Review Article Making Sense in Antisense: Therapeutic ...downloads.hindawi.com/journals/jdr/2013/834727.pdf · new diagnostic and therapeutic opportunities to treat diabetes-induced

Hindawi Publishing CorporationJournal of Diabetes ResearchVolume 2013 Article ID 834727 10 pageshttpdxdoiorg1011552013834727

Review ArticleMaking Sense in Antisense Therapeutic Potential of NoncodingRNAs in Diabetes-Induced Vascular Dysfunction

Suzanne M Eken Hong Jin Ekaterina Chernogubova and Lars Maegdefessel

Atherosclerosis Research Unit Department of Medicine Center for Molecular Medicine (CMM L8) Karolinska Institute17176 Stockholm Sweden

Correspondence should be addressed to Lars Maegdefessel larsmaegdefesselkise

Received 3 October 2013 Accepted 26 October 2013

Academic Editor Hanrui Zhang

Copyright copy 2013 Suzanne M Eken et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

The rapid rise of type II diabetes mellitus and its accompanying vascular complications call for novel approaches in unravellingits pathophysiological mechanisms and designing new treatment modalities Noncoding RNAs represent a class of previouslyunknown molecular modulators of this disease The most important features of diabetes-induced vascular disease which includemetabolic deregulation increased oxidative stress release of inflammatory mediators like adipokines and pathologic changes invascular cells all are depicted and governed by a certain set of noncoding RNAs While these mechanisms are being unravellednew diagnostic and therapeutic opportunities to treat diabetes-induced vascular disease emerge

1 Prevalence of Diabetes andVascular Complications

The prevalence of type II diabetes mellitus (T2DM) andrelated metabolic syndrome keeps rising at an alarmingrate and becomes a global health issue affecting childrenadolescents and adults According to the World HealthOrganization approximately 346 million people worldwidehave T2DM and this number is estimated to almost doubleby 2030 [1 2] T2DM is a progressive multisystem dis-ease accompanied by vascular dysfunction and a tremen-dous increase in cardiovascular mortality [3] Patients withdiabetes andor metabolic syndrome have a significantlyincreased risk of cardiovascular complications compared topeople with normal insulin sensitivity and production In thepast years many studies have tried to reveal the mechanismsfor diabetic vascular complications with varying degrees ofsuccess The recent discovery of noncoding RNA (ncRNAseg microRNAs and long noncoding RNAs) as well astheir influence on human pathophysiology provides us withnew opportunities to unravel and positively influence thedisease process In the present review we summarize thepathophysiology of T2DM associated vascular disease and

highlight the association of ncRNA with diabetic vasculardysfunction

2 Pathophysiology of DiabeticVascular Disease

The complications of T2DM encompass a diverse range ofpathologies of large and small arteries leading to diabeticmacrovascular occlusive disease andor microvascular dys-function which include coronary artery diseases cerebralartery diseases and peripheral vascular diseases amongstothers [1 4] Until now most common additive risk factorsfor vascular disease in peoplewith diabetes have beendemon-strated as hyperglycaemia insulin resistance dyslipidaemiahypertension tobacco use and obesity [2 5] however theinteraction of the factors and molecular signalling pathwayshave not been fully elucidated Established mechanismsfor vascular disease in diabetic patients are manifold andinclude the pathologic effects of advanced glycosylation endproduct (AGE) accumulation impaired vasodilator responseattributable to nitric oxide inhibition smooth muscle celldysfunction overproduction of endothelial growth factors

2 Journal of Diabetes Research

chronic inflammation hemodynamic deregulation impairedfibrinolytic ability and enhanced platelet aggregation [5]

21 Insulin Signalling and Hyperglycaemia Abnormalities invascular endothelial (EC) and vascular smooth muscle cell(VSMC) function as well as a propensity to thrombosisare important contributors to vascular complications [5]Hyperglycaemia and insulin resistance have been identifiedas key players in the development of diabetic atheroscle-rosis with metabolic insulin signalling being an importantcontributor to normal vascular function and homeostasis[5] Decreased insulin sensitivity in cardiovascular tissuesis an underlying abnormality in obesity hypertension andT2DM [3] In physiological conditions insulin promotesendothelium-dependent relaxation by a mechanism thatinvolves an increase of nitric oxide (NO) production viaactivation of phosphatidylinositol-3 kinase (PI3K) and Aktkinase pathways [6 7] NO has also been shown to preventendothelial apoptosis as well as neutrophil and plateletadhesion to the vascular wall [8] The initial trigger bywhich high glucose concentrations alter vascular functionis the imbalance between nitric oxide (NO) bioavailabilityand accumulation of reactive oxygen species (ROS) [9]Decrease in NO bioavailability is considered the hallmark ofendothelial dysfunction subsequently leading to attenuatedvascular relaxation and atherosclerosis [10]

Insulin signalling also plays a critical role in normalvascular function via modulation of calcium handling andsensitivity in VSMCs [3] When insulin signal transduc-tion is impaired bioavailability of NO in ECs decreaseswhile endothelin-1 production the inflammatory activityand smooth muscle cell proliferation increase [6]

22 Dyslipidaemia High circulating levels of triglyceride-rich particles reduced synthesis of HDL and enhanced pro-duction of atherogenic low-density lipoprotein (LDL) par-ticles characterize diabetic dyslipidaemia [11 12] Howeverthe regulation of lipid metabolism in diabetes is extremelycomplex and the mechanisms to trigger vascular dysfunctionare only partially explored The fading of the glycocalyx inlarge arteries exposed to hyperlipidaemic stress may be anearly characteristic of an increased vascular vulnerability [13]Also high levels of LDL and free fatty acids (FFAs) can causeincreased permeability of ECs induce EC-derived foam cellsformation and abnormal hyperplasia of extracellular matrixas well as disturbed secretion of NO and proinflammatorymolecules such as vascular cell adhesionmolecule 1 (VCAM-1) plateletendothelial cell adhesion molecule 1 (PECAM-1) intercellular adhesion molecule 1 (ICAM-1) P-selectinmonocyte chemoattractant protein 1 (MCP-1) interleukin 6(IL-6) Toll-like receptor 4 (TLR-4) CD40 PAI-1 and soforth [13] Although many clinical trials have demonstratedsignificant advantages of utilizing cholesterol-targeting drugsto reduce cardiovascular complications of diabetic patients[14ndash16] the remaining higher prevalence of vascular dysfunc-tion calls for further basic and clinical studies allowing forbetter prevention and treatment

23 Vascular Oxidative Stress T2DM resulting in vasculardysfunction may also occur through the increase in NAPDHoxidase-induced ROS production in the vasculature [17] InT2DM high intracellular glucose levels increase ROS pro-duction by triggering various cellular mechanisms and regu-lators such as protein kinase C (PKC) activation polyol andhexosamine flux AGEs and nuclear factor kappa B (NF-120581B)mediated vascular inflammation [9 10] Hyperglycaemia-mediated superoxide formation contributes to the patho-physiological complications in diabetic patients Not only isthe generation of reactive oxygen species (ROS) elevated indiabetes but the activity of the antioxidant defence systemalso declines [18] There are multiple targets of oxidativedamage in the diabetic vasculature with modifications ofproteins lipids and nucleic acids occurring in both ECsand VSMCs Enhanced oxygen radical production throughtumour necrosis factor 120572 (TNF-120572) and AGE or AGE receptor(RAGE) signalling reduces NO bioavailability and resultsin impairment of vascular function [19] Furthermore viascavenger receptor recognition ROS trigger recruitment ofmonocytes and their differentiation intomacrophages whichinitiate a vascular inflammation cascade [20]

24 Adipokines Obesity and T2DM are associated withadverse expression patterns of various adipose-derivedcytokines and chemokines and enhanced adipose inflamma-tory cell infiltration [21] Adipokines produced by adiposetissue may affect vascular function and insulin sensitivity[22] To date several adipokines have been identified andcharacterized to modify vascular function and the list is stillgrowing [23]

Adiponectin for example an anti-inflammatoryadipokine which is reduced in obesity and T2DM [24]has been shown to increase insulin sensitivity and to improvevascular function by reducing TNF-120572-stimulated expressionof endothelial adhesion molecules and monocyte attachment[25ndash27] In addition adiponectin attenuates the productionof ROS induced by high glucose oxidized LDL and palmitatein endothelial cells [27]

