REGULAR ARTICLEATP-binding cassette member A3 (E292V) gene mutation and pulmonarymorbidity in very-low-birth-weight infantsChristoph Hrtel (haertel@paedia.ukl.mu-luebeck.de)1, Ursula Felderhoff-Mser2, Corinna Gebauer3, Thomas Hoehn4, Angela Kribs5, Reinhard Laux6,Jens Mller7, Hugo Segerer8, Norbert Teig9, Axel von der Wense10, Christian Wieg11, Guido Stichtenoth1, Egbert Herting1, Wolfgang Gpel1for the German Neonatal Network (GNN)1.Department of Pediatrics, University at Lu beck, Lu beck, Germany2.Department of Pediatrics, University of Essen, Essen, Germany3.Department of Pediatrics, University of Leipzig, Leipzig, Germany4.Department of Pediatrics, University of Du sseldorf, Du sseldorf, Germany5.Department of Pediatrics, University of Cologne, Cologne, Germany6.Childrens Hospital Hamburg-Barmbek, Hamburg, Germany7.Childrens Hospital Saarbru cken, Saarbru cken, Germany8.Childrens Hospital St. Hedwig, Regensburg, Regensburg, Germany9.Department of Pediatrics, Childrens Hospital University of Bochum, Bochum, Germany10.Childrens Hospital Hamburg-Altona, Hamburg, Germany11.Childrens Hospital Aschaffenburg, Aschaffenburg, GermanyKeywordsATP-binding cassette member A3 (E292V) genemutation, Pulmonary morbidity, Very-low-birth-weight infantsCorrespondenceC Ha rtel, M.D., Department of Paediatrics, Universityof Lu beck, Ratzeburger Allee 160, 23538 Lu beck,Germany.Tel.: + 49-451-500 2685 |Fax: + 49-451-500 6222 |Email: haertel@paedia.ukl.mu-luebeck.deReceived18 August 2011; revised 10 October 2011;accepted 2 December 2011.DOI:10.1111/j.1651-2227.2011.02553.xStatementofnancial support:Thisstudywassup-portedby theGermanMinistry of EducationandResearch (BMBF).Category of study: Genetic association study.ABSTRACTAim: ATP-binding cassette member A 3 (ABCA3) plays a critical role for the transport ofsurfactant phospholipids into the lamellar bodies of type II alveolar epithelial cells. Terminfantscarrying the E292V missense mutation of the gene encoding ABCA3 are likely to developrespiratory distress syndrome, and the mutation has also been linked to interstitial lung diseasein paediatric patients. The aimof this study was to investigate the association of the E292Vgenotype with pulmonary morbidity in a large cohort of very-low-birth-weight (VLBW) infants.Methods: Weperformedageneticassociationstudywithaprospective, popula-tion-based multi-centre cohort of 3177 VLBW infants born in 16 German study centresbetween 2003 and 2009 (German Neonatal Network). The ABCA3 genotype wasdetermined by restriction fragment length polymorphismPCR in genomic DNA samplesderived from buccal swabs.Results: Inalargecohort of3177VLBWinfants, 11individualswerefoundtobeheterozygote for the E292V mutation (0.34%). After stratication according to ABCA3genotype, no differences were noted for clinical characteristics, necessary treatments andneonatal pulmonary outcomes.Conclusions: Within the size limits of our study cohort, the ABCA3 missense muta-tion E292V had no remarkable effect on pulmonary outcome in VLBW infants. Presentresults do not rule out the possibility that E292V phenotype is associated with minordifference in the morbidity.INTRODUCTIONDecient production of pulmonary surfactant is an essentialfactor for the development of respiratory distress syndrome(RDS),particularlyinpreterminfants.Surfactantisa com-plex mixture of specic proteins and phospholipids which isassembled, processedandstoredinthelamellarbodiesofalveolarepithelialtypeIIcells(AECII). ATP-bindingcas-sette member A 3 is located at the limiting membrane of thelamellar bodies andthereforeplays acritical rolefor thetransport of surfactant phospholipids into AECII. In an ani-mal model, targeted inactivation of the murine ABCA3 geneleads to respiratory failure in newborns with defectivelamellar bodies (1). Recently, several studies havelinkedABCA3genemutationswithlungdiseaseinneonatesand1Key notes ATP-bindingcassettemember A3(ABCA3) playsacritical rolefor thetransport of surfactant phospho-lipids, and the E292V missense mutation of theABCA3 gene has been linked to pulmonary mor-bidity. We performed a genetic association studywith a prospective, population-based multi-centrecohort of 3177 very-low-birth-weight (VLBW)infantsof theGermanNeonatal Network. 11 individuals were found to be heterozygote for theE292Vmutation(0.34%); however, noremarkableeffect on pulmonary outcome was noted.Acta Pdiatrica ISSN 08035253380 2011 The Author(s)/Acta Pdiatrica 2011 Foundation Acta Pdiatrica 2012 101, pp. 380383children (2,3; reviewed in 46). The E292V missense muta-tionwasfoundtobeover-representedinacohortofterminfants with RDS (7). This particular mutation consists of athymine for adenine substitutionat cDNAposition875(rst codonof exon9) andcauses substitutionof valine(mutation) for glutamic acid (wildtype) in codon 292(E292V, new recognition site for the restriction endonucle-aseBsrG1; 8). Bullardetal. (8)alsonotedanassociationbetween the E292V mutation and interstitial lung disease inchildren. Uchida etal. (9) demonstrated the clinical ndingof pulmonary nodules associated with ABCA3 mutations. Apotential pathophysiological linkhas beenestablishedbymeans of cell-culture-based studies demonstrating thatABCA3mutationsmayenhancestresspathwaysintheen-doplasmaticreticulumtherebyinducingapoptosisof lungepithelial cells(10). Ontheotherhand, fatal familial lungdiseasecausedbyABCA3deciencymayalsooccurwith-outidenticationofABCA3mutations(11). Inthisstudy,we hypothesized that the ABCA3 E292V is a potential can-didate mutationto inuence short-termpulmonary out-come in a multi-centre cohort of 3177 VLBW infants.METHODSGenetic association studyWe prospectively studied the inuence of the ABCA3E292Von pulmonary outcome in 3177 VLBWinfantsenrolled in a multi-centre trial involving 16 neonatalintensive care units in Germany fromApril 2003 untilDecember 2009. The inclusion criteria were as follows:birthweight