11 results on '"Manuela Scorza"'
Search Results
2. Cystic Fibrosis-Screening Positive Inconclusive Diagnosis: Newborn Screening and Long-Term Follow-Up Permits to Early Identify Patients with CFTR-Related Disorders
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Alice Castaldo, Chiara Cimbalo, Raimondo J. Castaldo, Marcella D’Antonio, Manuela Scorza, Laura Salvadori, Angela Sepe, Valeria Raia, and Antonella Tosco
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cystic fibrosis ,CF-SPID ,CFTR-RD ,newborn screening ,genotype–phenotype correlation ,Medicine (General) ,R5-920 - Abstract
Background: Newborn screening (NBS) early-identifies cystic fibrosis (CF), but in CF-screening positive inconclusive diagnosis (CF-SPID) the results of immunoreactive trypsinogen (IRT), molecular analysis and sweat test (ST) are discordant. A percentage of CF-SPID evolves to CF, but data on long-term monitoring are lacking. We describe the follow-up of all CF and CF-SPID identified between 2008 and 2019. Methods: NBS was performed by IRT followed by molecular analysis and ST between 2008 and 2014; double IRT followed by molecular analysis and ST after 2014. Results: NBS revealed 47 CF and 99 CF-SPID newborn, a ratio 1:2.1—the highest reported so far. This depends on the identification by gene sequencing of the second variant with undefined effect in 40 CF-SPID that otherwise would have been defined as carriers. Clinical complications and pulmonary infections occurred more frequently among CF patients than among CF-SPID. Two CF-SPID cases evolved to CF (at two years), while eight evolved to CFTR-related disorders (CFTR-RD), between one and eight years, with bronchiectasis (two), recurrent pneumonia (four, two with sinonasal complications), recurrent pancreatitis (two). No clinical, biochemical or imaging data predicted the evolution. Conclusion: Gene sequencing within the NBS reveals a higher number of CF-SPID and we first describe an approach to early identify CFTR-RD, with relevant impact on their outcome. more...
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- 2020
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Catalog
3. Genetic Diseases That Predispose to Early Liver Cirrhosis
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Manuela Scorza, Ausilia Elce, Federica Zarrilli, Renato Liguori, Felice Amato, and Giuseppe Castaldo
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Diseases of the digestive system. Gastroenterology ,RC799-869 - Abstract
Inherited liver diseases are a group of metabolic and genetic defects that typically cause early chronic liver involvement. Most are due to a defect of an enzyme/transport protein that alters a metabolic pathway and exerts a pathogenic role mainly in the liver. The prevalence is variable, but most are rare pathologies. We review the pathophysiology of such diseases and the diagnostic contribution of laboratory tests, focusing on the role of molecular genetics. In fact, thanks to recent advances in genetics, molecular analysis permits early and specific diagnosis for most disorders and helps to reduce the invasive approach of liver biopsy. more...
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- 2014
- Full Text
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4. Transcriptomics and Metabolomics Integration Reveals Redox-Dependent Metabolic Rewiring in Breast Cancer Cells
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Paola Paci, Marco Vanoni, Tatiana Volpari, Fabrizia Mastroianni, Stefano D'Errico, Valentina Bravatà, Anna Maria Colangelo, Marcella Bonanomi, Daniela Gaglio, Lilia Alberghina, Manuela Scorza, Federica Conte, Giulia Fiscon, Noemi Salmistraro, Elenio Avolio, Gennaro Piccialli, Giusi Irma Forte, Bonanomi, M., Salmistraro, N., Fiscon, G., Conte, F., Paci, P., Bravata, V., Forte, G. I., Volpari, T., Scorza, M., Mastroianni, F., D'Errico, S., Avolio, E., Piccialli, G., Colangelo, A. M., Vanoni, M., Gaglio, D., Alberghina, L., Bonanomi, M, Salmistraro, N, Fiscon, G, Conte, F, Paci, P, Bravata, V, Forte, G, Volpari, T, Scorza, M, Mastroianni, F, D'Errico, S, Avolio, E, Piccialli, G, Colangelo, A, Vanoni, M, Gaglio, D, and Alberghina, L more...
