The role of ERBB2 gene polymorphisms in leprosy susceptibility

braz j infect dis.2015;19(2):206–208

www .e l s e v i e r . c o m / l o c a t e / b j i d

The

Brazilian

Journal

of

INFECTIOUS

DISEASES

Brief

communication

The

role

of

ERBB2

gene

polymorphisms

in

leprosy

susceptibility

夽

Jamile

Leão

Rêgo

_,

_Joyce

_Moura

_Oliveira

_,

_Nadja

_de

_Lima

_Santana

_,

Paulo

Roberto

Lima

Machado

_,

_Léa

_Cristina

_Castellucci

a_{InstitutoNacionaldeCiênciaeTecnologiaemDoenc¸asTropicais,Brazil} b_{UniversidadeFederaldaBahia,Salvador,BA,Brazil}

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Received16October2014 Accepted1December2014 Availableonline27January2015

Keywords:

ERBB2 Geneanalysis Leprosy

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Mycobacteriumlepraeinfectsskinandperipheralnervescausingdeformitiesanddisability. TheM.lepraebacteriumbindstoErbB2ontheSchwanncellsurfacecausingdemyelination andfavoringspreadofthebacilliandcausingnerveinjury.PolymorphismsattheERBB2 genewerepreviouslyinvestigatedasgeneticriskfactorsforleprosyintwoBrazilian popu-lationsbutwithinconsistentresults.HereinweextendtheanalysisofERBB2variantstoa thirdgeographicallydistinctpopulationinBrazil.Ourresultsshowthatthereisno asso-ciationbetweenthegenotypedSNPsandthedisease(p>0.05)inthispopulation.Agene setorpathwayanalysisunderthegenomicregionofERBB2willbenecessarytoclarifyits regulationunderM.lepraestimulus.

©2015ElsevierEditoraLtda.Allrightsreserved.

Leprosyisa chronic infectiousdisease caused by Mycobac-terium leprae,andinfluenced bygeneticand environmental factors.Thisinfectionhasabroadclinicalandimmunological spectrum that causes high morbidity rates, with a major impact on public health. Although leprosy prevalence has beenextensivelyreducedaftertheintroductionofmultidrug therapyandvaccinationwithBCG1_{morethan 200,000new} cases are reported annually according to World Health Organization.2 _{The spectrum of clinical manifestations is} illustratedbytwopolarforms,tuberculoidandlepromatous leprosy, and various intermediate or borderline forms.3 Peripheralnervedamage,themostserious complicationof leprosy,isassociatedwithimmunologicalandinflammatory

夽_{Thisworkwasconductedwiththeeffortandcontributionofthefollowingpersons:JamileLeãoRêgo,JoyceMouraOliveira,Nadjade}

LimaSantana,ThaísLamêgoMagalhães,ThaillamarSilvaVieira,MayumeShibuya,LídiaMariaMachado,PauloRobertoLimaMachado andLéaCristinaCastellucci.

∗ _{Correspondingauthor.}

E-mailaddress:jamileao@hotmail.com(J.L.Rêgo).

events which evolve through time and haveconsequences that can extend for years after cure of the infection. The

ERBB2 gene lies on chromosome 17q11-q21, and encodes animportantclassofreceptortyrosinekinasesinvolvedin transmissionofbiochemicalsignals.4ThebindingofM.leprae

to myelinatedSchwann cells through ligationtothe ErbB2 receptorresultsinSchwanncelldemyelinationandincreases the population of de-differentiated Schwann cells through theErk1/2signalingpathway.5 _{Thiscreatesanenvironment} thatfavorsM.lepraeproliferationandleadstonervedamage. Recentdata haveshownthat polymorphismsinthe ERBB2 genewere associatedwithleprosydevelopmentinfamilies from Pará State, but not inapopulation from the stateof

http://dx.doi.org/10.1016/j.bjid.2014.12.008

brazj infect dis.2015;19(2):206–208

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Table1–Characteristicsofcase–controlsamplefromthe populationofBahia,Brazil.

(A)Patientscharacteristics

nIndividuals Meanage(years)±SDa _Male:female

Cases 362 42.32±12.88 199:163

Controls 368 34.80±10.24 258:110

(B)Clinicalcharacteristicsofthecasescohort

Clinicalphenotype n Tuberculoid(TT) 44 BorderlineTT(BT) 48 Borderline(BB) 38 Borderlinelepromatous(BL) 40 Lepromatous(LL) 124 Unclassifiedleprosy 63

Otherleprosyclinicalforms 5

RR 80

ENL 84

Totalb ₃₆₂

Note.

a _{SD,standard deviation; RRreversalreactions; ENL, erythema}

nodosumleprosum.

b _{RRandENL patientsarealsoclassifiedunderleprosyclinical}

spectrum.

