CDK1 inhibition reduces osteogenesis in endothelial cells in vascular calcification
Yan Zhao, … , Kristina I. Boström, Yucheng Yao
Yan Zhao, … , Kristina I. Boström, Yucheng Yao
Published January 23, 2024
Citation Information: JCI Insight. 2024;9(5):e176065. https://doi.org/10.1172/jci.insight.176065.
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Research Article Cell biology Vascular biology

CDK1 inhibition reduces osteogenesis in endothelial cells in vascular calcification

Abstract

Vascular calcification is a severe complication of cardiovascular diseases. Previous studies demonstrated that endothelial lineage cells transitioned into osteoblast-like cells and contributed to vascular calcification. Here, we found that inhibition of cyclin-dependent kinase (CDK) prevented endothelial lineage cells from transitioning to osteoblast-like cells and reduced vascular calcification. We identified a robust induction of CDK1 in endothelial cells (ECs) in calcified arteries and showed that EC–specific gene deletion of CDK1 decreased the calcification. We found that limiting CDK1 induced E-twenty-six specific sequence variant 2 (ETV2), which was responsible for blocking endothelial lineage cells from undergoing osteoblast differentiation. We also found that inhibition of CDK1 reduced vascular calcification in a diabetic mouse model. Together, the results highlight the importance of CDK1 suppression and suggest CDK1 inhibition as a potential option for treating vascular calcification.

Authors

Yan Zhao, Yang Yang, Xiuju Wu, Li Zhang, Xinjiang Cai, Jaden Ji, Sydney Chen, Abigail Vera, Kristina I. Boström, Yucheng Yao

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Figure 5

Limiting CDK1 upregulates ETV2 to prevent endothelial lineage cells from undergoing osteogenic differentiation.

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Limiting CDK1 upregulates ETV2 to prevent endothelial lineage cells from...
(A and B) Expression of ETV2 in tdTomato+ aortic cells isolated from VE-cadherincre/ERT2 RosatdTomato Mgp–/– mice and VE-cadherincre/ERT2 RosatdTomato mice, as shown by real-time PCR (A) and immunoblotting (B) (n = 6). Positive control (Pos Ctr) for ETV2 blotting: Mouse ECs induced from embryonic stem cells (66). Negative control (Neg Ctr) for ETV2 blotting: HAECs transfected with ETV2 siRNA. Each lane represents an independent experimental group. (C) Expression of ETV2 in CD31+CD45– aortic cells isolated from VE-cadherincre/ERT2 Cdk1fl/fl Mgp–/– mice and VE-cadherincre/ERT2 Cdk1fl/fl mice after tamoxifen administration (n = 6). (D) ETV2 expression in HAECs after transfection of MGP siRNA in combination with AT5719 treatment (1–10 μM), as shown by real-time PCR (n = 6). (E) Immunoblotting of ETV2, osterix, osteopontin (OPN), eNOS, and FLK1 in HAECs after transfection of MGP siRNA in combination with infection of lentiviral vectors containing CMV-driven ETV2 cDNA and treatment with osteogenic induction media. Positive and negative controls are the same as described in B and Figure 3. Each lane represents an independent experimental group. (F and G) Micro-CT imaging and Alizarin red and von Kossa staining (F), and total aortic calcium with ETV2 expression (G) of descending aortas of Mgp–/– mice treated with adeno-associated viral (AAV) vectors containing CMV-promoter-driven ETV2 cDNA (n = 8). Empty vectors were used as controls. Scale bars: 5 mm (micro-CT) and 100 μm (staining). (H) Expression of osteogenic and endothelial markers in CD31+CD45– aortic cells from Mgp–/– mice treated with AAV vectors containing CMV-promoter-driven ETV2 cDNA (n = 8). Empty vectors were used as controls. Data were analyzed for statistical significance by 1-way ANOVA with Tukey’s multiple-comparison test (A, C, D, and H) or unpaired, 2-tailed Student’s t test (G). The bounds of the boxes are upper and lower quartiles with data points, the line in the box is the median, and whiskers are maximal and minimal values. ***P < 0.0001.