Anti-LRP5/6 VHHs Promote Differentiation of Wnt-hypersensitive Intestinal Stem Cells

Overview

Journal Nat Commun

Specialty Biology

Date 2019 Jan 22

PMID 30664649

Citations 28

Authors

Nicola Fenderico

Revina C van Scherpenzeel

Michael Goldflam

Davide Proverbio

Ingrid Jordens

Tomica Kralj

Sarah Stryeck

Tarek Z Bass

Guy Hermans

Christopher Ullman

Teodor Aastrup

Piet Gros

Madelon M Maurice

Affiliations

Soon will be listed here.

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.

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