Elevated endothelial Sox2 causes lumen disruption and cerebral arteriovenous malformations
Jiayi Yao, … , Kristina I. Boström, Yucheng Yao
Jiayi Yao, … , Kristina I. Boström, Yucheng Yao
Published June 24, 2019
Citation Information: J Clin Invest. 2019;129(8):3121-3133. https://doi.org/10.1172/JCI125965.
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Research Article Vascular biology

Elevated endothelial Sox2 causes lumen disruption and cerebral arteriovenous malformations

Abstract

Lumen integrity in vascularization requires fully differentiated endothelial cells (ECs). Here, we report that endothelial-mesenchymal transitions (EndMTs) emerged in ECs of cerebral arteriovenous malformation (AVMs) and caused disruption of the lumen or lumen disorder. We show that excessive Sry-box 2 (Sox2) signaling was responsible for the EndMTs in cerebral AVMs. EC-specific suppression of Sox2 normalized endothelial differentiation and lumen formation and improved the cerebral AVMs. Epigenetic studies showed that induction of Sox2 altered the cerebral-endothelial transcriptional landscape and identified jumonji domain–containing protein 5 (JMJD5) as a direct target of Sox2. Sox2 interacted with JMJD5 to induce EndMTs in cerebral ECs. Furthermore, we utilized a high-throughput system to identify the β-adrenergic antagonist pronethalol as an inhibitor of Sox2 expression. Treatment with pronethalol stabilized endothelial differentiation and lumen formation, which limited the cerebral AVMs.

Authors

Jiayi Yao, Xiuju Wu, Daoqin Zhang, Lumin Wang, Li Zhang, Eric X. Reynolds, Carlos Hernandez, Kristina I. Boström, Yucheng Yao

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Figure 6

A high-throughput system identifies pronethalol as an inhibitor of Sox2 expression.

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A high-throughput system identifies pronethalol as an inhibitor of Sox2 ...
(A) Schematic diagram of the high-throughput system used for screening candidate compounds that may suppress Sox2 expression in ReNcell VM cells. (B and C) Expression of EGFP in ReNcell VM cells after treatment with increasing doses of pronethalol, as (B) detected by fluorescence microscope and (C) quantitated by ImageJ software (n = 5). Scale bars: 50 μm. (D) Time-course expression of EGFP in ReNcell VM cells treated with 10 μM pronethalol (n = 5). (E and F) Dose and time-course expression of Sox2 after treatment with pronethalol in ReNcell VM cells (n = 5). (G) Decreased expression of Sox2, JMJD5, N-cadherin, Par3, and Rasip1 in MGP CRISPR cells after treatment with 10 μM pronethalol for 48 hours, as shown by immunoblotting (n = 5). (H–J) Expression of (H) β1-, (I) β2- or (J) β3-adrenergic receptors (β1, β2, and β3) in ReNcell VM cells transfected with siRNA to individual β1-, β2-, or β3-adrenergic receptors (β1 si, β2 si, and β3 si) (n = 5). SCR, scrambled siRNA. (K) Expression of Sox2 in ReNcell VM cells treated with pronethalol and transfected with siRNA to individual β1-, β2-, or β3-adrenergic receptors (n = 5). SCR, scrambled siRNA. (L–N) Expression of (L) β1-, (M) β2-, or (N) β3-adrenergic receptors in MGP CRISPR cells transfected with siRNA to individual β1-, β2-, or β3-adrenergic receptors (n = 5). (O) Expression of Sox2 in MGP CRISPR cells treated and transfected with siRNA to individual β1-, β2-, or β3-adrenergic receptors (n = 5). Data shown in H–J and L–N were analyzed by Student’s t test. Data shown in E, F, K, and O were analyzed by 1-way ANOVA with Tukey’s multiple comparisons test. Data are shown by box and whisker plots. The bounds of the boxes represent upper and lower quartiles. The lines in the boxes represent the median, and the whiskers represent the maximum and minimal values. ***P < 0.001.