The forskolin-induced swelling assay (FIS) in patient-derived intestinal organoids (PDIOs), used to determine in vitro responsiveness to elexacaftor/tezacaftor/ivacaftor (ETI), showed variability in swelling among PDIOs obtained from people with CF (pwCF) carrying the same F508del/F508del CFTR genotype. We aimed to characterise the effect of ETI on the transcriptional activity of PDIOs-derived cells to understand the intracellular processes triggered by ETI and the differences in treatment response. Six high- and six low-responding PDIOs to ETI, derived from F508del/F508del pwCF, were incubated with or without ETI for 2 to 6 h. Gene expression was assessed using 3'-mRNA sequencing and modelled using negative binomial models. Incubation with ETI resulted in a significant upregulation of several biological processes: mostly related to chemokines and signalling, chemotaxis, and tissue development processes. No changes were observed in abundance of the CFTR transcripts or in CFTR-related gene sets and pathways. The genes and pathways associated with ETI did not overlap with those whose expression changed with time only. PDIOs with a high FIS response did not significantly differ in any interpretable gene from the FIS-low organoids. The changes in the PDIOs gene expression upon the exposure to ETI cannot explain differences in the magnitude of PDIOs FIS-measured response to ETI. In conclusion, on incubation with ETI, genes of the CFTR-related pathways do not change their transcriptional activity; instead, overexpression was observed in genes of inflammatory-like cytokine response and receptor activation pathways.

• Klíčová slova
• Cystic fibrosis, Differential expression, Elexacaftor, Intestinal organoid, Ivacaftor, Tezacaftor,
• MeSH
• aktivátory chloridových kanálů farmakologie   MeSH
• aminofenoly * farmakologie   MeSH
• benzodioxoly * farmakologie   MeSH
• chinoliny MeSH
• chinolony * farmakologie   MeSH
• cystická fibróza * genetika   farmakoterapie   MeSH
• fixní kombinace léků MeSH
• indoly * farmakologie   MeSH
• lidé MeSH
• organoidy * účinky léků   metabolismus   MeSH
• protein CFTR genetika   MeSH
• pyrazoly MeSH
• pyridiny * farmakologie   MeSH
• pyrrolidiny MeSH
• pyrroly * farmakologie   MeSH
• stanovení celkové genové exprese MeSH
• thiofeny farmakologie   MeSH
• transkriptom MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aktivátory chloridových kanálů MeSH
• aminofenoly * MeSH
• benzodioxoly * MeSH
• chinoliny MeSH
• chinolony * MeSH
• elexacaftor, ivacaftor, tezacaftor drug combination MeSH Prohlížeč
• elexacaftor MeSH Prohlížeč
• fixní kombinace léků MeSH
• indoly * MeSH
• ivacaftor MeSH Prohlížeč
• protein CFTR MeSH
• pyrazoly MeSH
• pyridiny * MeSH
• pyrrolidiny MeSH
• pyrroly * MeSH
• tezacaftor MeSH Prohlížeč
• thiofeny MeSH

INTRODUCTION: In silico tools capable of predicting the functional consequences of genomic differences between individuals, many of which are AI-driven, have been the most effective over the past two decades for non-synonymous single nucleotide variants (nsSNVs). When appropriately selected for the purpose of the study, a high predictive performance can be expected. In this feasibility study, we investigate the distribution of nsSNVs with an allele frequency below 5%. To classify the putative functional consequence, a tier-based filtration led by AI-driven predictors and scoring system was implemented to the overall decision-making process, resulting in a list of prioritised genes. METHODS: The study has been conducted on breast cancer patients of homogeneous ethnicity. Germline rare variants have been sequenced in genes that influence pharmacokinetic parameters of anticancer drugs or molecular signalling pathways in cancer. After AI-driven functional pathogenicity classification and data mining in pharmacogenomic (PGx) databases, variants were collapsed to the gene level and ranked according to their putative deleterious role. RESULTS: In breast cancer patients, seven of the twelve genes prioritised based on the predictions were found to be associated with response to oncotherapy, histological grade, and tumour subtype. Most importantly, we showed that the group of patients with at least one rare nsSNVs in cystic fibrosis transmembrane conductance regulator (CFTR) had significantly reduced disease-free (log rank, p = 0.002) and overall survival (log rank, p = 0.006). CONCLUSION: AI-driven in silico analysis with PGx data mining provided an effective approach navigating for functional consequences across germline genetic background, which can be easily integrated into the overall decision-making process for future studies. The study revealed a statistically significant association with numerous clinicopathological parameters, including treatment response. Our study indicates that CFTR may be involved in the processes influencing the effectiveness of oncotherapy or in the malignant progression of the disease itself.

