miR-30b-5p Downregulation as a Predictive Biomarker of Coronary In-Stent Restenosis 2021 Encarnación Gutierrez-Carretero.pdf

该【miR-30b-5p Downregulation as a Predictive Biomarker of Coronary In-Stent Restenosis 2021 Encarnación Gutierrez-Carretero 】是由【丰儿】上传分享，文档一共【15】页，该文档可以免费在线阅读，需要了解更多关于【miR-30b-5p Downregulation as a Predictive Biomarker of Coronary In-Stent Restenosis 2021 Encarnación Gutierrez-Carretero 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。:.
biomedicines
Article
miR-30b-5pDownregulationasaPredictiveBiomarkerof
CoronaryIn-StentRestenosis
EncarnaciónGutierrez-Carretero1,2,3,†,IsabelMayoral-González1,2,†,FranciscoJesúsMorón4,
MónicaFernández-Quero3,AlejandroDomínguez-Rodríguez1,AntonioOrdóñez1,2andTarikSmani1,5,*
1CardiovascularPathophysiology,InstituteofBiomedicineofSeville,UniversityHospitalofVirgendelRocío,
UniversityofSeville,CSIC,41013Seville,Spain;gutierrez.******@(.-C.);
******@(.-G.);******@(.-R.);******@(.)
2DepartmentofSurgery,FacultyofMedicine,UniversityofSeville,41009Sevilla,Spain
3UniversityHospitalVirgendelRocío,41013Sevilla,Spain;******@
4GenomicFacility,InstituteofBiomedicineofSeville,UniversityHospitalofVirgendelRocío,
UniversityofSeville,CSIC,41013Seville,Spain;mcivanto-******@
5DepartmentofMedicalPhysiologyandBiophysics,FacultyofMedicine,UniversityofSeville,
41009Seville,Spain
*Correspondence:******@
†Theseauthorscontributedequally.
Abstract:In-stentrestenosis(ISR)isoneofthemainlimitationsofpercutaneouscoronaryintervention
(PCI)therapywithdrug-elutingstents(DES)



ifcirculatingmicroRNAs(miRNAs)havediagnosticcapabilityfordeterminingISRinacohort



Citation:Gutierrez-Carretero,E.;
Mayoral-González,I.;JesúsMorón,F.;
comprisingpatientswhopresentedISRornot(non-ISR).Amicroarrayanalysisdeterminedthatupto
Fernández-Quero,M.;
Domínguez-Rodríguez,A.;Ordóñez,49miRNAsweredifferentiallyexpressedbetweenISRandnon-,10miRNAsare
A.;Smani,-30b-5prelatedtovascularsmoothmuscleandendothelialcellsproliferation,migration,anddifferentiation,
DownregulationasaPredictivewell-,weidentifledthattheexpressionof
BiomarkerofCoronaryIn-StentmiR-30b-5pissigniflcantlylowerinserumsamplesofISRpatients,ascomparedtonon-
,9,-30b-5pprovidesbettervaluesofthereceiveroperatorcharacteristic
/,thein-silicoanalysissuggeststhat
biomedicines9040354miR-30b-5pispredictedtotarget62genesinvolvedindifferentsignalingpathwaysinvolvedin
,wedeterminedfortheflrsttimethatcirculatingmi-R30b-5pcan
AcademicEditor:CelestinoSardureliablyprognoserestenosisinpatientwithimplantedDES,whichcouldbepotentiallyhelpfulinthe
establishmentofanearlydiagnosisandtherapyofISR.
Received:6March2021
Accepted:26March2021
Keywords:PrimaryCoronaryIntervention;in-stentrestenosis;miRNAs;biomarker
Published:30March2021
Publisher’sNote:MDPIstaysneutral
withregardtojurisdictionalclaimsin

