Madelon M. Maurice

The current treatment for refractory BRAF-V600E mutant metastatic colorectal cancer (mCRC) involves combined inhibition of BRAF and the epidermal growth factor receptor (EGFR). However, tumour responses are often short-lived due to a rebound in mitogen-activated protein kinase (MAPK) activity. In this study, we combined short-term cell viability assays with long-term regrowth assays following drug removal over a period of three weeks. This allowed assessment of regrowth after therapy discontinuation. We tested the effect of combined BRAF inhibition (encorafenib) and EGFR inhibition (afatinib) on BRAF-V600E mutant CRC patient-derived organoids (PDOs). Combined EGFR/BRAF inhibition initially caused a major reduction in PDO growth capacity in BRAF-V600E mutant PDOs. This was followed by rapid regrowth after drug removal, mirroring clinical outcomes. EGFR inhibition in BRAF-V600E mutant PDOs led to activation of the insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R). The IGF1R/IR inhibitor linsitinib prevented the rebound in MAPK activity following removal of afatinib and encorafenib, prevented regrowth of CRC PDOs, and improved the anti-tumour response in an in vivo model. PDO regrowth assays allow the identification of pathways driving tumour recurrence. IR/IGF1R-inhibition prevents regrowth following golden standard MAPK pathway-targeted therapy and provides a strategy to improve the treatment of BRAF-V600E mutant CRC

In multicellular organisms, Wnt proteins govern stem and progenitor cell renewal and differentiation to regulate embryonic development, adult tissue homeostasis and tissue regeneration. Defects in canonical Wnt signalling, which is transduced intracellularly by β-catenin, have been associated with developmental disorders, degenerative diseases and cancers. Although a simple model describing Wnt–β-catenin signalling is widely used to introduce this pathway and has largely remained unchanged over the past 30 years, in this Review we discuss recent studies that have provided important new insights into the mechanisms of Wnt production, receptor activation and intracellular signalling that advance our understanding of the molecular mechanisms that underlie this important cell–cell communication system. In addition, we review the recent development of molecules capable of activating the Wnt–β-catenin pathway with selectivity in vitro and in vivo that is enabling new lines of study to pave the way for the development of Wnt therapies for the treatment of human diseases.

Hyperactivation of WNT signaling is a well-established hallmark of cancer. Various epithelial cancers express high levels of WNT7B and WNT10A that are not commonly expressed during tissue homeostasis, but rather associate with tissue development and regeneration. Although increased WNT7B/10A expression correlates with aggressive disease and lower patient survival rates, the mechanism by which these WNTs influence cancer progression remains unknown. Here, we use patient-derived organoids to show that tumor-intrinsic expression of WNT7B/10A drives survival and growth of advanced pancreatic ductal adenocarcinoma (PDAC). Bulk and single-cell profiling reveal that WNT7B drives proliferation and promotes expression of a poor prognosis basal-like state by preventing expression of a more differentiated, classical PDAC signature. By generating WNT7B reporter organoids, we show that heterogeneously distributed WNT-high PDAC cells are shifted towards a more basal-like phenotype and stably co-exist with WNT-low/negative lineages. Furthermore, hybrid co-cultures of WNT7B-proficient and -knockout PDAC organoids demonstrate that WNT-sending cells drive survival and proliferation of neighboring WNT-negative cells within the cancer epithelium via short range, cell contact-dependent signaling. In summary, our work uncovers a prominent role of WNT7B/10A in driving PDAC subtype heterogeneity and argues that WNT inhibition may be applied to force a class switch to a more differentiated, less aggressive cancer subtype that correlates with improved therapeutical response.

In healthy cells, AXIN1 organizes assembly of a large destruction complex that mediates proteolysis of the transcriptional co-activator β-catenin to prevent inappropriate Wnt/β-catenin pathway activation. In hepatocellular carcinoma (HCC), AXIN1 mutations (11%) associate with a poor-prognosis subtype that is molecularly distinct from β-catenin-mutant HCC (28-40%). How AXIN1 deficiency drives HCC formation has remained highly debated. Here, we address this issue by introducing HCC-associated AXIN1 and CTNNB1 mutations in human liver cancer cells and liver-derived organoids. We show that different mutant AXIN1 classes activate varying degrees of Wnt signaling, although at lower overall levels than CTNNB1 mutations. Strikingly, premature stop codons in 5’ coding regions do not classify as knock-out mutations but drive alternative translation of an N-terminally truncated AXIN1 variant with partially retained suppressor activity. All AXIN1 variants endow liver progenitor organoids with the capacity to grow in the absence of R-spondin and Wnt, indicative of aberrant Wnt/β-catenin pathway activation. Additionally, induced Wnt/β-catenin pathway activation inversely correlates with YAP/TAZ-mediated signaling, thus leaving higher residual YAP/TAZ activity in AXIN1 -mutant versus CTNNB1 -mutant cells. We conclude that AXIN1 mutations drive physiologically relevant Wnt/β-catenin signaling in HCC, while providing a permissive environment for YAP/YAZ signaling, thereby distinguishing AXIN1 mutations from those in CTNNB1 .