Supplementary figures S1-7 from LOXL2 Is Highly Expressed in Cancer-Associated Fibroblasts and Associates to Poor Colon Cancer Survival

Torres, Sofía; Garcia-Palmero, Irene; Herrera, Mercedes; Bartolomé, Rubén A.; Peña, Cristina; Fernandez-Aceñero, M. Jesús; Padilla, Guillermo; Peláez-García, Alberto; Lopez-Lucendo, María; Rodriguez-Merlo, Rufo; de Herreros, Antonio García; Bonilla, Félix; Casal, J. Ignacio

doi:10.1158/1078-0432.22459233.v1

Supplementary figures S1-7 from LOXL2 Is Highly Expressed in Cancer-Associated Fibroblasts and Associates to Poor Colon Cancer Survival

journal contribution

posted on 2023-03-31, 18:49 authored by Sofía Torres, Irene Garcia-Palmero, Mercedes Herrera, Rubén A. Bartolomé, Cristina Peña, M. Jesús Fernandez-Aceñero, Guillermo Padilla, Alberto Peláez-García, María Lopez-Lucendo, Rufo Rodriguez-Merlo, Antonio García de Herreros, Félix Bonilla, J. Ignacio Casal

Supplementary figures S1-7. Figure S1. Characterization of fibroblasts isolated from patients with colon cancer. Figure S2. Proteins identified and quantified in both replicates of iTRAQ. Figure S3. Major signaling pathways affected by fibroblast activation. Figure S4 Expression of stromal biomarkers in tissues of colorectal cancer patients using HPA. Figure S5. Prognostic value of individual CAFs markers for overall survival according to GSE17538 dataset. Figure S6. Prognostic value of individual CAFs markers for disease-free survival according to GSE17538 in patients at stages I-III. Figure S7. Nuclear staining of LOXL2 in the stroma of human colon cancer

History

ARTICLE ABSTRACT

Purpose: Cancer-associated fibroblasts (CAF) are major mediators in tumor microenvironment. We investigated the changes in protein expression in colon cancer–associated fibroblasts compared with normal fibroblasts (NF) in the context of searching for prognostic biomarkers, particularly for stage II patients.Experimental Design: CAFs and NFs isolated from colon cancer patients were used to identify differentially expressed proteins using quantitative proteomics. Stromal expression of deregulated proteins was analyzed by IHC. Prognostic impact was studied using external gene-expression datasets for training, then quantitative PCR and IHC for validation in different cohorts of patients. Combined datasets were used for prediction of risk assessment at stages II and III.Results: A desmoplastic signature composed of 32 proteins, highly specific for stromal components in colon cancer, was identified. These proteins were enriched for extracellular matrix organization components, TGFβ signaling pathway, fibrosis, and wound-healing proteins. The expression in CAFs of 11 upregulated proteins and four downregulated proteins, selected for biomarker validation, was verified by orthogonal techniques. LOXL2 displayed a high prognostic impact by using external independent datasets and further validation in two different cohorts of patients. High expression of LOXL2 was associated with higher recurrence P = 0.001 HR, 5.38 [95% confidence interval (CI), 1.70–17.01] and overall survival P = 0.001 HR, 8.52 (95% CI, 1.90–38.29). IHC analysis revealed a prognostic value for LOXL2 in stage II patients.Conclusions: We identified LOXL2 to be associated with the outcome of colon cancer patients. Furthermore, it can be used to stratify patients at stages II and III for further therapeutic decisions. Clin Cancer Res; 21(21); 4892–902. ©2015 AACR.