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Pyruvate decarboxylation ,Cancer metabolic rewiring ,CtBP2 ,Epigenetics ,Metabolomics integration ,Transcriptomics ,Cancer Research ,epigenetics ,Chemistry ,cancer metabolic rewiring ,transcriptomics ,metabolomics integration ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Epigenetic ,Metabolism ,Article ,Cell biology ,Oncology ,Transcriptomic ,Anaerobic glycolysis ,Cancer cell ,Glycolysis ,NAD+ kinase ,Reprogramming ,RC254-282 - Abstract
Simple Summary Metabolic rewiring fuels cancer proliferation by enhanced glycolysis and the increased NADH/NAD+ ratio. In this study, we highlight the critical role of NADH in the epigenetic landscape mediated by CtBP2 (C-terminal binding protein 2) activation, linking metabolism to epigenetic transcriptional reprogramming. Moreover, using metabolomics and transcriptomics integration, we show that genetic and pharmacological down-regulation of CtBP2 strongly reduces cell proliferation by modulating the redox balance, nucleotide synthesis, reactive oxygen species (ROS) generation, and scavenging. Therefore, we provide evidence that metabolic rewiring plasticity regulates the crosstalk between metabolism and the transcriptional program that sustains energetic and anabolic demands in cancer cells. Abstract Rewiring glucose metabolism toward aerobic glycolysis provides cancer cells with a rapid generation of pyruvate, ATP, and NADH, while pyruvate oxidation to lactate guarantees refueling of oxidized NAD+ to sustain glycolysis. CtPB2, an NADH-dependent transcriptional co-regulator, has been proposed to work as an NADH sensor, linking metabolism to epigenetic transcriptional reprogramming. By integrating metabolomics and transcriptomics in a triple-negative human breast cancer cell line, we show that genetic and pharmacological down-regulation of CtBP2 strongly reduces cell proliferation by modulating the redox balance, nucleotide synthesis, ROS generation, and scavenging. Our data highlight the critical role of NADH in controlling the oncogene-dependent crosstalk between metabolism and the epigenetically mediated transcriptional program that sustains energetic and anabolic demands in cancer cells. more...
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- 2021
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5. Cystic Fibrosis-Screening Positive Inconclusive Diagnosis: Newborn Screening and Long-Term Follow-Up Permits to Early Identify Patients with CFTR-Related Disorders
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Valeria Raia, Chiara Cimbalo, Laura Salvadori, Manuela Scorza, Alice Castaldo, Angela Sepe, Marcella D’Antonio, Raimondo J. Castaldo, and Antonella Tosco
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medicine.medical_specialty ,Long term follow up ,Clinical Biochemistry ,Cystic fibrosis ,Gastroenterology ,Article ,cystic fibrosis ,03 medical and health sciences ,0302 clinical medicine ,Recurrent pancreatitis ,030225 pediatrics ,Internal medicine ,medicine ,Immunoreactive trypsinogen ,Sweat test ,CF-SPID ,Newborn screening ,lcsh:R5-920 ,Bronchiectasis ,medicine.diagnostic_test ,business.industry ,newborn screening ,Cystic fibrosis screening ,genotype–phenotype correlation ,medicine.disease ,030228 respiratory system ,CFTR-RD ,business ,lcsh:Medicine (General) - Abstract
Background: Newborn screening (NBS) early-identifies cystic fibrosis (CF), but in CF-screening positive inconclusive diagnosis (CF-SPID) the results of immunoreactive trypsinogen (IRT), molecular analysis and sweat test (ST) are discordant. A percentage of CF-SPID evolves to CF, but data on long-term monitoring are lacking. We describe the follow-up of all CF and CF-SPID identified between 2008 and 2019. Methods: NBS was performed by IRT followed by molecular analysis and ST between 2008 and 2014, double IRT followed by molecular analysis and ST after 2014. Results: NBS revealed 47 CF and 99 CF-SPID newborn, a ratio 1:2.1&mdash, the highest reported so far. This depends on the identification by gene sequencing of the second variant with undefined effect in 40 CF-SPID that otherwise would have been defined as carriers. Clinical complications and pulmonary infections occurred more frequently among CF patients than among CF-SPID. Two CF-SPID cases evolved to CF (at two years), while eight evolved to CFTR-related disorders (CFTR-RD), between one and eight years, with bronchiectasis (two), recurrent pneumonia (four, two with sinonasal complications), recurrent pancreatitis (two). No clinical, biochemical or imaging data predicted the evolution. Conclusion: Gene sequencing within the NBS reveals a higher number of CF-SPID and we first describe an approach to early identify CFTR-RD, with relevant impact on their outcome. more...
