bioRxiv [Preprint]. 2025 May 28:2025.05.28.656674. doi: 10.1101/2025.05.28.656674.

ABSTRACT

Cystic Fibrosis (CF) is a multiorgan disease caused by mutations in the CFTR gene, leading to chronic pulmonary infections and hyperinflammation. Among pathogens colonizing the CF lung, Pseudomonas aeruginosa is predominant, infecting over 50% of adults with CF, and becoming antibiotic-resistant over time. Current therapies for CF, while providing tremendous benefits, fail to eliminate persistent bacterial infections, chronic inflammation, and irreversible lung damage, necessitating novel therapeutic strategies. Our group engineered mesenchymal stromal cell derived extracellular vesicles (MSC EVs) to carry the microRNA let-7b-5p as a dual anti-infective and anti-inflammatory treatment. MSC EVs are low-immunogenicity platforms with innate antimicrobial and immunomodulatory properties, while let-7b-5p reduces biofilm formation and inflammation. In a preclinical CF mice model, we reported that let-7b-5p-loaded MSC EVs reduced P. aeruginosa burden, immune cells, and proinflammatory cytokines in the lungs. We hypothesize four complementary mechanisms for the observed in-vivo effects of the let-7b-5p loaded MSC EVs: antimicrobial activity, anti-inflammatory properties, inhibition of antibiotic-resistant P. aeruginosa biofilm formation in CF airways, and stimulation of anti-inflammatory macrophage behaviors. This study focused on the second and third mechanisms and demonstrates that MSC EVs engineered to contain let-7b-5p effectively blocked the formation of antibiotic-resistant P. aeruginosa biofilms on primary human bronchial epithelial cells (pHBECs) while also reducing P. aeruginosa -induced inflammation. This approach holds promise for improving outcomes for people with CF. Future work will focus on optimizing delivery strategies and expanding the clinical applicability of MSC EVs to target other CF-associated pathogens.

NEW AND NOTEWORTHY: This is the first study demonstrating that let-7b-5p loaded Mesenchymal Stromal Cell Extracellular Vesicles (MSC EVs) block antibiotic-resistant P. aeruginosa biofilm formation and reduce inflammation in CF primary human bronchial epithelial cells.

PMID:40501816 | PMC:PMC12154590 | DOI:10.1101/2025.05.28.656674

bioRxiv [Preprint]. 2025 Jun 8:2025.06.08.656500. doi: 10.1101/2025.06.08.656500.

ABSTRACT

Cystic fibrosis (CF), a common genetic disease, is caused by a defective CF-transmembrane conductance regulator (CFTR). People with CF (pwCF) are prone to develop infections by opportunistic pathogens, including Burkholderia cenocepacia, leading to chronic inflammation and lung function loss. Neutrophils, the most abundant cells in the chronically inflamed lungs of pwCF, release granular proteins and oxida-tive products that contribute to tissue damage. The CFTR modulators are a new treat-ment for pwCF aiming to correct the subcellular location and function of the CFTR ion channel. The triple modulator combination of Elexacaftor, Tezacaftor, and Ivacaftor (ETI) or Trikafta® has significantly improved clinical symptoms and overall provided a better quality of life for pwCF. The mechanism by which the CFTR modulators help to restore the antimicrobial functions of neutrophils is unknown. The present study demonstrates that neutrophils functionally express CFTR and reveals how ETI modi-fies subcellular CFTR trafficking in CF neutrophils. In addition, ETI treatment reduces intracellular chloride levels in human neutrophils, indicating activation of CFTR-dependent chloride efflux (outflow). Finally, ETI treatment also re-established the intra-cellular antimicrobial killing of CF neutrophils by potentiating NADPH oxidase activity and producing Neutrophil Extracellular Traps (NETs). Together, our findings suggest that CFTR has an essential role in controlling neutrophil functions and that the CFTR modulators improve the health of pwCF by restoring the antimicrobial functions of CF neutrophils.

