Endothelial YAP/TAZ activation promotes atherosclerosis in a mouse model of Hutchinson-Gilford progeria syndrome
Ana Barettino, … , Ignacio Benedicto, Vicente Andrés
Ana Barettino, … , Ignacio Benedicto, Vicente Andrés
Published October 1, 2024
Citation Information: J Clin Invest. 2024;134(22):e173448. https://doi.org/10.1172/JCI173448.
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Research Article Aging Vascular biology

Endothelial YAP/TAZ activation promotes atherosclerosis in a mouse model of Hutchinson-Gilford progeria syndrome

Abstract

Hutchinson-Gilford progeria syndrome (HGPS) is an extremely rare disease caused by the expression of progerin, an aberrant protein produced by a point mutation in the LMNA gene. HGPS patients show accelerated aging and die prematurely mainly from complications of atherosclerosis such as myocardial infarction, heart failure, or stroke. However, the mechanisms underlying HGPS vascular pathology remain ill-defined. We used single-cell RNA sequencing to characterize the aorta in progerin-expressing LmnaG609G/G609G mice and wild-type controls, with a special focus on endothelial cells (ECs). HGPS ECs showed gene expression changes associated with extracellular matrix alterations, increased leukocyte extravasation, and activation of the yes-associated protein 1/transcriptional activator with PDZ-binding domain (YAP/TAZ) mechanosensing pathway, all validated by different techniques. Atomic force microscopy experiments demonstrated stiffer subendothelial extracellular matrix in progeroid aortae, and ultrasound assessment of live HGPS mice revealed disturbed aortic blood flow, both key inducers of the YAP/TAZ pathway in ECs. YAP/TAZ inhibition with verteporfin reduced leukocyte accumulation in the aortic intimal layer and decreased atherosclerosis burden in progeroid mice. Our findings identify endothelial YAP/TAZ signaling as a key mechanism of HGPS vascular disease and open a new avenue for the development of YAP/TAZ-targeting drugs to ameliorate progerin-induced atherosclerosis.

Authors

Ana Barettino, Cristina González-Gómez, Pilar Gonzalo, María J. Andrés-Manzano, Carlos R. Guerrero, Francisco M. Espinosa, Rosa M. Carmona, Yaazan Blanco, Beatriz Dorado, Carlos Torroja, Fátima Sánchez-Cabo, Ana Quintas, Alberto Benguría, Ana Dopazo, Ricardo García, Ignacio Benedicto, Vicente Andrés

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Figure 7

YAP/TAZ inhibition with verteporfin reduces the accumulation of aortic intimal leukocytes in progeroid mice.

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YAP/TAZ inhibition with verteporfin reduces the accumulation of aortic i...
(A) Western blot analysis of thoracic aorta lysates, showing TAZ upregulation in vehicle-treated LmnaG6096/G609G versus Lmna+/+ mice, and its inhibition after verteporfin treatment (n = 3). Vinculin was used as loading control. (B) Representative en face immunofluorescence images of thoracic aortae showing ECs (CD31, green), TAZ (white), and nuclei (Hoechst 33342, blue), and quantification of percentage of cells in each quartile per mouse (n = 7). Boxed areas shown at higher magnification. (C) Representative en face immunofluorescence images of thoracic aortae showing ECs (CD31, green), EC nuclei (ERG, white), leukocytes (CD45.2, red), and nuclei (Hoechst 33342, blue), and quantification of intimal leukocytes. Mean values for individual mice (n = 10–12) were determined by averaging of cells from 9 fields from 3 aortic regions. (D) Circulating white blood cell counts (n = 17–18). (E) scRNA-Seq–determined relative expression of Selp, Vcam1, and Icam1 in different aortic cell types. (F) Selp, Vcam1, and Icam1 expression in aortic arches determined by RT-qPCR (n = 6–9). (G) Representative en face immunofluorescence images of aortae showing ECs (CD31, green) and SELP (top panel, aortic arch, red), VCAM1 (middle panel, thoracic aorta, red), or ICAM1 (bottom panel, thoracic aorta, red). The graph shows the percentage of SELP+ (n = 8), VCAM1+ (n = 7–9), and ICAM1+ (n = 7–8) area. Mean values for individual mice were determined by averaging of values from 3 fields. Data are presented as mean + SD. Statistical analysis was performed using 1-way ANOVA (A, F [Selp and Vcam1], and G), 1-tailed Student’s t test (B [Q1, Q2, Q4]), Mann-Whitney test (B [Q3]), 2-tailed Student’s t test (C and D), and Kruskal-Wallis test (F [Icam1]). Outliers assessed by ROUT test in F (Icam1) were excluded for analysis. Scale bars: 50 μm. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. FC, fold change; VP, verteporfin; WBC, white blood cells.