BAS/BSCR2 Analysis of microvessels within carotid plaques: comparison of symptomatic and asymptomatic patients to identify those at risk from plaque rupture

Natural antisense transcripts (NATs), a non-coding RNA subclass, being transcribed in antisense direction to protein coding genes, are an intriguing novel class of targetable modulators, exerting crucial effects on gene expression. Aim of the current study was to investigate the contribution of NATs to atherosclerotic plaque vulnerability. Using laser capture micro-dissection, we isolated fibrous caps tissue of carotid artery plaques from 20 symptomatic patients with ruptured lesions vs. 20 samples from asymptomatic patients with stable lesions. A human transcriptome array (HTA; GeneChip 2.0 ) was used to profile the expression of all currently annotated RNA transcripts. Nucleoside diphosphate-linked moiety X motif 6 (NUDT6) was identified as one of the most significantly up-regulated transcripts in fibrous caps of ruptured lesions. Interestingly, NUDT6 is an established antisense RNA targeting the fibroblast growth factor 2 (FGF2). Of importance, FGF2 was among the most significantly down-regulated transcripts in ruptured lesions, corresponding to elevated NUDT6 expression. In situ hybridization in both, human and mouse carotid atherosclerotic plaques, confirmed substantially higher expression levels of NUDT6 in ruptured lesions compared to stable. In addition, in situ hybridization revealed a distinct co-localization with smooth muscle cells (SMCs) in advanced plaques. Overexpression of NUDT6 in cultured human carotid artery SMCs effectively limited FGF2 on the mRNA as well as protein level. Furthermore, reduction of NUDT6 via siRNA stimulated proliferation and blocked apoptosis in SMCs. In an inducible atherosclerotic plaque rupture model using incomplete ligation and cuff placement on common carotid arteries of male apoE -/- mice, NUDT6 inhibition with gapmeRs was able to significantly improve SMC survival rates, leading to thicker fibrous caps, and to reduce the plaque rupture rate compared to scramble-gapmeR control-treated mice (22% vs . 63%, p = 0.03). The present study presents NUDT6 as a novel crucial antisense regulator of fibrous cap stability through steering SMC survival via targeting its sense RNA transcript FGF2.

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Prolonged QRS associated with left bundle branch conduction defect is a prognostic red flag in asymptomatic patients at risk for heart failure (ACCF/AHA stages A and B): Insights from the DAVID-Berg study

European Journal of Preventive Cardiology ◽

10.1177/2047487319896431 ◽

2019 ◽

Vol 27 (19) ◽

pp. 2326-2329

Author(s):

Andrea Pozzi ◽

Mauro Gori ◽

Annamaria Iorio ◽

Attilio Iacovoni ◽

Alessandra Carobbio ◽

...

Keyword(s):

Heart Failure ◽

At Risk ◽

Asymptomatic Patients ◽

Patients At Risk ◽

Red Flag ◽

Conduction Defect

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Intra-Plaque Vessels on Contrast-Enhanced Ultrasound Sonography Predict Carotid Plaque Histology

Cerebrovascular Diseases ◽

10.1159/000495299 ◽

2018 ◽

Vol 46 (5-6) ◽

pp. 265-269 ◽

Cited By ~ 2

Author(s):

Takaaki Amamoto ◽

Noriyuki Sakata ◽

Toshiyasu Ogata ◽

Hirofumi Shimada ◽

Tooru Inoue

Keyword(s):

Carotid Plaque ◽

Plaque Rupture ◽

Heart Association ◽

Receiver Operating Curve ◽

Rank Test ◽

Vessel Size ◽

Carotid Plaques ◽

Plaque Area ◽

Receiver Operating Curve Analysis ◽

Signed Rank Test

Background and Purpose: Although the echogenicity of carotid plaques on carotid enhanced ultrasound (CEUS) was reported to correlate with the historological density of intra-plaque neovessels, it is unclear whether the intra-plaque vessel size is associated with carotid plaque vulnerability. We evaluated the relationship of size of intra-plaque vessels on CEUS with carotid plaque histology. Methods: We prospectively registered patients with carotid stenosis who were hospitalized to receive carotid endarterectomy between 2012 and 2016. CEUS was performed by ultrasound specialists using a 7-MHz linear transducer (GE LOGIQ7; GE Healthcare, Milwaukee, WN, USA). Sonazoid® was used for conducting CEUS. The sizes of microbubbles inside the plaques were scored at 0, 1, 3, and 5 min after the injection of contrast agent, and were categorized according to our defined vascular score (Vas-S; 0: the effect of contrast was not recognized; 1: the microbubbles were visible, but so blurred and vague that their shape could not be recognized; 2: dot or string-like microbubbles with movement, localized in part or the whole site of the plaque). At histological examination, we simplified the modified American Heart Association classification and defined as an atherosclerotic category (Ath-cat; 1: unruptured plaque; 2: ruptured plaque; 3: healed plaque). We then assessed the correlation of Vas-S with Ath-cat, which describes the process of rupture and restoration of carotid plaques. Results: A total of 97 patients were included in this study. A higher Ath-cat was significantly associated with higher Vas-S at any time. Spearman signed-rank test indicated that Vas-S at 1 min was most strongly correlated with Ath-cat (ρ = 0.43, p = 0.001). Receiver operating curve analysis indicated that a Vas-S of 0 at 1 min was significantly associated with an unruptured plaque (area under curve [AUC] 0.72, p = 0.006), while a Vas-S of 2 at 1 min was significantly associated with a healed plaque (AUC 0.72, p = 0.001). Conclusion: Vas-S values of 0 and 2 at 1 min indicated unruptured and healed plaques respectively. Thus, a Vas-S of 1 at 1 min is an indicator of a ruptured plaque. The intra-plaque vessel size on CEUS was significantly associated with carotid plaque histology, and may predict the process of plaque rupture and restoration.