PMID:40501645 | PMC:PMC12157431 | DOI:10.1101/2025.06.08.656500

Methods Mol Biol. 2025;2942:199-215. doi: 10.1007/978-1-0716-4627-4_17.

ABSTRACT

The adhesion of Pseudomonas aeruginosa to submaxillary and airway mucins is subject to large intra- and interclonal bacterial diversity. Thus, a mucin adhesion study should include numerous bacterial strains that reflect the population biology of the taxon and/or the habitat of interest. Human airway mucins are isolated from induced sputa collected from healthy individuals or people with chronic lung diseases such as bronchiectasis, cystic fibrosis, or chronic obstructive pulmonary disease. In this protocol, bacteria are exposed to mucin-coated microplates. Adhesion is quantified by colony forming units.

PMID:40498318 | DOI:10.1007/978-1-0716-4627-4_17

Methods Mol Biol. 2025;2942:93-102. doi: 10.1007/978-1-0716-4627-4_8.

ABSTRACT

Confocal microscopy provides information on host-pathogen interactions. To examine infection at the respiratory epithelial barrier, primary bronchial cells differentiated at an air-liquid interface (ALI) can be imaged and reconstructed as a 3D model. In this chapter, we describe a method for immunostaining and fluorescence imaging of well-differentiated bronchial epithelial tissue that has been exposed to the cystic fibrosis respiratory pathogen Achromobacter xylosoxidans.

PMID:40498309 | DOI:10.1007/978-1-0716-4627-4_8

Expert Rev Gastroenterol Hepatol. 2025 Jun 11. doi: 10.1080/17474124.2025.2519160. Online ahead of print.

ABSTRACT

INTRODUCTION: Functional gastrointestinal disorders (FGIDs) and inflammatory bowel diseases (IBD) present significant challenges for both pediatric and adult populations. The low FODMAP diet (LFD) has gained prominence as an evidence-based dietary intervention, offering symptom relief for abdominal pain, bloating, and altered bowel habits. However, concerns about adherence, nutritional adequacy, and long-term safety remain critical, particularly in pediatric populations.

AREAS COVERED: This narrative review examines the nature and physiological impact of FODMAPs, focusing on the practical application of the low-FODMAP diet (LFD) in adults and pediatric gastroenterology. Particular attention is given to emerging insights into its impact on gut microbiota and long-term safety.

EXPERT OPINION: While LFD effectively alleviates symptoms in FGIDs and functional symptoms in IBD, its restrictive nature demands professional supervision to mitigate nutritional risks. For pediatric patients, adherence and reintroduction protocols require optimization. Further research into personalized dietary approaches and microbiome-targeted strategies could enhance the clinical utility of the LFD, ensuring its long-term safety and efficacy for diverse patient groups.

PMID:40497754 | DOI:10.1080/17474124.2025.2519160

Ann Clin Biochem. 2025 Jun 11:45632251350514. doi: 10.1177/00045632251350514. Online ahead of print.

ABSTRACT

BACKGROUND: The diagnosis of cystic fibrosis (CF) is challenging due to high quantity not sufficient (QNS) rates of sweat tests, leading to frequent retesting, increasing costs and adverse impacts on patient care. This study aimed to assess sweat test performance and implement a quality improvement project (QIP) to reduce QNS rates.

METHODS: A two-part retrospective audit was conducted. Part one spanned two-years reviewing the two-tiered testing with sweat conductivity as a screening tool, followed by chloride testing. Part two evaluated the QNS rates over two 6-month periods, separated by a QIP, which involved technologist training, clinician education, patient preparation protocols and revised testing procedures.

RESULTS: Over the two-year period: 425 sweat tests were performed on 291 patients. Sweat conductivity testing demonstrated a lower QNS rate, 13% (31/238), compared to sweat chloride testing's 31% (33/105). High QNS rates were observed in younger infants and in malnourished or acutely ill patients. Post-QIP, the QNS rates for the total study population decreased by 5%, from an initial 30% to 25% in the sweat chloride cohort, while the acceptable QNS rate of 12% remained unchanged in the sweat conductivity cohort Conclusion: Achieving target QNS rates remains challenging, especially in younger infants, with improved QNS rates in older infants and children. Recommendations include limiting sweat testing to experienced technologists and ensuring patient readiness.

