CDK1 inhibition reduces osteogenesis in endothelial cells in vascular calcification
Yan Zhao, Yang Yang, Xiuju Wu, Li Zhang, Xinjiang Cai, Jaden Ji, Sydney Chen, Abigail Vera, Kristina I. Boström, Yucheng Yao
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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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 3

Inhibition of CDK1 prevents osteogenic differentiation in MGP-depleted ECs.

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Inhibition of CDK1 prevents osteogenic differentiation in MGP-depleted E...
(A) Expression of CDKs in tdTomato+ aortic cells isolated from VE-cadherincre/ERT2 RosatdTomato Mgp–/– mice and VE-cadherincre/ERT2 RosatdTomato mice, as determined by real-time PCR (n = 6). (B) Immunostaining of CDK1 in the aortic tissues of VE-cadherincre/ERT2 RosatdTomato Mgp–/– mice and VE-cadherincre/ERT2 RosatdTomato mice (n = 6). Scale bars: 50 μm. (C) Expression of MGP in HAECs after transfection of MGP siRNAs (n = 8). SCR, scramble siRNA. (D) Immunoblotting of CDK1, osteogenic, and endothelial markers in HAECs after transfection of MGP siRNAs and treatment with osteogenic induction media. Positive control (Pos Ctr) for CDK1 blotting: MCF-7 cells as verified by Thermo Fisher Scientific. Negative control (Neg Ctr) for CDK1 blotting: MCF-7 cells transfected with CDK1 siRNA. Positive control for osterix and osteopontin (OPN) blotting: MC3T3 osteoblast cells. Negative control for osterix and OPN blotting: HAECs. Positive control for eNOS and FLK1 blotting: HAECs. Negative control for eNOS and FLK1 blotting: MC3T3 osteoblast cells. Each lane represents an independent experimental group. (E) Immunoblotting of CDK1, OPN, and eNOS in HAECS after transfection of MGP siRNA in combination of with CDK1 siRNA and treatment with osteogenic induction media. Positive and negative controls are the same as in D. Each lane represents an independent experimental group. (F) Time-course expression of osteogenic and endothelial markers in HAECs after transfection of MGP siRNA and treatment with AT7519 or saline control (n = 3). Data were analyzed for statistical significance by unpaired, 2-tailed Student’s t test (A and C) or 1-way ANOVA with Tukey’s multiple-comparison test (F). In A and C, 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. Data in F are presented as mean ± SD. ***P < 0.0001.