Another well studied adipokine leptin has been sug-gested to induce vascular endothelial dysfunction [28] andVSMC proliferation [29] Also resistin which is elevatedin obesity and T2DM was found to be a strong risk factorfor acute coronary syndrome in different clinical studies[30] Resistin promotes atherogenic changes in VSMCs suchas increased proliferation migration and contractility [31]Many other adipokines and cytokines such as visfatin TNF-120572 plasminogen activator inhibitor 1 (PAI-1) and so on mayall contribute to vascular dysfunction by affecting vasculartone and infiltration of inflammatory cells [31 32]

3 Noncoding RNAs

In the last two decades our view of genetic biologicalregulation has changed dramatically Transcriptomic analysesrevealed that only a surprisingly minor portion of 1-2of the human genome is protein-coding transcripts andthe remaining 98 is largely transcribed into noncoding

Journal of Diabetes Research 3

RNA which has been identified as an immensely complexand important regulatory machinery Among the differentnoncoding RNAs that exist microRNAs (miRNAs) and longnoncoding RNAs (lncRNAs) currently receive most interestbecause their regulating capabilities have been shown to playa significant role in human disease [33]

miRNAs are 20- to 22-nucleotides long RNA moleculesinitially discovered in the nematode worm C elegans asposttranscriptional negative regulators of gene expressionvia antisense RNA-RNA interaction [34 35] Up to 60of all mammalian genes are reported to be under miRNAinfluence [36]OnemiRNA is capable of targeting a collectionof messenger RNA (mRNA) molecules and therefore a setof targets for example in a stress cascade can be entirelymiRNA governed Disease states are typical examples ofmiRNA deregulation [37] Together these facts imply adisease modifying potential for miRNA mimics (premiRs)and inhibitors (antagomiRs) and indeed antagomiRs havealready proven their therapeutic potential in preclinical andclinical trials [38 39] miRNAs size allows them to leave thenucleus and exert their actions at a distance from where theywere transcribed Other thanmRNA thatmdashupon entering thecirculationmdashis prone to degradation miRNAs associate withdiverse types of carriers such asmicroparticles and exosomeswhich allow them to be detected peripherally [40] As a resultin addition to their therapeutic potential a lot of research hasshifted towards prospective utilization as disease biomarkersin plasma and other body fluids

The functional roles of long noncoding RNAs (lncRNAs)are less well known and span a wider range of regulatorymechanisms lncRNA release is triggered by a number of cel-lular responses Transcription and processing of lncRNAs aresimilar to those ofmiRNAs lncRNAs influence the regulationof gene expression by driving the formation of ribonucleic-protein complexes [41 42] To explain these functions severalmodels have been proposed [41] but the exact mechanismshave yet to be unravelled Recent findings show that lncRNAscan function as host transcripts for miRNAs [43]

In this review we aim at summarizing recent advances inthe role of miRNAs and lncRNAs in diabetes-related vasculardysfunction

4 Human Arrays Investigating miRNAAssociations with Diabetes

In 2010 a human plasma microarray was the first to iden-tify miRNAs associated with incident or manifest T2DMmiRNAs that were differentially regulated between T2DMcases and controls were miR-15a -20b -21 -24 -28-3p -29b-126 -150 -191 -197 -223 -320 and -486 In particularlya loss of endothelial miR-126 was characteristic for theT2DM signature as confirmed by the exposure of humanumbilical vein endothelial cells (HUVECs) to high glucosewhich led to a drop in the cellsrsquo release of miR-126 [44] Inaddition when looking specifically for miR-146a because ofits relation with heme oxygenase-1 (HO-1) expression Ronget al recently showed that this miRNA was also significantlyincreased in the circulation of patients with newly diagnosed

Skeletal muscle

miR-24miR-27abmiR-26abmiR-29amiR-29cmiR-30dmiR-107miR-126miR-181miR-210

Adipocytes

Plasma

miR-126amiR-146

Human miRNA profiling arrays

miR-378378lowast

Figure 1 The most well-established miRNAs identified by humanprofiling arrays in diabetic disease

T2DM as compared to age- and sex-matched controls [45]These different findings illustrate the importance of mecha-nistic studies in miRNA research to reveal the biochemicalbackground of differences in miRNA expression With thisknowledge lacking it is not likely that there will soon bea clinically available miRNA signature of vascular risk inT2DM

A genome-wide association study (GWAS) of humanskeletal muscle from subjects with and without T2DMrevealed different expression levels of miR-15a -15b -98-99a -100a -106b -133a -133b -143 -152 -185 and -190[46] Insulin administration in human skeletal muscle wasdemonstrated causal for the decreased expression of 39 miR-NAs [47] The targets of these miRNAs are associated withinsulin signalling and ubiquitination-mediated proteolysisT2DM was shown to be associated with impaired regulationof miRNAs by insulin Among these 39 miRNAs miR-24-26a -26b -27a -27b -29a -29c -30d -107 -126 -181 and-210 indeed do affect vascular and diabetic parameters in invitro and in vivo disease models as will be discussed in thefollowing sections Figure 1 depicts the most well-establishedmiRNAs identified by human profiling arrays in diabetes

5 miRNA Involvement inMetabolic Pathophysiology

A recent comprehensive description of miRNAs in humanmetabolism captured the majority of diabetes-associatedmiRNAs [48] and not surprisingly many of them over-lap with those influencing endothelial dysfunction-relatedatherosclerosis We will discuss the most-established as well


miR-33ab

miR-122

miR-370

Let-7

miR-802

miR-21miR-34amiR-146amiR-24miR-26miR-148miR-182

miR-30d

miR-210

Lipid metabolism

Glucose metabolism

Responsive Genes

PI3K-mTOR pathway

ABCA1

SREBP1 HNF1B

NF-120581B

Figure 2 miRNAs involved in glucose and lipid homeostasisABCA1 a subfamily ATP-binding cassette 1 SREBP1 sterol regu-latory element-binding protein 1 HNF1B hepatocyte nuclear factor1120573 NF-120581B nuclear factor kappa B PI3K-mTOR phosphoinositide3-kinase-mTOR

as the most recently discovered miRNAs A summary isprovided in Figure 2

51 Glucose Metabolism The Let-7 miRNA family is a groupof miRNAs acting as tumour suppressors inhibiting a set ofoncogenes and cell cycle regulators [49] Let-7 function isinhibited by the RNA-binding proteins Lin28a and Lin28ba regulatory capacity associated with developmental pro-gression in nematodes [50] and also Let-7 has been shownto be crucial for physiologic glucose homeostasis glucosetolerance and insulin signalling by inhibiting a varietyof targets in the phosphoinositide 3-kinase-mTOR (PI3K-mTOR) pathway in mouse models of obesity and T2DM[51 52]

One of the miRNAs identified by human skeletal muscleGWAS as being associated with type II diabetes [46] miR-106b has been further investigated regarding its role inmitofusin-2 (MFN2) mediated mitochondrial dysfunctionIn a series of in vitro gain-of-function and loss-of-functionstudies in mouse C2C12 myoblasts it was shown that byinhibitingMfn2 miR-106b negatively affected mitochondrialmorphology and function and increased ROS production[53]

miR-802 is upregulated in liver tissue of obese humanindividuals and has been shown to negatively regulate thegene encoding hepatocyte nuclear factor 1120573 (Hnf1b) in mice[54] HNF1B is causally linkedwithmaturity onset of diabetesin the young (MODY) type 5 and loss of function of thisgene activates pathways involved in gluconeogenesis 120573-oxidation of fatty acids oxidative phosphorylation and thetricarbonic acid cycle By affecting HNF1B function miR-802 counteracted glucose tolerance and insulin sensitivity asshown in vitro as well as in vivo [54]

miR-21 -24 -126 and -146a are significant modulators ofglucose metabolism in different in vitro and in vivo modelsof diabetes By targeting NF-120581B responsive genes miR-21-34a and -146a regulate cytokine-mediated 120573-cell dysfunc-tion during the initial phases of type I diabetes in nonobesediabetic mice [55] In mouse pancreas miR-24 -26 -182and -148 inhibit insulin biosynthesis via SRY-box 6 (Sox6)and e22 basic helix-loop-helix transcription factor (Bhlhe22)transcriptional repressors of insulin production [56]