PMID:40497297 | DOI:10.1177/00045632251350514

Heart Rhythm O2. 2025 Feb 25;6(5):720-732. doi: 10.1016/j.hroo.2025.02.015. eCollection 2025 May.

ABSTRACT

Serum and glucocorticoid-regulated kinase 1 (SGK1) is a serine/threonine kinase that is involved in various cellular pathways, such as ion transport, cell survival, proliferation, and immune responses. Dysregulation of this enzyme is increasingly being associated with the progression of 2 prominent types of diseases, namely viral infections, such as COVID-19, and cardiovascular disorders, such as atrial fibrillation (AF), positioning it as a potential therapeutic target. With regard to coronavirus 2019 (COVID-19), SGK1 detrimentally affects inflammatory pathways and modulates the cytokine storm, leading to lung tissue damage. Considering this dysregulation, researchers are exploring SGK1 inhibition as a potential strategy for mitigating severe COVID-19 outcomes. SGK1 also regulates pumps and ion channels, significantly affecting cardiac performance in AF. This protein is responsible for promoting fibrosis and inflammation in the cardiac tissue, making it a potential target for reducing atrial fibrillation. SGK1 inhibition offers a new avenue for therapeutic targets against both COVID-19 and AF. This review is aimed at providing a comprehensive overview of SGK1 dysregulation in both diseases, underscoring the urgent need for more preclinical and clinical trials to evaluate effective SGK1 inhibitors for patients with coexisting COVID-19 and AF.

PMID:40496592 | PMC:PMC12147618 | DOI:10.1016/j.hroo.2025.02.015

Front Endocrinol (Lausanne). 2025 May 27;16:1586941. doi: 10.3389/fendo.2025.1586941. eCollection 2025.

ABSTRACT

INTRODUCTION: Data about efficacy and safety of GLP1 receptor agonists in liver-transplanted patients are lacking.

METHODS: Among a population of liver-transplanted individuals with diabetes, we evaluated 68 patients before, 6, 12 and 18 months after starting a GLP1RA-based therapy, as add on to metformin or insulin. We assessed glycemic control, body weight and composition (with bio-impedance analysis), liver fibrosis and steatosis (with transient elastography). Amylase, lipase levels and concomitant therapies were recorded at basal and follow up evaluations. Patients had an e-mail contact to report any adverse events.

RESULTS: We observed a significant decrease in fasting plasma glucose, HbA1c, weight, BMI, waist circumference. We demonstrated a reduction in total and LDL cholesterol. Liver stiffness decreased during the first 6 months. The rate of adverse events was low and the symptoms reported didn't require any medical measures: 26.9% reported mild nausea, only 3 patients (7.69%) discontinued the drug dose due to gastrointestinal intolerance. No pancreatitis episodes were detected, amylase and lipase levels didn't increase (despite concomitant calcineurin inhibitors). No adjustments in immunosuppressant therapy were reported. Among the 45 patients requiring insulin when a GLP1RA therapy was added on, 20 (33.2%) and 31 (45.5%) could suspend insulin therapy at, respectively, 6 and 18 months.

DISCUSSION: In conclusion, GLP1RA-based therapy can be considered safe and effective in a short-term follow up in liver-transplanted patients. Further studies are needed to assess the effects of this drugs on long term complications, such as renal impairment, cardiovascular events and all-cause mortality.

PMID:40496570 | PMC:PMC12148859 | DOI:10.3389/fendo.2025.1586941

Public Health Chall. 2023 Apr 9;2(2):e74. doi: 10.1002/puh2.74. eCollection 2023 Jun.

ABSTRACT

BACKGROUND: Cystic fibrosis (CF) is the most common autosomal recessive condition in Caucasian populations globally. To date, there has been very little research conducted into the oral health of adults with CF possibly due to historic premature mortality. The purpose of this survey was to ascertain knowledge, attitude and practices among dental professionals regarding the oral health of people with cystic fibrosis (PWCF) as part of a larger study being conducted in Ireland.

