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12 Twenty years of newborn screening for cystic fibrosis in Eastern Saxony in Germany
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Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
10 Gene therapy to rescue of premature stop mutations R1162X and W1282X L. Cebotaru1 , C. Boinot1 , I. Sabirzhanova1 , W.B. Guggino1 . 1 Johns Hopkins University, Baltimore, United States Stop codon mutations are common in CF. To address if function can be restored by transcomplementation we infected CFBE41o- cell lines containing either R1162X or W1282X with AAV1 containing D264, or D27–264 or CFTRDR-missing residues 708–759. We found that transcomplementation with D264 or D27–264 generated CFTR currents that were approximately 2 fold greater for R1162X and 1.5 fold for W1282X compared to control. DRD transduction generated 3.5 and 4.5 more current than control for R1162X and W1282X, respectively. Conclusion: R1162X and R1282X can be rescued to a small extent with transcomplementation. However, the better option will most likely be achieved that contain AAV1-CFTR vectors which generates currents on their own.
2. Screening and Diagnosis 11 Early severe anemia as the first sign of cystic fibrosis 1 , A.T. Aslan2 , M. Kose ¨ 3 , S. Pekcan4 , F.S. Ezgu¨ 5 , I.I˙ . Budako˘glu6 , T. Si ¸ smanlar ¸ I˙ . Yenicesu7 . 1 Gazi University Medicine Faculty, Ankara, Turkey; 2 Gazi University Medicine Faculty, Pediatric Pulmonology, Ankara, Turkey; 3 Erciyes University Medicine Faculty, Pediatric Pulmonology, Ankara, Turkey; 4 Necmettin Erbakan University, Meram Medicine Faculty, Pediatric Pulmonology, Konya, Turkey; 5 Gazi University Medicine Faculty, Pediatric Metabolism and Genetic, Ankara, Turkey; 6 Gazi University Medicine Faculty, Medical Education, Ankara, Turkey; 7 Gazi University Medicine Faculty, Pediatric Hematology, Ankara, Turkey
Objectives: Severe anemia is reported to occur rarely in patients with cystic fibrosis (CF). This study aimed to determine the factors associated with early severe anemia in infants with CF. Methods: This study included 231 infants with CF from 3 pediatric CF centers between 2006 and 2015 that were retrospectively reviewed in terms of severe anemia as the first sign of CF. Factors that could affect anemia, such as age, feeding pattern, pancreatic sufficiency/insufficiency, genetic mutations, vitamin A and E levels, and the albumin level were evaluated. Clinical and laboratory findings in CF patients that presented with severe anemia and no respiratory symptoms were compared to those in CF patients that did not present with severe anemia. Results: Severe anemia as the first sign of CF was noted in 17 of 231 patients. Patient age and the albumin level differed significantly between the 2 groups of patients (P < 0.001). Vitamin E and A levels, and the types of genetic mutations did not differ between the 2 groups. The mean hemoglobin level was 5.59±0.21 g/dL and respiratory symptoms began a mean 6.3 months after diagnosis of CF in the anemia group. Conclusion: Interestingly, no differences were found associated with anemia and feeding pattern, vitamin A and E levels, pancreatic sufficiency/insufficiency, or mutations. In early infancy severe anemia in the absence of respiratory symptoms can be the first sign of CF. CF should be considered in the differential diagnosis of severe anemia in infants. Anemia can occur several months before respiratory symptoms in patients with CF. 12 Twenty years of newborn screening for cystic fibrosis in Eastern Saxony in Germany J. Hammermann1 , M. Stopsack2 . 1 University Hospital Carl Gustav Cars, TU Dresden, Department of Pediatrics, Dresden, Germany; 2 University Hospital Carl Gustav Cars, TU Dresden, Institute for Clinical Chemistry and Laboratory Medicine, Dresden, Germany Objectives: In June 1996 a regional cystic fibrosis newborn screening program started in Eastern Saxony. By now, we screened around 270 000 babies, and detected 61 children with cystic fibrosis.
