Search

Introduction: Interstitial lung diseases (ILD), such as idiopathic pulmonary fibrosis (IPF), are diseases of elderly people that are characterised by pulmonary deposition of fibrous tissue and often have a poor prognosis. Interstitial lung abnormalities (ILA) are radiologic changes that are similar in appearance to ILD but are characterised in cohort study participants without known ILD. Prior research findings, including risk factors parallels such as advanced age, suggest that ILA are likely a precursor to ILD in some cases and are themselves associated with poor health outcomes. Due to the strong links that ILA and ILD share with advanced aging, the aims of the thesis were to assess the relationship of ILA with aging-related biological markers and outcomes in four papers. These were functional status, pulmonary and extra-pulmonary malignancies, a variety of blood proteins and leukocyte telomere length. Methods: Data from three cohort studies, the Age/Gene-Environment Susceptibility-Reykjavik (AGES-Reykjavik) study, the Genetic epidemiology of COPD (COPDGene) study and the Framingham Heart study were used. CT images of participants had been manually read with regards to ILA status. Outcomes were ascertained from the cohorts’ phenotyping data and from electronic medical records. The associations of three self-reported measures of health and functional status with ILA were assessed with logistic regression. The relationship of ILA with diagnoses of and mortality from pulmonary and non-pulmonary malignancies was assessed with Gray’s tests and proportional hazards models. The associations of ILA, ILA progression and single nucleotide polymorphisms associated with pulmonary fibrosis with protein markers were assessed with adjusted regression models of single proteins. Adaptive LASSO modelling of bootstrap data samples was used to find sets of proteins predictive of ILA and ILA progression. The relations of ILA with leukocyte telomere length and telomere length with mortality among those with ILA were modelled with regression and Cox proportional hazards models. Results: In the AGES-Reykjavik cohort, participants with ILA were less likely to be independent in activities of daily living (odds ratio (OR) 0.70, 95% confidence interval (CI) 0.55-0.90), to perceive their health as good or better (OR 0.66, CI 0.52-0.82) and to be regularly physically active (OR 0.72, CI 0.56-0.91). The cumulative incidences of lung cancer diagnoses and lung cancer mortality were higher among participants with ILA than others (p < 0.001), but such differences were not found for other cancers. In adjusted Cox proportional hazards models, ILA were associated with diagnoses of lung cancer (hazard ratio (HR) 2.77, CI 1.76-4.36) and mortality from lung cancer (HR 2.89, CI 1.80-4.66) but not with diagnoses of, or mortality from, cancers excluding lung cancer. In adjusted analyses of single serum proteins and ILA in AGES-Reykjavik, 287 proteins were significantly associated with ILA. The most significant associations were for Surfactant protein B (SFTPB), Secretoglobin family 3A member 1 (SCGB3A1) and WAP four-disulfide core domain protein 2 (WFDC2). An eight-protein model of ILA status was created based on inclusion in 200 adaptive LASSO models in bootstrap data samples which had an area under the receiving operator characteristic curve (AUROC) value of 0.880 after bootstrap validation. Single-protein results for SFTPB, SCGB3A1 and WFDC2 and the eight-protein model were validated in COPDGene (validated AUROC 0.826). Similarly, 121 proteins were associated with ILA progression in single protein models in AGES-Reykjavik and multi-protein modelling yielded a four-protein model which had a validated AUROC of 0.824. In analyses of fibrosis-associated genetic polymorphisms, the rs35705950 polymorphism near the MUC5B gene was found to be associated with levels of SFTPB (β 0.26, p = 8×10-18). As for analyses of ILA and leukocyte telomere length, shorter telomere length was associated with increased odds of ILA in COPDGene (OR 2.2, CI 1.5-3.4) and AGES-Reykjavik (OR 2.6, CI 1.4-4.9) and ILA was associated with shorter telomere length in FHS (β -767 base pairs, CI -76 - -1584). Adjusted Cox proportional hazards models did not demonstrate a significant association of telomere length with mortality among those with ILA in COPDGene nor in AGES-Reykjavik. Conclusions: These cohort study-based data demonstrate the associations of ILA with several aging-related markers and outcomes. These are subjective markers of functional status, diagnoses of and mortality from pulmonary malignancies and telomere length in leukocytes. Additionally, many novel protein biomarkers of ILA are proposed, some of them previously related to aging. These findings improve knowledge of biological markers and epidemiological outcomes associated with ILA as a possible marker of early pulmonary fibrosis., Eimskipasjóður Háskóla Íslands