In mouse insulinoma (MIN6) cells stimulated with glu-cose miR-30d enhances insulin gene transcription indicat-ing that miR-30d could be responsible for downregulatinginsulin transcription repressors [57] miR-34a -132 -184-199a-3p -203 -210 -338-3p and -383 deregulation has beenshown to induce 120573-cell apoptosis in MIN6 cells dispersedrat islet cells and dissociated human pancreatic island cells[58 59]

52 Lipid Metabolism miR-33ab and miR-122 are liver-specific miRNAs directly regulating lipid metabolism ThemiR-33a and -33b sequences are hosted by the sterol reg-ulatory element-binding protein (SREBP) genes 1 and 2They negatively regulate high-density lipoprotein (HDL)cholesterol synthesis and reverse cholesterol transport viain-hibition of the A subfamily ATP-binding cassette (ABCA1) inhuman liver cells In a preclinical trial miR-33ab antagonismsuccessfully lowered plasma triglycerides in non-humanprimates [38] In human liver miR-122 is the most abun-dantly expressed miRNA and it has important liver-specificfunctions that can be modulated in vivo with antagomiRs[39 60] miR-122 affects fatty acid synthesis and oxidation aswell as triglyceride synthesis via AMP-activated 1205721 catalyticsubunit protein kinase (Prkaa1) Srebp1 and diacylglycerol O-acyltransferase 2 (Dgat2) inmouse hepatic cells [61] miR-370increases miR-122 expression in HepG2 cells [62] miR-17-5p -99a -132 -134 -145 181a and -197 are associated withadipose tissue morphology and key metabolic parametersin human overweight and obese individuals [63] miR-122a miRNA that is essential for hepatitis C virus (HCV)stability and propagation in the liver has proven to be aneffective target in HCV infection miR-122 inhibitors arecurrently being used in clinical trials now entering Phase 3[39] miR-33 inhibition raises atheroprotective plasma highdensity lipoprotein (HDL) cholesterol while lowering verylow density lipoprotein (VLDL) cholesterol in non-humanprimates [38] This suggests that anti-miR-33 therapies nowalso entering human clinical trials are an effective approachin ameliorating plasma cholesterol profiles in patients

53 Vascular Oxidative Stress Regarding the importance ofcellular responses to redox imbalance in vascular diseasecertain miRNAs are crucially modulated Magenta et al[64] recently reviewed the role of different miRNAs in ECand VSMC oxidative pathophysiology In ECs responsiveto oxidative stress induced by hydrogen peroxide (H

2O2)

stimulation miR-200 family members were upregulatedIts role in redox signalling is likely exerted through ZincFinger E-Box Binding Homeobox (ZEB1) inhibition NO


stimulation also increased miR-200 family miRNAs andinhibited another ZEB splice variant ZEB2 Modulation ofthese proteins could indicate a role for miR-200 in ROS-induced apoptosis and senescence (ZEB1) and cardiovasculardevelopment (ZEB2) Silent mating type information regu-lation 2 homolog (SIRT1) is a longevity-associated enzymeimportant in cellular metabolism in general and specificallyin EC response to oxidative stress [64] miR-200a [65] miR-34a [66] miR-92a [67] miR-199a [68] andmiR-217 [69] havebeen shown to affect SIRT1 function in vitro

Vascular occlusion resulting in ischemia triggers ahypoxic response in affected cells Under hypoxic conditionsmitochondrial ROS production is increased generating anoxidative environment To date miR-210 is the most promi-nentmiRNA in hypoxia It is produced in response to hypoxiainducible factor (HIF) transcription factor activation andaffects a number of target genes involved in many differentcellular pathways [70] It is suggested that miR-210 deregu-lation might have a detrimental role in the cellular responseto hypoxia-induced oxidative stress but the mechanismsresponsible are not yet entirely clear [64]

Deletion in mice of miR-378378lowast two miRNAs derivedfrom the same hairpin precursor produces animals protectedagainst diet-induced obesity [71] It was shown that bothmiRNAs are involved in energy homeostasis via carnitine O-acetyltransferase (CRAT) targeted by miR-378 and mediatorsubunit complex 13 (MED13) targeted by its passenger strandmiR-378lowast Mice with miR-378378lowast knocked out displayedincreased energy expenditure and mitochondrial oxidativecapacity in insulin target tissues such as adipose and skeletalmuscle tissue These findings make miR-378378lowast interestingdrug targets

Very few studies have until now focused on the role ofmiRNAs in cardiovascular AGERAGE signalling Togliattoet al showed in HUVECs that miR-221222 downregula-tion is important in AGE- and high glucose mediated cellcycle arrest by downregulation of cyclin-dependent kinaseinhibitors 1B and 1C (CDKN1B or p27Kip1 and CDKN1C orp57Kip2 resp) [72] In colon cancer miR-155 is regulatedin a RAGE-responsive manner [73] In vitro exposure ofhuman monocytes to AGEs induced miR-214 productionand subsequent phosphatase and tensin homolog (PTEN)downregulation in these cells By luciferase reporter assayPTEN was validated as a miR-214 target [74]

6 Circulating Inflammatory Mediators andmiRNA Involvement

The proinflammatory function of adipokines is an importantgeneral mechanism in diabetes-related vascular dysfunction

61 Adipokines These adipose tissue-derived cytokines showa complex interplay with different miRNAs InvestigatingmiRNAs in adipose tissue from subjects of Indian descentMeerson et al found that miR-221 abundance was correlatedwith obesity miR expression was negatively regulated bythe adipokine leptin as well as by TNF-120572 [75] The authorsfound that miR-221 suppressed the adiponectin receptor 1

(ADIPOR1) and the transcription factor v-ets erythroblastosisvirus E26 oncogene homolog 1 (ETS1) in HEK293 cells Onthe mRNA level this function was not observed in adiposetissue but at the protein level both ADIPOR1 and ETS1 werereducedThis reduction could lead to changes in insulin sen-sitivity and promote obesity-associated inflammation [75]

7 Vascular Cell-Specific miRNAStress Responses

Different cell types may respond to and themselves releasedifferent miRNAs Certain miRNAs for example that arescarce in nucleated cells might have profound actions inplatelets [76] Different cell types might present differentmiRNA targets which demands an even more carefulapproach towards miRNA expression interpretation We willpresent the most influential cell types that constitute thevasculature as well as the miRNAs proven to have a role intheir pathophysiology (Figure 3)

71 Endothelial Cells miR-17asymp92 -21 -23asymp27asymp24 -126-143 -145 and -146a have the most extensive record inendothelial cell physiology and pathology [77ndash82] Inter-action of these miRNAs with ECs affects the cellsrsquo angio-genesis sprouting and vascular remodelling capabilities viaSIRT1 integrin subunit 1205725 (ITGA5) and Janus kinase 1(JAK1) (miR-17 asymp 92) [67 77 79 80] Sprouty protein 2(SPROUTY2) and Semaphorin-6A (SEMA6A) (miR-23 and-27) [80] and Sprouty-related EVH1 domain-containingprotein 1 (SPRED1) phosphatidylinositide 3-kinase regu-latory subunit 2 (PI3KR2p85120573) and vascular cell adhe-sion molecule 1 (VCAM1) (miR-126) [44] miR-21 inhibitsEC inflammation through peroxisome proliferator-activatedreceptor 120572 (PPAR120572) [81] miR-24 mediates EC apoptosis viaGATA-binding protein 2 (GATA2) and p21 protein-activatedkinase 4 (PAK4) [82] Endothelial cell-derived miR-143145can repress ETS domain-containing protein Elk1 (ELK1)Kruppel-like factor 4 (KLF4) and calciumcalmodulin-dependent protein kinase II delta (CAMK2d) in VSMCs [77]Exactly how this kind of communication is established hasdeveloped into its own independent small area of research

EC injury triggers a release of endothelial cell derivedmicroparticles (EMPs) [83] EMPs have a range of functionsin vascular homeostasis such as coagulation inflammationendothelial function and angiogenesis [83] and are rich inmiRNAs particularly miR-126 [84] Recent work by Jansenet al shows that miR-126 is reduced in circulating EMPsof patients with T2DM versus non-diabetic controls andcontributes to EMP-mediated regeneration of target cells invitro and in vivo [84] What remains to be investigated isif therapeutic reconstitution of miR-126 containing EMPsin patients with T2DM can reverse the vascular pathologyobserved in this disease Other groups have also showedthat loss of endothelial miR-126 is part of the T2DM plasmamiRNA signature as mentioned previously [44]