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- 2020
- Full Text
- View/download PDF
6. Twelve Novel Mutations in the SLC26A3 Gene in 17 Sporadic Cases of Congenital Chloride Diarrhea
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Sabrina Cardile, Vincenza Pezzella, Licia Lugli, Giuseppe Castaldo, Ausilia Elce, Roberto Berni Canani, Sonia Giordano, Manuela Scorza, Marika Comegna, Claudio Romano, Renato Liguori, Giuseppe Cardillo, Laura Lucaccioni, Felice Amato, Amato, Felice, Cardillo, Giuseppe, Liguori, Renato, Scorza, Manuela, Comegna, Marika, Elce, Ausilia, Giordano, Sonia, Lucaccioni, Laura, Lugli, Licia, Cardile, Sabrina, Romano, Claudio, Pezzella, Vincenza, Castaldo, Giuseppe, and BERNI CANANI, Roberto more...
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Diarrhea ,Genetic Markers ,0301 basic medicine ,Genotype ,Genotyping Techniques ,Congenital chloride diarrhea ,In silico ,SLC26A3 ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Missense mutation ,Chloride-Bicarbonate Antiporters ,Genetic Testing ,Gene ,Genetics ,biology ,Gastroenterology ,medicine.disease ,030104 developmental biology ,Sulfate Transporters ,Case-Control Studies ,Mutation ,chloride losing diarrhea ,diarrhea anion exchanger ,genotype ,member 3 of solute carrier family 26 ,mutations ,splicing effect ,Metabolism, Inborn Errors ,Pediatrics, Perinatology and Child Health ,RNA splicing ,biology.protein ,Gastroenterology, chloride losing diarrhea, diarrhea anion exchanger, mutations, SLC26A3, splicing effect ,030211 gastroenterology & hepatology ,Minigene - Abstract
Objectives: We aimed to improve the knowledge of pathogenic mutations in sporadic cases of congenital chloride diarrhea (CCD) and emphasize the importance of functional studies to define the effect of novel mutations. Methods: All member 3 of solute carrier family 26 (SLC26A3) coding regions were sequenced in 17 sporadic patients with CCD. Moreover, the minigene system was used to analyze the effect of 2 novel splicing mutations. Results: We defined the SLC26A3 genotype of all 17 patients with CDD and identified 12 novel mutations. Using the minigene system, we confirmed the in silico prediction of a complete disruption of splicing pattern caused by 2 of these novel mutations: the c.971þ3_971þ4delAA and c.735þ4_c.735þ7delAGTA. Moreover, several prediction tools and a structure-function prediction defined the pathogenic role of 6 novel missense mutations. Conclusions: We confirm the molecular heterogeneity of sporadic CDD adding 12 novel mutations to the list of known pathogenic mutations. Moreover, we underline the importance, for laboratories that offer molecular diagnosis and genetic counseling, to perform fast functional analysis of novel mutations more...
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- 2017
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7. Genotype–phenotype correlation and functional studies in patients with cystic fibrosis bearing CFTR complex alleles
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Giuseppe Castaldo, Ausilia Elce, Natalia Cirilli, Marika Comegna, Antonella Miriam Di Lullo, Manuela Scorza, Adriano Angioni, Valeria Raia, Paola Iacotucci, Valentina Maria Sofia, Anna Perfetti, Roberta Cimino, Rosaria Casciaro, Carla Colombo, Felice Amato, Vincenzo Carnovale, Vincenzina Lucidi, Manuela Seia, Donatello Salvatore, Marco Lucarelli, Vito Terlizzi, Serena Quattrucci, Federica Zarrilli, Terlizzi, Vito, Castaldo, Giuseppe, Salvatore, Donatello, Lucarelli, Marco, Raia, Valeria, Angioni, Adriano, Carnovale, Vincenzo, Cirilli, Natalia, Casciaro, Rosaria, Colombo, Carla, DI LULLO, ANTONELLA MIRIAM, Elce, Ausilia, Iacotucci, Paola, Comegna, Marika, Scorza, Manuela, Lucidi, Vincenzina, Perfetti, Anna, Cimino, Roberta, Quattrucci, Serena, Seia, Manuela, Sofia, Valentina Maria, Zarrilli, Federica, and Amato, Felice more...
