. Open Access. This article is licensed under a Creative Commons CC-BY International License. |