• Klíčová slova
• Breast cancer, Cystic fibrosis transmembrane conductance regulator, Gene prioritisation, Machine learning, Survival,
• MeSH
• dospělí MeSH
• frekvence genu MeSH
• jednonukleotidový polymorfismus MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu * genetika   farmakoterapie   patologie   MeSH
• protein CFTR * genetika   MeSH
• senioři MeSH
• studie proveditelnosti * MeSH
• umělá inteligence * MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• CFTR protein, human MeSH Prohlížeč
• protein CFTR * MeSH

• Klíčová slova
• Cystic fibrosis, Elexacaftor/tezacaftor/ivacaftor, Exocrine pancreatic insufficiency,
• MeSH
• aktivátory chloridových kanálů terapeutické užití   MeSH
• aminofenoly * terapeutické užití   MeSH
• benzodioxoly * terapeutické užití   MeSH
• chinoliny terapeutické užití   MeSH
• chinolony * terapeutické užití   MeSH
• cystická fibróza * farmakoterapie   genetika   MeSH
• dospělí MeSH
• exokrinní pankreatická insuficience farmakoterapie   etiologie   genetika   MeSH
• fixní kombinace léků MeSH
• genotyp MeSH
• indoly * terapeutické užití   MeSH
• lidé MeSH
• protein CFTR * genetika   MeSH
• pyrazoly * terapeutické užití   MeSH
• pyridiny terapeutické užití   MeSH
• pyrrolidiny terapeutické užití   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• dopisy MeSH
• kazuistiky MeSH
• Názvy látek
• aktivátory chloridových kanálů MeSH
• aminofenoly * MeSH
• benzodioxoly * MeSH
• chinoliny MeSH
• chinolony * MeSH
• elexacaftor, ivacaftor, tezacaftor drug combination MeSH Prohlížeč
• elexacaftor MeSH Prohlížeč
• fixní kombinace léků MeSH
• indoly * MeSH
• ivacaftor MeSH Prohlížeč
• protein CFTR * MeSH
• pyrazoly * MeSH
• pyridiny MeSH
• pyrrolidiny MeSH
• tezacaftor MeSH Prohlížeč

BACKGROUND: Cystic fibrosis (CF) is a rare multi-systemic recessive disorder. The spectrum and the frequencies of CFTR mutations causing CF vary amongst different populations in Europe and the Middle East. In this study, we characterised the distribution of CF-causing mutations (i.e. pathogenic variants in the CFTR gene) in a representative CF cohort from the Kingdom of Bahrain based on a three-decade-long analysis at a single tertiary centre. We aim to improve CF genetic diagnostics, introduce of CF neonatal screening and provide CFTR modulator therapy (CFTRm). METHODS: CFTR genotyping and associated clinical information were drawn from a longitudinal cohort. We sequenced 56 people with CF (pwCF) that had one or both CFTR mutations unidentified and carried out comprehensive bioinformatic- and family-based segregation analyses of detected variants, including genotype-phenotype correlations and disease incidence estimates. The study methodology could serve as a basis for other non-European CF populations with a high degree of consanguinity. RESULTS: Altogether 18 CF-causing mutations were identified, 15 of which were not previously detected in Bahrain, accounting for close to 100% of all population-specific alleles. The most common alleles comprise c.1911delG [2043delG; 22.8%], c.2988+1G > A [3120+1G>A; 16.3%], c.2989-1G>A [3121-1G>A; 14.1%], c.3909C>G [N1303K; 13.0%], and c.1521_1523delCTT [p.PheF508del; 7.6%]. Although the proportion of 1st cousin marriages has decreased to 50%, the frequency of homozygosity in our pwCF is 67.4%, thereby indicating that CF still occurs in large, often related, families. pwCF in Bahrain present with faltering growth, pancreatic insufficiency and classical sino-pulmonary manifestations. Interestingly, two pwCF also suffer from sickle cell disease. The estimated incidence of CF in Bahrain based on data from the last three decades is 1 in 9,880 live births. CONCLUSION: The most commonCF-causing mutations in Bahraini pwCF were identified, enabling more precise diagnosis, introduction of two-tier neonatal screening and fostering administration of CFTRm.

