Multi-analyte profiling in human carotid atherosclerosis uncovers pro-inflammatory macrophage programming in plaques

Abstract

SummaryMolecular characterisation of vulnerable atherosclerosis is necessary for targeting functional imaging and plaque-stabilising therapeutics. Inflammation has been linked to atherogenesis and the development of high-risk plaques. We set to quantify cytokine, chemokine and matrix metalloproteinase (MMP) protein production in cells derived from carotid plaques to map the inflammatory milieu responsible for instability. Carotid endarterectomies from carefully characterised symptomatic (n=35) and asymptomatic (n=32) patients were enzymatically dissociated producing mixed cell type atheroma cell suspensions which were cultured for 24 hours. Supernatants were interrogated for 45 analytes using the Luminex 100 platform. Twenty-nine of the 45 analytes were reproducibly detectable in the majority of donors. The in vitro production of a specific network of mediators was found to be significantly higher in symptomatic than asymptomatic plaques, including: tumour necrosis factor α, interleukin (IL) 1β, IL-6, granulocytemacrophage colony-stimulating factor (GM-CSF), macrophage colonystimulating factor (M-CSF), CCL5, CCL20, CXCL9, matrix metalloproteinase (MMP)-3 and MMP-9. Ingenuity pathway analysis of differentially expressed analytes between symptomatic and asymptomatic patients identified a number of key biological pathways (p< 10 -25 ). In conclusion, the carotid artery plaque culprit of ischaemic neurological symptoms is characterised by an inflammatory milieu favouring inflammatory cell recruitment and pro-inflammatory macrophage polarisation.

Inflammatory Differences in Plaque Erosion and Rupture in Patients With ST‐Segment Elevation Myocardial Infarction

“…Array analysis confirmed that multiple molecules thought to be important in plaque rupture were preferentially expressed in the RFC group, including matrix metalloproteinase 9, lipocalin 2, and RANTES . We also confirmed significantly increased I‐TAC/CXCL11 in coronary plasma samples in RFC, which is a new finding.…”

Molecular imaging of MMP activity discriminates unstable from stable plaque phenotypes in shear-stress induced murine atherosclerosis

“…However, MMPs are particularly attractive since they are involved in a multitude of molecular mechanisms directly promoting plaque instability, including matrix degradation of the fibrous cap [ 19 ] [ 20 ] [ 21 ] [ 22 ]. For human carotid endatherectomy samples, it has been shown that MMPs are overexpressed and activated in rather vulnerable and symptomatic plaque states [ 2 ] [ 3 ]. Interestingly, MMP activity was associated with a thin fibrous cap and a large lipid core phenotype of carotid plaques making MMP activity a highly promising molecular target for molecular imaging [ 2 ].…”

“…Here we aim at specifically image the activity of matrix metalloproteinases (MMPs) to discriminate stable from unstable plaques–a novel approach in the later described human like atherosclerosis model. This approach is based on findings, that in human carotid plaques MMPs, primarily MMP-9 and MMP-2, are overexpressed and activated in rather vulnerable and symptomatic plaque states as analysed by immunohistochemistry of endatherectomy samples [ 2 ] [ 3 ]. Interestingly, MMP activity was associated with a thin fibrous cap and a large lipid core phenotype of carotid plaques, making MMP activity a promising molecular target for molecular imaging.…”

Cytokines and atherosclerosis – new research directions

“…Этим же методом в супернатантах культур Т-лимфоцитов из нестабильных АСБ был определен более высокий уровень TNFα и более низкий уровень IL-4, чем из стабильных бляшек [40]. Установлена достоверно более высокая продукция IL-1β и IL-6 при культивировании образцов АСБ сонной артерии при выраженных симптомах заболевания, чем при бессимптомном течении [42]. В ýкспериментах ex vivo было проанализировано образование TGFβ и IL-11 Т-лимфоцитами АСБ, установлено снижение их синтеза в АСБ, даже при стимуляции поликлональным активатором пролиферации [43].…”