Identification of novel proteins and mechanistic pathways associated with early-onset hypertension by deep proteomic mapping of resistance arteries
Joakim Bastrup, Christian Aalkjær, Thomas A. Jepps
Abstract
Resistance arteries are small blood vessels that create resistance to blood flow. In hypertension, resistance arteries undergo remodeling, affecting their ability to contract and relax appropriately. To date, no study has mapped the hypertension-related proteomic changes in resistance arteries. Using a novel data-independent acquisition–mass spectrometry (DIA-MS) approach, we determined the proteomic changes in small mesenteric and renal arteries in pre- and early-onset hypertension from the spontaneously hypertensive rat (SHR) model, which represents human primary hypertension. Compared with normotensive controls, mesenteric arteries from 12-week-old SHRs had 286 proteins that were significantly up- or downregulated, whereas 52 proteins were identified as up- or downregulated in mesenteric arteries from 6-week-old SHRs. Of these proteins, 18 were also similarly regulated in SHR renal arteries. Our pathway analyses reveal several novel pathways in the pathogenesis of hypertension. Finally, using a matrisome database, we identified 38 altered extracellular-matrix-associated proteins, many of which have never previously been associated with hypertension. Taken together, this study reveals novel proteins and mechanisms that are associated with early-onset hypertension, thereby providing novel insights into disease progression. Resistance arteries are small blood vessels that create resistance to blood flow. In hypertension, resistance arteries undergo remodeling, affecting their ability to contract and relax appropriately. To date, no study has mapped the hypertension-related proteomic changes in resistance arteries. Using a novel data-independent acquisition–mass spectrometry (DIA-MS) approach, we determined the proteomic changes in small mesenteric and renal arteries in pre- and early-onset hypertension from the spontaneously hypertensive rat (SHR) model, which represents human primary hypertension. Compared with normotensive controls, mesenteric arteries from 12-week-old SHRs had 286 proteins that were significantly up- or downregulated, whereas 52 proteins were identified as up- or downregulated in mesenteric arteries from 6-week-old SHRs. Of these proteins, 18 were also similarly regulated in SHR renal arteries. Our pathway analyses reveal several novel pathways in the pathogenesis of hypertension. Finally, using a matrisome database, we identified 38 altered extracellular-matrix-associated proteins, many of which have never previously been associated with hypertension. Taken together, this study reveals novel proteins and mechanisms that are associated with early-onset hypertension, thereby providing novel insights into disease progression. Hypertension is the main risk factor for cardiovascular diseases and is a major global health burden, with increasing prevalence (1NCD Risk Factor Collaboration (NCD-RisC)Worldwide trends in blood pressure from 1975 to 2015: A pooled analysis of 1479 population-based measurement studies with 19·1 million participants.Lancet. 2017; 389: 37-55Abstract Full Text Full Text PDF PubMed Scopus (1466) Google Scholar). Although many studies have investigated specific genes, proteins, and pathways that are altered in arteries from hypertensive animals and humans, there is no overview of the changes that occur in arteries during hypertension. As such, the pathophysiology of essential hypertension remains unclear. To advance research in the field of hypertension, we need a better overview of the changes occurring in arteries, which will promote new research ideas and potential therapeutic targets (2Dzau V.J. Balatbat C.A. Future of hypertension.Hypertension. 2019; 74: 450-457Crossref PubMed Scopus (75) Google Scholar). In hypertension, resistance arteries undergo eutrophic and/or hypertrophic remodeling, which contributes to increased peripheral resistance (3Heagerty A.M. Aalkjaer C. Bund S.J. Korsgaard N. Mulvany M.J. Small artery structure in hypertension. Dual processes of remodeling and growth.Hypertension. 1993; 21: 391-397Crossref PubMed Scopus (591) Google Scholar). In patients with essential hypertension and the spontaneously hypertensive rat (SHR), inward eutrophic remodeling predominates (3Heagerty A.M. Aalkjaer C. Bund S.J. Korsgaard N. Mulvany M.J. Small artery structure in hypertension. Dual processes of remodeling and growth.Hypertension. 1993; 21: 391-397Crossref PubMed Scopus (591) Google Scholar). Several mechanisms are proposed to influence vascular remodeling in hypertension (4Intengan H.D. Schiffrin E.L. Vascular remodeling in hypertension.Hypertension. 2001; 38: 581-587Crossref PubMed Scopus (815) Google Scholar), including apoptosis (5González A. López B. Ravassa S. Querejeta R. Larman M. Díez J. Fortuño M.A. Stimulation of cardiac apoptosis in essential hypertension.Hypertension. 2002; 39: 75-80Crossref PubMed Scopus (101) Google Scholar), expanded extracellular matrix (ECM) (6Cai Z. Gong Z. Li Z. Li L. Kong W. Vascular extracellular matrix remodeling and hypertension.Antioxid. Redox Signal. 2021; 34: 765-783Crossref PubMed Scopus (27) Google Scholar), vascular inflammation (7Virdis A. Schiffrin E.L. Vascular inflammation: A role in vascular disease in hypertension?.Curr. Opin. Nephrol. Hypertens. 