12(S)-HETE mediates diabetes-induced endothelial dysfunction by activating intracellular endothelial cell TRPV1
Mandy Otto, … , Eric R. Gross, Nana-Maria Wagner
Mandy Otto, … , Eric R. Gross, Nana-Maria Wagner
Published June 25, 2020
Citation Information: J Clin Invest. 2020;130(9):4999-5010. https://doi.org/10.1172/JCI136621.
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Research Article Vascular biology

12(S)-HETE mediates diabetes-induced endothelial dysfunction by activating intracellular endothelial cell TRPV1

Abstract

Patients with diabetes develop endothelial dysfunction shortly after diabetes onset that progresses to vascular disease underlying the majority of diabetes-associated comorbidities. Increased lipid peroxidation, mitochondrial calcium overload, and mitochondrial dysfunction are characteristics of dysfunctional endothelial cells in diabetic patients. We here identified that targeting the lipid peroxidation product 12(S)-hydroxyeicosatetraenoic acid–induced [12(S)-HETE–induced] activation of the intracellularly located cation channel transient receptor potential vanilloid 1 (TRPV1) in endothelial cells is a means to causally control early-stage vascular disease in type I diabetic mice. Mice with an inducible, endothelium-specific 12/15-lipoxygenase (12/15Lo) knockout were protected similarly to TRPV1-knockout mice from type 1 diabetes–induced endothelial dysfunction and impaired vascular regeneration following arterial injury. Both 12(S)-HETE in concentrations found in diabetic patients and TRPV1 agonists triggered mitochondrial calcium influx and mitochondrial dysfunction in endothelial cells, and 12(S)-HETE effects were absent in endothelial cells from TRPV1-knockout mice. As a therapeutic consequence, we found that a peptide targeting 12(S)-HETE–induced TRPV1 interaction at the TRPV1 TRP box ameliorated diabetes-induced endothelial dysfunction and augmented vascular regeneration in diabetic mice. Our findings suggest that pharmacological targeting of increased endothelial lipid peroxidation can attenuate diabetes-induced comorbidities related to vascular disease.

Authors

Mandy Otto, Clarissa Bucher, Wantao Liu, Melanie Müller, Tobias Schmidt, Marina Kardell, Marvin Noel Driessen, Jan Rossaint, Eric R. Gross, Nana-Maria Wagner

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Figure 4

12(S)-HpETE effects on mitochondrial and endothelial function are mediated by TRPV1.

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12(S)-HpETE effects on mitochondrial and endothelial function are mediat...
(A) Patch clamp experiments with human endothelial cells show that 700 nM 12(S)-HpETE induces currents comparable to those induced by 10 μM capsaicin and that these currents are antagonized by 10 μM of the TRPV1 antagonist BCTC. (B) Mitochondrial calcium increase induced by 1 μM 12(S)-HpETE added as indicated by the arrow and assessed as increase in fluorescence intensity of Rhod-2–loaded murine endothelial cells isolated from WT (dashed lines) versus Trpv1–/– mice. **P < 0.01 vs. baseline, 2-way ANOVA/Bonferroni, summary of n = 5 independent experiments. (C) Mitochondrial OCR is reduced by 100 nM 12(S)-HpETE in endothelial cells from WT but not Trpv1–/– mice. *P < 0.05, **P < 0.01 vs. EtOH or as indicated, 1-way ANOVA/Bonferroni, n = 5–10 independent experiments. (D and E) Effects of 12(S)-HpETE on endothelial capillary-like tube formation on Matrigel. Effects of 100 nM 12(S)-HpETE are absent in murine endothelial cells isolated from Trpv1–/– mice and endothelial cells isolated from WT mice in the presence of 10 μM of the TRPV1 antagonist BCTC. *P <0.05, ***P < 0.001 vs. EtOH or as indicated, 1-way ANOVA/Bonferroni, n = 5–10 independent experiments. Scale bar: 500 μm. All data are presented as mean ± SEM.