Journal Article

Anti-LRP5/6 VHHs promote differentiation of Wnt-hypersensitive intestinal stem cells

Fenderico, Nicola, Aastrup, Teodor, Maurice, Madelon M., Stryeck, Sarah, Hermans, Guy, van Scherpenzeel, Revina C., Bass, Tarek Z., Gros, Piet, Jordens, Ingrid, Proverbio, Davide, Kralj, Tomica, Goldflam, Michael, Ullman, Christopher

2019

Overview

Wnt-induced β-catenin-mediated transcription is a driving force for stem cell self-renewal during adult tissue homeostasis. Enhanced Wnt receptor expression due to mutational inactivation of the ubiquitin ligases RNF43 / ZNRF3 recently emerged as a leading cause for cancer development. Consequently, targeting canonical Wnt receptors such as LRP5/6 holds great promise for treatment of such cancer subsets. Here, we employ CIS display technology to identify single-domain antibody fragments (VHH) that bind the LRP6 P3E3P4E4 region with nanomolar affinity and strongly inhibit Wnt3/3a-induced β-catenin-mediated transcription in cells, while leaving Wnt1 responses unaffected. Structural analysis reveal that individual VHHs variably employ divergent antigen-binding regions to bind a similar surface in the third β-propeller of LRP5/6, sterically interfering with Wnt3/3a binding. Importantly, anti-LRP5/6 VHHs block the growth of Wnt-hypersensitive Rnf43 / Znrf3 -mutant intestinal organoids through stem cell exhaustion and collective terminal differentiation. Thus, VHH-mediated targeting of LRP5/6 provides a promising differentiation-inducing strategy for treatment of Wnt-hypersensitive tumors. Enhanced Wnt receptor activity is a major cause of cancer development. Here the authors identify camelid single-domain antibody fragments (VHHs) that bind to the Wnt receptor LRP5/6 ectodomain, determine the crystal structures and show that these VHHs selectively inhibit Wnt3- mediated cellular responses and block the growth of mutant Wnt-hypersensitive intestinal tumor organoids.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13/1 / 13/100 / 13/106 / 13/109 / 13/95 / 14 / 14/19 / 38 / 49 / 49/47 / 631/154/51/2318 / 631/45/535/1266 / 631/532/1360 / 631/532/2437 / 631/67/1059/602 / 64 / 82/80 / 82/83 / 9/10 / 96 / Animal behavior / Animals / beta Catenin - genetics / beta Catenin - metabolism / Binding Sites / Cell Differentiation / Cell Line, Tumor / Cell Proliferation / Cell self-renewal / Circuits / Crystallography, X-Ray / Fibroblasts - cytology / Fibroblasts - drug effects / Fibroblasts - metabolism / Gene Expression Regulation / HEK293 Cells / Humanities and Social Sciences / Humans / Inactivation / Intestine, Small - cytology / Intestine, Small - drug effects / Intestine, Small - metabolism / Low Density Lipoprotein Receptor-Related Protein-5 - antagonists & inhibitors / Low Density Lipoprotein Receptor-Related Protein-5 - chemistry / Low Density Lipoprotein Receptor-Related Protein-5 - genetics / Low Density Lipoprotein Receptor-Related Protein-5 - metabolism / Low Density Lipoprotein Receptor-Related Protein-6 - antagonists & inhibitors / Low Density Lipoprotein Receptor-Related Protein-6 - chemistry / Low Density Lipoprotein Receptor-Related Protein-6 - genetics / Low Density Lipoprotein Receptor-Related Protein-6 - metabolism / Mice / Models, Molecular / multidisciplinary / Nanobodies / Neural networks / Organoids / Organoids - cytology / Organoids - drug effects / Organoids - metabolism / Protein Binding / Protein Conformation, beta-Strand / Protein Interaction Domains and Motifs / Science / Science (multidisciplinary) / Single-Domain Antibodies - chemistry / Single-Domain Antibodies - genetics / Single-Domain Antibodies - metabolism / Stem cells / Stem Cells - cytology / Stem Cells - drug effects / Stem Cells - metabolism / Structural analysis / Transcription, Genetic / Ubiquitin-Protein Ligases - genetics / Ubiquitin-Protein Ligases - metabolism / Uncertainty / Wnt3A Protein - genetics / Wnt3A Protein - metabolism