Methods: In June 1996 a two tier IRT/DNA-protocol had been started, looking for the three mutations F508del, G551D and R553X. In February 2008 we changed to an IRT/PAP-protocol. All patients with pathological screening results have been contacted by our cystic fibrosis center for sweat testing and further diagnostics. Results: In the whole time we know only about four patients with a false negative screening, one of them with a meconium ileus, one asymptomatic, and one born in the 31st week of pregnancy with the screening done only at the first day of life. Four patients had a false negative sweat test, one with also negative screening, the other three patients were pancreatic sufficient and detected by pulmonary symptoms later on. 46 percent of the patients are homozygous for the mutation F508del, 48 percent heterozygous for F508del. We found 29 other mutations. 16 percent of the patients are pancreatic sufficient, 18 percent ha a meconium ileus. Birth weight and length where significant lower than in the normal german population and there was a higher rate of premature infants. Conclusion: We could show over the last twenty years that newborn screening for cystic fibrosis is practicable in Germany. Both methods IRT/DNA and IRT/PAP showed good sensitivity and specify. Clinical findings correlate good with national data, the mutation spectrum in our region differs from the all over German spectrum. 13 Cases of false negative cystic fibrosis newborn screening in Greece M. Noni1 , A. Katelari1 , A. Petrocheilou1 , E. Papakonstantinou2 , I. Loukas3 , S.-E. Doudounakis1 . 1 ‘Aghia Sophia’ Children’s Hospital, Department of Cystic Fibrosis, Athens, Greece; 2 Biochemistry, Athens, Greece; 3 University of Athens, Athens, Greece Objectives: Cystic fibrosis (CF) has not been added yet to the national newborn screening (NBS) in Greece. During the last eight years, the test is performed only in private diagnostic centers based on the immunoreactive trypsinogen (IRT) protocol. Our aim was to explain the reasons which led to an early CF diagnosis in infants with false negative IRT. Methods: Our CF Center’s population was studied retrospectively in order to identify patients that were diagnosed with CF from January 1st 2007 to December 31st 2014. Then, medical records were reviewed in order to detect those patients who have undergone a NBS program for CF. A number of patients’ characteristics such as gender, genotype, sweat test results, pancreatic function and weight of birth were also collected. Results: During the 8-year period, approximately 160,000 infants were screened with the IRT-IRT method in Greece. Nineteen infants were reported as NBS positive and 10 had false negative results. Infants with false negative results presented mainly with symptoms of dehydration (80%) and poor weight gain (20%) which led to CF diagnosis. Six of them had exocrine pancreatic sufficiency and all expressed symptoms of dehydration. Among the rest 4 patients who had pancreatic insufficiency, two presented with poor weight gain and two with symptoms of dehydration. None of them expressed symptoms from the respiratory system. Conclusion: Despite the mild genotype, the diagnosis of CF in Greek infants with false negative NBS results was set in early infancy due to warm climatic conditions. 14 Evaluating the first year following diagnosis of cystic fibrosis via screening J. Cowlard1 , C. Nwokoro1 , C. Pao1 , A. Schmidt1 . 1 Barts Health, Paediatric Respiratory Medicine, London, United Kingdom Objectives: To describe characteristics of infants identified during the first 6 yrs of screening and analyse contact with the CF team against clinical status in the year from diagnosis. Methods: A retrospective audit of children diagnosed between July 2007 and June 2013. Associations were analysed using linear regression. Primary outcome was amount of contact with the CF nurse. Results: n = 37, 18 male, 27 caucasian, 92% pancreatic insufficient, 11% meconium ileus and 41% homozygous p.Phe508del. Mean age at
Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
10 Gene therapy to rescue of premature stop mutations R1162X and W1282X L. Cebotaru1 , C. Boinot1 , I. Sabirzhanova1 , W.B. Guggino1 . 1 Johns Hopkins University, Baltimore, United States Stop codon mutations are common in CF. To address if function can be restored by transcomplementation we infected CFBE41o- cell lines containing either R1162X or W1282X with AAV1 containing D264, or D27–264 or CFTRDR-missing residues 708–759. We found that transcomplementation with D264 or D27–264 generated CFTR currents that were approximately 2 fold greater for R1162X and 1.5 fold for W1282X compared to control. DRD transduction generated 3.5 and 4.5 more current than control for R1162X and W1282X, respectively. Conclusion: R1162X and R1282X can be rescued to a small extent with transcomplementation. However, the better option will most likely be achieved that contain AAV1-CFTR vectors which generates currents on their own.
2. Screening and Diagnosis 11 Early severe anemia as the first sign of cystic fibrosis 1 , A.T. Aslan2 , M. Kose ¨ 3 , S. Pekcan4 , F.S. Ezgu¨ 5 , I.I˙ . Budako˘glu6 , T. Si ¸ smanlar ¸ I˙ . Yenicesu7 . 1 Gazi University Medicine Faculty, Ankara, Turkey; 2 Gazi University Medicine Faculty, Pediatric Pulmonology, Ankara, Turkey; 3 Erciyes University Medicine Faculty, Pediatric Pulmonology, Ankara, Turkey; 4 Necmettin Erbakan University, Meram Medicine Faculty, Pediatric Pulmonology, Konya, Turkey; 5 Gazi University Medicine Faculty, Pediatric Metabolism and Genetic, Ankara, Turkey; 6 Gazi University Medicine Faculty, Medical Education, Ankara, Turkey; 7 Gazi University Medicine Faculty, Pediatric Hematology, Ankara, Turkey
Objectives: Severe anemia is reported to occur rarely in patients with cystic fibrosis (CF). This study aimed to determine the factors associated with early severe anemia in infants with CF. Methods: This study included 231 infants with CF from 3 pediatric CF centers between 2006 and 2015 that were retrospectively reviewed in terms of severe anemia as the first sign of CF. Factors that could affect anemia, such as age, feeding pattern, pancreatic sufficiency/insufficiency, genetic mutations, vitamin A and E levels, and the albumin level were evaluated. Clinical and laboratory findings in CF patients that presented with severe anemia and no respiratory symptoms were compared to those in CF patients that did not present with severe anemia. Results: Severe anemia as the first sign of CF was noted in 17 of 231 patients. Patient age and the albumin level differed significantly between the 2 groups of patients (P < 0.001). Vitamin E and A levels, and the types of genetic mutations did not differ between the 2 groups. The mean hemoglobin level was 5.59±0.21 g/dL and respiratory symptoms began a mean 6.3 months after diagnosis of CF in the anemia group. Conclusion: Interestingly, no differences were found associated with anemia and feeding pattern, vitamin A and E levels, pancreatic sufficiency/insufficiency, or mutations. In early infancy severe anemia in the absence of respiratory symptoms can be the first sign of CF. CF should be considered in the differential diagnosis of severe anemia in infants. Anemia can occur several months before respiratory symptoms in patients with CF. 12 Twenty years of newborn screening for cystic fibrosis in Eastern Saxony in Germany J. Hammermann1 , M. Stopsack2 . 1 University Hospital Carl Gustav Cars, TU Dresden, Department of Pediatrics, Dresden, Germany; 2 University Hospital Carl Gustav Cars, TU Dresden, Institute for Clinical Chemistry and Laboratory Medicine, Dresden, Germany Objectives: In June 1996 a regional cystic fibrosis newborn screening program started in Eastern Saxony. By now, we screened around 270 000 babies, and detected 61 children with cystic fibrosis.
