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WS09.5 The association between disease severity and telomere length in lung tissue and peripheral blood leukocytes of cystic fibrosis patients
Workshops / Journal of Cystic Fibrosis 16S1 (2017) S1–S62
heritability of phenotypic traits was evaluated we confirmed the influence of genetic factors rather than environmental factors among the CC lines during P. aeruginosa infection. Quantitative trait locus (QTL) mapping detect a significant locus on Chromosome 3 associated to the severity of infection after seven days post infection (mortality). We refined QTL localization, exploiting the sequence variations of the eight inbred strain founders of the CC and gene/protein public databases, and we identified 14 genes as new candidate diseasemodifiers of the airways infection. Conclusion: Novel disease models (e.g. CC mice) have been instrumental for demonstrating that P. aeruginosa pneumonia is linked to host genetic determinants and may be used to complement human studies. WS09.5 The association between disease severity and telomere length in lung tissue and peripheral blood leukocytes of cystic fibrosis patients E.J. Lammertyn1, D.S. Martens2, K. Colpaert3, P.C. Goeminne3,4, B.M. Vanaudenaerde1, T.S. Nawrot2,5, L.J. Dupont3. 1KU Leuven, Department of Clinical and Experimental Medicine, Laboratory of Respiratory Diseases, Leuven, Belgium; 2Hasselt University, Centre for Environmental Sciences, Hasselt, Belgium; 3University Hospitals Leuven, Department of Respiratory Diseases, Cystic Fibrosis Unit, Leuven, Belgium; 4AZ Nikolaas, Department of Respiratory Diseases, Sint-Niklaas, Belgium; 5KU Leuven, Department of Public Health and Primary Care, Leuven, Belgium Objectives: As 57% of Belgian CF patients is currently older than 18 y, clinicians become increasingly challenged by aging and age-related diseases. The molecular mechanisms behind aging remain unclear but cellular senescence triggered by telomere shortening is emerging as research target. For the first time, we investigated both lung tissue telomere length (TL) and leukocyte telomere length (LTL) as a predictor of disease severity. Methods: 12 CF explant lungs and 10 unused donor lungs (controls) were frozen at total lung capacity and cut into slices of 2 cm. 4 samples of each lung (equally from upper and lower lobe, per lobe 1 sample from normal looking and abnormal looking tissue as evaluated on HRCT, randomly for controls) were analysed. Secondly, peripheral blood from 176 CF patients (aged 0–72) was collected and buffy coat DNA was extracted. Relative tissue TL and LTL was measured using a real-time qPCR method. Results: Tissue TL demonstrated high variability within the same lung both in CF (mean log T/S ratio = 1.29 ± 11%) and in controls (mean log T/S ratio = 1.11 ± 17%). There was no difference in TL between CF and control lungs after correcting for age or between normal and abnormal looking CF tissue. Using multivariate models, peripheral blood LTL was correlated with several clinical measures of disease severity: for every 20% decrease in LTL, FEV1 decreased with 6.1% ( p = 0.020) in male but not in female CF patients. Diagnosis of CF asthma and treatment with inhaled corticosteroids was associated with a -8.5% ( p = 0.040) shorter LTL in male CF patients. Furthermore, male homozygous CF patients demonstrated a −8.2% ( p = 0.054) shorter LTL than heterozygotes. Conclusion: Our results suggest increased cellular senescence through shorter LTL in male CF patients with more severe lung disease, ΔF508 homozygosity and CF asthma. In contrast, tissue TL analysis did not confirm the presence of increased cellular senescence in severely damaged regions of the end-stage CF lung. WS09.6 Telomere length in cystic fibrosis patients – are patients with CF ageing too quickly? I. Ashworth1, J. Dodge2, K. Norris1, M. Kelson3, D. Baird1, J. Duckers4. 1Cardiff University, School of Medicine, Division of Cancer and Genetics, Cardiff, United Kingdom; 2Swansea University, Swansea, United Kingdom; 3Cardiff University, Cardiff, United Kingdom; 4University Hospital Llandough, All Wales Adult CF Centre, Cardiff, United Kingdom Introduction: Telomeres cap the ends of eukaryotic chromosomes. As cells divide, telomeres in normal human somatic cells progressively shorten, ultimately reaching a length that triggers cell-cycle arrest, known as replicative senescence. Telomere erosion and senescence is considered to underpin the ageing process in humans and provides a useful marker of