METHODS: A cross-sectional survey of dental practitioners in Ireland was conducted via an online questionnaire. The survey contained close-ended questions, and clinical scenarios which allowed for both close-ended and free text responses in relation to the provision of dental treatment for PWCF. Data was subject to descriptive data analysis using IBM SPSS 29.

RESULTS: The results from the survey indicate a wide variety in the knowledge, attitudes and practices of dental practitioners regarding the oral health of PWCF. There is significant variation in the provision of dental treatment and the conditions under which such treatment is provided.

CONCLUSION: There is a paucity of information and guidelines specific to this vulnerable population which can make the provision of dental treatment challenging for practitioners. Dental professionals may benefit from continuing professional development and further education targeting patient-specific populations.

PMID:40495875 | PMC:PMC12039724 | DOI:10.1002/puh2.74

Mol Med. 2025 Jun 11;31(1):231. doi: 10.1186/s10020-025-01266-7.

ABSTRACT

BACKGROUND: The Lung is the major focus of therapeutic approaches for the inherited disorder cystic fibrosis (CF) as without treatment lung disease is life-limiting. However, the initiating events that predispose the CF lung to cycles of infection, inflammation and resultant tissue damage are still unclear. Inflammation may occur in the CF lung prior to birth in human and several large animal models suggesting an in utero origin for the disease and encouraging further studies prior to birth.

METHODS: Here we used the sheep model of CF (CFTR-/-) and age-matched wild-type (WT) sheep of the same breed to investigate the single cell transcriptomes of proximal and distal lung tissue at 80 days and 120 days of gestation and at term (147 days). Single cell RNA-seq was performed on tissues from 4 to 7 animals of each genotype (WT and CFTR-/-) at each time point.

RESULTS: At term, FOXJ1-expressing ciliated cells are overrepresented in both lung regions from CFTR-/- lambs, while secretory epithelial and basal cells are underrepresented in proximal lung, as are T cells and monocytes in distal lung. The imbalance in ciliated and basal cells was confirmed by immunohistochemistry. At 120 days of gestation, lymphoid cells are slightly more abundant in proximal and distal lung from CFTR-/- animals compared to WT, consistent with the transient CF-associated inflammatory response in utero. At 80 days of gestation, T and B cells are underrepresented in both lung regions.

CONCLUSIONS: The differences in epithelial cell abundance observed in the CFTR-/- lambs at term may reflect sequelae from the loss of CFTR on lung development and differentiation in utero. These findings provide novel insights into the cellular mechanisms of pathology and may be relevant to the design of new therapeutic approaches for CF lung disease.

PMID:40495135 | DOI:10.1186/s10020-025-01266-7

J Cyst Fibros. 2025 Jun 9:S1569-1993(25)01490-0. doi: 10.1016/j.jcf.2025.05.007. Online ahead of print.

NO ABSTRACT

PMID:40494748 | DOI:10.1016/j.jcf.2025.05.007

Dig Liver Dis. 2025 Jun 9:S1590-8658(25)00793-5. doi: 10.1016/j.dld.2025.05.020. Online ahead of print.

ABSTRACT

Mucosal healing is the main treatment goal in the management of pediatric inflammatory bowel diseases (IBD), and accurate endoscopic assessment is essential for evaluating disease severity, monitoring progression, and assessing therapeutic responses. However, the wide variability of mucosal lesions, combined with the lack of standardized guidelines and limited training opportunities, complicates the scoring process and contributes to discrepancies in scoring accuracy. To address these issues, a panel of Italian pediatric endoscopists with expertise in IBD, convened by the Italian Society of Pediatric Gastroenterology, Hepatology, and Nutrition (SIGENP), developed practical recommendations for assessing endoscopic lesions and applying endoscopic scoring systems in pediatric IBD. These guidelines, based on a review of current literature and expert consensus, aim to improve the consistency, reliability, and accuracy of endoscopic evaluations in children with IBD. By providing clear recommendations, the guidelines seek to enhance the reliability of scoring systems, ultimately supporting more effective disease management and treatment outcomes for pediatric patients.