2003; 12: 181-187Crossref PubMed Scopus (168) Google Scholar), and dysfunctional endothelium (8Dharmashankar K. Widlansky M.E. Vascular endothelial function and hypertension: Insights and directions.Curr. Hypertens. Rep. 2010; 12: 448-455Crossref PubMed Scopus (315) Google Scholar). In addition, several contractile and dilatory mechanisms are compromised in arteries from hypertensive animals and humans, which also contribute to the development and persistence of hypertension. These maladaptive changes in the vessel wall influence the development and cardiovascular complications of hypertension. Although proteins have been implicated in vascular remodeling in hypertension, these proteins do not work in isolation. Mass spectrometry (MS) analysis has advanced rapidly over the past 2 decades and demonstrated clear advantages in mapping complex biological systems with high reproducibility (9Aebersold R. Mann M. Mass-spectrometric exploration of proteome structure and function.Nature. 2016; 537: 347-355Crossref PubMed Scopus (1226) Google Scholar, 10Gillet L.C. Navarro P. Tate S. Röst H. Selevsek N. L. R. R. of the data-independent A new for and proteome Full Text Full Text PDF PubMed Scopus Google Scholar). analysis identified proteomic changes in the J. H. A. in the of from the spontaneously hypertensive from proteomic PubMed Scopus Google Scholar), J. B. of is a for hypertensive PubMed Scopus Google Scholar), and L. L. in hypertrophic with and hypertension in spontaneously hypertensive PubMed Scopus Google in the SHR blood pressure and of and the changes in human essential hypertension, as cardiac and vascular remodeling K. K. of a of spontaneously hypertensive J. PubMed Scopus Google Scholar). To date, no study has mapped the proteomic changes in the resistance arteries of the SHR or in patients with essential hypertension. of this study to changes and pathways in mesenteric resistance and renal arteries from the which is for human essential hypertension and to proteomic we data-independent analysis to proteomic of resistance arteries to novel proteins and the mechanisms to vascular remodeling and early-onset hypertension. investigated the of mesenteric artery from the SHR we small mesenteric arteries from SHR and and of in the SHR to of to a from K. K. of a of spontaneously hypertensive J. PubMed Scopus Google Scholar). Our study to the changes that occur in the wall during the early-onset of high these were to and early-onset hypertensive and changes associated with hypertension. To proteomic we a that high mesenteric artery and the of to the J. L. R. L. identified and proteins in a and analysis 2019; PubMed Google Scholar). 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To for this factor in the model, we analysis of mesenteric artery from to we a in the a significantly regulated proteins the normotensive of SHR and analyses of proteomic in and early-onset hypertensive Using we identified 52 and significantly regulated proteins the mesenteric arteries of the SHRs to the and of and proteins 2 and and were and the significantly regulated proteins with identified in the and proteins that in the in the of proteins from the SHR as these to or in the in with 286 significantly regulated proteins and these proteins in the analysis to changes and the and downregulated proteins the and as and and proteins, including and These proteins are associated with and the changes in of the in regulated proteins in mesenteric proteins A of factor downregulated proteins of of of the up- and downregulated proteins identified the proteomic of 12-week-old mesenteric arteries from spontaneously hypertensive (SHR) and normotensive of identified is in a new of the up- and downregulated proteins identified the proteomic of 12-week-old mesenteric arteries from spontaneously hypertensive (SHR) and normotensive of identified is regulated proteins had in which and in the To in pathway in the we pathway analysis the of significantly regulated proteins using B. H. P. M. A. Z. J. A to and pathway PubMed Scopus Google and analysis identified of and of and and to proteins and had and using the analysis of proteins associated with hypertension of and of of to analysis of significantly regulated proteins identified in mesenteric arteries from 12-week-old and spontaneously hypertensive or of and as is with the the in a new analysis of significantly regulated proteins identified in mesenteric arteries from 12-week-old and spontaneously hypertensive or of and as is with the the from the SHR undergo vascular remodeling with of the wall in (3Heagerty A.M. Aalkjaer C. Bund S.J. Korsgaard N. Mulvany M.J. Small artery structure in hypertension. Dual processes of remodeling and growth.Hypertension. 1993; 21: 391-397Crossref PubMed Scopus (591) Google Scholar). Using we the of vascular remodeling in 12-week-old SHRs with A and In a we in in mesenteric arteries from the SHR with controls, the of hypertrophic remodeling of the has been associated with vascular remodeling (6Cai Z. Gong Z. Li Z. Li L. Kong W. Vascular extracellular matrix remodeling and hypertension.Antioxid. Redox Signal. 2021; 34: 765-783Crossref PubMed Scopus (27) Google Scholar). To we of of significantly regulated proteins and for in pathway analysis using which identified the extracellular matrix pathway and To this we of the to a and proteins A. S. H. In and in of and extracellular Full Text Full Text PDF PubMed Scopus Google Scholar, A. H. 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