Anti-LRP5/6 VHHs promote differentiation of Wnt-hypersensitive intestinal stem cells
Fenderico, Nicola; van Scherpenzeel, Revina C.; Goldflam, Michael; Proverbio, Davide; Jordens, Ingrid; Kralj, Tomica; Stryeck, Sarah; Bass, Tarek Z.; Hermans, Guy; Ullman, Christopher; Aastrup, Teodor; Gros, Piet; Maurice, Madelon M.
(2019) Nature Communications, volume 10, issue 1, pp. 365
(Article)
Abstract
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
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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.
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Keywords: Animals, Binding Sites, Cell Differentiation, Cell Line, Tumor, Cell Proliferation, Crystallography, X-Ray, Fibroblasts/cytology, Gene Expression Regulation, HEK293 Cells, Humans, Intestine, Small/cytology, Low Density Lipoprotein Receptor-Related Protein-5/antagonists & inhibitors, Low Density Lipoprotein Receptor-Related Protein-6/antagonists & inhibitors, Mice, Models, Molecular, Organoids/cytology, Protein Binding, Protein Conformation, beta-Strand, Protein Interaction Domains and Motifs, Single-Domain Antibodies/chemistry, Stem Cells/cytology, Transcription, Genetic, Ubiquitin-Protein Ligases/genetics, Wnt3A Protein/genetics, beta Catenin/genetics, Journal Article, Research Support, Non-U.S. Gov't
ISSN: 2041-1723
Publisher: Nature Publishing Group
(Peer reviewed)