Enhancing a Wnt-Telomere Feedback Loop Restores Intestinal Stem Cell Function in a Human Organotypic Model of Dyskeratosis Congenita

Cell Stem Cell. 2016 Sep 1;19(3):397-405. doi: 10.1016/j.stem.2016.05.024. Epub 2016 Aug 18.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Differentiation / drug effects
  • Dyskeratosis Congenita / metabolism
  • Dyskeratosis Congenita / pathology*
  • Feedback, Physiological* / drug effects
  • HEK293 Cells
  • Humans
  • Intestines / cytology*
  • Lithium / pharmacology
  • Mice
  • Models, Biological*
  • Organoids / drug effects
  • Organoids / metabolism*
  • Phenotype
  • Stem Cells / drug effects
  • Stem Cells / metabolism*
  • Telomere / metabolism*
  • Telomeric Repeat Binding Protein 2 / metabolism
  • Wnt Signaling Pathway* / drug effects

Substances

  • TERF2 protein, human
  • Telomeric Repeat Binding Protein 2
  • Lithium