Another component in vascular repair are endothelialprogenitor cells (EPCs) Via paracrine routes these circu-lating CD34+CD133+ VEGFR2+ immature hematopoietic


Early stage (ECs)miR-10a

miR-21

miR-126miR-143miR-145miR-146a

Advanced stage (VSMCs)miR-21miR-143miR-145miR-133miR-221miR-222

Oxidative stressmiR-34amiR-92amiR-199amiR-200amiR-217miR-210

SIRT1 ZEB1

HIF

KLF45 ELK1CAMK2D PDGFACE

Atherosclerosis

SIRT1ITGA5JAK1

KLF45ELK1

SPROUTY2SEMA6A SPRED1GATA2 PAK4

PI3KR2VCAM1

PTEN PDCD4

KIT CDKN1BPPAR120572

miR-17asymp29

miR-23asymp27asymp24

Figure 3 miRNAs involved in vascular pathophysiology ECs endothelial cells VSMCs vascular smooth muscle cells SIRT1 silent matingtype information regulation 2 homolog ITGA5 integrin subunit 1205725 JAK1 Janus kinase 1 KLF Kruppel-like factor ELK1 ETS domain-containing protein Elk1 PPAR120572 peroxisome proliferator-activated receptor 120572 SPROUTY2 sprouty protein 2 SEMA6A semaphorin-6ASPRED1 sprouty-related EVH1 domain-containing protein 1 GATA2 GATA-binding protein 2 PAK4 p21 protein-activated kinase 4PI3KR2 phosphatidylinositide 3-kinase regulatory subunit 2 VCAM1 vascular cell adhesion molecule 1 HIF hypoxia-inducible factor 1ZEB1 Zinc Finger E-Box Binding Homeobox 1 PTEN phosphatase and tensin homolog PDCD4 programmed cell death 4 CAMK2Dcalciumcalmodulin-dependent protein kinase II delta PDGF platelet-derived growth factor ACE angiotensin-converting enzyme KITv-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog CDKN1B cyclin-dependent kinase inhibitor 1B

cells could orchestrate the reaction of the endothelium toinjury [85 86] and EPC function might explain in part thebeneficial effects of statins in cardiovascular disease [87] Incirculating EPCs from T2DM patients miR-21 -27a -27b-126 and -130a are downregulated [88] miR-130a inhibitionin EPCs reduces the proliferation migration and colonyformation of these cells in vitro Runt-related transcriptionfactor 3 (RUNX3) is a direct target of miR-130a and reductionof Runx3 protein level rescues EPC proliferation colonyformation and migration In these particular cells these areeffects likely to be evoked by miR-130a [88]

Lesswell-knownmodulators of endothelial cell behaviourare miR-10a promoting EC inflammation via inhibition ofhomeoboxA1 (HOXA1) mitogen-activated protein 3 kinase 7(MAP3K7) and 120573-transducin repeat containing E3 ubiquitinprotein ligase (BTRC) andmiR-210 a proangiogenic miRNAinhibiting ephrin-A3 (EFNA3) [70 89] On the oppositeside miRNAs miR-221 and -222 acting through v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT)-ligand (KITLG) and signal transducer and activator of tran-scription 5A (STAT5A) inhibit angiogenesis [90 91] In vitroexperiments in HUVECs recently identified miR-503 as aregulator of EC cycle progression via the phosphatase celldivision cycle 25A (CDC25A) and cyclin E1 (CCNE1) [92] Inearly phases of endothelial dysfunctionmiR-146a is primarilyinvolved in the regulation of inflammation targeting toll-likereceptor pathway signals [93] Interestingly miR-146a is alsoinvolved initial phases of type 1 diabetes in nonobese diabeticmice acting on NF-120581B responsive genes in 120573 cells [55]

72 Vascular Smooth Muscle Cells Progressing atheroscle-rotic plaques are characterised by migration of VSMCstowards the lesion [20] The fibrous cap they create promotes

plaque stability VSMC interplay with the inflammatoryenvironment leads to LDL cholesterol accumulation andROSgeneration as well as VSMC apoptosis which both aggra-vate the inflammatory reaction and lead to plaque ruptureVSMC survival proliferationmigration and remodelling aregoverned by miR-143 and -145 via their targets KLF4 and5 ELK1 CAMK2D platelet-derived growth factor (PDGF)and angiotensin I converting enzyme (ACE) miR-21 inhibitsPTEN and programmed cell death 4 (PDCD4) and inhibitsthe apoptosis regulator B-cell CLLlymphoma 2 (BCL2)thereby promoting VSMC proliferation a mechanism vali-dated in murine models of vascular disease [94 95] miR-221 and miR-222 contribute to VSMC dedifferentiation andproliferation by targeting the tyrosine-protein kinase KITas well as CDKN1B and CDKN1C [96 97] In silico analysisshowed that miR-133 could have the potential to block VSMCphenotypic switching through the transcription factor Sp-1[98]

8 lncRNAs and Diabetes

Genome-wide association studies have identified antisensenoncoding RNA in the INK4 locus (ANRIL) a lncRNA dis-covered in 2007 and associated with neural system tumours[99] as a genetic locus of susceptibility for coronary diseaseintracranial aneurysm and T2DM [100 101] Its host locicyclin-dependent kinase inhibitors 2A and 2B (CDKN2AB)encode for tumour suppressor genes This implicates thatANRILmight affect cellular senescence and replicative func-tion and thus influences the molecular mechanisms involvedin these diseases [102] In human peripheral blood mononu-clear cells (PBMCs) and the monocytic cell line MonoMacANRIL guided by its core sequence called the Alu motif


was shown to bind to different proteins contained in thechromatin modifying complexes polycomb repressive com-plex (PRC) genes The inhibitory and activation functionsaffected atherosclerosis-related cell functions such as adhe-sion proliferation and apoptosis and perhaps surprisinglynot CDKN2AB [103]

There are gt1100 human 120573-cell lncRNAs They are anintegral component of the 120573-cell differentiation and matura-tion programHI-LNC25 for example regulates GLI-similarzinc fincer protein 3 (GLIS3) mRNA an islet transcriptionfactor Two other lncRNAs KCNQ1OT1 and HI-LNC45 areup- respectively downregulated in human T2DM pancreaticislets Two lncRNAs map within the established T2DM sus-ceptibility loci prospero homeobox 1 (PROX1) and Wolframsyndrome 1 (WFS1) [104]

lncRNAs in the rat genome play a role in the response ofrat VSMCs to angiotensin II Associated lncRNAs are Lnc-Ang26 Lnc-Ang383 Lnc-Ang58 Lnc-Ang219 Lnc-Ang202Lnc-Ang249 and Lnc-Ang362 Lnc-Ang362 is a host genefor miR-221 and miR-222 which means that these miRNAsare cotranscribed with and excised from the lncRNA miR-221 and miR-222 are miRNAs known to regulate VSMCproliferation [43]

9 Summary and Conclusion

The rapid rise of T2DM and its accompanying cardiovas-cular complications demand new treatment strategies Thewidespread possibilities of modulating gene expression usingvarious subtypes of ncRNAs present great opportunities tofight the burden of the associated diseases However beforeinitiating treatment of patients by modulating ncRNA thein-depth mechanisms of their action and regulation need tobe completely understood ncRNA interaction translation tohumans drug delivery off-target side effects andmany otherchallenges still require thorough basic and translational stud-ies However utilizingmodern technology (eg microarraysRNAseq) has enabled us to discover ncRNA regulation ofdisease which makes the identification of novel treatmentoptions in cardiovascular disease more rapid compared totraditional avenues of drug design This may allow us toget closer to a solution for the rapidly expanding worldwidehealth problem of T2DM and its related pathologies

Acknowledgment

The authors would like to acknowledge and thank thiersupporting funding agencies theKarolinska InstituteCardio-vascular Program Career Development Grant the SwedishHeart-Lung-Foundation (20120615) and the Ake WibergFoundation (all to Lars Maegdefessel)

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[2] W T Cade ldquoDiabetes-relatedmicrovascular andmacrovasculardiseases in the physical therapy settingrdquo Physical Therapy vol88 no 11 pp 1322ndash1335 2008

[3] S B Bender A P McGraw I Z Jaffe and J R SowersldquoMineralocorticoid receptor-mediated vascular insulin resis-tance an early contributor to diabetes-related vascular diseaserdquoDiabetes vol 62 no 2 pp 313ndash319 2013