Rio Grande do Norte in Brazil.6 _{Herein we investigate a} geographicallydistinct populationfromNortheasternBrazil tofurtherevaluatetheroleofthisgeneinleprosy suscepti-bility.ThestateofBahiaisconsideredofmediumendemicity, accordingtotheparameterofprevalence.7_{Threehundredand} sixty-twoleprosycaseswereselectedattheHospitalsEdgard Santosand Dom Rodrigode Menezes.Both are considered referencecentersfortreatmentofthediseasewithpatients diagnosedaccordingtothe guidelinesoftheBrazilian Min-istryofHealth.Additionally,368localbloodbankdonorswere recruitedascontrols. Informedconsentwasobtainedfrom allparticipants.Approvalfortheuseofthesamplesinthis studywasobtainedfromtheFederalUniversityofBahia (CEP-50/2010)andtheBrazilianNationalEthicalCommittee(CONEP 11019).Detailedcomplementarydataaboutcasesandcontrols are described in Table 1. Five single nucleotide polymor-phisms (SNPs) (rs2517955, rs2517956, rs1810132, rs2952156, rs1136201) were genotyped by TaqMan RT-PCR technology (AppliedBiosystems®_{)usingPre-designedgenotypingassays.}

Table3–Resultsoflogisticregressionanalysesforthe

genotypedERBB2polymorphisms.

ERBB2alleles Globaltest OR(95%CI) p-Value

2df 1df rs2517955(Tvs.C) 0.239 0.227 1.14(0.92–1.41) 0.228 rs2517956(Avs.G) 0.051 0.364 1.10(0.88–1.37) 0.365 rs1810132(Tvs.C) 0.179 0.406 1.10(0.87–1.40) 0.407 rs2952156(Gvs.A) 0.098 0.368 1.10(0.88–1.38) 0.369 rs1136201(Avs.G) 0.660 0.550 1.10(0.79–1.54) 0.550

Note.The2dftestrepresentsthegenotype-wisetest,andthe1df

testrepresentstheallele-wisetest.CI,confidenceinterval;OR,odds ratio.TheORandp-valuesreferto1dftests.

Table2describescharacteristicsoftheSNPsusedinthisstudy andcomparestheminorallelefrequenciesbetweenthethree Northeasternpopulations.TestsforHardy-Weinberg equilib-riumandunconditionallogisticregressionanalysisweredone usingSTATA8.2withthefreelyavailableGenAssocpackage (http://www-gene.cimr.cam.ac.uk/clayton/software/stata/)to determineallele-wise(1dftest)andgenotype-wise(2dftest) associationsbetweenERBB2SNPs andleprosy.Ouranalysis did not show any significant association between the five genotypedmarkersandleprosyperse(p>0.05),aspresented inTable3.Norwere thereany associationsbetweenERBB2

polymorphismsandeitherlepromatousortuberculoidforms ofdisease(datanotshown).

Thedata showedhere supportprevious observationsof a lack of association between ERBB2 genetic variants and susceptibility to leprosy per se in a similar population-basedcase-controldatasetfromRioGrandedoNorte.6_Our population-basedstudy inBahiaand thatinRioGrandedo Nortehadlargersamplesizesthanthesmallerfamily-based study in Pará,suggesting that the association observed in the latterpopulation may representa falsepositive result. However,Araújoetal.6_{drawattentiontothefactthatERBB2} liesunderalinkagepeak8,9_{forleprosyobservedinthesame} large pedigrees employedfor the study in Pará State. The regionofthelinkagepeakatchromosome17q11-q22isagene denseregionofpotentialimmunologicalcandidategenes con-tributingtoleprosysusceptibility.8,9_{Thisraisesthepossibility} that variants at ERBB2 might contribute to larger chromo-somehaplotypestransmittedtoaffectedindividualsinthese Paráfamilies. Furtherinvestigationisnecessarytoclarifyif

ERBB2contributesgeneticallyand/orfunctionallytoleprosy

Table2–DetailsofERBB2singlenucleotidepolymorphisms(SNPs)genotypedinthethreepopulations.

SNPrs# Location(BuildHg19) Cohortsgenotyped Minorallele Minorallelefrequency(Pará/RN/Bahia) Position/function rs2517955 37843931 RN/Bahia C 0.49/0.41 Upstreamregion rs2517956a _{37844109 Pará/RN/Bahia G 0.38/0.38/0.42 Upstreamregion}

rs1810132 37866255 RN/Bahia C 0.39/0.40 Intronic

rs2952156a _{37877085 Pará/RN/Bahia A 0.38/0.38/0.38 Intronic}

rs1136201a _{37879838 RN/Bahia G 0.15/0.12 Coding,non-syn(I655V)}

AdaptedfromAraújoetal.6

Note.CohortsarefromPará,RNandBahiastatesinnortheasternBrazil.

