Assessment of telomere length in the combined pulmonary fibrosis and emphysema syndrome AV Cardoso 1, PC Mota 1,2, A Palmeiro 3, P Rendeiro 3, N Melo 1, JM Pereira 4, CS Moura 5, O Sokhatska 2,6, A Morais 1,2 1 Department of Pneumology, 2 Faculty of Medicine of University of Porto, 3 Centre of Clinical Genetic of Porto, 4 Department of Radiology, 5 Departmente of Anatomical Pathology, 6 Department and Laboratory of Immunology Centro Hospitalar de São João, Porto, Portugal

INTRODUTION Telomeres Telomerase (2 components: telomerase reverse transcriptase - TERT; telomerase RNA -TR) Telomere length shortens progressively during cell division Critically short telomeres signal a DNA damage response that can lead to apoptosis or induce to senescence Role of telomeres in age-related diseases

INTRODUTION Telomeres shortening (TS) has been associated to idiopathic pulmonary fibrosis (IPF) with or without mutations in the telomerase components (TERT and TR) TS was observed in lungs of patients with emphysema Is TS more prevalent and significant in the combined pulmonary fibrosis and emphysema (CPFE) syndrome?

OBJECTIVE Assessment of TS in CPFE patients in comparison with IPF patients without emphysema

MATERIAL AND METHODS Extraction of DNA from peripheral blood samples of patients with IPF followed in Diffuse Lung Diseases outpatient clinic Assessment of telomeres length: qPCR with SYBR® Green Ratio between the number of copies of the telomeres repeat (T) and single copy gene (S) on the samples, relatively to a standard sample (T/S ratio) T/S ratio < 1: telomere length shorter than of the standard DNA

MATERIAL AND METHODS Telomere length was compared between IPF without emphysema patients and CPFE patients The extent of emphysema in CPFE patients was quantified in > 20% and < 20%, and telomere length was evaluated in these two groups Statistical analysis – IBM SPSS Statistics (version 19) t test or Mann-Whitney U test (continuous variables) and Chi-square test or Fischer’s exact test (categorial variables) P value of less than 0.05 was considered significant

RESULTS IPF n = 33 CPFE n = 17 (51%) emphysema > 20% n = 10 (59%) emphysema < 20% n = 7 (41%) IPF without emphysema n = 16 (49%)

RESULTS FPI without emphysema (n=16) CPFE (n=17) P value Male, n (%)11 (69%)16 (94%)NS Age (years), mean ± SD71.9 ± ± 11.1NS Smoking n (%)9 (56%)15(88%)0.045 PFT, mean ± SD FEV1 (%) FVC (%) FEV1/FVC TLC (%) DLCO (%) 80.1 ± ± ± ± ± ± ± ± ± ± 22.7 NS PaO2 (mmHg), mean ± SD78.1 ± ± 11.4NS Table 1. Socio-demographic and functional characteristics of patients with IPF without emphysema (n=16) and CPFE (n=17)

RESULTS Table 2. Assessment and comparison of telomeres length in two groups of patients (CPFE and IPF without emphysema) CPFE (n=17) IPF without emphysema (n=16) P value T/S ratio, median (IQ range) 0.7 ( )1.0 ( )0,22 T/S ratio < 1, n (%)13 (77%)8 (50%)0,11

RESULTS Table 3. Assessment and comparison of telomeres length in two groups of patients with CPFE (> 20% and < 20% of emphysema) CPFE >20% emphysema (n=10) CPFE <20% emphysema (n=7) P value T/S ratio, median (IQ range) 0.8 ( )0.6 ( )0.96 T/S ratio < 1, n (%)8 (80%)5(71%)1.00

CONCLUSION Despite the lack of statistical significance, TS is more frequent in CPFE patients compared to IPF patients without emphysema, suggesting that this change is more significant in patients with this syndrome However, studies with larger sample sizes are required to better determine the role of this genetic mechanism

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