Publisher's version (útgefin grein), In lung adenocarcinoma, low lamin A expression in pleural metastatic cells has been proposed as a pejorative factor. miR-9 physiologically inhibits the expression of lamin A in neural cells and seems to be a central actor in the carcinogenesis and the metastatic process in lung cancer. Thus, it could be a good candidate to explain the reduction of lamin A expression in lung adenocarcinoma cells. miR-9 expression was analyzed in 16 pleural effusions containing metastatic cells from lung adenocarcinoma and was significantly reduced in patients from the ‘Low lamin A expression’ group compared to patients from the ‘High lamin A expression’ group. Then, carcinoma cells selection by fluorescence-activated cell sorting (FACS) was performed according to epithelial membrane antigen (EMA) expression, reflecting lamin A expression. miR-9 was underexpressed in lamin A– carcinoma cells compared to lamin A+ carcinoma cells in patients from the ‘Low lamin A expression’ group, whereas there was no difference of miR-9 expression between lamin A+ and lamin A– carcinoma cells in patients from the ‘High lamin A expression’ group. These results suggest that miR-9 does not regulate lamin A expression in metastatic cells from lung adenocarcinoma. On the contrary, miR-9 expression was shown to be reduced in lamin A-negative carcinoma cells., This research was funded by an ARARD (Association Régionale d’Aide Respiratoire à Domicile) grant, Parc d’activités de Napollon, 100 avenue des Templiers, Aubagne Cedex, France.

Publisher's version (útgefin grein), Rationale: Idiopathic pulmonary fibrosis (IPF) is a complex lung disease characterized by scarring of the lung that is believed to result from an atypical response to injury of the epithelium. Genome-wide association studies have reported signals of association implicating multiple pathways including host defense, telomere maintenance, signaling, and cell-cell adhesion. Objectives: To improve our understanding of factors that increase IPF susceptibility by identifying previously unreported genetic associations. Methods: We conducted genome-wide analyses across three independent studies and meta-analyzed these results to generate the largest genome-wide association study of IPF to date (2,668 IPF cases and 8,591 controls). We performed replication in two independent studies (1,456 IPF cases and 11,874 controls) and functional analyses (including statistical fine-mapping, investigations into gene expression, and testing for enrichment of IPF susceptibility signals in regulatory regions) to determine putatively causal genes. Polygenic risk scores were used to assess the collective effect of variants not reported as associated with IPF. Measurements and Main Results: We identified and replicated threenewgenome-wide significant (P, R.J.A. is an Action for Pulmonary Fibrosis Research Fellow. L.V.W. holds a GSK/British Lung Foundation Chair in Respiratory Research. R.G.J. is supported by a National Institute for Health Research (NIHR) Research Professorship (NIHR reference RP-2017-08-ST2-014). I.N. is supported by the NHLBI (R01HL130796). B.G.-G. is funded by Agencia Canaria de Investigación, Innovación y Sociedad de la Información (TESIS2015010057) cofunded by European Social Fund. J.M.O. is supported by the NHLBI (K23HL138190). C.F. is supported by the Spanish Ministry of Science, Innovation and Universities (grant RTC-2017-6471-1; Ministerio de Ciencia e Innovacion/Agencia Estatal de Investigación/Fondo Europeo de Desarrollo Regional, Unión Europea) cofinanced by the European Regional Development Funds “A way of making Europe” from the European Union and by agreement OA17/008 with Instituto Tecnológico y de Energías Renovables to strengthen scientific and technological education, training, research, development and innovation in Genomics, Personalized Medicine and Biotechnology. The Spain Biobank array genotyping service was performed at CEGEN-PRB3-ISCIII, which is supported by PT17/0019, of the PE I+D+i 2013–2016, funded by Instituto de Salud Carlos III, and cofinanced by the European Regional Development Funds. P.L.M. is an Action for Pulmonary Fibrosis Research Fellow. M.O. is a fellow of the Parker B. Francis Foundation and a Scholar of the Michael Smith Foundation for Health Research. B.D.H. is supported by NIH K08 HL136928, Parker B. Francis Research Opportunity Award. M.H.C. and G.M.H. are supported by NHLBI grants R01HL113264 (M.H.C.), R01HL137927 (M.H.C.), R01HL135142 (M.H.C. and G.M.H.), R01111024 (G.M.H.), and R01130974 (G.M.H.