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biological age. We aimed to see if telomere length is associated with the age of the patient and the severity of their CF condition. Methods: Twenty patients, aged 17–43 years, attending annual review at the AWACFC consented for blood to be taken, in addition to the clinical sample taken as part of their standard care. Telomere length profiles were assessed from the blood samples, using the high-resolution single telomere length analysis technique (STELA). Telomere length profiles were correlated with patient demographics and measures of severity of CF. Patient telomere lengths were also compared to those of healthy individuals of a range of ages. Results: Patients with CF had significantly shorter telomere lengths than healthy individuals, when adjusting for age ( p = <0.001). CF patients were found to have telomere lengths equivalent to those observed in normal individuals around 20 years older than them. There was no clear correlation between patients FEV1% predicted, BMI and telomere length. Conclusions: In conclusion, patients with CF tend to have shorter telomeres than healthy individuals of the same age. This suggests that there may be some premature ageing in CF patients. A larger sample size would be useful to determine any correlations between telomere length, age and the severity of CF.
CF diagnosis: new tests explored and old tests re-appraised WS10.1 Measuring the angulation of the uncinate process: a new specific tool for cystic fibrosis diagnosis? V. Escabasse1,2,3, R. Hervochon4, J.R. Blondeau5, N. Remus6, L. Bassinet6, F. Canoui Poitrine7, T. Van Den Abbeele8. 1Centre Hospitalier Intercommunal de Créteil, Ear Nose and Throat Department and Cystic Fibrosis Centre, Créteil, France; 2INSERM 955, équipe 5, Créteil, France; 3Université Paris Est Créteil, Créteil, France; 4Centre Hospitalier Intercommunal de Creteil, ENT, Créteil, France; 5Centre Hospitalier Intercommunal de Créteil, Créteil, France; 6Centre Hospitalier Intercommunal de Créteil, CRCM, Créteil, France; 7CHU H Mondor, Santé Publique, Créteil, France; 8Robert Debré Pediatric Hospital, Ear Nose and Throat Department, Paris, France Objectives: Computed Tomography (CT) criteria of Cystic Fibrosis (CF) chronic rhinosinusitis combine: sinuses filling, bulging of the maxillary sinus inner wall and hypoplasia of frontal sinuses. In CFpatients, the uncinate process (UP) is deflected inwards, protruding in the nasal cavity. The purpose of the study is to objectify by angles measurements on CT-scan the deviation of UP and to compare CF to other chronic rhinosinusitis and control patients. Methods: We included retrospectively in two university hospitals with CF centre care: 30 CF patients, 30 primary ciliary dyskinesia (PCD), 30 nasal polyposis (NP) and 30 control patients. We measured bilaterally: angles A, B and C on coronal sections perpendicular to the hard palate, and angles D and E on axial sections parallel to the hard palate. Angle A is formed by the UP and the inner wall of orbit, the others by the UP and the midline. Student’s test was used, with the null hypothesis of equal means for each angle between two groups. Results: There is no significant difference for each angle between groups: control, PCD and NP. CF patients had 3 statistically different angles with controls, 5 with NP, and 4 with PCD patients. The angle A has an average value of 126 (±16)° in CF patients, 138 (±19)° in controls ( p = 0.007), 145 (±15)° in PCD ( p = 0.001), and 138 (±14)° in NP ( p = 0.001). The angle E has an average value of 35 (±10)° in CF patients, 20 (±6)° in controls ( p < 0.001), 21 (±4)° in PCD ( p < 0.001), and 22 (±6)° in NP ( p < 0.001). Conclusion: This work highlights that UP’s anatomy is significantly different in CF from other chronic rhinosinusitis. In CF patients, the angle between PU and the inner wall of orbit in coronal sections is closed, and angles between PU and midline are opened. These measures support endoscopic observations and could be a new element for CF diagnosis.