publishedmapsandinstitutionalaffll-
(CAD)isconsideredthemajorcauseofdeathanddisabilityin
developedcountries[1].CADismainlyassociatedwithatherosclerosisplaquedeposition
intheintimaofcoronaryarteries,whereitnarrowsthearterydiameters,whichlimitsor
evenblocksthebloodflow[2].Theatheroscleroticprogressissilentuntiltheappearance
Copyright:©,acutemyocardialinfarction(AMI),oreven
LicenseeMDPI,Basel,[3].Percutaneouscoronaryintervention(PCI)hassigniflcantly
ThisarticleisanopenaccessarticleimprovedtheprognosisforpatientswithCAD,,
distributedunderthetermsandrestenosis,deflnedastherepeatednarrowingofthevesseldiameter,remainsaserious
conditionsoftheCreativeCommonslimitationofthisprocedure[4].Themaincauseofrestenosiswasattributedtotheexcessive
Attribution(CCBY)license(https://proliferationandmigrationofvascularsmoothmusclecells(VSMC)totheintima[5]and
,drug-elutingstents(DES),
/).mainlytargetingVSMCproliferationandmigration,arewidelyused,whichdrastically
Biomedicines2021,9,:///biomedicines9040354:.
Biomedicines2021,9,3542of15
reducestherateandtheincidenceofin-stentrestenosis(IRS).Actually,theuseofDES
achievedsigniflcantreliefofclinicalsymptomsandreducedCADpatientsmortality,
asreviewedrecently[6].Currently,conventionalandcomputedcoronarytomography
angiographiesaretheonlyeffectivestrategiesofdetectingISRinpatientspresentingAMI-
,theconsequencesofsuccessivecoronaryangiography:itshigh
cost,itsinvasivecharacter,anditstestcontradictionsmakeurgenttheneedofasensitive
andreliablemarkertodiagnoserestenosis,whichcouldinfluencepositivelyapreventive
treatment,especiallyinhigh-riskpatients.
MicroRNAs(miRNAs)aresmallnoncodingRNAswithapproximately19–25nucleotides
involvedinregulatingawiderangeofdevelopmentalandphysiologicalprocesses[7].MiR-
NAsplaycriticalroleintheregulationofgeneexpressionatthepost-transcriptionallevel[8]
andinthemodulationofthecellularmechanismimplicatedincardiovasculardiseases
(CVDs),suchascardiachypertrophy,cardiacarrhythmias,orheartfailure[9].MiRNAs
couldbereleasedaftercellularactivation,stress,orinjury[10],sotheyareconsidered
asvaluableandstablebiomarkersforheartdisease,suchasrestenosis,asreviewedelse-
where[2].However,littleisknownregardingtheexpressionofmiRNAsinpatientswho
underwentPCIanddeveloped,ornot,in-stentrestenosis(ISR).Inthepresentstudy,we
evaluatedthediagnosticperformanceofcirculatingmiRNAsasserumbiomarkersfor
patientwhohavedevelopedISR.

Thissingle-centerstudywasconductedaccordingtotheEthicalPrinciplesofthe

theUniversityHospital“VirgendelRocio”ofSeville(-0313-2016;dateof
Biomedicines2021,9,xFORPEERREVIEWapproval:2February2017).TheStrengtheningthereportingofobservationalstudiesin3of15
epidemiology(STROBE)guidelineswerefollowedtoreportourflndings(Figure1).
(STROBE)diagramofStrengtheningthereportingofobservationalstudiesinepidemiology(STROBE)diagram
:drug-elutingstent,ISR:patientwithin-stentrestenosis,non-ISR::drug-elutingstent,ISR:patientwithin-stentrestenosis,non-
ISR:patientwithnonin-stentrestenosis,miRNA:microRNA,andPCI:percutaneouscoronary
patientwithnonin-stentrestenosis,miRNA:microRNA,andPCI:,.

Coronaryarterylesionscharacteristicswereanalyzedaftercoronaryangiography,
Thisretrospectivestudywasconductedin55patientsundergoingcoronaryangiogra-
phywhohadpriorcoronaryangiographywithDESimplantationandmettheeligibilitysuchasmultivesselarterylesionsandtargetlesionattheleftanteriordescending(LAD)
artery,attheleftcircumflex(LCX)artery,andattherightcoronaryartery(RCA).Baseline,
postoperative,andfollow-upcoronaryangiogramsweredigitallyrecorded,andthequan-
titativecoronaryangiography(QCA)analyseswereperformedwithanautomatededge-
detectionsystemCarestream(MedisMedicalImagingSystems,Leiden,TheNetherland).
Wealsocollectedandanalyzedclinicaldata,suchasage,gender,smokingconditions,
hypertension,diabetesmellitus(DM),insulintherapy,chronickidneydisease(CKD),he-
moglobin,creatinine,creatinephosphokinase(CPK),C-reactiveprotein(CRP),troponin,
N-terminalnatriureticpeptide(NT-proBNP),totalcholesterol(TC),high-densitylipopro-
teincholesterol(HDL-C),low-densitylipoproteincholesterol(LDL-C),triglycerides(TG),
erythrocytes,leucocytes,andneutrophils.
,miRNAIsolation,andQuantification
Bloodsampleswerecollectedfrompatientsbeforethesecondangiographymoti-

