Replication Study: Wnt activity defines colon cancer stem cells and is regulated by the microenvironment

Abstract

As part of the Reproducibility Project: Cancer Biology we published a Registered Report (Evans et al., 2015), that described how we intended to replicate selected experiments from the paper ‘Wnt activity defines colon cancer stem cells and is regulated by the microenvironment’ (Vermeulen et al., 2010). Here, we report the results. Using three independent primary spheroidal colon cancer cultures that expressed a Wnt reporter construct we observed high Wnt activity was associated with the cell surface markers CD133, CD166, and CD29, but not CD24 and CD44, while the original study found all five markers were correlated with high Wnt activity (Figure 2F; Vermeulen et al., 2010). Clonogenicity was highest in cells with high Wnt activity and clonogenic potential of cells with low Wnt activity were increased by myofibroblast-secreted factors, including HGF. While the effects were in the same direction as the original study (Figure 6D; Vermeulen et al., 2010) whether statistical significance was reached among the different conditions varied. When tested in vivo, we did not find a difference in tumorigenicity between high and low Wnt activity, while the original study found cells with high Wnt activity were more effective in inducing tumors (Figure 7E; Vermeulen et al., 2010). Tumorigenicity, however, was increased with myofibroblast-secreted factors, which was in the same direction as the original study (Figure 7E; Vermeulen et al., 2010), but not statistically significant. Finally, we report meta-analyses for each results where possible.

Fibroblast-Derived Extracellular Vesicles Induce Colorectal Cancer Progression by Transmitting Amphiregulin

“…Interestingly, CAFs critically contribute to the cellular heterogeneity of CRC and to the acquision of the aggressive cancer stem cell phenotype ( Vermeulen et al, 2010 ; Essex et al, 2019 ). In addition, we found that intestinal fibroblast-derived EVs carry amphiregulin (AREG), a member of the EGF ligand family, and EVs have a central role in shaping the intestinal stem cell niche ( Oszvald et al, 2020 ).…”

How the interplay among the tumor microenvironment and the gut microbiota influences the stemness of colorectal cancer cells

“…This research had a great impact on the study of CSC and recently, Essex and collaborators replicated these studies and obtained similar results. They found that TME regulates the activation of the Wnt signaling pathway, increases CCSC characteristics and enhances tumor-initiating potential[ 54 ]. Regarding this, it is known that several Wnt ligands are secreted mostly by CAFs[ 53 - 56 ].…”

Tumor microenvironment involvement in colorectal cancer progression via Wnt/β-catenin pathway: Providing understanding of the complex mechanisms of chemoresistance

“…Vermeulen and colleagues showed that CAF-derived hepatocyte growth factor (HGF) activates Wnt/β-catenin signaling and subsequently the clonogenicity in cancer stem cells (CSCs) isolated from CRC patients that were transiently transfected with TCF/LEF luciferase reporter vector. Moreover, HGF restored the CSC phenotype in more differentiated tumor cells both in vitro and in vivo [ 52 ]. Essex and collaborators recently replicated these studies, emphasizing the relevance of this pathway and the interaction of tumor cells with their stroma, and in the promotion and development of CRC[ 53 ].…”

“…Activation of the c-MET pathway through HGF signaling is related to drug resistance in many types of tumors[ 111 ]. Several works have already shown that HGF is associated with both Met and β-catenin pathways[ 52 , 54 ]. Studies published by Woo and collaborators showed that this factor is related to resistance to CPT-11 in CRC cells[ 112 ].…”