heritability of phenotypic traits was evaluated we confirmed the influence of genetic factors rather than environmental factors among the CC lines during P. aeruginosa infection. Quantitative trait locus (QTL) mapping detect a significant locus on Chromosome 3 associated to the severity of infection after seven days post infection (mortality). We refined QTL localization, exploiting the sequence variations of the eight inbred strain founders of the CC and gene/protein public databases, and we identified 14 genes as new candidate diseasemodifiers of the airways infection. Conclusion: Novel disease models (e.g. CC mice) have been instrumental for demonstrating that P. aeruginosa pneumonia is linked to host genetic determinants and may be used to complement human studies. WS09.5 The association between disease severity and telomere length in lung tissue and peripheral blood leukocytes of cystic fibrosis patients E.J. Lammertyn1, D.S. Martens2, K. Colpaert3, P.C. Goeminne3,4, B.M. Vanaudenaerde1, T.S. Nawrot2,5, L.J. Dupont3. 1KU Leuven, Department of Clinical and Experimental Medicine, Laboratory of Respiratory Diseases, Leuven, Belgium; 2Hasselt University, Centre for Environmental Sciences, Hasselt, Belgium; 3University Hospitals Leuven, Department of Respiratory Diseases, Cystic Fibrosis Unit, Leuven, Belgium; 4AZ Nikolaas, Department of Respiratory Diseases, Sint-Niklaas, Belgium; 5KU Leuven, Department of Public Health and Primary Care, Leuven, Belgium Objectives: As 57% of Belgian CF patients is currently older than 18 y, clinicians become increasingly challenged by aging and age-related diseases. The molecular mechanisms behind aging remain unclear but cellular senescence triggered by telomere shortening is emerging as research target. For the first time, we investigated both lung tissue telomere length (TL) and leukocyte telomere length (LTL) as a predictor of disease severity. Methods: 12 CF explant lungs and 10 unused donor lungs (controls) were frozen at total lung capacity and cut into slices of 2 cm. 4 samples of each lung (equally from upper and lower lobe, per lobe 1 sample from normal looking and abnormal looking tissue as evaluated on HRCT, randomly for controls) were analysed. Secondly, peripheral blood from 176 CF patients (aged 0–72) was collected and buffy coat DNA was extracted. Relative tissue TL and LTL was measured using a real-time qPCR method. Results: Tissue TL demonstrated high variability within the same lung both in CF (mean log T/S ratio = 1.29 ± 11%) and in controls (mean log T/S ratio = 1.11 ± 17%). There was no difference in TL between CF and control lungs after correcting for age or between normal and abnormal looking CF tissue. Using multivariate models, peripheral blood LTL was correlated with several clinical measures of disease severity: for every 20% decrease in LTL, FEV1 decreased with 6.1% ( p = 0.020) in male but not in female CF patients. Diagnosis of CF asthma and treatment with inhaled corticosteroids was associated with a -8.5% ( p = 0.040) shorter LTL in male CF patients. Furthermore, male homozygous CF patients demonstrated a −8.2% ( p = 0.054) shorter LTL than heterozygotes. Conclusion: Our results suggest increased cellular senescence through shorter LTL in male CF patients with more severe lung disease, ΔF508 homozygosity and CF asthma. In contrast, tissue TL analysis did not confirm the presence of increased cellular senescence in severely damaged regions of the end-stage CF lung. WS09.6 Telomere length in cystic fibrosis patients – are patients with CF ageing too quickly? I. Ashworth1, J. Dodge2, K. Norris1, M. Kelson3, D. Baird1, J. Duckers4. 1Cardiff University, School of Medicine, Division of Cancer and Genetics, Cardiff, United Kingdom; 2Swansea University, Swansea, United Kingdom; 3Cardiff University, Cardiff, United Kingdom; 4University Hospital Llandough, All Wales Adult CF Centre, Cardiff, United Kingdom Introduction: Telomeres cap the ends of eukaryotic chromosomes. As cells divide, telomeres in normal human somatic cells progressively shorten, ultimately reaching a length that triggers cell-cycle arrest, known as replicative senescence. Telomere erosion and senescence is considered to underpin the ageing process in humans and provides a useful marker of