• Klíčová slova
• CFTR gene, Bahrain, CFTR modulator therapy (CFTRm), Cystic fibrosis, Mutations, Sequencing,
• MeSH
• alely MeSH
• cystická fibróza * genetika   MeSH
• dítě MeSH
• dospělí MeSH
• genetické asociační studie metody   MeSH
• genotyp MeSH
• kohortové studie MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mutace * MeSH
• novorozenec MeSH
• novorozenecký screening MeSH
• předškolní dítě MeSH
• protein CFTR * genetika   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Bahrajn MeSH

The major cause of mortality in people with cystic fibrosis (pwCF) is progressive lung disease characterised by acute and chronic infections, the accumulation of mucus, airway inflammation, structural damage and pulmonary exacerbations. The prevalence of Pseudomonas aeruginosa rises rapidly in the teenage years, and this organism is the most common cause of chronic lung infection in adults with cystic fibrosis (CF). It is associated with an accelerated decline in lung function and premature death. New P. aeruginosa infections are treated with antibiotics to eradicate the organism, while chronic infections require long-term inhaled antibiotic therapy. The prevalence of P. aeruginosa infections has decreased in CF registries since the introduction of CF transmembrane conductance regulator modulators (CFTRm), but clinical observations suggest that chronic P. aeruginosa infections usually persist in patients receiving CFTRm. This indicates that pwCF may still need inhaled antibiotics in the CFTRm era to maintain long-term control of P. aeruginosa infections. Here, we provide an overview of the changing perceptions of P. aeruginosa infection management, including considerations on detection and treatment, the therapy burden associated with inhaled antibiotics and the potential effects of CFTRm on the lung microbiome. We conclude that updated guidance is required on the diagnosis and management of P. aeruginosa infection. In particular, we highlight a need for prospective studies to evaluate the consequences of stopping inhaled antibiotic therapy in pwCF who have chronic P. aeruginosa infection and are receiving CFTRm. This will help inform new guidelines on the use of antibiotics alongside CFTRm.

• Klíčová slova
• Bacterial Infection, Bronchiectasis, Cystic Fibrosis, Respiratory Infection,
• MeSH
• antibakteriální látky * aplikace a dávkování   terapeutické užití   MeSH
• aplikace inhalační MeSH
• cystická fibróza * komplikace   mikrobiologie   farmakoterapie   MeSH
• lidé MeSH
• protein CFTR * genetika   MeSH
• pseudomonádové infekce * farmakoterapie   MeSH
• Pseudomonas aeruginosa * účinky léků   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antibakteriální látky * MeSH
• protein CFTR * MeSH

• MeSH
• cystická fibróza * MeSH
• individualizovaná medicína MeSH
• kolforsin farmakologie   MeSH
• lidé MeSH
• mutace MeSH
• protein CFTR genetika   MeSH
• střevní sliznice MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• Názvy látek
• kolforsin MeSH
• protein CFTR MeSH

CFTR is a membrane protein that functions as an ion channel. Mutations that disrupt its biosynthesis, trafficking or function cause cystic fibrosis (CF). Here, we present a novel in vitro model system prepared using CRISPR/Cas9 genome editing with endogenously expressed WT-CFTR tagged with a HiBiT peptide. To enable the detection of CFTR in the plasma membrane of live cells, we inserted the HiBiT tag in the fourth extracellular loop of WT-CFTR. The 11-amino acid HiBiT tag binds with high affinity to a large inactive subunit (LgBiT), generating a reporter luciferase with bright luminescence. Nine homozygous clones with the HiBiT knock-in were identified from the 182 screened clones; two were genetically and functionally validated. In summary, this work describes the preparation and validation of a novel reporter cell line with the potential to be used as an ultimate building block for developing unique cellular CF models by CRISPR-mediated insertion of CF-causing mutations.

• MeSH
• buněčná membrána metabolismus   MeSH
• buněčné linie MeSH
• CRISPR-Cas systémy genetika   MeSH
• cystická fibróza * genetika   metabolismus   MeSH
• lidé MeSH
• protein CFTR * genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CFTR protein, human MeSH Prohlížeč
• protein CFTR * MeSH

This is the second in a series of four papers updating the European Cystic Fibrosis Society (ECFS) standards for the care of people with CF. This paper focuses on establishing and maintaining health. The guidance is produced using an evidence-based framework and with wide stakeholder engagement, including people from the CF community. Authors provided a narrative description of their topic and statements, which were more directive. These statements were reviewed by a Delphi exercise, achieving good levels of agreement from a wide group for all statements. This guidance reinforces the importance of a multi-disciplinary CF team, but also describes developing models of care including virtual consultations. The framework for health is reinforced, including the need for a physically active lifestyle and the strict avoidance of all recreational inhalations, including e-cigarettes. Progress with cystic fibrosis transmembrane conductance regulator (CFTR) modulator therapy is reviewed, including emerging adverse events and advice for dose reduction and interruption. This paper contains guidance that is pertinent to all people with CF regardless of age and eligibility for and access to modulator therapy.

