Microarray gene profiling of laser-captured cells: A new tool to study atherosclerosis in mice

Paul, Antoni; Yechoor, Vijay; Raja, Rajiv; Li, Lan

doi:10.1016/j.atherosclerosis.2007.12.056

Cited by 15 publications

(12 citation statements)

References 34 publications

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“…RNA was extracted with the PicoPure RNA Isolation Kit (Applied Biosystems). mRNA was submitted to 2 rounds of amplification with the RiboAmp HS RNA Amplification Kit (Applied Biosystems), each of which consisted of synthesis of double‐stranded cDNA using oligo(dT) primers tagged with the T7 promoter sequence, followed by in vitro transcription (IVT) with T7 polymerase 22. A 260 /A 280 ratios and size distribution analysis of amplified cRNAs were assessed with an Agilent 2100 bioanalyzer (Agilent Technologies Inc., Santa Clara, CA).…”

Section: Methodsmentioning

confidence: 99%

“…LCM of macrophages and RNA processing were performed as we previously described. 22 Briefly, %2000 laser shots (power, %65 mV; pulse, %2500 ls) were performed with a Veritas Microdissection System (Applied Biosystems), and cells were collected on CapSure HS LCM Caps (Applied Biosystems). RNA was extracted with the PicoPure RNA Isolation Kit (Applied Biosystems).…”

Section: Lcm and Rna Amplificationmentioning

confidence: 99%

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Transcriptional Profiling of Foam Cells Reveals Induction of Guanylate‐Binding Proteins Following Western Diet Acceleration of Atherosclerosis in the Absence of Global Changes in Inflammation

Goo

Son

Yechoor

et al. 2016

JAHA

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BackgroundFoam cells are central to two major pathogenic processes in atherogenesis: cholesterol buildup in arteries and inflammation. The main underlying cause of cholesterol deposition in arteries is hypercholesterolemia. This study aimed to assess, in vivo, whether elevated plasma cholesterol also alters the inflammatory balance of foam cells.Methods and ResultsApolipoprotein E–deficient mice were fed regular mouse chow through the study or were switched to a Western‐type diet (WD) 2 or 14 weeks before death. Consecutive sections of the aortic sinus were used for lesion quantification or to isolate RNA from foam cells by laser‐capture microdissection (LCM) for microarray and quantitative polymerase chain reaction analyses. WD feeding for 2 or 14 weeks significantly increased plasma cholesterol, but the size of atherosclerotic lesions increased only in the 14‐week WD group. Expression of more genes was affected in foam cells of mice under prolonged hypercholesterolemia than in mice fed WD for 2 weeks. However, most transcripts coding for inflammatory mediators remained unchanged in both WD groups. Among the main players in inflammatory or immune responses, chemokine (C‐X‐C motif) ligand 13 was induced in foam cells of mice under WD for 2 weeks. The interferon‐inducible GTPases, guanylate‐binding proteins (GBP)3 and GBP6, were induced in the 14‐week WD group, and other GBP family members were moderately increased.ConclusionsOur results indicate that acceleration of atherosclerosis by hypercholesterolemia is not linked to global changes in the inflammatory balance of foam cells. However, induction of GBPs uncovers a novel family of immune modulators with a potential role in atherogenesis.

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Section: Methodsmentioning

confidence: 99%

Section: Lcm and Rna Amplificationmentioning

confidence: 99%

Transcriptional Profiling of Foam Cells Reveals Induction of Guanylate‐Binding Proteins Following Western Diet Acceleration of Atherosclerosis in the Absence of Global Changes in Inflammation

Goo

Son

Yechoor

et al. 2016

JAHA

Self Cite

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“…28,30 This observation is consistent with the fact that human CRP concentrations can cover a 10 000-fold range (from Ͻ50 g/L to Ͼ500 mg/L) in the acute-phase response, and this is unlikely to be compatible with significant effects of CRP on vascular tone, activation of inflammatory cells, triggering of coagulation, or any of the other purported signaling functions lately ascribed to CRP on the basis of in vitro studies with commercial CRP preparations. Although there is 1 report of enhanced atherosclerosis in apolipoprotein E knockout mice expressing transgenic human CRP, 31 other and larger studies show no such effect [32][33][34] nor any proinflammatory or prothrombotic action even in aged atherosclerotic animals. 35 Furthermore, in the more humanized model of atherosclerosis in LDL receptor knockout mice expressing apolipoprotein B100, transgenic human CRP was found to be atheroprotective.…”

