Abstract
RUNX3, an important regulator of T-cell differentiation plays a crucial role in activation and maintenance of various T-cells including cytotoxic-, helper- and memory-cells. Former studies have highlighted RUNX3 as a prognostic factor across several cancer types, but few have studied its cell-type specific expression patterns. We used multiplex immunohistochemistry to assess and quantify cells co-expressing CD3 and RUNX3 in three cohorts consisting of 452 colon adenocarcinoma (COAD), 239 lung adenocarcinoma (LUAD) and 307 lung squamous cell carcinoma (LUSC) patients. Further, we correlated the expression of CD3/RUNX3 to disease-specific survival and to previously investigated immune markers in these cohorts. We found that a high density of CD3+RUNX3+ cells was an independent prognostic factor for disease-specific survival in COAD (HR 0.37, 95% CI 0.21–0.64) and LUAD (HR 0.61, 95% CI (0.39–0.97), but not in LUSC. Interestingly, RUNX3 expression was positively correlated with CD8 in all cohorts, CD20 in LUAD and FOXP3 in LUSC. Univariate subgroup analyses revealed that COAD patients with high numbers of both CD3+RUNX3+ and CD8+ cells rarely experienced a DSS event (HR 0.24, 95% CI 0.15–0.39). Contrasting previous studies, we did not observe RUNX3 expression in epithelial cells. A high level of CD3+RUNX3+ density is an independent prognostic factor in COAD and LUAD, but not in LUSC. In COAD, a subset of patients in stage II/III with CD3+RUNX3+high/CD8 + high may be spared adjuvant treatment due to excellent prognosis. However, further studies are needed to confirm and elucidate the protective role of CD3+RUNX3+ cells in COAD and LUAD.
Data availability
The data presented in this study are available upon reasonable request to the corresponding author. The data are not publicly available due to privacy regulations.
Abbreviations
- CD:
-
Cluster of differentiation
- CRC:
-
Colorectal cancer
- COAD:
-
Colon adenocarcinoma
- DSS:
-
Disease-specific survival
- H&E:
-
Hematoxylin and eosin
- IHC:
-
Immunohistochemistry
- LUAD:
-
Lung adenocarcinoma
- LUSC:
-
Lung squamous cell carcinoma
- ML:
-
Machine learning
- NSCLC:
-
Non-small cell lung cancer
- nStage:
-
Node stage
- pStage:
-
Pathological stage
- REK:
-
Regional committee for medical and health research ethics
- REMARK:
-
Reporting recommendations for tumour MARKer prognostic studies
- RUNX:
-
Runt related transcription factors
- TMA:
-
Tissue microarray
- TNM:
-
Tumor node metastasis
- tStage:
-
Tumor stage
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Acknowledgements
We would like to thank Magnus L. Persson for preparing the TMA-slides.
Funding
Open access funding provided by UiT The Arctic University of Norway (incl University Hospital of North Norway). The works was funded by the North Norwegian Health Authority grant no. HNF1521-20. The publication charges for this article have been funded by a grant from the publication fund of UiT The Arctic University of Norway.
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Contributions
All authors contributed to study design. H.S., E.E.P., T.D, and S.A., collected the clinical data.L.-T.R.B. collected and revised the pathological specimens. M.I.P. and A.P.L. conducted the IHC experiments. T.K.K and D.F.N devised the ML pipeline. T.K.K. analyzed and interpreted the data. T.K.K wrote the manuscript. All authors participated during revisions and approved the final manuscript.
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Competing interests
T.K.K. has received a honorary fee for participation in an advisory board for Sun Pharma, M.R. has received lecture fees from AstraZeneca. All other authors declare no competing interests.
Ethics approval and consent to participate
The study was conducted according to the guidelines of the Declaration of Helsinki, and was approved by the Regional Committee for Medical and Health Research Ethics North (REK Nord, protocol IDs: 2011/2503 and 2011/2151). The need for informed consent was waived by REK Nord due to the retrospective nature of the study. The reporting of clinicopathological variables, survival data and biomarker expression was conducted in accordance with the REMARK guidelines.
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Kilvaer, T.K., Førde, D., Paulsen, EE. et al. CD3+RUNX3+ lymphocyte density; an independent prognostic factor in colon and lung adenocarcinoma but not in lung squamous cell carcinoma. Sci Rep (2026). https://doi.org/10.1038/s41598-026-38765-4
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DOI: https://doi.org/10.1038/s41598-026-38765-4