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Male ,0301 basic medicine ,Cystic Fibrosis ,[L997F ,Cystic Fibrosis Transmembrane Conductance Regulator ,Compound heterozygosity ,medicine.disease_cause ,Cystic fibrosis ,nasal brushing ,0302 clinical medicine ,Genotype ,[I148T ,3199del6bp] ,R117L] ,[R74W ,V201M ,D1270N] ,gating activity ,Child ,Genetics (clinical) ,Mutation ,Homozygote ,Middle Aged ,Cystic fibrosis transmembrane conductance regulator ,Phenotype ,Child, Preschool ,Female ,cystic fibrosis, genotype phenotype correlation, CFTR mutations, complex alleles, functional characterization, gating activity, chloride transport ,Adult ,Heterozygote ,medicine.medical_specialty ,Adolescent ,Biology ,Young Adult ,03 medical and health sciences ,Molecular genetics ,Genetics ,medicine ,Humans ,Genetic Predisposition to Disease ,Allele ,Alleles ,Genetic Association Studies ,Heterozygote advantage ,medicine.disease ,Molecular biology ,digestive system diseases ,Nasal Mucosa ,030104 developmental biology ,030228 respiratory system ,Immunology ,biology.protein - 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 (pT] 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. more...
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- 2016
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8. Genetic Diseases That Predispose to Early Liver Cirrhosis
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Ausilia Elce, Felice Amato, Renato Liguori, Federica Zarrilli, Giuseppe Castaldo, Manuela Scorza, Manuela, Scorza, Ausilia, Elce, Zarrilli, Federica, Renato, Liguori, Amato, Felice, and Castaldo, Giuseppe more...
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medicine.medical_specialty ,Cirrhosis ,Hepatology ,medicine.diagnostic_test ,business.industry ,Liver Cirrhosi ,HEMOCHROMATOSIS ,Review Article ,medicine.disease ,Bioinformatics ,Pathophysiology ,AAT ,Molecular analysis ,Liver biopsy ,Molecular genetics ,Medicine ,lcsh:Diseases of the digestive system. Gastroenterology ,lcsh:RC799-869 ,business ,cystic fibrosi ,Wilson disease - Abstract
Inherited liver diseases are a group of metabolic and genetic defects that typically cause early chronic liver involvement. Most are due to a defect of an enzyme/transport protein that alters a metabolic pathway and exerts a pathogenic role mainly in the liver. The prevalence is variable, but most are rare pathologies. We review the pathophysiology of such diseases and the diagnostic contribution of laboratory tests, focusing on the role of molecular genetics. In fact, thanks to recent advances in genetics, molecular analysis permits early and specific diagnosis for most disorders and helps to reduce the invasive approach of liver biopsy. more...
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- 2014
- Full Text
- View/download PDF
9. Biological role of mannose binding lectin: From newborns to centenarians
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Francesco Salvatore, Giuseppe Castaldo, Manuela Scorza, Renato Liguori, Ausilia Elce, Scorza, Manuela, Liguori, Renato, Elce, Ausilia, Salvatore, Francesco, and Castaldo, Giuseppe
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Clinical Biochemistry ,Mannose ,chemical and pharmacologic phenomena ,MBL ,Biology ,medicine.disease_cause ,Mannose-Binding Lectin ,Biochemistry ,Autoimmune Diseases ,chemistry.chemical_compound ,Haplotype ,medicine ,Humans ,Polymorphism ,Lung cancer ,Mannan-binding lectin ,Innate immunity ,Aged, 80 and over ,Polymorphism, Genetic ,Innate immune system ,Biochemistry (medical) ,Infant, Newborn ,Lectin ,Biological activity ,General Medicine ,bacterial infections and mycoses ,medicine.disease ,Immunity, Innate ,chemistry ,Immunology ,biology.protein ,Carcinogenesis - Abstract
Mannose binding lectin (MBL) is a protein of innate immunity that activates the complement and promotes opsonophagocytosis. The deficiency of MBL due to several common gene polymorphisms significantly enhances the risk of severe infections, particularly in the neonatal age and in childhood. On the contrary, the role of the protein in carcinogenesis and atherogenesis is still debated: MBL has a relevant role against neoplastic cells, but some studies described a protective effect of low levels of MBL toward breast cancer and a longer survival of lung cancer patients with a reduced MBL activity. Similarly, some studies concluded on the protective role of low levels of MBL toward cardiovascular diseases while other focused on a higher risk of myocardial infarction in subjects with a deficient activity of the protein. More recently, a role of MBL in the clearance of senescent cells emerged, and a study in two large cohorts of centenarians demonstrated that a high biological activity of the protein enhances the risk of autoimmune diseases. This body of data strongly suggests that the optimal levels of MBL activity depend on the age and on the environmental context of each subject. more...
