Cancer-Associated Fibroblasts Are Key Determinants of Cancer Cell Invasion in the Earliest Stage of Colorectal Cancer

Hao Dang; Tom J Harryvan; Chen-Yi Liao; Erik H J Danen; Vienna N L N Spalburg; Szymon M Kielbasa; Hailiang Mei; Jelle J Goeman; Eveline S de Jonge-Muller; Stefanus G T Janson; Johan J van der Reijden; Stijn Crobach; James C H Hardwick; Jurjen J Boonstra; Noel F C C de Miranda; Lukas J A C Hawinkels

doi:10.1016/j.jcmgh.2023.04.004

Cancer-Associated Fibroblasts Are Key Determinants of Cancer Cell Invasion in the Earliest Stage of Colorectal Cancer

Cell Mol Gastroenterol Hepatol. 2023;16(1):107-131. doi: 10.1016/j.jcmgh.2023.04.004. Epub 2023 Apr 20.

Authors

Hao Dang¹, Tom J Harryvan¹, Chen-Yi Liao², Erik H J Danen², Vienna N L N Spalburg¹, Szymon M Kielbasa³, Hailiang Mei³, Jelle J Goeman³, Eveline S de Jonge-Muller¹, Stefanus G T Janson¹, Johan J van der Reijden¹, Stijn Crobach⁴, James C H Hardwick¹, Jurjen J Boonstra¹, Noel F C C de Miranda⁴, Lukas J A C Hawinkels⁵

Affiliations

¹ Department of Gastroenterology and Hepatology, Leiden University Medical Center, Leiden, The Netherlands.
² Division of Drug Discovery and Safety, Leiden Academic Centre of Drug Research, Leiden University, Leiden, The Netherlands.
³ Department of Biomedical Data Sciences, Leiden University Medical Center, Leiden, The Netherlands.
⁴ Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
⁵ Department of Gastroenterology and Hepatology, Leiden University Medical Center, Leiden, The Netherlands. Electronic address: l.j.a.c.hawinkels@lumc.nl.

Abstract

Background & aims: Improving clinical management of early stage colorectal cancers (T1CRCs) requires a better understanding of their underlying biology. Accumulating evidence shows that cancer-associated fibroblasts (CAFs) are important determinants of tumor progression in advanced colorectal cancer (CRC), but their role in the initial stages of CRC tumorigenesis is unknown. Therefore, we investigated the contribution of T1CAFs to early CRC progression.

Methods: Primary T1CAFs and patient-matched normal fibroblasts (NFs) were isolated from endoscopic biopsy specimens of histologically confirmed T1CRCs and normal mucosa, respectively. The impact of T1CAFs and NFs on tumor behavior was studied using 3-dimensional co-culture systems with primary T1CRC organoids and extracellular matrix (ECM) remodeling assays. Whole-transcriptome sequencing and gene silencing were used to pinpoint mediators of T1CAF functions.

Results: In 3-dimensional multicellular cultures, matrix invasion of T1CRC organoids was induced by T1CAFs, but not by matched NFs. Enhanced T1CRC invasion was accompanied by T1CAF-induced ECM remodeling and up-regulation of CD44 in epithelial cells. RNA sequencing of 10 NF-T1CAF pairs revealed 404 differentially expressed genes, with significant enrichment for ECM-related pathways in T1CAFs. Cathepsin H, a cysteine-type protease that was specifically up-regulated in T1CAFs but not in fibroblasts from premalignant lesions or advanced CRCs, was identified as a key factor driving matrix remodeling by T1CAFs. Finally, we showed high abundance of cathepsin H-expressing T1CAFs at the invasive front of primary T1CRC sections.

Conclusions: Already in the earliest stage of CRC, cancer cell invasion is promoted by CAFs via direct interactions with epithelial cancer cells and stage-specific, cathepsin H-dependent ECM remodeling. RNA sequencing data of the 10 NF-T1CAF pairs can be found under GEO accession number GSE200660.

Keywords: Cancer-Associated Fibroblast; T1 Colorectal Cancer; Tumor Microenvironment.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cancer-Associated Fibroblasts* / metabolism
Cathepsin H / metabolism
Colorectal Neoplasms* / pathology
Fibroblasts / metabolism
Humans
Neoplasm Invasiveness / genetics
Neoplasm Invasiveness / pathology

Substances

Cathepsin H