Section: Article See P 2088mentioning

confidence: 99%

C-Reactive Protein and Coronary Disease

Danesh

Pepys

2009

Circulation

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“…7,9 Transgenic expression of human CRP protects mice against endotoxin shock and microbial pathogens. 2 Transgenic expression of CRP in apolipoprotein E-null mice has been reported to accelerate, 16,18 to show no effect, 19,20 or even to slow atherosclerosis development. 21 Such divergent results are not easily explained and may reflect inherent difficulties in interpreting results from mouse models of atherogenesis.…”

mentioning

confidence: 99%

Platelets Affect the Structure and Function of C-Reactive Protein

Filep

2009

Circulation Research

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C-reactive protein (CRP) is an acute-phase reactant and an active regulator of the innate immune system. 1,2 Prospective clinical studies have shown that elevations in baseline CRP levels confer, albeit to varying degrees, additional prognostic value for predicting future cardiovascular events and death across all levels of the Framingham risk score. 3 CRP has been implicated in multiple aspects of atherogenesis and plaque vulnerability; however, a direct pathogenetic role for CRP in these events is presently controversial. 4,5 CRP has at least 2 conformationally distinct forms, native pentameric (p)CRP and monomeric (m)CRP. Native CRP consists of 5 identical subunits arranged in a cyclic pentamer. Loss of pentameric symmetry in pCRP, yielding mCRP, is associated with expression of distinct bioactivities. 6 -9 To date, little is known about the in vivo source(s) of mCRP.Platelets represent an important interface between thrombosis, innate and adaptive immunity, and atherogenesis. 10 Platelet-triggered inflammatory pathways contribute to atherosclerotic lesion formation and atherothrombosis. 10 In this issue of Circulation Research, Eisenhardt et al 11 provide new insights into the relationship of platelets, CRP, and inflammation. They show that activated platelets dissociate pCRP into mCRP and provide evidence, suggesting that mCRP, rather than pCRP, localizes monocyte-mediated inflammation to the atherosclerotic plaque.Native CRP is thought to be very stable and is not expected to dissociate into separate subunits without denaturation. 12 However, calcium-dependent binding of pCRP to liposomes or cell membranes results in separation into subunits. 13 Eisenhardt et al 11 show that binding of pCRP to lysophosphatidylcholine expressed on the surface of activated platelets and apoptotic monocytic THP-1 cells triggers its rapid dissociation into mCRP. Formation of mCRP involves dissociation of the CRP pentameric disk, resulting in a structural change from predominantly ␤-sheet structure to an ␣-helical structure and expression of intersubunit contact residues, in particular residues 197 to 202, the predominant epitope only expressed on mCRP. 14 mCRP-specific antigen has been detected in inflamed tissues. 15 Immunohistological staining of human atherosclerotic lesions consistently places CRP within the lesion, frequently along with the terminal complement complex. 4,5 Because most anti-CRP antibodies, including the widely used antibody clone 8, recognize both pCRP and mCRP, 16 it is uncertain whether lesions express mCRP, pCRP, or both. Eisenhardt et al 11 show that human aortic and coronary artery atherosclerotic plaques stained with the clone 8 antibody and a specific anti-mCRP antibody but not with a specific anti-pCRP antibody, indicating accumulation of mCRP rather than pCRP in atherosclerotic lesions. mCRP-staining correlated with areas that stained for macrophages or platelets, suggesting generation of mCRP by these cells. Formal colocalization studies would lend additional support to this notion. Future stud...

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Microarray gene profiling of laser-captured cells: A new tool to study atherosclerosis in mice

Cited by 15 publications

References 34 publications

Transcriptional Profiling of Foam Cells Reveals Induction of Guanylate‐Binding Proteins Following Western Diet Acceleration of Atherosclerosis in the Absence of Global Changes in Inflammation

Transcriptional Profiling of Foam Cells Reveals Induction of Guanylate‐Binding Proteins Following Western Diet Acceleration of Atherosclerosis in the Absence of Global Changes in Inflammation

C-Reactive Protein and Coronary Disease

Platelets Affect the Structure and Function of C-Reactive Protein

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