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- 2015
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10. An Update on Laboratory Diagnosis of Liver Inherited Diseases
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Felice Amato, Ausilia Elce, Federica Zarrilli, Giuseppe Castaldo, Sonia Giordano, Manuela Scorza, Zarrilli, Federica, Ausilia, Elce, Manuela, Scorza, Sonia, Giordano, Amato, Felice, and Castaldo, Giuseppe more...
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Pathology ,medicine.medical_specialty ,Wilson’s disease ,lcsh:Medicine ,Review Article ,Degeneration (medical) ,Disease ,Cystic fibrosis ,General Biochemistry, Genetics and Molecular Biology ,cystic fibrosis ,Liver disease ,Hepatolenticular Degeneration ,alpha 1-Antitrypsin Deficiency ,Liver inherited diseases, Wilson’s disease, hepatolenticular degeneration, hereditary hemochromatosis, alpha-1 antitrypsin deficiency, cystic fibrosis ,medicine ,Humans ,Hemochromatosis ,alpha-1 antitrypsin deficiency ,Alpha 1-antitrypsin deficiency ,General Immunology and Microbiology ,alpha 1 antitrypsin deficiency, cystic fibrosis, genetic counseling, genetic disorder, genotype, hemochromatosis, molecular diagnosis, Wilson disease ,Clinical Laboratory Techniques ,business.industry ,lcsh:R ,Liver inherited diseases ,General Medicine ,hereditary hemochromatosis ,medicine.disease ,Wilson's disease ,Liver ,Hereditary hemochromatosis ,business - Abstract
Liver inherited diseases are a group of genetically determined clinical entities that appear with an early chronic liver involvement. They include Wilson’s disease (hepatolenticular degeneration), hereditary hemochromatosis, and alpha-1-antitrypsin deficiency. In addition, cystic fibrosis, although it is not specifically a liver disease, may cause a severe liver involvement in a significant percentage of cases. For all these pathologies, the disease gene is known, and molecular analysis may contribute to the unequivocal diagnosis. This approach could avoid the patient invasive procedures and limit complications associated with a delay in diagnosis. We review liver inherited diseases on the basis of the genetic defect, focusing on the contribution of molecular analysis in the multistep diagnostic workup. more...
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- 2013
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11. An 'ex vivo model' contributing to the diagnosis and evaluation of new drugs in cystic fibrosis
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G. Ilardi, Elena Cantone, A. M. Di Lullo, Giuseppe Castaldo, Manuela Scorza, Maurizio Iengo, Marika Comegna, Valeria Raia, Luigi Maiuri, Felice Amato, DI LULLO, ANTONELLA MIRIAM, Scorza, M, Amato, Felice, Comegna, Marika, Raia, Valeria, Maiuri, L, Ilardi, Gennaro, Cantone, E, Castaldo, Giuseppe, and Iengo, Maurizio more...
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Pathology ,medicine.medical_specialty ,Cystic Fibrosis ,Cystic Fibrosis Transmembrane Conductance Regulator ,Butyrate ,medicine.disease_cause ,Cystic fibrosis ,03 medical and health sciences ,0302 clinical medicine ,Genotype ,Medicine ,Humans ,CFTR ,030223 otorhinolaryngology ,Cystic Fibrosi ,Gene ,Cells, Cultured ,Mutation ,business.industry ,Nasal brushing ,CF ,Rhinology ,medicine.disease ,Transmembrane protein ,Nasal Mucosa ,General Energy ,Otorhinolaryngology ,030220 oncology & carcinogenesis ,RNA splicing ,Cancer research ,business ,Ex vivo ,Mutations - Abstract
Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. About 2000 mutations have been described so far. We setup an ex vivo model of human nasal epithelial cells (HNECs) to study CF patients testing the effect of novel mutations and molecular therapies. We performed sampling (by brushing), followed by culture and analysis of HNECs using a series of molecular techniques. We performed 50 brushings from CF patients and controls. Using cultured cells, we: i) demonstrated the widely heterogeneous CFTR expression in patients and in controls; ii) defined the splicing effect of a CFTR mutation; iii) assessed the CFTR gating activity in patients bearing different mutations; iv) demonstrated that butyrate significantly enhances CFTR expression. Based on our data, we can conclude: 1) HNEC brushing is performed without anaesthesia and is well tolerated in all CF patients (children and adults); 2) HNECs can be preserved for up to 48 hours before culture allowings multicentre studies; 3) HNECs culture can be considered a suitable model to study the molecular effects of new CFTR gene mutations