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The funding body has no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript. T.M.M. is supported by an NIHR Clinician Scientist Fellowship (NIHR Ref: CS-2013-13-017) and a British Lung Foundation Chair in Respiratory Research (C17-3). M.D.T. is supported by a Wellcome Trust Investigator Award (WT202849/Z/16/Z). The research was partially supported by the NIHR Leicester Biomedical Research Centre; the views expressed are those of the author(s) and not necessarily those of the National Health Service (NHS), the NIHR, or the Department of Health. I.P.H. was partially supported by the NIHR Nottingham Biomedical Research Centre; the views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, or the Department of Health. I.S. is supported by Medical Research Council (G1000861) and Asthma UK (AUK-PG-2013-188). D.F. was supported by an Intermediate Fellowship from the Wellcome Trust (097152/Z/11/Z). This work was partially supported by the National Institute for Health Research (NIHR) Oxford Biomedical Research Centre. V.N. is funded by an NIHR Clinical Lectureship. G.G. is supported by project grant 141513-051 from the Icelandic Research Fund and Landspitali Scientific Fund A-2016-023, A-2017-029, and A-2018-025. D.J.L. and A.M. are supported by Multi-Ethnic Study of Atherosclerosis (MESA) and the MESA SNP Health Association Resource (SHARe) project are conducted and supported by the NHLBI in collaboration with MESA investigators. Support for MESA is provided by contracts HHSN268201500003I, N01-HC-95159, N01-HC-95160, N01-HC-95161, N01-HC-95162, N01-HC-95163, N01-HC-95164, N01-HC-95165, N01-HC-95166, N01-HC-95167, N01-HC-95168, N01-HC-95169, UL1-TR-000040, UL1-TR-001079, UL1-TR-001420, UL1-TR-001881, and DK063491. Funding for SHARe genotyping was provided by NHLBI Contract N02-HL-64278. Genotyping was performed at Affymetrix (Santa Clara, California) and the Broad Institute of Harvard and Massachusetts Institute of Technology (Boston, Massachusetts) using the Affymetrix Genome-Wide Human SNP Array 6.0. This work was supported by NIH grants R01 HL131565 (A.M.), R01 HL103676 (D.J.L.), and R01 HL137234 (D.J.L.).

Publisher's version (útgefin grein), Introduction: Airway micro-aspiration might contribute to the proposed associations between gastroesophageal reflux disease (GERD) and some lung diseases, including lung cancer. This study aimed to examine the hypothesis that antireflux surgery decreases the risk of small cell carcinoma, squamous cell carcinoma and adenocarcinoma of the lung differently depending on their location in relation to micro-aspiration. Methods: Population-based cohort study including patients having undergone antireflux surgery during 1980–2014 in Denmark, Finland, Iceland, Norway or Sweden. Patients having undergone antireflux surgery were compared with two groups: 1) the corresponding background population, by calculating standardised incidence ratios (SIRs) with 95% confidence intervals (CIs) and 2) non-operated GERD-patients, by calculating hazard ratios (HRs) with 95% CIs using multivariable Cox regression with adjustment for sex, age, calendar period, country, chronic obstructive pulmonary disease and obesity diagnosis or type 2 diabetes. Results: Among all 812,617 GERD-patients, 46,996 (5.8%) had undergone antireflux surgery. The SIRs were statistically significantly decreased for small cell carcinoma (SIR = 0.57, 95% CI 0.41–0.77) and squamous cell carcinoma (SIR = 0.75, 95% CI 0.60–0.92), but not for adenocarcinoma of the lung (SIR = 0.90, 95% CI 0.76–1.06). The HRs were also below unity for small cell carcinoma (HR = 0.63, 95% CI 0.44–0.90) and squamous cell carcinoma (HR = 0.80, 95% CI 0.62–1.03), but not for adenocarcinoma of the lung (HR = 1.03, 95% CI 0.84–1.26). Analyses restricted to patients with objective GERD (reflux oesophagitis or Barrett's oesophagus) showed similar results. Conclusions: This all-Nordic study indicates that patients who undergo antireflux surgery are at decreased risk of small cell carcinoma and squamous cell carcinoma of the lung, but not of adenocarcinoma of the lung., This work was supported by the Nordic Cancer Union (grant number 186058), Swedish Cancer Society (grant number 180684), and Swedish Research Council (grant number 340-2013-5478). Jesper Lagergren was supported by the United European Gastroenterology Research Prize and the Distinguished Professor Award at Karolinska Institutet.