208

susceptibilityinthesefamilies.RecentdatashowsthatM. lep-raehijackstheplasticityofadultSchwanncells,toreprogram infectedcellstoaprogenitor/stemcell-likecellsasa bacte-rialstrategytospreadinfectiontoothertissues.10_Inaddition, reprogrammedcellscanattractmacrophages,providing evi-dence for a functional role of innate immune response genesduringreprogramming.11_{Finally,althoughgene} analy-sesbasedonasingledatatype,suchasgeneexpressiondataor SNPdata,havesuccessfullyrevealedalteredcellularprocesses associated with different complex diseases12–15 _{it is also} knownthatsinglevariantorsinglegeneanalysesgenerally accountforonlyasmallproportionofthephenotypic varia-tionincomplextraits.Inthissense,wehavetoconsiderthat

ERBB2islocatedwithinageneticlocusthatcontainsanumber ofgenesdirectlyinvolvedintheimmuneresponseand patho-genesisofinfectiousdiseases.Awell-poweredgenome-wide associationstudy would beneededto determinearole for ERBB2relativetoothergenesthathavebeenshowntoachieve genome-widesignificanceinotherpopulations.

Conflicts

of

interest

Theauthorsdeclarenoconflictsofinterest.

Financial

support

CNPq,INCT(http://www.cnpq.br).

Acknowledgements

WethankthestaffofMagalhãesNetoLeprosyService, Hospi-talDomRodrigodeMenezesandHEMOBAforthelogistical supportinthe samplecollection. WealsothankDr.Selma Jeronimo forthe scientific support. Wethank Prof.Jenefer Blackwellforhelpfulcommentsonthemanuscript.

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1. KarongaPTG.RandomisedcontrolledtrialofsingleBCG, repeatedBCG,orcombinedBCGandkilledMycobacterium

lepraevaccineforpreventionofleprosyandtuberculosisin Malawi.Lancet.1996;348:17–24.

2.Globalleprosy:updateonthe2012situation–WorldHealth Organization,WHO.WklyEpidemiolRec.2013;88:365–80.

3.MontoyaD,ModlinRL.LearningfromLeprosy.Insightinto thehumaninnateimmuneresponse.AdvImmunol.2010; 10:1–24.

4.SchlessingerJ.Cellsignalingbyreceptortyrosinekinases. Cell.2000;103:211–25.

5.TapinosN,OhnishiM,RambukkanaA.ErbB2receptor tyrosinekinasesignalingmediatesearlydemyelination inducedbyleprosybacilli.NatMed.2006;12:961–6.

6.AraújoSRF,JamiesonSE,DupnikKM,etal.ExaminingERBB2 asacandidategeneforsusceptibilitytoleprosy(Hansen’s disease)inBrazil.2014MemInstOswaldoCruz.

2014;109:182–8.

7.OliveiraVM,AssisCRD,SilvaKCC.Levantamento

epidemiológicodahanseníasenonordestebrasileirodurante operíodode2001-2010.ScireSalutis.2013;3:16–27.

8.JamiesonSE,MillerEN,BlackGF,etal.Evidenceforacluster ofgenesonchromosome17q11-q21controllingsusceptibility totuberculosisandleprosyinBrazilians.GenesImmun. 2004;5:46–57.

9.MillerEN,JamiesonSE,JobertyC,etal.Genome-widescans forleprosyandtuberculosissusceptibilitygenesinBrazilians. GenesImmun.2004;5:63–7.

10.MasakiT,QuJ,Cholewa-WaclawJ,BurrK,RaaumR,

RambukkanaA.ReprogrammingadultSchwanncellstostem cell-likecellsbyleprosybacillipromotesdisseminationof infection.Cell.2013;152:51–67.

11.MasakiT,McGlincheyA,Cholewa-WaclawJ,QuJ,Tomlinson SR,RambukkanaA.Innateimmuneresponseprecedes

Mycobacteriumleprae-inducedreprogrammingofadult SchwannCells.CellReprogram.2014;16:

9–17.

12.BaranziniSE,GalweyNW,WangJ,etal.Pathwayand network-basedanalysisofgenome-wideassociationstudies inmultiplesclerosis.HumMolGenet.2009;18:2078–90.

13.OoiCH,IvanovaT,WuJ,etal.Oncogenicpathway combinationspredictclinicalprognosisingastriccancer. PLoSGenet.2009;5:e1000676.

14.LiuYJ,GuoYF,ZhangLS,etal.Biologicalpathway-based genome-wideassociationanalysisidentifiedthevasoactive intestinalpeptide(VIP)pathwayimportantforobesity. Obesity(SilverSpring).2010;18:2339–46.

15.ZhangL,GuoYF,LiuYZ,etal.Pathway-basedgenome-wide associationanalysisidentifiedtheimportanceof

regulation-of-autophagypathwayforultradistalradiusBMD.J BoneMinerRes.2010;25:1572–80.