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Modulation of Neuropeptide‐Y receptor expression by IGF‐1 and atheroma‐associated cytokines in carotid plaques of symptomatic and asymptomatic patients

The FASEB Journal ◽

10.1096/fasebj.24.1_supplement.1028.2 ◽

2010 ◽

Vol 24 (S1) ◽

Author(s):

Divya Pankajakshan ◽

Guanghong Jia ◽

Iraklis Pipinos ◽

Devendra Agrawal

Keyword(s):

Neuropeptide Y ◽

Receptor Expression ◽

Carotid Plaques ◽

Neuropeptide Y Receptor ◽

Asymptomatic Patients

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Evolution of Myocardial Dysfunction in Asymptomatic Patients at Risk of Heart Failure

JACC Cardiovascular Imaging ◽

10.1016/j.jcmg.2020.09.032 ◽

2020 ◽

Author(s):

Amera Halabi ◽

Hilda Yang ◽

Leah Wright ◽

Elizabeth Potter ◽

Quan Huynh ◽

...

Keyword(s):

Heart Failure ◽

At Risk ◽

Myocardial Dysfunction ◽

Asymptomatic Patients ◽

Patients At Risk

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Abstract 127: Induction of miR-21 Increases Fibrous Cap Stability in Vulnerable Atherosclerotic Lesions

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.36.suppl_1.127 ◽

2016 ◽

Vol 36 (suppl_1) ◽

Author(s):

Hong Jin ◽

Yuhuang Li ◽

Alexandra Bäcklund ◽

Albert Busch ◽

Suzanne M Eken ◽

...

Keyword(s):

Carotid Artery ◽

Cellular Proliferation ◽

Plaque Rupture ◽

Oxidized Ldl ◽

Survival Rates ◽

Experimental Models ◽

Vascular Remodelling ◽

Plaque Vulnerability ◽

Plaque Instability ◽

Asymptomatic Patients

The aim of the present study was to explore the role of miRNAs as potential regulators in patients with carotid artery stenosis and concordant vulnerable atherosclerotic plaques. A pre-determined miRNA-array of laser captured micro-dissected (LCM) tissue specimen from fibrous caps of 10 symptomatic patients (stroke or TIA within > 14 days and histo-morphologically identified ruptured lesion) compared to fibrous caps from 10 asymptomatic patients (stable lesions; high grade stenosis) discovered miR-21 as one of the two miRNAs (miRs-21 and -210) being substantially down-regulated in symptomatic patients. To functionally evaluate the contribution of miR-21 to plaque vulnerability, we created miR21 -/- / ApoE -/- mice on a C57BL/6 background. We explored the phenotype of these newly developed miR21 -/- /ApoE -/- mice in experimental models of vascular remodelling and plaque vulnerability. First, miR21 -/- mice revealed a complete lack of SMC proliferation in response to carotid ligation injury. In the second inducible plaque rupture model, using incomplete ligation of the common carotid artery with consecutive cuff placement proximal to the ligated region indicated that all miR21 -/- /ApoE -/- mice ( n =10) presented atherothombotic events and signs of severe plaque instability. The rupture rate in control miR21 +/+ /ApoE -/- mice was significantly lower at 56% ( n =9). In addition, miR21 -/- /ApoE -/- showed an increase in lesion area in the aortic root and substantially higher levels of macrophage infiltration in injured carotid arteries. Dynamic live cell imaging, using isolated aortic SMCs and peritoneal macrophages from miR21 -/- /ApoE -/- vs. miR2 +/+ /ApoE -/- mice displayed substantial lower cellular proliferation and survival rates (for SMCs), and distinct advanced inflammatory activity upon oxidized LDL (oxLDL) stimulation of macrophages. In the present study we identified miR-21 as a key modulator of pathologic processes in atherosclerosis-related vascular remodelling. Targeted, lesion-site specific overexpression of miR-21 could be a novel and powerful future strategy to stabilize vulnerable plaques by inducing pro-proliferative mechanisms in SMC-enriched fibrous caps.