[4] GW Gibbons and PM Shaw ldquoDiabetic vascular disease char-acteristics of vascular disease unique to the diabetic patientrdquoSeminars in Vascular Surgery vol 25 no 2 pp 89ndash92 2012

[5] J-A Kim M Montagnani K K Kwang and M J QuonldquoReciprocal relationships between insulin resistance andendothelial dysfunction molecular and pathophysiologicalmechanismsrdquo Circulation vol 113 no 15 pp 1888ndash1904 2006

[6] A P Dantas Z B Fortes and M H de Carvalho ldquoVasculardisease in diabetic women why do they miss the femaleprotectionrdquo Experimental Diabetes Research vol 2012 ArticleID 570598 10 pages 2012

[7] Q Li K Park C Li et al ldquoInduction of vascular insulinresistance and endothelin-1 expression and acceleration ofatherosclerosis by the overexpression of protein kinase C-120573isoform in the endotheliumrdquo Circulation Research vol 113 no4 pp 418ndash427 2013

[8] U Forstermann and W C Sessa ldquoNitric oxide synthases regu-lation and functionrdquo European Heart Journal vol 33 no 7 pp829ndash837 2012

[9] F Paneni J A Beckman M A Creager and F CosentinoldquoDiabetes and vascular disease pathophysiology clinical conse-quences andmedical therapymdashpart Irdquo European Heart Journalvol 34 no 31 pp 2436ndash2443 2013

[10] H Brunner J R Cockcroft J Deanfield et al ldquoEndothelialfunction and dysfunctionmdashpart II association with cardiovas-cular risk factors and diseases A statement by the WorkingGroup on Endothelins and Endothelial Factors of the EuropeanSociety of Hypertensionrdquo Journal of Hypertension vol 23 no 2pp 233ndash246 2005

[11] A D Mooradian ldquoDyslipidemia in type 2 diabetes mellitusrdquoNature Clinical Practice EndocrinologyampMetabolism vol 5 no3 pp 150ndash159 2009

[12] M PHermans S A Ahn andM F Rousseau ldquoThe atherogenicdyslipidemia ratio [log(TG)HDL-C] is associated with residualvascular risk beta-cell function loss and microangiopathy intype 2 diabetes femalesrdquo Lipids in Health and Disease vol 11article 132 2012

[13] C S Stancu L Toma and A V Sima ldquoDual role of lipoproteinsin endothelial cell dysfunction in atherosclerosisrdquo Cell andTissue Research vol 349 no 2 pp 433ndash446 2012

[14] S J Hamilton G T Chew T M E Davis and G F WattsldquoFenofibrate improves endothelial function in the brachialartery and forearm resistance arterioles of statin-treated type 2diabetic patientsrdquo Clinical Science vol 118 no 10 pp 607ndash6152010

[15] A Ceriello R Assaloni R da Ros et al ldquoEffect of atorvastatinand irbesartan alone and in combination on postprandialendothelial dysfunction oxidative stress and inflammation intype 2 diabetic patientsrdquo Circulation vol 111 no 19 pp 2518ndash2524 2005

[16] D C Chan A T Y Wong S Yamashita and G F WattsldquoApolipoprotein B-48 as a determinant of endothelial functionin obese subjects with type 2 diabetes mellitus effect offenofibrate treatmentrdquo Atherosclerosis vol 221 no 2 pp 484ndash489 2012

[17] R A Cohen and X Tong ldquoVascular oxidative stress thecommon link in hypertensive and diabetic vascular diseaserdquo


Journal of Cardiovascular Pharmacology vol 55 no 4 pp 308ndash316 2010

[18] S W Schaffer C J Jong and M Mozaffari ldquoRole of oxida-tive stress in diabetes-mediated vascular dysfunction unifyinghypothesis of diabetes revisitedrdquoVascular Pharmacology vol 57no 5-6 pp 139ndash149 2012

[19] A Goldin J A Beckman A M Schmidt and M A CreagerldquoAdvanced glycation end products sparking the developmentof diabetic vascular injuryrdquo Circulation vol 114 no 6 pp 597ndash605 2006

[20] C K Glass and J LWitztum ldquoAtherosclerosis the road aheadrdquoCell vol 104 no 4 pp 503ndash516 2001

[21] Y Park J Wu H Zhang Y Wang and C Zhang ldquoVasculardysfunction in type 2 diabetes emerging targets for therapyrdquoExpert Review of Cardiovascular Therapy vol 7 no 3 pp 209ndash213 2009

[22] B Chandrasekar W H Boylston K Venkatachalam N J GWebster S D Prabhu and A J Valente ldquoAdiponectin blocksinterleukin-18-mediated endothelial cell death via APPL1-dependent AMP-activated protein kinase (AMPK) activationand IKKNF-120581BPTEN suppressionrdquo The Journal of BiologicalChemistry vol 283 no 36 pp 24889ndash24898 2008

[23] N Ouchi J L Parker J J Lugus and K Walsh ldquoAdipokines ininflammation andmetabolic diseaserdquoNature Reviews Immunol-ogy vol 11 no 2 pp 85ndash97 2011

[24] Y Arita S Kihara N Ouchi et al ldquoParadoxical decrease of anadipose-specific protein adiponectin in obesityrdquo Biochemicaland Biophysical Research Communications vol 257 no 1 pp79ndash83 1999

[25] W Koenig N Khuseyinova J Baumert C Meisinger and HLowel ldquoSerum concentrations of adiponectin and risk of type2 diabetes mellitus and coronary heart disease in apparentlyhealthy middle-aged men results from the 18-year follow-up ofa large cohort from southernGermanyrdquo Journal of the AmericanCollege of Cardiology vol 48 no 7 pp 1369ndash1377 2006

[26] B B Duncan M I Schmidt J S Pankow et al ldquoAdiponectinand the development of type 2 diabetes the atherosclerosis riskin communities studyrdquo Diabetes vol 53 no 9 pp 2473ndash24782004

[27] J van de Voorde B Pauwels C Boydens and K DecaluweldquoAdipocytokines in relation to cardiovascular diseaserdquo Meta-bolism vol 62 no 11 pp 1513ndash1521 2013

[28] L Manuel-Apolinar R Lopez-Romero A Zarate et al ldquoLeptinmediated ObRb receptor increases expression of adhesionintercellular molecules and cyclooxygenase 2 on murine aortatissue inducing endothelial dysfunctionrdquo International Journalof Clinical and Experimental Medicine vol 6 no 3 pp 192ndash1962013

[29] A Elkalioubie C Zawadzki C Roma-Lavisse et al ldquoFreeleptin carotid plaque phenotype and relevance to related symp-tomatology insights from the OPAL-Lille carotid endarterec-tomy studyrdquo International Journal of Cardiology vol 168 no 5pp 4879ndash4881 2013

[30] M S Jamaluddin S M Weakley Q Yao and C ChenldquoResistin functional roles and therapeutic considerations forcardiovascular diseaserdquo British Journal of Pharmacology vol165 no 3 pp 622ndash632 2012

[31] J M Northcott A Yeganeh C G Taylor P Zahradka and J TWigle ldquoAdipokines and the cardiovascular systemmechanismsmediating health and diseaserdquo Canadian Journal of Physiologyand Pharmacology vol 90 no 8 pp 1029ndash1059 2012

[32] I Falcao-Pires P Castro-Chaves D Miranda-Silva A PLourenco and A F Leite-Moreira ldquoPhysiological pathologicaland potential therapeutic roles of adipokinesrdquo Drug DiscoveryToday vol 17 no 15-16 pp 880ndash889 2012

[33] J S Mattick and I V Makunin ldquoNon-coding RNArdquo HumanMolecular Genetics vol 15 supplement 1 pp R17ndashR29 2006

[34] B Wightman T R Burglin J Gatto P Arasu and G RuvkunldquoNegative regulatory sequences in the lin-14 31015840-untranslatedregion are necessary to generate a temporal switch duringCaenorhabditis elegans developmentrdquo Genes and Developmentvol 5 no 10 pp 1813ndash1824 1991

[35] R C Lee R L Feinbaum and V Ambros ldquoThe C elegansheterochronic gene lin-4 encodes small RNAs with antisensecomplementarity to lin-14rdquoCell vol 75 no 5 pp 843ndash854 1993