PMID:40494721 | DOI:10.1016/j.dld.2025.05.020

Proc Natl Acad Sci U S A. 2025 Jun 17;122(24):e2418731122. doi: 10.1073/pnas.2418731122. Epub 2025 Jun 10.

ABSTRACT

In utero gene editing has the potential to modify disease-causing genes in multiple developing tissues before birth, possibly allowing for normal organ development, disease improvement, and conceivably, cure. In cystic fibrosis (CF), a disease that arises from mutations in the CF transmembrane conductance regulator (CFTR) gene, there are signs of multiorgan disease affecting the function of the respiratory, gastrointestinal, and reproductive systems already present at birth. Thus, treating CF patients early is crucial for preventing or delaying irreversible organ damage. Here, we demonstrate proof-of-concept of multiorgan mutation correction in CF using peptide nucleic acids encapsulated in polymeric nanoparticles and delivered systemically in utero. In utero editing was associated with sustained postnatal CFTR activity, at a level similar to that of wild-type mice, in both respiratory and gastrointestinal tissues, without detection of off-target mutations in partially homologous loci. This work suggests that systemic in utero gene editing represents a viable strategy for treating monogenic diseases before birth that impact multiple tissue types.

PMID:40493185 | DOI:10.1073/pnas.2418731122

Microbiol Spectr. 2025 Jun 10:e0288224. doi: 10.1128/spectrum.02882-24. Online ahead of print.

ABSTRACT

The rise of antibiotic-resistant bacteria has necessitated the development of alternative therapeutic strategies, such as bacteriophage therapy, where viruses infect bacteria, reducing bacterial burden. However, rapid bacterial resistance to phage treatment remains a critical challenge, potentially leading to failure. Phage steering, which leverages the evolutionary dynamics between phage and bacteria, offers a novel solution by driving bacteria to evolve away from virulence factors or resistance mechanisms. In this study, we examined whether phage steering using bacteriophage Luz19 could function in the presence of a competing pathogen, Staphylococcus aureus (SA) (USA300), while targeting Pseudomonas aeruginosa (PAO1). Through in vitro co-evolution experiments with and without the competitor, we observed that Luz19 consistently steered P. aeruginosa away from the Type IV pilus (T4P), a key virulence factor, without interference from SA. Genomic analyses revealed mutations in T4P-associated genes, including pilR and pilZ, which conferred phage resistance. Our findings suggest that phage steering remains effective even in polymicrobial environments, providing a promising avenue for enhancing bacteriophage therapy efficacy in complex infections.IMPORTANCEPhage steering-using phages that bind essential virulence or resistance-associated structures-offers a promising solution by selecting for resistance mutations that attenuate pathogenic traits. However, it remains unclear whether this strategy remains effective in polymicrobial contexts, where interspecies interactions may alter selective pressures. Here, we demonstrate that Pseudomonas aeruginosa evolves phage resistance via loss-of-function mutations in Type IV pilus (T4P) when challenged with the T4P-binding phage Luz19 and that this evolutionary trajectory is preserved even in the presence of a competing pathogen, Staphylococcus aureus. Phage resistance was phenotypically confirmed via twitching motility assays and genotypically via whole-genome sequencing. These findings support the robustness of phage steering under interspecies competition, underscoring its translational potential for managing complex infections-such as those seen in cystic fibrosis-where microbial diversity is the norm.

PMID:40492711 | DOI:10.1128/spectrum.02882-24

Food Sci Biotechnol. 2025 May 15;34(11):2625-2634. doi: 10.1007/s10068-025-01881-4. eCollection 2025 Jul.

