Vito Terlizzi1, Giuseppe Castaldo2,3, Donatello Salvatore4, Marco Lucarelli5, Valeria Raia6, Adriano Angioni7, Vincenzo Carnovale8, Natalia Cirilli9, Rosaria Casciaro10, Carla Colombo11, Antonella Miriam Di Lullo2,3,12, Ausilia Elce13, Paola Iacotucci8, Marika Comegna2,3, Manuela Scorza2,14, Vincenzina Lucidi15, Anna Perfetti2, Roberta Cimino16, Serena Quattrucci17, Manuela Seia18, Valentina Maria Sofia7, Federica Zarrilli19, Felice Amato2,3. 1. Dipartimento di Pediatria, Centro Regionale Toscano per la Fibrosi Cistica, Azienda Ospedaliero-Universitaria Meyer, Florence, Italy. 2. CEINGE-Biotecnologie Avanzate, Naples, Italy. 3. Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli Federico II, Naples, Italy. 4. Centro Fibrosi Cistica, AOR San Carlo, Potenza, Italy. 5. Dipartimento di Biotecnologie Cellulari ed Ematologia, Istituto Pasteur Fondazione Cenci Bolognetti, Sapienza Università e Policlinico Umberto I, Rome, Italy. 6. Centro Regionale Fibrosi Cistica, Sezione Pediatrica, Dipartimento di Scienze Mediche Traslazionali, Università di Napoli Federico II, Naples, Italy. 7. Laboratorio di Genetica Medica, Ospedale Pediatrico Bambino Gesù, Rome, Italy. 8. Dipartimento di Scienze Mediche Traslazionali, Centro Regionale Fibrosi Cistica, Sezione Adulti, Università di Napoli Federico II, Naples, Italy. 9. Dipartimento Materno-Infantile, Ospedali Riuniti Ancona, Centro Regionale Fibrosi Cistica, Ancona, Italy. 10. Dipartimento di Pediatria, Centro Regionale Fibrosi Cistica, IRCCS G. Gaslini, Genova, Italy. 11. Centro Regionale Fibrosi Cistica, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, Università degli Studi di Milano, Milan, Italy. 12. Dipartimento di Neuroscienze, Sezione di ORL, Università di Napoli Federico II, Naples, Italy. 13. Università Telematica Pegaso, Napoli, Italy. 14. Dipartimento di Biotecnologie e Bioscienze, Università di Milano Bicocca, Milan, Italy. 15. Unità di Fibrosi Cistica, IRCCS Ospedale Pediatrico Bambin Gesù, Rome, Italy. 16. Dipartimento di Scienze Neurologiche, Riproduttive ed Odontostomatologiche, Università di Napoli Federico II, Naples, Italy. 17. Centro Fibrosi Cistica, Sapienza Università e Policlinico Umberto I, Rome, Italy. 18. Laboratorio Genetica Medica, Ospedale Maggiore Policlinico, Milano, Italy. 19. Dipartimento di Bioscienze e Territorio, Università del Molise, Isernia, Italy.
Abstract
BACKGROUND: The effect of complex alleles in cystic fibrosis (CF) is poorly defined for the lack of functional studies. OBJECTIVES: 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 carrying cystic fibrosis transmembrane conductance regulator (CFTR) complex alleles. METHODS: We studied 70 homozygous, compound heterozygous or heterozygous for CFTR mutations: 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. RESULTS: 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 mutation in trans, or CFTR-related disorders (CFTR-RD) in three having in trans a 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 trans with a class I-II CFTR mutation, 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%). CONCLUSIONS: The effect of complex alleles partially depends on the mutation in trans. Although larger studies are necessary, the CFTR activity on NEC is a rapid contributory tool to classify patients with CFTR dysfunction. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.
BACKGROUND: The effect of complex alleles in cystic fibrosis (CF) is poorly defined for the lack of functional studies. OBJECTIVES: 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 carrying cystic fibrosis transmembrane conductance regulator (CFTR) complex alleles. METHODS: We studied 70 homozygous, compound heterozygous or heterozygous for CFTR mutations: 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. RESULTS: 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 mutation in trans, or CFTR-related disorders (CFTR-RD) in three having in trans a 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 trans with a class I-II CFTR mutation, 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%). CONCLUSIONS: The effect of complex alleles partially depends on the mutation in trans. Although larger studies are necessary, the CFTR activity on NEC is a rapid contributory tool to classify patients with CFTR dysfunction. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.
Authors: Krittika Teerapuncharoen; J Michael Wells; S Vamsee Raju; Karen S Raraigh; Melis Atalar Aksit; Garry R Cutting; Lawrence Rasmussen; P Hrudaya Nath; Surya P Bhatt; George M Solomon; Mark T Dransfield; Steven M Rowe Journal: Ann Am Thorac Soc Date: 2019-01
Authors: Kavisha Arora; Fanmuyi Yang; John Brewington; Gary McPhail; Alexander R Cortez; Nambirajan Sundaram; Yashaswini Ramananda; Herbert Ogden; Michael Helmrath; John P Clancy; Anjaparavanda P Naren Journal: Am J Physiol Gastrointest Liver Physiol Date: 2021-05-05 Impact factor: 4.871
Authors: Mao Wang; Alex G Gauthier; Thomas P Kennedy; Haichao Wang; Uday Kiran Velagapudi; Tanaji T Talele; Mosi Lin; Jiaqi Wu; LeeAnne Daley; Xiaojing Yang; Vivek Patel; Sung Soo Mun; Charles R Ashby; Lin L Mantell Journal: Mol Med Date: 2021-07-16 Impact factor: 6.354