rpmfor15mintoseparatetheserumthatwerelaterprocessedinthelaboratoryforRNA
extractionusingthemiRNeasySerum/Plasmakit(Qiagen,Hilden,Germany)toextract
®lysis
-
tionbytheQubitmiRNAassay(ThermoFisherScientific,Waltham,MA,USA).

ThetotalRNAwaslabeledusingtheFlashTag®BiotinHSRlabelingKit(Thermo
FisherScientific,Inc.,Waltham,MA,USA)followingthemanufacturer’
usedGeneChip®(ThermoFisherScientific,Inc.,Waltham,MA,USA)
,staining(GeneChip®FluidicsStation450),
andscanning(GeneChip®Scanner3000,ThermoFisherScientific,Inc.,Waltham,MA,
USA)wereperformedfollowingtheprotocolsoutlinedintheusermanualforcartridge:.
Biomedicines2021,9,3543of15
criteriabasedonsymptomssuggestingischemia,suchaschestpainradiationtoarm,neck,
-stentrestenosis(ISR)andnon-ISRgroupswerederivedfrompatientsundergoing
%inatleast


consecutiveseriesinthisstudy.
Coronaryarterylesionscharacteristicswereanalyzedaftercoronaryangiography,
suchasmultivesselarterylesionsandtargetlesionattheleftanteriordescending(LAD)
artery,attheleftcircumflex(LCX)artery,andattherightcoronaryartery(RCA).Base-
line,postoperative,andfollow-upcoronaryangiogramsweredigitallyrecorded,and
thequantitativecoronaryangiography(QCA)analyseswereperformedwithanauto-
matededge-detectionsystemCarestream(MedisMedicalImagingSystems,Leiden,The
Netherland).Wealsocollectedandanalyzedclinicaldata,suchasage,gender,smoking
conditions,hypertension,diabetesmellitus(DM),insulintherapy,chronickidneydisease
(CKD),hemoglobin,creatinine,creatinephosphokinase(CPK),C-reactiveprotein(CRP),
troponin,N-terminalnatriureticpeptide(NT-proBNP),totalcholesterol(TC),high-density
lipoproteincholesterol(HDL-C),low-densitylipoproteincholesterol(LDL-C),triglycerides
(TG),erythrocytes,leucocytes,andneutrophils.
,miRNAIsolation,andQuantiflcation
Bloodsampleswerecollectedfrompatientsbeforethesecondangiographymotivated

15mintoseparatetheserumthatwerelaterprocessedinthelaboratoryforRNAextraction
usingthemiRNeasySerum/Plasmakit(Qiagen,Hilden,Germany)toextractsmallRNAs.
Twohundredmicrolitersofserumwasmixedwith700LofQIAzol®lysisreagentincluded

miRNAassay(ThermoFisherScientiflc,Waltham,MA,USA).

ThetotalRNAwaslabeledusingtheFlashTag®BiotinHSRlabelingKit(Thermo
FisherScientiflc,Inc.,Waltham,MA,USA)followingthemanufacturer’
usedGeneChip®(ThermoFisherScientiflc,Inc.,Waltham,MA,USA)to
,staining(GeneChip®FluidicsStation450),
andscanning(GeneChip®Scanner3000,ThermoFisherScientiflc,Inc.,Waltham,MA,
USA)wereperformedfollowingtheprotocolsoutlinedintheusermanualforcartridge
,CELflleimportandmiRNAlevelwithrobustmultiarrayaverage(RMA)
normalizationanddetectionabovethebackground(DABG)wereperformedusingTran-
scriptomeAnalysisConsole(TAC)(ThermoFisherScientiflc,Inc.,Waltham,
MA,USA).
-ISR
patientswascarriedoutusingafoldchangeofover
withap-value<,inwhichthenullhypothesiswas“thereisnodifferencebetweenthe
groups”.Inaddition,aprobesetwasconsideredexpressedif50%sampleshadDABG
valuesbelowthethreshold(DABG<).Hierarchicalclusteringwasperforme