Lymphocytic infiltration in stage II microsatellite stable colorectal tumors: A retrospective prognosis biomarker analysis

Autoři: Rebeca Sanz-Pamplona aff001;  Marilena Melas aff004;  Asaf Maoz aff005;  Stephanie L. Schmit aff006;  Hedy Rennert aff007;  Flavio Lejbkowicz aff007;  Joel K. Greenson aff008;  Xavier Sanjuan aff009;  Maria Lopez-Zambrano aff009;  M. Henar Alonso aff001;  Chenxu Qu aff011;  Kevin J. McDonnell aff011;  Gregory E. Idos aff011;  Marissa Vignali aff012;  Ryan Emerson aff012;  Paul Fields aff012;  Elisabet Guinó aff001;  Cristina Santos aff001;  Ramon Salazar aff001;  Harlan S. Robins aff012;  Gad Rennert aff007;  Stephen B. Gruber aff011;  Victor Moreno aff001
Působiště autorů: Catalan Institute of Oncology (ICO), Hospitalet de Llobregat, Barcelona, Spain aff001;  ONCOBELL Program, Bellvitge Biomedical Research Institute (IDIBELL), Hospitalet de Llobregat, Barcelona, Spain aff002;  Consortium for Biomedical Research in Epidemiology and Public Health (CIBERESP), Spain aff003;  Nationwide Children’s Hospital, Columbus, Ohio, United States of America aff004;  Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, United States of America aff005;  H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, United States of America aff006;  Carmel Medical Center, and Technion, Haifa, Israel aff007;  University of Michigan Medical School, Ann Arbor, Michigan, United States of America aff008;  University Hospital Bellvitge (HUB-IDIBELL), L'Hospitalet de Llobregat, Barcelona, Spain aff009;  Department of Clinical Sciences, Faculty of Medicine, University of Barcelona, Barcelona, Spain aff010;  City of Hope National Medical Center, Duarte, California, United States of America aff011;  Adaptive Biotechnologies, Seattle, Washington, United States of America aff012;  Consortium for Biomedical Research in Oncology (CIBERONC), Spain aff013;  Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America aff014
Vyšlo v časopise: Lymphocytic infiltration in stage II microsatellite stable colorectal tumors: A retrospective prognosis biomarker analysis. PLoS Med 17(9): e32767. doi:10.1371/journal.pmed.1003292
Kategorie: Research Article
doi: 10.1371/journal.pmed.1003292



Identifying stage II patients with colorectal cancer (CRC) at higher risk of progression is a clinical priority in order to optimize the advantages of adjuvant chemotherapy while avoiding unnecessary toxicity. Recently, the intensity and the quality of the host immune response in the tumor microenvironment have been reported to have an important role in tumorigenesis and an inverse association with tumor progression. This association is well established in microsatellite instable CRC. In this work, we aim to assess the usefulness of measures of T-cell infiltration as prognostic biomarkers in 640 stage II, CRC tumors, 582 of them confirmed microsatellite stable.

Methods and findings

We measured both the quantity and clonality index of T cells by means of T-cell receptor (TCR) immunosequencing in a discovery dataset (95 patients with colon cancer diagnosed at stage II and microsatellite stable, median age 67, 30% women) and replicated the results in 3 additional series of stage II patients from 2 countries. Series 1 and 2 were recruited in Barcelona, Spain and included 112 fresh frozen (FF, median age 69, 44% women) and 163 formalin-fixed paraffin-embedded (FFPE, median age 67, 39% women) samples, respectively. Series 3 included 270 FFPE samples from patients recruited in Haifa, Northern Israel, as part of a large case-control study of CRC (median age 73, 46% women). Median follow-up time was 81.1 months. Cox regression models were fitted to evaluate the prognostic value of T-cell abundance and Simpson clonality of TCR variants adjusting by sex, age, tumor location, and stage (IIA and IIB). In the discovery dataset, higher TCR abundance was associated with better prognosis (hazard ratio [HR] for ≥Q1 = 0.25, 95% CI 0.10–0.63, P = 0.003). A functional analysis of gene expression on these tumors revealed enrichment in pathways related to immune response. Higher values of clonality index (lower diversity) were not associated with worse disease-free survival, though the HR for ≥Q3 was 2.32 (95% CI 0.90–5.97, P = 0.08). These results were replicated in an independent FF dataset (TCR abundance: HR = 0.30, 95% CI 0.12–0.72, P = 0.007; clonality: HR = 3.32, 95% CI 1.38–7.94, P = 0.007). Also, the association with prognosis was tested in 2 independent FFPE datasets. The same association was observed with TCR abundance (HR = 0.41, 95% CI 0.18–0.93, P = 0.03 and HR = 0.56, 95% CI 0.31–1, P = 0.042, respectively, for each FFPE dataset). However, the clonality index was associated with prognosis only in the FFPE dataset from Israel (HR = 2.45, 95% CI 1.39–4.32, P = 0.002). Finally, a combined analysis combining all microsatellite stable (MSS) samples demonstrated a clear prognosis value both for TCR abundance (HR = 0.39, 95% CI 0.26–0.57, P = 1.3e-06) and the clonality index (HR = 2.13, 95% CI 1.44–3.15, P = 0.0002). These associations were also observed when variables were considered continuous in the models (HR per log2 of TCR abundance = 0.85, 95% CI 0.78–0.93, P = 0.0002; HR per log2 or clonality index = 1.16, 95% CI 1.03–1.31, P = 0.016).


This is a retrospective study, and samples had been preserved with different methods. Validation series lack complete information about microsatellite instability (MSI) status and pathology assessment. The Molecular Epidemiology of Colorectal Cancer (MECC) study had information about overall survival instead of progression-free survival.


Results from this study demonstrate that tumor lymphocytes, assessed by TCR repertoire quantification based on a sequencing method, are an independent prognostic factor in microsatellite stable stage II CRC.

Klíčová slova:

Biomarkers – Cancers and neoplasms – Colorectal cancer – Lymphocytes – Prognosis – T cell receptors – T cells – Tumor-infiltrating lymphocytes


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