The mammalian intestine is one of the most rapidly self-renewing tissues, driven by stem cells residing at the crypt bottom. Paneth cells form a major element of the niche microenvironment providing various growth factors to orchestrate intestinal stem cell homeostasis, such as Wnt3. Different Wnt ligands can selectively activate β-catenin-dependent (canonical) or -independent (noncanonical) signaling. Here, we report that the Dishevelled-associated activator of morphogenesis 1 (Daam1) and its paralogue Daam2 asymmetrically regulate canonical and noncanonical Wnt (Wnt/PCP) signaling. Daam1/2 interacts with the Wnt inhibitor RNF43, and Daam1/2 double knockout stimulates canonical Wnt signaling by preventing RNF43-dependent degradation of the Wnt receptor, Frizzled (Fzd). Single-cell RNA sequencing analysis revealed that Paneth cell differentiation is impaired by Daam1/2 depletion because of defective Wnt/PCP signaling. Together, we identified Daam1/2 as an unexpected hub molecule coordinating both canonical and noncanonical Wnt, which is fundamental for specifying an adequate number of Paneth cells.

Center for Genome Engineering Institute for Basic Science 55 Expo ro Yuseong gu Daejeon 34126 Republic of Korea

Department of Biosystems Science and Engineering ETH Zurich Mattenstrasse 26 4058 Basel Switzerland

Department of Cytokinetics Institute of Biophysics Academy of Sciences of the Czech Republic Brno Czech Republic

Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic

Department of Life Sciences Pohang University of Science and Technology Pohang Republic of Korea

Department of Medical and Biological Sciences The Catholic University of Korea Bucheon Republic of Korea

Division of Metabolism and Division of Oncology University Children's Hospital Zurich and Children's Research Center University of Zurich 8032 Zurich Switzerland

HUB Organoids Utrecht Netherlands

Institute of Molecular Biotechnology of the Austrian Academy of Sciences 1030 Vienna Austria

Oncode Institute and Centre for Molecular Medicine University Medical Centre Utrecht Utrecht Netherlands

UKM Medical Molecular Biology Institute Jalan Yaacob Latiff Bandar Tun Razak 56000 Kuala Lumpur Malaysia

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