S17
biological age. We aimed to see if telomere length is associated with the age of the patient and the severity of their CF condition. Methods: Twenty patients, aged 17–43 years, attending annual review at the AWACFC consented for blood to be taken, in addition to the clinical sample taken as part of their standard care. Telomere length profiles were assessed from the blood samples, using the high-resolution single telomere length analysis technique (STELA). Telomere length profiles were correlated with patient demographics and measures of severity of CF. Patient telomere lengths were also compared to those of healthy individuals of a range of ages. Results: Patients with CF had significantly shorter telomere lengths than healthy individuals, when adjusting for age ( p = <0.001). CF patients were found to have telomere lengths equivalent to those observed in normal individuals around 20 years older than them. There was no clear correlation between patients FEV1% predicted, BMI and telomere length. Conclusions: In conclusion, patients with CF tend to have shorter telomeres than healthy individuals of the same age. This suggests that there may be some premature ageing in CF patients. A larger sample size would be useful to determine any correlations between telomere length, age and the severity of CF.
CF diagnosis: new tests explored and old tests re-appraised WS10.1 Measuring the angulation of the uncinate process: a new specific tool for cystic fibrosis diagnosis? V. Escabasse1,2,3, R. Hervochon4, J.R. Blondeau5, N. Remus6, L. Bassinet6, F. Canoui Poitrine7, T. Van Den Abbeele8. 1Centre Hospitalier Intercommunal de Créteil, Ear Nose and Throat Department and Cystic Fibrosis Centre, Créteil, France; 2INSERM 955, équipe 5, Créteil, France; 3Université Paris Est Créteil, Créteil, France; 4Centre Hospitalier Intercommunal de Creteil, ENT, Créteil, France; 5Centre Hospitalier Intercommunal de Créteil, Créteil, France; 6Centre Hospitalier Intercommunal de Créteil, CRCM, Créteil, France; 7CHU H Mondor, Santé Publique, Créteil, France; 8Robert Debré Pediatric Hospital, Ear Nose and Throat Department, Paris, France Objectives: Computed Tomography (CT) criteria of Cystic Fibrosis (CF) chronic rhinosinusitis combine: sinuses filling, bulging of the maxillary sinus inner wall and hypoplasia of frontal sinuses. In CFpatients, the uncinate process (UP) is deflected inwards, protruding in the nasal cavity. The purpose of the study is to objectify by angles measurements on CT-scan the deviation of UP and to compare CF to other chronic rhinosinusitis and control patients. Methods: We included retrospectively in two university hospitals with CF centre care: 30 CF patients, 30 primary ciliary dyskinesia (PCD), 30 nasal polyposis (NP) and 30 control patients. We measured bilaterally: angles A, B and C on coronal sections perpendicular to the hard palate, and angles D and E on axial sections parallel to the hard palate. Angle A is formed by the UP and the inner wall of orbit, the others by the UP and the midline. Student’s test was used, with the null hypothesis of equal means for each angle between two groups. Results: There is no significant difference for each angle between groups: control, PCD and NP. CF patients had 3 statistically different angles with controls, 5 with NP, and 4 with PCD patients. The angle A has an average value of 126 (±16)° in CF patients, 138 (±19)° in controls ( p = 0.007), 145 (±15)° in PCD ( p = 0.001), and 138 (±14)° in NP ( p = 0.001). The angle E has an average value of 35 (±10)° in CF patients, 20 (±6)° in controls ( p < 0.001), 21 (±4)° in PCD ( p < 0.001), and 22 (±6)° in NP ( p < 0.001). Conclusion: This work highlights that UP’s anatomy is significantly different in CF from other chronic rhinosinusitis. In CF patients, the angle between PU and the inner wall of orbit in coronal sections is closed, and angles between PU and midline are opened. These measures support endoscopic observations and could be a new element for CF diagnosis.