Background: COPD is a common cause of morbidity and mortality. The aim of this study was to explore patients’ experiences, self-reported needs, and needs-driven strategies to cope with self-management of COPD. Patients and methods: In this phenomenological study, 10 participants with mild to severe COPD were interviewed 1–2 times, until data saturation was reached. In total, 15 indepth interviews were conducted, recorded, transcribed, and analyzed. Results: COPD negatively affected participants’ physical and psychosocial well-being, their family relationships, and social life. They described their experiences of COPD like fighting a war without weapons in an ever-shrinking world with a loss of freedom at most levels, always fearing possible breathlessness. Fourteen needs were identified and eight clusters of needs-driven strategies that participants used to cope with self-management of COPD. Coping with the reality of COPD, a life-threatening disease, meant coping with dyspnea, feelings of suffocation, indescribable smoking addiction, anxiety, and lack of knowledge about the disease. Reduced participation in family and social life meant loss of ability to perform usual and treasured activities. Having a positive mindset, accepting help and assuming healthy lifestyle was important, as well as receiving continuous professional health care services. The participants’ needs-driven strategies comprised conducting financial arrangements, maintaining hope, and fighting their smoking addiction, seeking knowledge about COPD, thinking differently, facing the broken chain of health care, and struggling with accepting support. Procrastination and avoidance were also evident. Finally, the study also found that participants experienced a perpetuating cycle of dyspnea, anxiety, and fear of breathlessness due to COPD which could lead to more severe dyspnea and even panic attacks. Conclusion: COPD negatively affects patients’ physical and psychosocial well-being, family relationships and, social life. Identifying patients’ self-reported needs and needsdriven strategies can enable clinicians to empower patients by educating them to improve their self-management. Keywords: COPD, patients’ needs, self-management, pulmonary rehabilitation, qualitative research, We are thankful to the Oddur Ólafsson Fund and the B-Section of the Scientific Fund of INA for their financial support.

Thesis for a doctoral degree at the University of Iceland, Idiopathic pulmonary fibrosis is a severe lung disease with unknown aetiology. The lifespan is approximately 3 to 5 years after diagnosis, and the treatments available such as lung transplant are high risk and are not suitable for all patients. In recent years, new drugs such as pirferidone and nintedanib have proved to slow down IPF progression by targeting profibrotic signaling pathways. However, IPF is usually detected in advanced stages, diminishing the beneficial effect of new treatments. Therefore, to find novel risk factors and preclinical characteristics of IPF has become an important task in the field. A recent genome wide association study (GWAS) performed in IPF patients compared to controls revealed a common MUC5B cis-regulatory region polymorphism (rs35705950), where a G is replaced by a T allele in around 45% of IPF patient compared to an 11% allele frequency in controls. This polymorphism has become the strongest risk factor (genetic and otherwise) associated with IPF. Furthermore, the presence of the minor T allele has been associated to subclinical lung fibrosis, suggesting an important role of the MUC5B polymorphism early in IPF development. However, the mechanism underlying the effect of the MUC5B cis-regulatory polymorphism remains unknown. In this study we have focused on the molecular consequences of the polymorphism on the regulation of MUC5B expression in human bronchial epithelial cells. The results shown in this project corroborate the gain of function role of the minor T allele. Also, we have identified a combined mechanism involving both methylation and direct transcriptional regulation mediated by the polymorphic variant on MUC5B overexpression. On the one hand, the minor T allele destroys a CpG methylation site, previously associated with MUC5B inhibition. On the other hand, the replacement of the wild type G allele with a T allele generates a novel C/EBPβ activating transcription binding site. Using two different approach models that include the use of luciferase reporter vectors and CRISPR-Cas9 editing technique we show the differential methylation pattern between both alleles and analyze the effect on