ABSTRACT

Excessive antibiotic use can lead to gut microbiota dysbiosis, resulting in diarrhea. This study aimed to investigate the effects of Bifidobacterium animalis QC08 (QC08) on the antibiotic-induced diarrhea mouse model using lincomycin hydrochloride. Mice were divided into five groups: normal, diarrhea, a drug group, high-dose QC08 (QC08-H), low-dose QC08 (QC08-L). Compared with the diarrhea group, the QC08-H and QC08-L groups showed a decrease in serum serotonin, interleukin (IL)-17A, IL-6, and malondialdehyde levels, whereas total antioxidant capacity levels increased. In the colon and small intestine tissues of QC08-H and QC08-L treated mice, mRNA expression of cystic fibrosis transmembrane conductance regulator (CFTR), epidermal growth factor receptor (EGFR), and transforming growth factor β1 (TGFβ1) was significantly downregulated, whereas expression of sodium-hydrogen exchanger 1 (NHE1) and NHE4 was upregulated compared to the AAD model group. Altogether, this in vivo study demonstrates the potential of QC08 in alleviating AAD in mice, providing valuable insights for understanding and potentially treating antibiotic-related gut issues.

PMID:40492055 | PMC:PMC12145372 | DOI:10.1007/s10068-025-01881-4

ERJ Open Res. 2025 Jun 9;11(3):00747-2024. doi: 10.1183/23120541.00747-2024. eCollection 2025 May.

ABSTRACT

Neonicotinoid pesticides suppress CFTR function and mucociliary clearance via α-7 nicotinic acetylcholine receptors in airway epithelium, as shown by studies in human cells and rat lungs, elucidating their link to respiratory disease https://bit.ly/3Z0yuas.

PMID:40491467 | PMC:PMC12147129 | DOI:10.1183/23120541.00747-2024

ERJ Open Res. 2025 Jun 9;11(3):00962-2024. doi: 10.1183/23120541.00962-2024. eCollection 2025 May.

ABSTRACT

Patients with bronchiectasis treated with long-term high-flow nasal therapy showed a significant improvement in mucus plug score https://bit.ly/3NV39zI.

PMID:40491465 | PMC:PMC12147106 | DOI:10.1183/23120541.00962-2024

ERJ Open Res. 2025 Jun 9;11(3):00996-2024. doi: 10.1183/23120541.00996-2024. eCollection 2025 May.

ABSTRACT

Pneumonia and parapneumonic effusion are uncommon in adults with CF. Clinical presentation and recovery may change following introduction of CFTR modulators (HEMT). Further prospective studies are needed. https://bit.ly/3BktB2N.

PMID:40491459 | PMC:PMC12147165 | DOI:10.1183/23120541.00996-2024

Diabetes Obes Metab. 2025 Jun 9. doi: 10.1111/dom.16516. Online ahead of print.

NO ABSTRACT

PMID:40490408 | DOI:10.1111/dom.16516

J Vis Exp. 2025 May 23;(219). doi: 10.3791/67982.

ABSTRACT

The CRISPR-Cas9 system has been harnessed and repurposed into a powerful genome editing tool. By leveraging this technology, researchers can precisely cut, paste, and even rewrite DNA sequences within living cells. Nevertheless, the application of CRISPR screen technology goes far beyond mere experimentation. It serves as a pivotal tool in the fight against genetic diseases, systematically dissecting complex genetic landscapes, empowering researchers to unravel the molecular mechanisms underlying biological phenomena, and enabling scientists to identify and target the root causes of illnesses such as cancer, cystic fibrosis, and sickle cell anemia. Among all, cancer poses a formidable challenge for medicine, spurring eradication efforts. Radiotherapy, as a traditional treatment, yields results but has limitations. It eradicates cancer cells but also damages healthy tissues, causing adverse effects that reduce quality of life. Additionally, not all cancer cells respond to radiotherapy, and some may develop resistance, worsening the condition. To address this, a comprehensive whole-genome CRISPR screen technology is introduced, as it enables the efficient identification of radiosensitive and radioresistant genes, thereby advancing the field of cancer research and treatment. A genome-wide CRISPR screen was conducted in lung adenocarcinoma cells exposed to irradiation following the described protocol, through which both radioresistance- and radiosensitivity-associated genes were identified.

PMID:40489409 | DOI:10.3791/67982