Peripheral arteriopathy caused by Notch3 gain-of-function mutation involves ER and oxidative stress and blunting of NO∕sGC∕cGMP pathway

Neves, K. B. , Morris, H., Alves-Lopes, R., Muir, K. W. , Moreton, F., Delles, C. , Montezano, A. C. and Touyz, R. M. (2021) Peripheral arteriopathy caused by Notch3 gain-of-function mutation involves ER and oxidative stress and blunting of NO∕sGC∕cGMP pathway. Clinical Science, 135(6), pp. 753-773. (doi: 10.1042/CS20201412) (PMID:33681964)

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Abstract

Notch3 mutations cause Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoenephalopathy (CADASIL), which predisposes to stroke and dementia. CADASIL is characterized by vascular dysfunction and granular osmiophilic material (GOM) accumulation in cerebral small vessels. Systemic vessels may also be impacted by Notch3 mutations. However vascular characteristics and pathophysiological processes remain elusive. We investigated mechanisms underlying the peripheral vasculopathy mediated by CADASIL-causing Notch3 gain-of-function mutation. We studied: i) small arteries and vascular smooth muscle cells (VSMC) from TgNotch3R169C mice (CADASIL model), ii) VSMCs from peripheral arteries from CADASIL patients, and iii) post-mortem brains from CADASIL individuals. TgNotch3R169C vessels exhibited GOM deposits, increased vasoreactivity and impaired vasorelaxation. Hypercontractile responses were normalized by fasudil (Rho kinase inhibitor) and 4-PBA (endoplasmic-reticulum (ER) stress inhibitor). Ca2+ transients and Ca2+ channel expression were increased in CADASIL VSMCs, with increased expression of Rho GEFs and ER stress proteins. Vasorelaxation mechanisms were impaired in CADASIL, evidenced by decreased eNOS phosphorylation and reduced cGMP levels, with associated increased guanylate cyclase (sGC) oxidation, decreased sGC activity and reduced levels of the vasodilator H2O2. In VSMCs from CADASIL patients, sGC oxidation was increased and cGMP levels decreased, effects normalized by fasudil and 4-PBA. Cerebral vessels in CADASIL patients exhibited significant oxidative damage. In conclusion, peripheral vascular dysfunction in CADASIL is associated with altered Ca2+ homeostasis, oxidative stress and blunted eNOS/sGC/cGMP signaling, processes involving Rho kinase and ER stress. We identify novel pathways underlying the peripheral arteriopathy induced by Notch3 gain-of-function mutation, phenomena that may also be important in cerebral vessels.

Item Type:Articles
Additional Information:This study was supported by grants from the British Heart Foundation (BHF) (RE/13/5/30177; 18/6/34217) and the MRC (MC-PC-15076). The CADASIL biopsy samples were funded by grants from The Neurosciences Foundation and The Stroke Association (TSA 2013/02) and the control biopsy samples were funded by grants from the Department of Medicine, University of Ottawa and the Canadian Institutes of Health Research, Canada. RMT is supported through a BHF Chair award (CH/12/29762) and ACM is supported through a Walton Foundation fellowship, University of Glasgow.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Moreton, Dr Fiona and Montezano, Dr Augusto and Delles, Professor Christian and Muir, Professor Keith and Morris, Hannah and Touyz, Professor Rhian and Neves, Dr Karla
Authors: Neves, K. B., Morris, H., Alves-Lopes, R., Muir, K. W., Moreton, F., Delles, C., Montezano, A. C., and Touyz, R. M.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Neuroscience and Psychology
Journal Name:Clinical Science
Publisher:Portland Press
ISSN:0143-5221
ISSN (Online):1470-8736
Published Online:08 March 2021

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
190814BHF centre of excellenceRhian TouyzBritish Heart Foundation (BHF)RE/13/5/30177Institute of Cardiovascular & Medical Sciences
303944BHF Centre of ExcellenceRhian TouyzBritish Heart Foundation (BHF)RE/18/6/34217CAMS - Cardiovascular Science
172586Thiol Modification and Redox SignallingNeil BulleidMedical Research Council (MRC)MC_PC_15076Institute of Molecular, Cell & Systems Biology
168941Characterising vascular pathophysiology in CADASIL (cerebral autosomal dominant arteriopathy with subacute infarcts and leukoencephalopathy) by evaluation of gluteal resistance vesselsKeith MuirStroke Association (STROKEAS)TSA 2013/02Institute of Neuroscience & Psychology
190615Vascular Noxs as therapeutic targets and biomarkers in hypertensionRhian TouyzBritish Heart Foundation (BHF)CH/12/4/29762Institute of Cardiovascular & Medical Sciences