Cyclin-dependent kinase inhibitor 2A (CDKN2A) is a key regulator of cell cycle arrest and senescence in endothelial colony-forming cells (ECFCs) in moyamoya disease (MMD)

• Published in: Journal of Korean Neurosurgical Society Vol. 66; no. 6; pp. 642 - 651
• Main Authors: Choi, Seung Ah, Moon, Youn Joo, Koh, Eun Jung, Phi, Ji Hoon, Lee, Ji Yeoun, Kim, Kyung Hyun, Kim, Seung-Ki
• Format: Journal Article
• Language: English
• Published: Korea (South) Korean Neurosurgical Society 01.11.2023; 대한신경외과학회
• Subjects: Laboratory Research; 신경외과학; Senescence; Endothelial colony-forming cells; Moyamoya disease; Cyclin-dependent kinase inhibitor 2A; Cell cycle
• ISSN: 2005-3711; 1598-7876
• DOI: 10.3340/jkns.2023.0005

Endothelial colony-forming cells (ECFCs) have been reported to play an important role in the pathogenesis of moyamoya disease (MMD). We have previously observed stagnant growth in MMD ECFCs with functional impairment of tubule formation. We aimed to verify the key regulators and related signaling pathways involved in the functional defects of MMD ECFCs. ECFCs were cultured from peripheral blood mononuclear cells (PBMNCs) of healthy volunteers (normal) and MMD patients. Low-density lipoproteins (LDL) uptake, flow cytometry, high content screening (HCS), senescence-associated β-galactosidase, immunofluorescence, cell cycle, tubule formation, microarray, RT-qPCR, siRNA transfection, and western blot analyses were performed. The acquisition of cells that can be cultured for a long time with the characteristics of late ECFCs was significantly lower in the MMD patients than the normal. Importantly, the MMD ECFCs showed decreased cellular proliferation with G1 cell cycle arrest and cellular senescence compared to the normal ECFCs. A pathway enrichment analysis demonstrated that the cell cycle pathway was the major enriched pathway, which is consistent with the results of the functional analysis of ECFCs. Among the genes associated with the cell cycle, cyclin-dependent kinase inhibitor 2A (CDKN2A) showed the highest expression in MMD ECFCs. Knockdown of CDKN2A in MMD ECFCs enhanced proliferation by reducing G1 cell cycle arrest and inhibiting senescence through the regulation of CDK4 and phospho retinoblastoma protein (pRB). Our study suggests that CDKN2A plays an important role in the growth retardation of MMD ECFCs by inducing cell cycle arrest and senescence.