TCF7L2 silencing results in altered gene expression patterns accompanied by local genomic reorganization

Canonical Wnt signaling is crucial for intestinal homeostasis as TCF4, the major Wnt signaling effector in the intestines, is required for stem cell maintenance. The capability of TCF4 to maintain the stem cell phenotype is contingent upon beta-catenin, a potent transcriptional activator, which interacts with histone acetyltransferases and chromatin remodeling complexes. We used RNAi to explore the influence of TCF4 on chromatin structure (Hi-C) and gene expression (RNA sequencing) across a 72-hour time series in colon cancer. We found that TCF4 reduction results in a disproportionate up-regulation of gene expression, including a powerful induction of SOX2. Integration of RNA sequencing and Hi-C data revealed a TAD boundary loss, which occurred concomitantly with the over-expression of a cluster of CEACAM genes on chromosome 19. We identified EMT and E2F as the 2 most deregulated pathways upon TCF4 depletion and LUM, TMPO, and AURKA as highly influential genes in these networks using measures of centrality. Results from gene expression, chromatin structure, and centrality analyses were integrated to generate a list of candidate transcription factors crucial for colon cancer cell homeostasis. The top ranked factor was c-JUN, an oncoprotein known to interact with TCF4 and beta-catenin, confirming the usefulness of this approach.

Automated summary

Canonical Wnt signaling, mediated by TCF4, plays a crucial role in intestinal homeostasis by maintaining stem cell phenotype. Reduction of TCF4 leads to disproportionate up-regulation of gene expression and loss of TAD boundaries, with over-expression of CEACAM gene cluster. EMT and E2F pathways are most deregulated upon TCF4 depletion, with LUM, TMPO, and AURKA identified as highly influential genes in these networks.