MUC5B expression. Also, based on CRISPR edited cell lines we show the binding of C/EBPβ in MUC5B cis-regulatory domain only in presence of the minor T allele. Finally, we show preliminary data that correlates the MUC5B overexpression in T allele cells to characteristics of the cellular stress, as reduced proliferation ratio, overexpression of the ER stress marker C/EBPζ transcription factor and increased number of mitochondria. Altogether, the findings presented in this project assessed the bases for a better understanding of the rs35705950 T allele effect in MUC5B overexpression that would lead to specific treatments in the future as well as validate the use of rs35705950 polymorphism as a risk marker for IPF development., Lungnatrefjun af óþekktum uppruna (IPF) er alvarlegur og ólæknandi sjúkdómur. Lifun eftir greiningu er um 3-5 ár og eina læknandi meðferðin er lungnaígræðsla, sem ekki allir þola. Nýverið hafa komið fram lyf á borð við pirferidone og nintedanib sem hafa náð að hægja á framgangi sjúkdómsins með því að hamla boðferla sem leiða til trefjunar. Þó er það þannig að IPF greinist oftast seint í sjúkdómsferlinu þannig að líkur á læknandi meðferð eru minni. Því er afar mikilvægt að uppgötva nýja áhættuþætti sjúkdómsins sem hægt er að nota til greiningar fyrr í ferlinu. Nýleg erfðamengis – tengslagreining á IPF sjúklingum samanborið við heilbrigða einstaklinga leiddi í ljós að einbasabreytileiki (G-T) (rs35705950) í stýrisvæði MUC5B gensins var til staðar í 45% IPF sjúklinga samanborið við 11% heilbrigðra. Þessi breytileiki er í dag stærsti þekkti áhættuþáttur fyrir IPF. G-T breytingin hefur einnig verið tengd við snemkomna trefjun í lungum, sem bendir til tengsla stökkbreytinga í MUC5B og lungnatrefjunar. Fram til þessa hefur ekki verið ljóst hvernig þessi einbasabreytileiki hefur þessi áhrif. Í þessu verkefni eru sameindalíffræðilegar afleiðingar stökkbreytingarinnar á tjáningarstjórn MUC5B í þekjufrumum lungna útskýrðar, en meðal annars er notast við CRISPR-Cas9 erfðabreytingaraðferðina. Niðurstöður rannsóknarinnar sýna fram á aukna tjáningu í viðurvist T- einbasabreytileikans. Þessi tjáningaraukning virðist drifin af tveimur atburðum; CpG metyleringar eyja eyðileggst (sem áður hefur verið tengd við bælingu á tjáningu) og nýr bindistaður myndast fyrir hvetjandi umritunarþáttinn C/EBPβ. Ennfremur sýnum við gögn sem tengja T erfðabreytileika MUC5B við lykilþætti frumuálags, svo sem minnkaða frumufjölgun, aukna tjáningu á umritunarþættinum C/EBPζ og aukinn fjölda hvatbera. Þetta verkefni hefur varpað nýju ljósi á rs35705950 stökkbreytinguna í IPF og tengt oftjáningu á MUC5B við sjúkdóminn með beinum hætti. Þessar niðurstöður tengja MUC5B beint við IPF og leggja grunn að nýrri sýn á því hvernig þessi einbasabreytileiki tengist tilurð IPF., Supported by the Icelandic Research Council (RANNÍS) project grant.

Background: Rare missense mutations in the gene encoding coatomer subunit alpha (COPA) have recently been shown to cause autoimmune interstitial lung, joint and kidney disease, also known as COPA syndrome, under a dominant mode of inheritance. Case presentation: Here we describe an Icelandic family with three affected individuals over two generations with a rare clinical presentation of lung and joint disease and a histological diagnosis of follicular bronchiolitis. We performed whole-genome sequencing (WGS) of the three affected as well as three unaffected members of the family, and searched for rare genotypes associated with disease using 30,067 sequenced Icelanders as a reference population. We assessed all coding and splicing variants, prioritizing variants in genes known to cause interstitial lung disease. We detected a heterozygous missense mutation, p.Glu241Lys, in the COPA gene, private to the affected family members. The mutation occurred de novo in the paternal germline of the index case and was absent from 30,067 Icelandic genomes and 141,353 individuals from the genome Aggregation Database (gnomAD). The mutation occurs within the conserved and functionally important WD40 domain of the COPA protein. Conclusions: This is the second report of the p.Glu241Lys mutation in COPA, indicating the recurrent nature of the mutation. The mutation was reported to co-segregate with COPA syndrome in a large family from the USA with five affected members, and classified as pathogenic. The two separate occurrences of the p.Glu241Lys mutation in cases and its absence from a large number of sequenced genomes confirms its role in the pathogenesis of the COPA syndrome. Keywords: COPA syndrome, Lung disease, Arthritis, Immune dysregulation, Case report