and/or uncertain meaning specific mutations of carriers; 4) an ex vivo model of HNECs may be used to evaluate, before human use, the effect of new drugs on patients' cells bearing specific CFTR mutations; 5) the methodology is adequate for a quantitative measurement, by fluorescence, of the CFTR gating activity of the HNECs from patients with different genotypes identifying: a) CF patients bearing two severe mutations with an activity10% (compared to controls - 100%); b) CF patients bearing at least a mild mutation with an activity of 10-20%; c) CF carriers (heterozygous subjects) with an activity between 40-70%.La fibrosi cistica (FC) è una malattia autosomica recessiva causata da mutazioni nel gene CFTR (Cystic Fibrosis Transmembrane conductance Regulator). Finora sono state descritte circa 2000 mutazioni, ma per la maggior parte di esse è difficile definirne l’effetto senza complesse procedure in vitro. Abbiamo effettuato il campionamento (mediante brushing), la cultura e l’analisi di cellule epiteliali nasali umane (HNEC) utilizzando una serie di tecniche che possono aiutare a testare l’effetto delle mutazioni CFTR. Abbiamo eseguito 50 brushing da pazienti FC e controlli, e in 45 casi si è ottenuta una coltura positiva. Utilizzando cellule in coltura: i) abbiamo dimostrato l’espressione ampiamente eterogenea del CFTR nei pazienti e nei controlli; ii) abbiamo definito l’effetto di splicing di una mutazione sul gene CFTR; iii) abbiamo valutato l’attività di gating di CFTR in pazienti portatori di differenti mutazioni; iv) abbiamo dimostrato che il butirrato migliora in modo significativo l’espressione di CFTR. I dati provenienti dal nostro studio sperimentale dimostrano che l’uso del modello ex-vivo di cellule epiteliali nasali è un importante e valido strumento di ricerca e di diagnosi nella studio della FC e può anche essere mirato alla sperimentazione ed alla verifica di nuovi farmaci. In definitiva, in base ai nostri dati è possibile esprimere le seguenti conclusioni: 1) il prelievo delle cellule epiteliali nasali mediante brushing è applicabile senza alcuna anestesia ed è ben tollerato da tutti i pazienti affetti da FC (bambini e adulti), è scarsamente invasivo e facilmente ripetibile, è anche in grado di ottenere una sufficiente quantità di HNECs rappresentative, ben conservate, idonee allo studio della funzionalità di CFTR; 2) la conservazione delle cellule prelevate è possibile fino a 48 ore prima che si provveda all’allestimento della coltura e ciò permette di avviare studi multicentrici con prelievi in ogni sede e quindi di ottenere una ampia numerosità campionaria; 3) la coltura di cellule epiteliali nasali può essere considerata un modello adatto a studiare l’effetto molecolare di nuove mutazioni del gene CFTR e/o mutazioni specifiche di pazienti “carriers” dal significato incerto; 4) il modello ex-vivo delle HNECs consente inoltre di valutare, prima dell’impiego nell’uomo, l’effetto di farmaci (potenziatori e/o correttori) sulle cellule di pazienti portatori di mutazioni specifiche di CFTR; tali farmaci possono modulare l’espressione genica del canale CFTR aprendo così nuove frontiere terapeutiche e migliori prospettive di vita per pazienti affetti da una patologia cronica come la Fibrosi Cistica; 5) la metodologia da noi istituita risulta essere idonea alla misura quantitativa, mediante fluorescenza, dell’attività di gating del canale CFTR presente nelle membrane delle cellule epiteliali nasali prelevate da pazienti portatori di differenti genotipi; in tal modo è possibile individuare: a) pazienti FC portatori di 2 mutazioni gravi con un’attività10% (in rapporto ai controlli -100%), b) soggetti FC portatori contemporaneamente di una mutazione grave e di una lieve con un’attività tra 10-30%, c) i cosiddetti portatori “carriers”- eterozigoti - con un’attività tra 40-70%. In conclusione la possibilità di misurare l’attività del canale CFTR in HNECs fornisce un importante contributo alla diagnosi di FC, mediante individuazione di un “cut-off diagnostico”, ed anche alla previsione della gravità fenotipica della malattia; quindi quanto rilevabile dalla misura del suddetto canale permette di prospettare per il futuro la possibilità di valutare meglio i pazienti per i quali il test del sudore ha dato risultati ambigui (borderline o negativi). La metodica da noi sperimentata consente anche di monitorare i pazienti durante il trattamento farmacologico, valutando in tal modo i reali effetti delle nuove terapie. more...
- Published
- 2016
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