title: Genotype-phenotype correlation and functional studies in patients with cystic fibrosis bearing CFTR complex alleles.
abstract: The effect of complex alleles in cystic fibrosis (CF) is poorly defined for the lack of functional studies.
  
  
  
  To describe the genotype-phenotype correlation and the results of either in vitro and ex vivo studies performed on nasal epithelial cells (NEC) in a cohort of patients with CF carryingcystic fibrosis transmembrane conductance regulator(CFTR) complex alleles.
  
  
  
  We studied 70 homozygous, compound heterozygous or heterozygous forCFTRmutations: p.[Arg74Trp;Val201Met;Asp1270Asn], n=8; p.[Ile148Thr;Ile1023_Val1024del], n=5; p.[Arg117Leu;Leu997Phe], n=6; c.[1210-34TG[12];1210-12T[5];2930C>T], n=3; p.[Arg74Trp;Asp1270Asn], n=4; p.Asp1270Asn, n=2; p.Ile148Thr, n=6; p.Leu997Phe, n=36. In 39 patients, we analysed the CFTR gating activity on NEC in comparison with patients with CF (n=8) and carriers (n=4). Finally, we analysed in vitro the p.[Arg74Trp;Val201Met;Asp1270Asn] complex allele.
  
  
  
  The p.[Ile148Thr;Ile1023_Val1024del] caused severe CF in five compound heterozygous with a class I-II mutation. Their CFTR activity on NEC was comparable with patients with two class I-II mutations (mean 7.3% vs 6.9%). The p.[Arg74Trp;Asp1270Asn] and the p.Asp1270Asn have scarce functional effects, while p.[Arg74Trp;Val201Met;Asp1270Asn] caused mild CF in four of five subjects carrying a class I-II mutationin trans, or CFTR-related disorders (CFTR-RD) in three havingin transa class IV-V mutation. The p.[Arg74Trp;Val201Met;Asp1270Asn] causes significantly (p<0.001) higher CFTR activity compared with compound heterozygous for class I-II mutations. Furthermore, five of six compounds heterozygous with the p.[Arg117Leu;Leu997Phe] had mild CF, whereas the p.Leu997Phe,in transwith a class I-IICFTRmutation, caused CFTR-RD or a healthy status (CFTR activity: 21.3-36.9%). Finally, compounds heterozygous for the c.[1210-34TG[12];1210-12T[5];2930C>T] and a class I-II mutation had mild CF or CFTR-RD (gating activity: 18.5-19.0%).
  
  
  
  The effect of complex alleles partially depends on the mutationin trans. Although larger studies are necessary, the CFTR activity on NEC is a rapid contributory tool to classify patients with CFTR dysfunction.
authors.affiliation: Dipartimento di Pediatria, Centro Regionale Toscano per la Fibrosi Cistica, Azienda Ospedaliero-Universitaria Meyer, Florence, Italy.
authors.forename: Vito
authors.initials: V
authors.lastname: Terlizzi
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.|Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, Naples, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Castaldo
authors.affiliation: Centro Fibrosi Cistica, AOR San Carlo, Potenza, Italy.
authors.forename: Donatello
authors.initials: D
authors.lastname: Salvatore
authors.affiliation: Dipartimento di Biotecnologie Cellulari ed Ematologia, Istituto Pasteur Fondazione Cenci Bolognetti, Sapienza Università e Policlinico Umberto I, Rome, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Lucarelli
authors.affiliation: Centro Regionale Fibrosi Cistica, Sezione Pediatrica, Dipartimento di Scienze Mediche Traslazionali, Università di Napoli Federico II, Naples, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Raia
authors.affiliation: Laboratorio di Genetica Medica, Ospedale Pediatrico Bambino Gesù, Rome, Italy.
authors.forename: Adriano
authors.initials: A
authors.lastname: Angioni
authors.affiliation: Dipartimento di Scienze Mediche Traslazionali, Centro Regionale Fibrosi Cistica, Sezione Adulti, Università di Napoli Federico II, Naples, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Carnovale
authors.affiliation: Dipartimento Materno-Infantile, Ospedali Riuniti Ancona, Centro Regionale Fibrosi Cistica, Ancona, Italy.
authors.forename: Natalia
authors.initials: N
authors.lastname: Cirilli
authors.affiliation: Dipartimento di Pediatria, Centro Regionale Fibrosi Cistica, IRCCS G. Gaslini, Genova, Italy.
authors.forename: Rosaria
authors.initials: R
authors.lastname: Casciaro
authors.affiliation: Centro Regionale Fibrosi Cistica, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, Università degli Studi di Milano, Milan, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Colombo
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.|Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, Naples, Italy.|Dipartimento di Neuroscienze, Sezione di ORL, Università di Napoli Federico II, Naples, Italy.
authors.forename: Antonella Miriam
authors.initials: AM
authors.lastname: Di Lullo
authors.affiliation: Università Telematica Pegaso, Napoli, Italy.
authors.forename: Ausilia
authors.initials: A
authors.lastname: Elce
authors.affiliation: Dipartimento di Scienze Mediche Traslazionali, Centro Regionale Fibrosi Cistica, Sezione Adulti, Università di Napoli Federico II, Naples, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Iacotucci
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.|Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, Naples, Italy.
authors.forename: Marika
authors.initials: M
authors.lastname: Comegna
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.|Dipartimento di Biotecnologie e Bioscienze, Università di Milano Bicocca, Milan, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Scorza
authors.affiliation: Unità di Fibrosi Cistica, IRCCS Ospedale Pediatrico Bambin Gesù, Rome, Italy.
authors.forename: Vincenzina
authors.initials: V
authors.lastname: Lucidi
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Perfetti
authors.affiliation: Dipartimento di Scienze Neurologiche, Riproduttive ed Odontostomatologiche, Università di Napoli Federico II, Naples, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Cimino
authors.affiliation: Centro Fibrosi Cistica, Sapienza Università e Policlinico Umberto I, Rome, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Quattrucci
authors.affiliation: Laboratorio Genetica Medica, Ospedale Maggiore Policlinico, Milano, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Seia
authors.affiliation: Laboratorio di Genetica Medica, Ospedale Pediatrico Bambino Gesù, Rome, Italy.
authors.forename: Valentina Maria
authors.initials: VM
authors.lastname: Sofia
authors.affiliation: Dipartimento di Bioscienze e Territorio, Università del Molise, Isernia, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Zarrilli
authors.affiliation: CEINGE-Biotecnologie Avanzate, Naples, Italy.|Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, Naples, Italy.
authors.forename: Felice
authors.initials: F
authors.lastname: Amato
mesh_terms: D000293:Adolescent; D000328:Adult; D000483:Alleles; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000235:genetics* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005260:Female; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009297:Nasal Mucosa / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D055815:Young Adult
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: [I148T;3199del6bp]; [L997F;R117L]; [R74W;V201M;D1270N]; gating activity; nasal brushing