• Klíčová slova
• Cystic fibrosis, ECFS, Guidance, Health, Standards,
• MeSH
• cystická fibróza * farmakoterapie   MeSH
• látky ovlivňující dýchací systém * terapeutické užití   MeSH
• lidé MeSH
• mutace MeSH
• protein CFTR genetika   MeSH
• systémy dodávající nikotin elektronicky * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• látky ovlivňující dýchací systém * MeSH
• protein CFTR MeSH

There is considerable activity with respect to diagnosis in the field of cystic fibrosis (CF). This relates primarily to developments in newborn bloodspot screening (NBS), more extensive gene analysis and improved characterisation of CFTR-related disorder (CFTR-RD). This is particularly pertinent with respect to accessibility to variant-specific therapy (VST), a transformational intervention for people with CF with eligible CFTR gene variants. This advance reinforces the need for a timely and accurate diagnosis. In the future, there is potential for trials to assess effectiveness of variant-specific therapy for CFTR-RD. The guidance in this paper reaffirms previous standards, clarifies a number of issues, and integrates emerging evidence. Timely and accurate diagnosis has never been more important for people with CF.

• Klíčová slova
• CFSPID, CFTR, CFTR-related disorder, CRMS/CFSPID, Cystic fibrosis, Extended Gene Analysis (EGA), Newborn Bloodspot Screening (NBS),
• MeSH
• cystická fibróza * diagnóza   genetika   terapie   MeSH
• lidé MeSH
• novorozenec MeSH
• novorozenecký screening metody   MeSH
• protein CFTR genetika   MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• protein CFTR MeSH

BACKGROUND: We previously documented that elevated HE4 plasma concentration decreased in people with CF (pwCF) bearing the p.Gly551Asp-CFTR variant in response to CFTR modulator (CFTRm) ivacaftor (IVA), and this level was inversely correlated with the FEV1% predicted values (ppFEV1). Although the effectiveness of lumacaftor (LUM)/IVA in pwCF homozygous for the p.Phe508del-CFTR variant has been evaluated, plasma biomarkers were not used to monitor treatment efficacy thus far. METHODS: Plasma HE4 concentration was examined in 68 pwCF drawn from the PROSPECT study who were homozygous for the p.Phe508del-CFTR variant before treatment and at 1, 3, 6 and 12 months after administration of LUM/IVA therapy. Plasma HE4 was correlated with ppFEV1 using their absolute and delta values. The discriminatory power of delta HE4 was evaluated for the detection of lung function improvements based on ROC-AUC analysis and multiple regression test. RESULTS: HE4 plasma concentration was significantly reduced below baseline following LUM/IVA administration during the entire study period. The mean change of ppFEV1 was 2.6% (95% CI, 0.6 to 4.5) by 6 months of therapy in this sub-cohort. A significant inverse correlation between delta values of HE4 and ppFEV1 was observed especially in children with CF (r=-0.7053; p<0.0001). Delta HE4 predicted a 2.6% mean change in ppFEV1 (AUC: 0.7898 [95% CI 0.6823-0.8972]; P < 0.0001) at a cut-off value of -10.7 pmol/L. Moreover, delta HE4 independently represented the likelihood of being a responder with ≥ 5% delta ppFEV1 at 6 months (OR: 0.89, 95% CI: 0.82-0.95; P = 0.001). CONCLUSIONS: Plasma HE4 level negatively correlates with lung function improvement assessed by ppFEV1 in pwCF undergoing LUM/IVA CFTRm treatment.

• Klíčová slova
• Biomarker, Cystic fibrosis, HE4, Lumacaftor/ivacaftor, Sweat chloride, ppFEV1,
• MeSH
• aktivátory chloridových kanálů terapeutické užití   MeSH
• aminofenoly terapeutické užití   MeSH
• aminopyridiny terapeutické užití   MeSH
• benzodioxoly terapeutické užití   MeSH
• cystická fibróza * diagnóza   farmakoterapie   genetika   MeSH
• dítě MeSH
• fixní kombinace léků MeSH
• homozygot MeSH
• lidé MeSH
• mutace MeSH
• protein CFTR genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktivátory chloridových kanálů MeSH
• aminofenoly MeSH
• aminopyridiny MeSH
• benzodioxoly MeSH
• CFTR protein, human MeSH Prohlížeč
• fixní kombinace léků MeSH
• ivacaftor MeSH Prohlížeč
• lumacaftor MeSH Prohlížeč
• protein CFTR MeSH