Self-renewal and high proliferative colony forming capacity of late-outgrowth endothelial progenitors is regulated by cyclin-dependent kinase inhibitors driven by notch signaling

Patel, Jatin, Wong, Ho Yi, Wang, Weili, Alexis, Josue, Shafiee, Abbas, Stevenson, Alexander J., Gabrielli, Brian, Fisk, Nicholas M. and Khosrotehrani, Kiarash (2016) Self-renewal and high proliferative colony forming capacity of late-outgrowth endothelial progenitors is regulated by cyclin-dependent kinase inhibitors driven by notch signaling. Stem Cells, 34 4: 902-912. doi:10.1002/stem.2262


Author Patel, Jatin
Wong, Ho Yi
Wang, Weili
Alexis, Josue
Shafiee, Abbas
Stevenson, Alexander J.
Gabrielli, Brian
Fisk, Nicholas M.
Khosrotehrani, Kiarash
Title Self-renewal and high proliferative colony forming capacity of late-outgrowth endothelial progenitors is regulated by cyclin-dependent kinase inhibitors driven by notch signaling
Journal name Stem Cells   Check publisher's open access policy
ISSN 1549-4918
1066-5099
Publication date 2016-04
Sub-type Article (original research)
DOI 10.1002/stem.2262
Volume 34
Issue 4
Start page 902
End page 912
Total pages 11
Place of publication Durham, United States
Publisher AlphaMed Press
Collection year 2017
Language eng
Formatted abstract
Since the discovery of endothelial colony forming cells (ECFC), there has been significant interest in their therapeutic potential to treat vascular injuries. ECFC cultures display significant heterogeneity and a hierarchy among cells able to give rise to high proliferative versus low proliferative colonies. Here we aimed to define molecularly this in vitro hierarchy. Based on flow cytometry, CD34 expression levels distinguished two populations. Only CD34 + ECFC had the capacity to reproduce high proliferative potential (HPP) colonies on replating, whereas CD34− ECFCs formed only small clusters. CD34 + ECFCs were the only ones to self-renew in stringent single-cell cultures and gave rise to both CD34 + and CD34− cells. Upon replating, CD34 + ECFCs were always found at the centre of HPP colonies and were more likely in G0/1 phase of cell cycling. Functionally, CD34 + ECFC were superior at restoring perfusion and better engrafted when injected into ischemic hind limbs. Transcriptomic analysis identified cyclin-dependent kinase (CDK) cell cycle inhibiting genes (p16, p21, and p57), the Notch signaling pathway (dll1, dll4, hes1, and hey1), and the endothelial cytokine il33 as highly expressed in CD34 + ECFC. Blocking the Notch pathway using a γ-secretase inhibitor (DAPT) led to reduced expression of cell cycle inhibitors, increased cell proliferation followed by a loss of self-renewal, and HPP colony formation capacity reflecting progenitor exhaustion. Similarly shRNA knockdown of p57 strongly affected self-renewal of ECFC colonies. ECFC hierarchy is defined by Notch signalling driving cell cycle regulators, progenitor quiescence and self-renewal potential.
Keyword Endothelial progenitor cells
Hierarchy
Notch
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
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UQ Diamantina Institute Publications
 
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