BACKGROUND: The choroid plexus (ChP) is the secretory epithelial structure located in the brain ventricles. Choroid plexus tumors (CPTs) are rare neoplasms predominantly occurring in young patients with intensified malignancy in children. CPT treatment is hindered by insufficient knowledge of tumor pathology and the limited availability of valid models. METHODS: Genomic and transcriptomic data from CPT patients were analyzed to identify the putative pathological pathway. Cellular and molecular techniques were employed to validate bioinformatic results in CPT patient samples. Pharmacologic inhibition of Wnt/β-catenin signaling was assessed in CPT cells. Cell-based assays of ChP cell lines were performed following CRISPR-Cas9-derived knockout and overexpression of Wnt/β-catenin pathway genes. A 3D CPT model was generated through CRISPR-Cas9-derived knockout of APC. RESULTS: We discovered that Wnt/β-catenin signaling is activated in human CPTs, likely as a consequence of large-scale chromosomal instability events of the CPT genomes. We demonstrated that CPT-derived cells depend on autocrine Wnt/β-catenin signaling for survival. Constitutive Wnt/β-catenin pathway activation, either through knockout of the negative regulator APC or overexpression of the ligand WNT3A, induced tumorigenic properties in ChP 2D in vitro models. Increased activation of the Wnt/β-catenin pathway in ChP organoids, through treatment with a potent GSK3β inhibitor, reduced the differentiation of mature ChP epithelial cells. Remarkably, the depletion of APC was sufficient to induce the oncogenic transformation of ChP organoids. CONCLUSIONS: Our research identifies Wnt/β-catenin signaling as a critical driver of CPT tumorigenesis and provides the first 3D in vitro model for future pathological and therapeutic studies of CPT.

Central Institute of Mental Health Medical Faculty Mannheim Heidelberg University Mannheim Germany

Centre for Organismal Studies Heidelberg University Heidelberg Germany

Danish Research Institute of Translational Neuroscience DANDRITE and Center of Excellence PROMEMO Department of Biomedicine Aarhus University Aarhus Denmark

Department of Neurosciences Rita Levi Montalcini Turin University Turin Italy

Department of Pediatrics Pediatric Infectious Diseases Medical Faculty Mannheim Heidelberg University Heidelberg Germany

Department of Veterinary Sciences Turin University Grugliasco Italy

Division of Signal Transduction and Growth Control DKFZ ZMBH Alliance Heidelberg Germany

Faculty of Biosciences Heidelberg University Heidelberg Germany

German Cancer Research Center Heidelberg Germany

Hector Institute for Translational Brain Research Mannheim Germany

Institute of Neuropathology University Hospital Münster Münster Germany

Interdisciplinary Center for Neuroscience Heidelberg University Heidelberg Germany

Laboratory of Cilia and Centrosome Biology Department of Histology and Embryology Faculty of Medicine Masaryk University Brno Czechia

Laboratory of Clinical Regenerative Medicine Department of Neurosurgery Faculty of Medicine University of Tsukuba Tsukuba Japan

Schaller Research Group German Cancer Research Center Heidelberg Germany

Section of Animal Physiology and Immunology Department of Experimental Biology Faculty of Science Masaryk University Brno Czechia

Zentrum für Molekulare Biologie der Universität Heidelberg DKFZ ZMBH Alliance Heidelberg Germany

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