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DYRK1A Is a Regulator of S-Phase Entry in Hepatic Progenitor Cells
- Source :
- Stem Cells and Development, 27(2), 133. Mary Ann Liebert inc
- Publication Year :
- 2017
-
Abstract
- Hepatic progenitor cells (HPCs) are adult liver stem cells that act as second line of defense in liver regeneration. They are normally quiescent, but in case of severe liver damage, HPC proliferation is triggered by external activation mechanisms from their niche. Although several important proproliferative mechanisms have been described, it is not known which key intracellular regulators govern the switch between HPC quiescence and active cell cycle. We performed a high-throughput kinome small interfering RNA (siRNA) screen in HepaRG cells, a HPC-like cell line, and evaluated the effect on proliferation with a 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay. One hit increased the percentage of EdU-positive cells after knockdown: dual specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A). Although upon DYRK1A silencing, the percentage of EdU- and phosphorylated histone H3 (pH3)-positive cells was increased, and total cell numbers were not increased, possibly through a subsequent delay in cell cycle progression. This phenotype was confirmed with chemical inhibition of DYRK1A using harmine and with primary HPCs cultured as liver organoids. DYRK1A inhibition impaired Dimerization Partner, RB-like, E2F, and multivulva class B (DREAM) complex formation in HPCs and abolished its transcriptional repression on cell cycle progression. To further analyze DYRK1A function in HPC proliferation, liver organoid cultures were established from mBACtgDyrk1A mice, which harbor one extra copy of the murine Dyrk1a gene (Dyrk+++). Dyrk+++ organoids had both a reduced percentage of EdU-positive cells and reduced proliferation compared with wild-type organoids. This study provides evidence for an essential role of DYRK1A as balanced regulator of S-phase entry in HPCs. An exact gene dosage is crucial, as both DYRK1A deficiency and overexpression affect HPC cell cycle progression.
- Subjects :
- 0301 basic medicine
Small interfering RNA
Transcription, Genetic
proliferation
Biology
Protein Serine-Threonine Kinases
Cell Line
S Phase
03 medical and health sciences
RNA interference screen
Organoid
Humans
Progenitor cell
E2F
hepatic progenitor cells
Cell Biology
Hematology
DYRK1A
Cell cycle
Protein-Tyrosine Kinases
Liver regeneration
Cell biology
Adult Stem Cells
030104 developmental biology
Liver
Cell culture
cell cycle
Stem cell
Developmental Biology
Subjects
Details
- ISSN :
- 15578534 and 15473287
- Volume :
- 27
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Stem cells and development
- Accession number :
- edsair.doi.dedup.....76f51311da3b6e0109830265a43e70f7