Anti-LRP5/6 VHHs promote differentiation of Wnt-hypersensitive intestinal stem cells
Files
Publication date
2019-01-21
Authors
Fenderico, Nicola
van Scherpenzeel, Revina C.
Goldflam, Michael
Proverbio, Davide
Jordens, Ingrid
Kralj, Tomica
Stryeck, Sarah
Bass, Tarek Z.
Hermans, Guy
Ullman, Christopher
Editors
Advisors
Supervisors
Document Type
Article
Metadata
Show full item recordCollections
License
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 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.
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
Citation
Fenderico, N, van Scherpenzeel, R C, Goldflam, M, Proverbio, D, Jordens, I, Kralj, T, Stryeck, S, Bass, T Z, Hermans, G, Ullman, C, Aastrup, T, Gros, P & Maurice, M M 2019, 'Anti-LRP5/6 VHHs promote differentiation of Wnt-hypersensitive intestinal stem cells', Nature Communications, vol. 10, no. 1, 365, pp. 365. https://doi.org/10.1038/s41467-018-08172-z