[36] R C Friedman K K-H Farh C B Burge and D P BartelldquoMost mammalian mRNAs are conserved targets of microR-NAsrdquo Genome Research vol 19 no 1 pp 92ndash105 2009

[37] D P Bartel ldquoMicroRNAs target recognition and regulatoryfunctionsrdquo Cell vol 136 no 2 pp 215ndash233 2009

[38] K J Rayner C C Esau F N Hussain et al ldquoInhibition of miR-33ab in non-human primates raises plasma HDL and lowersVLDL triglyceridesrdquo Nature vol 478 no 7369 pp 404ndash4072011

[39] H L Janssen H W Reesink E J Lawitz et al ldquoTreatmentof HCV infection by targeting microRNArdquo The New EnglandJournal of Medicine vol 368 no 18 pp 1685ndash1694 2013

[40] S F Mause and C Weber ldquoMicroparticles protagonists ofa novel communication network for intercellular informationexchangerdquo Circulation Research vol 107 no 9 pp 1047ndash10572010

[41] J L Rinn and H Y Chang ldquoGenome regulation by long non-coding RNAsrdquo The Annual Review of Biochemistry vol 81 pp145ndash166 2012

[42] S Djebali C A Davis AMerkel et al ldquoLandscape of transcrip-tion in human cellsrdquoNature vol 489 no 7414 pp 101ndash108 2012

[43] A Leung C Trac W Jin et al ldquoNovel long noncoding RNAsare regulated by angiotensin II in vascular smoothmuscle cellsrdquoCirculation Research vol 113 no 3 pp 266ndash278 2013

[44] A Zampetaki S Kiechl I Drozdov et al ldquoPlasma MicroRNAprofiling reveals loss of endothelial MiR-126 and other MicroR-NAs in type 2 diabetesrdquo Circulation Research vol 107 no 6 pp810ndash817 2010

[45] Y Rong W Bao Z Shan et al ldquoIncreased microRNA-146alevels in plasmaof patientswith newly diagnosed type 2 diabetesmellitusrdquo PLoS ONE vol 8 no 9 Article ID e73272 2013

[46] I J Gallagher C Scheele P Keller et al ldquoIntegration ofmicroRNA changes in vivo identifies novel molecular featuresof muscle insulin resistance in type 2 diabetesrdquo GenomeMedicine vol 2 no 2 article 9 2010

[47] A Granjon M-P Gustin J Rieusset et al ldquoThe microRNAsignature in response to insulin reveals its implication in thetranscriptional action of insulin in human skeletal muscleand the role of a sterol regulatory element-binding protein-1cmyocyte enhancer factor 2C pathwayrdquo Diabetes vol 58 no11 pp 2555ndash2564 2009

[48] O Dumortier C Hinault and E van Obberghen ldquoMicroR-NAs and metabolism crosstalk in energy homeostasisrdquo CellMetabolism vol 18 no 3 pp 312ndash324 2013

[49] C Mayr M T Hemann and D P Bartel ldquoDisrupting thepairing between let-7 andHmga2 enhances oncogenic transfor-mationrdquo Science vol 315 no 5818 pp 1576ndash1579 2007


[50] V Ambros and H R Horvitz ldquoHeterochronic mutants of thenematode Caenorhabditis elegansrdquo Science vol 226 no 4673pp 409ndash416 1984

[51] R J A Frost and E N Olson ldquoControl of glucose homeostasisand insulin sensitivity by the Let-7 family of microRNAsrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 108 no 52 pp 21075ndash21080 2011

[52] H Zhu S-CNg A V Segr et al ldquoTheLin28let-7 axis regulatesglucose metabolismrdquo Cell vol 147 no 1 pp 81ndash94 2011

[53] Y Zhang L Yang Y F Gao et al ldquoMicroRNA-106b inducesmitochondrial dysfunction and insulin resistance in C2C12myotubes by targeting mitofusin-2rdquo Molecular and CellularEndocrinology vol 381 no 1-2 pp 230ndash240 2013

[54] J W Kornfeld C Baitzel A C Konner et al ldquoObesity-inducedoverexpression of miR-802 impairs glucose metabolismthrough silencing of Hnf1brdquo Nature vol 494 no 7435 pp111ndash115 2013

[55] E Roggli A Britan S Gattesco et al ldquoInvolvement ofmicroRNAs in the cytotoxic effects exerted by proinflammatorycytokines on pancreatic120573-cellsrdquoDiabetes vol 59 no 4 pp 978ndash986 2010

[56] T Melkman-Zehavi R Oren S Kredo-Russo et al ldquomiRNAscontrol insulin content in pancreatic 120573 2-cells via downregula-tion of transcriptional repressorsrdquo The EMBO Journal vol 30no 5 pp 835ndash845 2011

[57] X Tang L Muniappan G Tang and S Ozcan ldquoIdentificationof glucose-regulated miRNAs from pancreatic 120573 cells reveals arole for miR-30d in insulin transcriptionrdquo RNA vol 15 no 2pp 287ndash293 2009

[58] E ZhaoM P Keller M E Rabaglia et al ldquoObesity and geneticsregulate microRNAs in islets liver and adipose of diabeticmicerdquoMammalian Genome vol 20 no 8 pp 476ndash485 2009

[59] V Nesca C Guay C Jacovetti et al ldquoIdentification of particulargroups of microRNAs that positively or negatively impact onbeta cell function in obese models of type 2 diabetesrdquo Diabeto-logia vol 56 no 10 pp 2203ndash2212 2013

[60] J Krutzfeldt N Rajewsky R Braich et al ldquoSilencing ofmicroRNAs in vivowith lsquoantagomirsrsquordquoNature vol 438 no 7068pp 685ndash689 2005

[61] C Esau S Davis S F Murray et al ldquomiR-122 regulation oflipid metabolism revealed by in vivo antisense targetingrdquo CellMetabolism vol 3 no 2 pp 87ndash98 2006

[62] D Iliopoulos K Drosatos Y Hiyama I J Goldberg and V IZannis ldquoMicroRNA-370 controls the expression ofMicroRNA-122 and Cpt1120572 and affects lipid metabolismrdquo Journal of LipidResearch vol 51 no 6 pp 1513ndash1523 2010

[63] N Kloting S Berthold P Kovacs et al ldquoMicroRNA expressionin human omental and subcutaneous adipose tissuerdquo PLoSONE vol 4 no 3 Article ID e4699 2009

[64] A Magenta S Greco C Gaetano and F Martelli ldquoOxidativestress and MicroRNAs in vascular diseasesrdquo International Jour-nal of Molecular Sciences vol 14 no 9 pp 17319ndash17346 2013

[65] G Eades Y Yao M Yang Y Zhang S Chumsri and Q ZhouldquomiR-200a regulates SIRT1 expression and Epithelial to Mes-enchymal Transition (EMT)-like transformation in mammaryepithelial cellsrdquoThe Journal of Biological Chemistry vol 286 no29 pp 25992ndash26002 2011

[66] M Yamakuchi M Ferlito and C J Lowenstein ldquomiR-34arepression of SIRT1 regulates apoptosisrdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 105 no 36 pp 13421ndash13426 2008

[67] A Bonauer G Carmona M Iwasaki et al ldquoMicroRNA-92acontrols angiogenesis and functional recovery of ischemictissues in micerdquo Science vol 324 no 5935 pp 1710ndash1713 2009

[68] S Rane M He D Sayed et al ldquoDownregulation of MiR-199a derepresses hypoxia-inducible factor-1120572 and sirtuin 1 andrecapitulates hypoxia preconditioning in cardiac myocytesrdquoCirculation Research vol 104 no 7 pp 879ndash886 2009

[69] R Menghini V Casagrande M Cardellini et al ldquoMicroRNA217modulates endothelial cell senescence via silent informationregulator 1rdquo Circulation vol 120 no 15 pp 1524ndash1532 2009

[70] C Devlin S Greco F Martelli and M Ivan ldquoMiR-210 morethan a silent player in hypoxiardquo IUBMB Life vol 63 no 2 pp94ndash100 2011

[71] M Carrer N Liu C E Grueter et al ldquoControl ofmitochondrialmetabolism and systemic energy homeostasis by microRNAs378 and 378lowastrdquo Proceedings of the National Academy of Sciencesof the United States of America vol 109 no 38 pp 15330ndash153352012

