Targeting the ASMase/S1P pathway protects from sortilin-evoked vascular damage in hypertension
Paola Di Pietro, … , Annibale A. Puca, Carmine Vecchione
Paola Di Pietro, … , Annibale A. Puca, Carmine Vecchione
Published February 1, 2022
Citation Information: J Clin Invest. 2022;132(3):e146343. https://doi.org/10.1172/JCI146343.
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Research Article Vascular biology

Targeting the ASMase/S1P pathway protects from sortilin-evoked vascular damage in hypertension

Abstract

Sortilin has been positively correlated with vascular disorders in humans. No study has yet evaluated the possible direct effect of sortilin on vascular function. We used pharmacological and genetic approaches coupled with study of murine and human samples to unravel the mechanisms recruited by sortilin in the vascular system. Sortilin induced endothelial dysfunction of mesenteric arteries through NADPH oxidase 2 (NOX2) isoform activation, dysfunction that was prevented by knockdown of acid sphingomyelinase (ASMase) or sphingosine kinase 1. In vivo, recombinant sortilin administration induced arterial hypertension in WT mice. In contrast, genetic deletion of sphingosine-1-phosphate receptor 3 (S1P3) and gp91phox/NOX2 resulted in preservation of endothelial function and blood pressure homeostasis after 14 days of systemic sortilin administration. Translating these research findings into the clinical setting, we detected elevated sortilin levels in hypertensive patients with endothelial dysfunction. Furthermore, in a population-based cohort of 270 subjects, we showed increased plasma ASMase activity and increased plasma levels of sortilin, S1P, and soluble NOX2-derived peptide (sNOX2-dp) in hypertensive subjects, and the increase was more pronounced in hypertensive subjects with uncontrolled blood pressure. Our studies reveal what we believe is a previously unrecognized role of sortilin in the impairment of vascular function and in blood pressure homeostasis and suggest the potential of sortilin and its mediators as biomarkers for the prediction of vascular dysfunction and high blood pressure.

Authors

Paola Di Pietro, Albino Carrizzo, Eduardo Sommella, Marco Oliveti, Licia Iacoviello, Augusto Di Castelnuovo, Fausto Acernese, Antonio Damato, Massimiliano De Lucia, Fabrizio Merciai, Paola Iesu, Eleonora Venturini, Raffaele Izzo, Valentina Trimarco, Michele Ciccarelli, Giuseppe Giugliano, Roberto Carnevale, Vittoria Cammisotto, Serena Migliarino, Nicola Virtuoso, Andrea Strianese, Viviana Izzo, Pietro Campiglia, Elena Ciaglia, Bodo Levkau, Annibale A. Puca, Carmine Vecchione

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Figure 2

Sortilin evokes endothelial dysfunction through the S1P-signaling pathway.

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Sortilin evokes endothelial dysfunction through the S1P-signaling pathwa...
(A) NOX activity (n = 4) and (B) ASMase activity (n = 8) in HUVECs treated with vehicle (Ctrl) or sortilin. RFU, relative fluorescence units. (C) Acetylcholine-evoked vasorelaxation in WT mesenteric arteries exposed to vehicle or pretransfected with siRNA against ASMase (Si-ASMase) before sortilin; scrambled siRNA (Si-SCR) was used as control (n = 6). ASMase protein levels after siRNA silencing. (D) Effect of si-ASMase on NOX activity in WT mesenteric arteries exposed to sortilin (n = 4). (E) Acetylcholine-evoked vasorelaxation in WT mesenteric arteries exposed to vehicle or sortilin or pretreated with either K145 or SK1-I before sortilin (n = 5). (F) Effects of K145 and SK1-I on NOX activity in sortilin-stimulated HUVECs (n = 3). (G) LC-MS/MS quantification of extracellular S1P levels in vehicle- or sortilin-treated HUVECs (n = 7). (H) Representative immunoblots and densitometric analyses of 3 independent experiments evaluating S1P1 and S1P3 expression in WT mesenteric arteries exposed to vehicle or sortilin. (I) Acetylcholine-evoked vasorelaxation in WT mesenteric arteries exposed to vehicle or sortilin or pretreated with W146 or TY52156 before sortilin (n = 6). (J and K) NOX activity in (J) HUVECs treated with vehicle or sortilin or pretreated with W146 or TY52156 and in (K) S1P3–/– mesenteric arteries exposed to vehicle or sortilin (n = 4–5 replicates from 3 independent experiments). (L) Acetylcholine-evoked vasorelaxation in mesenteric arteries from WT and S1P3–/– mice exposed to vehicle or sortilin (n = 5). Data are represented as mean ± SD. Unpaired Student’s t test (A, B, G, and H); 1-way (D, F, J, and K) or 2-way (C, E, I, and L) ANOVA followed by Bonferroni’s post hoc test. *P < 0.0001 versus vehicle or SK1-I plus sortilin at the same acetylcholine concentration (as indicated by color code) (E); *P < 0.0001 versus vehicle or TY52156 plus sortilin at the same acetylcholine concentration (as indicated by color code) (I).