title: Enhancing a Wnt-Telomere Feedback Loop Restores Intestinal Stem Cell Function in a Human Organotypic Model of Dyskeratosis Congenita.
abstract: Patients with dyskeratosis congenita (DC) suffer from stem cell failure in highly proliferative tissues, including the intestinal epithelium. Few therapeutic options exist for this disorder, and patients are treated primarily with bone marrow transplantation to restore hematopoietic function. Here, we generate isogenic DC patient and disease allele-corrected intestinal tissue using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene correction in induced pluripotent stem cells and directed differentiation. We show that DC tissue has suboptimal Wnt pathway activity causing intestinal stem cell failure and that enhanced expression of the telomere-capping protein TRF2, a Wnt target gene, can alleviate DC phenotypes. Treatment with the clinically relevant Wnt agonists LiCl or CHIR99021 restored TRF2 expression and reversed gastrointestinal DC phenotypes, including organoid formation in vitro, and maturation of intestinal tissue and xenografted organoids in vivo. Thus, the isogenic DC cell model provides a platform for therapeutic discovery and identifies Wnt modulation as a potential strategy for treatment of DC patients.
authors.affiliation: Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Dong-Hun
authors.initials: DH
authors.lastname: Woo
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Qijun
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Cell and Molecular Biology Program, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Ting-Lin B
authors.initials: TL
authors.lastname: Yang
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Cell and Molecular Biology Program, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: M Rebecca
authors.initials: MR
authors.lastname: Glineburg
authors.affiliation: Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Vagelos Scholars Program, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Carla
authors.initials: C
authors.lastname: Hoge
authors.affiliation: Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Nicolae A
authors.initials: NA
authors.lastname: Leu
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Cell and Molecular Biology Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute on Aging, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: johnsonb@mail.med.upenn.edu.
authors.forename: F Brad
authors.initials: FB
authors.lastname: Johnson
authors.affiliation: Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Cell and Molecular Biology Program, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: lengner@vet.upenn.edu.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lengner
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D019871:Dyskeratosis Congenita / Q000378:metabolism / Q000473:pathology*; D025461:Feedback, Physiological / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D007422:Intestines / Q000166:cytology*; D008094:Lithium / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D009940:Organoids / Q000187:drug effects / Q000378:metabolism*; D010641:Phenotype; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism*; D016615:Telomere / Q000378:metabolism*; D035341:Telomeric Repeat Binding Protein 2 / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects
chemical_list: C504753:TERF2 protein, human; D035341:Telomeric Repeat Binding Protein 2; D008094:Lithium