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Abstract WP434: Serum MicroRNA Panel as a Candidate Biomarker of Carotid Plaque Rupture and Stroke

Stroke ◽

10.1161/str.48.suppl_1.wp434 ◽

2017 ◽

Vol 48 (suppl_1) ◽

Author(s):

Aaron M Brug ◽

Sullivan Smith ◽

Ashton J Brooks ◽

Daniel J Lightell ◽

Hernan A Bazan ◽

...

Keyword(s):

Ischemic Stroke ◽

Acute Ischemic Stroke ◽

Carotid Plaque ◽

Plaque Rupture ◽

Rna Seq ◽

Circulating Mirnas ◽

Ischemic Event ◽

Biomarker Panel ◽

Serum Microrna ◽

Asymptomatic Patients

Introduction: Atherosclerotic plaque vulnerability and subsequent rupture are mediated by alterations in miRNAs that promote inflammation and fibrous cap thinning, leading to an acute ischemic stroke. Release of these miRNAs from the plaque tissue into the circulation may provide a miRNA profile that serves as a biomarker panel to identify the vulnerable patient at greatest risk of plaque rupture. Methods: Using RNA-Seq methodology, serum microRNA content was compared between patients undergoing carotid endarterectomy (CEA) for 2 distinct clinical phenotypes: patients without previous neurological events but high-grade carotid stenosis (asymptomatic, n=5), and patients with an acute neurological event within 5 days of the CEA (urgent, n=7). Differential expression of miRNAs was performed using the R statistical software and the DESeq2 package. Results: A total of 695 miRNAs detected in sera on RNA-Seq, 54 were significantly different between the asymptomatic and urgent groups. 22 miRNAs were decreased and 32 were increased in the serum of urgent patient’s compared to asymptomatic patients (adjusted p-value < 0.05). Of these miRNAs, miR-21, miR-486, miR-23b, miR-27b, and let-7f were selected for validation as a biomarker panel based on a prior association with vascular disease. A pilot study (n= 6 asymptomatic and 6 urgent) measuring these miRNAs demonstrated that while miR-486 was detectable in all samples and exhibited a 9.4-fold increase (p < 0.10) in the urgent group, the other miRNAs were only detected in a fraction of the samples (33 - 75%). Furthermore, miR-486 alone did not exhibit characteristics of a good indicator of ischemic stroke. A score normalizing the expression of miR-486 to the mean expression of the detectable miRNAs from this panel, did yield a promising C-statistic (0.97 ± 0.08, p < 0.01), suggesting its potential as an indicator of an acute ischemic event. Conclusion: Carotid plaque rupture resulting in an acute ischemic event is associated with alterations in the profile of circulating miRNAs and these may be useful as biomarkers of ischemic stroke. Our pilot data suggest that the development of a score based on a panel of circulating miRNAs may allow for the diagnosis of patients experiencing plaque rupture and an acute ischemic stroke.

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Predicting Human Carotid Plaque Site of Rupture Using 3D Critical Plaque Wall Stress and Flow Shear Stress: A 3D Multi-Patient FSI Study Based on In Vivo MRI of Plaques With and Without Prior Rupture

ASME 2010 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2010-19080 ◽

2010 ◽

Author(s):

Zhongzhao Teng ◽

Gador Canton ◽

Chun Yuan ◽

Marina Ferguson ◽

Chun Yang ◽

...

Keyword(s):

Shear Stress ◽

Critical Stress ◽

Plaque Rupture ◽

Wall Stress ◽

Global Maximum ◽

Flow Shear ◽

Carotid Plaques ◽

Flow Shear Stress ◽

Human Carotid

Atherosclerotic plaque rupture is the primary cause of cardiovascular clinical events such as heart attack and stroke. Image-based computational models of vulnerable plaques have been introduced seeking critical mechanical indicators which may be used to identify potential sites of rupture [1–5]. Models derived from 2D ex vivo and in vivo magnetic resonance images (MRI) have shown that 2D local critical stress values rather than global maximum stress values correlated better with plaque vulnerability, as defined by histopathological and morphological analyses [5]. A recent study by Tang et al. [4] using in vivo MRI-based 3D fluid-structure interaction (FSI) models for ruptured human carotid plaques, reported that mean plaque wall stress (PWS) values from ulcer nodes were 86% higher than mean PWS values from all non-ulcer nodes (p<0.0001). This study extends the “critical stress” concept to 3D and uses 3D FSI models based on in vivo MRI data of human atherosclerotic carotid plaques with and without prior rupture to identify 3D critical plaque wall stress (CPWS), critical flow shear stress (CFSS), and to investigate their associations with plaque rupture.

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