Development of Potent, Selective Surrogate WNT Molecules and Their Application in Defining Frizzled Requirements

Highlights

• Developed a flexible system to generate potent and selective surrogate WNTs

• Multivalent binding to FZD and LRP is a requirement for maximal activation

• Active receptor complexes can contain different ratios of receptors

• Recruitment of two different FZDs together with LRP induces efficient signaling

Summary

WNTs regulate myriad biological processes during embryonic development and are key regulators of stem cell function, tissue homeostasis, and injury repair in adults. The creation of WNT-based therapies has been hampered by challenges in developing soluble, potent, and selective WNT molecules. Soluble WNT surrogates have been reported, but they demonstrate relatively weak WNT signaling activity. Here, we describe a platform for potent, selective WNT surrogate generation. We identify multivalent binding to Frizzleds (FZDs) and low-density lipoprotein receptor-related proteins (LRPs) to be a requirement for maximal WNT/β-catenin activation. Furthermore, we show that recruitment of two different FZDs together with LRP causes efficient signaling. Surrogate WNT targeting either FZD_1,2,7 or FZD_5,8 induces expansive growth of intestinal organoids. This flexible WNT surrogate platform yields potent agonists with any desired receptor specificity and will be useful for research and therapeutic applications for tissue regeneration.