[72] G Togliatto A Trombetta P Dentelli A Rosso and MF Brizzi ldquoMIR221MIR222-driven post-transcriptional regula-tion of P27KIP1 and P57KIP2 is crucial for high-glucose- andAGE-mediated vascular cell damagerdquo Diabetologia vol 54 no7 pp 1930ndash1940 2011

[73] B C Onyeagucha M E Mercado-Pimentel J Hutchison EK Flemington and M A Nelson ldquoS100PRAGE signalingregulatesmicroRNA-155 expression via AP-1 activation in coloncancerrdquo Experimental Cell Research vol 319 no 13 pp 2081ndash2090 2013

[74] L-M Li D-X Hou Y-L Guo et al ldquoRole of microRNA-214-targeting phosphatase and tensin homolog in advancedglycation end product-induced apoptosis delay in monocytesrdquoJournal of Immunology vol 186 no 4 pp 2552ndash2560 2011

[75] AMeersonM Traurig VOssowski JM FlemingMMullinsand L J Baier ldquoHuman adipose microRNA-221 is upregulatedin obesity and affects fat metabolism downstream of leptin andTNF-120572rdquo Diabetologia vol 56 no 9 pp 1971ndash1979 2013

[76] D A Stakos A Gatsiou K Stamatelopoulos A D Tselepisand K Stellos ldquoPlatelet microRNAs from platelet biology topossible disease biomarkers and therapeutic targetsrdquo Plateletsvol 24 no 8 pp 579ndash589 2013

[77] E Hergenreider S Heydt K Treguer et al ldquoAtheroprotectivecommunication between endothelial cells and smooth musclecells through miRNAsrdquo Nature Cell Biology vol 14 no 3 pp249ndash256 2012

[78] C Fernandez-Hernando C M Ramırez L Goedeke andY Suarez ldquoMicroRNAs in metabolic diseaserdquo ArteriosclerosisThrombosis and Vascular Biology vol 33 no 2 pp 178ndash1852013

[79] R A Boon E Hergenreider and S Dimmeler ldquoAtheroprotec-tive mechanisms of shear stress-regulated microRNAsrdquoThrom-bosis and Haemostasis vol 108 no 4 pp 616ndash620 2012

[80] A Zampetaki and M Mayr ldquoMicroRNAs in vascular andmetabolic diseaserdquo Circulation Research vol 110 no 3 pp 508ndash522 2012

[81] J Zhou K-C Wang W Wu et al ldquoMicroRNA-21 targetsperoxisome proliferators-activated receptor-120572 in an autoregula-tory loop to modulate flow-induced endothelial inflammationrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 108 no 25 pp 10355ndash10360 2011

[82] J Fiedler V Jazbutyte B C Kirchmaier et al ldquoMicroRNA-24regulates vascularity after myocardial infarctionrdquo Circulationvol 124 no 6 pp 720ndash730 2011


[83] F Dignat-George and C M Boulanger ldquoThe many faces ofendothelial microparticlesrdquo Arteriosclerosis Thrombosis andVascular Biology vol 31 no 1 pp 27ndash33 2011

[84] F Jansen X Yang M Hoelscher et al ldquoEndothelial micro-particle-mediated transfer ofMicroRNA-126 promotes vascularendothelial cell repair via SPRED1 and is abrogated in glucose-damaged endothelial microparticlesrdquo Circulation vol 128 no18 pp 2026ndash2038 2013

[85] C Urbich and S Dimmeler ldquoEndothelial progenitor cells char-acterization and role in vascular biologyrdquo Circulation Researchvol 95 no 4 pp 343ndash353 2004

[86] SWassmann NWerner T Czech and G Nickenig ldquoImprove-ment of endothelial function by systemic transfusion of vascularprogenitor cellsrdquo Circulation Research vol 99 no 8 pp e74ndashe83 2006

[87] M Steinmetz C Brouwers G Nickenig and S WassmannldquoSynergistic effects of telmisartan and simvastatin on endothe-lial progenitor cellsrdquo Journal of Cellular andMolecularMedicinevol 14 no 6B pp 1645ndash1656 2010

[88] SMeng J Cao X Zhang et al ldquoDownregulation ofmicroRNA-130a contributes to endothelial progenitor cell dysfunction indiabetic patients via its target Runx3rdquo PLoS ONE vol 8 no 7Article ID e68611 2013

[89] S Hu M Huang Z Li et al ldquoMicroRNA-210 as a novel therapyfor treatment of ischemic heart diseaserdquo Circulation vol 122no 11 supplement pp S124ndashS131 2010

[90] L Poliseno A Tuccoli L Mariani et al ldquoMicroRNAsmodulatethe angiogenic properties of HUVECsrdquo Blood vol 108 no 9pp 3068ndash3071 2006

[91] P Dentelli A Rosso F Orso C Olgasi D Taverna andM F Brizzi ldquomicroRNA-222 controls neovascularization byregulating signal transducer and activator of transcription 5Aexpressionrdquo Arteriosclerosis Thrombosis and Vascular Biologyvol 30 no 8 pp 1562ndash1568 2010

[92] A Caporali M Meloni C Vollenkle et al ldquoDeregulationof microRNA-503 contributes to diabetes mellitus-inducedimpairment of endothelial function and reparative angiogenesisafter Limb Ischemiardquo Circulation vol 123 no 3 pp 282ndash2912011

[93] M Hulsmans D de Keyzer and P Holvoet ldquoMicroRNAsregulating oxidative stress and inflammation in relation toobesity and atherosclerosisrdquo The FASEB Journal vol 25 no 8pp 2515ndash2527 2011

[94] B N Davis A C Hilyard G Lagna and A Hata ldquoSMADproteins control DROSHA-mediated microRNA maturationrdquoNature vol 454 no 7200 pp 56ndash61 2008

[95] L Maegdefessel J Azuma R Toh et al ldquoMicroRNA-21 blocksabdominal aortic aneurysm development and nicotine-aug-mented expansionrdquo Science Translational Medicine vol 4 no122 Article ID 122ra22 2012

[96] X Liu Y Cheng S Zhang Y Lin J Yang and C Zhang ldquoAnecessary role ofmiR-221 andmiR-222 in vascular smoothmus-cle cell proliferation and neointimal hyperplasiardquo CirculationResearch vol 104 no 4 pp 476ndash486 2009

[97] B N Davis A C Hilyard P H Nguyen G Lagna and A HataldquoInduction of MicroRNA-221 by platelet-derived growth factorsignaling is critical for modulation of vascular smooth musclephenotyperdquoThe Journal of Biological Chemistry vol 284 no 6pp 3728ndash3738 2009

[98] D Torella C Iaconetti D Catalucci et al ldquoMicroRNA-133controls vascular smoothmuscle cell phenotypic switch in vitro

and vascular remodeling in vivordquo Circulation Research vol 109no 8 pp 880ndash893 2011

[99] E Pasmant I Laurendeau D Heron M Vidaud D Vidaudand I Bieche ldquoCharacterization of a germ-line deletion includ-ing the entire INK4ARF locus in a melanoma-neural systemtumor family identification of ANRIL an antisense noncodingRNA whose expression coclusters with ARFrdquo Cancer Researchvol 67 no 8 pp 3963ndash3969 2007

[100] E Pasmant A Sabbagh M Vidaud and I Bieche ldquoANRILa long noncoding RNA is an unexpected major hotspot inGWASrdquoThe FASEB Journal vol 25 no 2 pp 444ndash448 2011

[101] C Wahlestedt ldquoTargeting long non-coding RNA to thera-peutically upregulate gene expressionrdquo Nature Reviews DrugDiscovery vol 12 no 6 pp 433ndash446 2013

[102] N E Sharpless and R A DePinho ldquoHow stem cells age andwhythis makes us grow oldrdquoNature Reviews Molecular Cell Biologyvol 8 no 9 pp 703ndash713 2007

[103] LMHoldt S Hoffmann K Sass et al ldquoAlu elements in ANRILnon-coding RNA at chromosome 9p21 modulate atherogeniccell functions through trans-regulation of gene networksrdquo PLoSGenetics vol 9 no 7 Article ID e1003588 2013

[104] I Moran I Akerman M van de Bunt et al ldquoHuman 120573cell transcriptome analysis uncovers lncRNAs that are tissue-specific dynamically regulated and abnormally expressed intype 2 diabetesrdquo Cell Metabolism vol 16 no 4 pp 435ndash4482012