Methods: In June 1996 a two tier IRT/DNA-protocol had been started, looking for the three mutations F508del, G551D and R553X. In February 2008 we changed to an IRT/PAP-protocol. All patients with pathological screening results have been contacted by our cystic fibrosis center for sweat testing and further diagnostics. Results: In the whole time we know only about four patients with a false negative screening, one of them with a meconium ileus, one asymptomatic, and one born in the 31st week of pregnancy with the screening done only at the first day of life. Four patients had a false negative sweat test, one with also negative screening, the other three patients were pancreatic sufficient and detected by pulmonary symptoms later on. 46 percent of the patients are homozygous for the mutation F508del, 48 percent heterozygous for F508del. We found 29 other mutations. 16 percent of the patients are pancreatic sufficient, 18 percent ha a meconium ileus. Birth weight and length where significant lower than in the normal german population and there was a higher rate of premature infants. Conclusion: We could show over the last twenty years that newborn screening for cystic fibrosis is practicable in Germany. Both methods IRT/DNA and IRT/PAP showed good sensitivity and specify. Clinical findings correlate good with national data, the mutation spectrum in our region differs from the all over German spectrum. 13 Cases of false negative cystic fibrosis newborn screening in Greece M. Noni1 , A. Katelari1 , A. Petrocheilou1 , E. Papakonstantinou2 , I. Loukas3 , S.-E. Doudounakis1 . 1 ‘Aghia Sophia’ Children’s Hospital, Department of Cystic Fibrosis, Athens, Greece; 2 Biochemistry, Athens, Greece; 3 University of Athens, Athens, Greece Objectives: Cystic fibrosis (CF) has not been added yet to the national newborn screening (NBS) in Greece. During the last eight years, the test is performed only in private diagnostic centers based on the immunoreactive trypsinogen (IRT) protocol. Our aim was to explain the reasons which led to an early CF diagnosis in infants with false negative IRT. Methods: Our CF Center’s population was studied retrospectively in order to identify patients that were diagnosed with CF from January 1st 2007 to December 31st 2014. Then, medical records were reviewed in order to detect those patients who have undergone a NBS program for CF. A number of patients’ characteristics such as gender, genotype, sweat test results, pancreatic function and weight of birth were also collected. Results: During the 8-year period, approximately 160,000 infants were screened with the IRT-IRT method in Greece. Nineteen infants were reported as NBS positive and 10 had false negative results. Infants with false negative results presented mainly with symptoms of dehydration (80%) and poor weight gain (20%) which led to CF diagnosis. Six of them had exocrine pancreatic sufficiency and all expressed symptoms of dehydration. Among the rest 4 patients who had pancreatic insufficiency, two presented with poor weight gain and two with symptoms of dehydration. None of them expressed symptoms from the respiratory system. Conclusion: Despite the mild genotype, the diagnosis of CF in Greek infants with false negative NBS results was set in early infancy due to warm climatic conditions. 14 Evaluating the first year following diagnosis of cystic fibrosis via screening J. Cowlard1 , C. Nwokoro1 , C. Pao1 , A. Schmidt1 . 1 Barts Health, Paediatric Respiratory Medicine, London, United Kingdom Objectives: To describe characteristics of infants identified during the first 6 yrs of screening and analyse contact with the CF team against clinical status in the year from diagnosis. Methods: A retrospective audit of children diagnosed between July 2007 and June 2013. Associations were analysed using linear regression. Primary outcome was amount of contact with the CF nurse. Results: n = 37, 18 male, 27 caucasian, 92% pancreatic insufficient, 11% meconium ileus and 41% homozygous p.Phe508del. Mean age at