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


chronic inflammation hemodynamic deregulation impairedfibrinolytic ability and enhanced platelet aggregation [5]

21 Insulin Signalling and Hyperglycaemia Abnormalities invascular endothelial (EC) and vascular smooth muscle cell(VSMC) function as well as a propensity to thrombosisare important contributors to vascular complications [5]Hyperglycaemia and insulin resistance have been identifiedas key players in the development of diabetic atheroscle-rosis with metabolic insulin signalling being an importantcontributor to normal vascular function and homeostasis[5] Decreased insulin sensitivity in cardiovascular tissuesis an underlying abnormality in obesity hypertension andT2DM [3] In physiological conditions insulin promotesendothelium-dependent relaxation by a mechanism thatinvolves an increase of nitric oxide (NO) production viaactivation of phosphatidylinositol-3 kinase (PI3K) and Aktkinase pathways [6 7] NO has also been shown to preventendothelial apoptosis as well as neutrophil and plateletadhesion to the vascular wall [8] The initial trigger bywhich high glucose concentrations alter vascular functionis the imbalance between nitric oxide (NO) bioavailabilityand accumulation of reactive oxygen species (ROS) [9]Decrease in NO bioavailability is considered the hallmark ofendothelial dysfunction subsequently leading to attenuatedvascular relaxation and atherosclerosis [10]

Insulin signalling also plays a critical role in normalvascular function via modulation of calcium handling andsensitivity in VSMCs [3] When insulin signal transduc-tion is impaired bioavailability of NO in ECs decreaseswhile endothelin-1 production the inflammatory activityand smooth muscle cell proliferation increase [6]

22 Dyslipidaemia High circulating levels of triglyceride-rich particles reduced synthesis of HDL and enhanced pro-duction of atherogenic low-density lipoprotein (LDL) par-ticles characterize diabetic dyslipidaemia [11 12] Howeverthe regulation of lipid metabolism in diabetes is extremelycomplex and the mechanisms to trigger vascular dysfunctionare only partially explored The fading of the glycocalyx inlarge arteries exposed to hyperlipidaemic stress may be anearly characteristic of an increased vascular vulnerability [13]Also high levels of LDL and free fatty acids (FFAs) can causeincreased permeability of ECs induce EC-derived foam cellsformation and abnormal hyperplasia of extracellular matrixas well as disturbed secretion of NO and proinflammatorymolecules such as vascular cell adhesionmolecule 1 (VCAM-1) plateletendothelial cell adhesion molecule 1 (PECAM-1) intercellular adhesion molecule 1 (ICAM-1) P-selectinmonocyte chemoattractant protein 1 (MCP-1) interleukin 6(IL-6) Toll-like receptor 4 (TLR-4) CD40 PAI-1 and soforth [13] Although many clinical trials have demonstratedsignificant advantages of utilizing cholesterol-targeting drugsto reduce cardiovascular complications of diabetic patients[14ndash16] the remaining higher prevalence of vascular dysfunc-tion calls for further basic and clinical studies allowing forbetter prevention and treatment

23 Vascular Oxidative Stress T2DM resulting in vasculardysfunction may also occur through the increase in NAPDHoxidase-induced ROS production in the vasculature [17] InT2DM high intracellular glucose levels increase ROS pro-duction by triggering various cellular mechanisms and regu-lators such as protein kinase C (PKC) activation polyol andhexosamine flux AGEs and nuclear factor kappa B (NF-120581B)mediated vascular inflammation [9 10] Hyperglycaemia-mediated superoxide formation contributes to the patho-physiological complications in diabetic patients Not only isthe generation of reactive oxygen species (ROS) elevated indiabetes but the activity of the antioxidant defence systemalso declines [18] There are multiple targets of oxidativedamage in the diabetic vasculature with modifications ofproteins lipids and nucleic acids occurring in both ECsand VSMCs Enhanced oxygen radical production throughtumour necrosis factor 120572 (TNF-120572) and AGE or AGE receptor(RAGE) signalling reduces NO bioavailability and resultsin impairment of vascular function [19] Furthermore viascavenger receptor recognition ROS trigger recruitment ofmonocytes and their differentiation intomacrophages whichinitiate a vascular inflammation cascade [20]

24 Adipokines Obesity and T2DM are associated withadverse expression patterns of various adipose-derivedcytokines and chemokines and enhanced adipose inflamma-tory cell infiltration [21] Adipokines produced by adiposetissue may affect vascular function and insulin sensitivity[22] To date several adipokines have been identified andcharacterized to modify vascular function and the list is stillgrowing [23]

Adiponectin for example an anti-inflammatoryadipokine which is reduced in obesity and T2DM [24]has been shown to increase insulin sensitivity and to improvevascular function by reducing TNF-120572-stimulated expressionof endothelial adhesion molecules and monocyte attachment[25ndash27] In addition adiponectin attenuates the productionof ROS induced by high glucose oxidized LDL and palmitatein endothelial cells [27]

Another well studied adipokine leptin has been sug-gested to induce vascular endothelial dysfunction [28] andVSMC proliferation [29] Also resistin which is elevatedin obesity and T2DM was found to be a strong risk factorfor acute coronary syndrome in different clinical studies[30] Resistin promotes atherogenic changes in VSMCs suchas increased proliferation migration and contractility [31]Many other adipokines and cytokines such as visfatin TNF-120572 plasminogen activator inhibitor 1 (PAI-1) and so on mayall contribute to vascular dysfunction by affecting vasculartone and infiltration of inflammatory cells [31 32]

3 Noncoding RNAs

In the last two decades our view of genetic biologicalregulation has changed dramatically Transcriptomic analysesrevealed that only a surprisingly minor portion of 1-2of the human genome is protein-coding transcripts andthe remaining 98 is largely transcribed into noncoding








Skeletal muscle


Adipocytes

Plasma

miR-126amiR-146


miR-378378lowast







miR-33ab

miR-122

miR-370

Let-7

miR-802


miR-30d

miR-210

Lipid metabolism

Glucose metabolism

Responsive Genes

PI3K-mTOR pathway

ABCA1

SREBP1 HNF1B

NF-120581B










2O2)


















miR-21




SIRT1 ZEB1

HIF


Atherosclerosis

SIRT1ITGA5JAK1

KLF45ELK1


PI3KR2VCAM1

PTEN PDCD4


miR-17asymp29















Acknowledgment


References



















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of























Skeletal muscle


Adipocytes

Plasma

miR-126amiR-146


miR-378378lowast







miR-33ab

miR-122

miR-370

Let-7

miR-802


miR-30d

miR-210

Lipid metabolism

Glucose metabolism

Responsive Genes

PI3K-mTOR pathway

ABCA1

SREBP1 HNF1B

NF-120581B










2O2)


















miR-21




SIRT1 ZEB1

HIF


Atherosclerosis

SIRT1ITGA5JAK1

KLF45ELK1


PI3KR2VCAM1

PTEN PDCD4


miR-17asymp29















Acknowledgment


References



















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















miR-33ab

miR-122

miR-370

Let-7

miR-802


miR-30d

miR-210

Lipid metabolism

Glucose metabolism

Responsive Genes

PI3K-mTOR pathway

ABCA1

SREBP1 HNF1B

NF-120581B










2O2)


















miR-21




SIRT1 ZEB1

HIF


Atherosclerosis

SIRT1ITGA5JAK1

KLF45ELK1


PI3KR2VCAM1

PTEN PDCD4


miR-17asymp29















Acknowledgment


References



















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
































miR-21




SIRT1 ZEB1

HIF


Atherosclerosis

SIRT1ITGA5JAK1

KLF45ELK1


PI3KR2VCAM1

PTEN PDCD4


miR-17asymp29















Acknowledgment


References



















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















miR-21




SIRT1 ZEB1

HIF


Atherosclerosis

SIRT1ITGA5JAK1

KLF45ELK1


PI3KR2VCAM1

PTEN PDCD4


miR-17asymp29















Acknowledgment


References



















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






















Acknowledgment


References



















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of















































































of






Disease Markers



OncologyJournal of





PPAR Research



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Documents

Review Article Making Sense in Antisense: Therapeutic ...downloads.hindawi.com/journals/jdr/2013/834727.pdf · new diagnostic and therapeutic opportunities to treat diabetes-induced