Stat Med. 2025 May;44(10-12):e70082. doi: 10.1002/sim.70082.

ABSTRACT

Group sequential designs in clinical trials allow for interim efficacy and futility monitoring. Adjustment for baseline covariates can increase power and precision of estimated effects. However, inconsistently applying covariate adjustment throughout the stages of a group sequential trial can result in inflation of type I error, biased point estimates, and anticonservative confidence intervals. We propose methods for performing correct interim monitoring, estimation, and inference in this setting that avoid these issues. We focus on two-arm trials with simple, balanced randomization and continuous outcomes. We study the performance of our boundary, estimation, and inference adjustments in simulation studies. We end with recommendations about the application of covariate adjustment in group sequential designs.

PMID:40377247 | DOI:10.1002/sim.70082

J Respir Biol Transl Med. 2025 Mar;2(1):10002. doi: 10.70322/jrbtm.2025.10002. Epub 2025 Mar 24.

ABSTRACT

Pulmonary arterial hypertension (PAH) is a progressive, lethal, and incurable disease of the pulmonary vasculature. A previous genome-wide association study (GWAS) with Affymetrix microarray analysis data exhibited elevated histidine triad nucleotide-binding protein 3 (HINT3) in the lung samples of PAH compared to control subjects (failed donors, FD) and the positive correlations of HINT3 with deubiquitinase USP11 and B-cell lymphoma 2 (BCL2). In this study, we aim to investigate the roles and interplay of USP11 and HINT3 in the apoptosis resistance of PAH. The levels of USP11 and HINT3 were increased in the lungs of idiopathic PAH (IPAH) patients and Hypoxia/Sugen-treated mice. USP11 and HINT3 interacted physically, as shown by co-immunoprecipitation (co-IP) assay in human pulmonary arterial endothelial cells (HPAECs). HINT3 was degraded by polyubiquitination, which was reversed by USP11. Furthermore, HINT3 interacted with the anti-apoptotic mediator, BCL2. Overexpression of USP11 increased BCL2 content, congruent to elevated lung tissue levels seen in IPAH patients and Hypoxia/Sugen-treated mice. Conversely, the knockdown of HINT3 function led to a depletion of BCL2. Thus, we conclude that USP11 stabilizes HINT3 activation, which contributes to endothelial apoptosis-resistance of pulmonary arterial endothelial cells in PAH. This can potentially be a novel therapeutic target for ubiquitination modulators for PAH.

PMID:40376595 | PMC:PMC12080269 | DOI:10.70322/jrbtm.2025.10002

Front Pharmacol. 2025 May 1;16:1559399. doi: 10.3389/fphar.2025.1559399. eCollection 2025.

ABSTRACT

BACKGROUND: Pharmacogenomic (PGx) variants can significantly impact drug response, but limited data exists on their prevalence in Middle Eastern populations. This study aimed to investigate the inheritance of certain markers in candidate pharmacogenes among healthy Saudis.

METHODS: DNA samples from 95 unrelated healthy Saudi participants were genotyped using the Affymetrix Axiom Precision Medicine Diversity Array. Thirty-eight variants in 15 pharmacogenes were analyzed based on their clinical relevance and lack of previous reporting in Saudi populations.

RESULTS: Twenty-six of the 37 tested markers were undetected in the cohort. The selected variants in six genes [DPYD (rs1801268), CACNA1S (rs772226819), EGFR (rs121434568), RYR1 (rs193922816), CYP2B6 (rs3826711), and MT-RNR1 (rs267606617, rs267606618, rs267606619)] were found to be non-existing among Saudis. In contrast, 11 variants and alleles in nine pharmacogenes were detected at varying frequencies. Notable findings included high frequencies of variants in ATIC [rs4673993, minor allele frequency (MAF) = 0.71)] and SLC19A1 (rs1051266, MAF = 0.48) affecting methotrexate efficacy. Three alleles were identified in CYP3A4, including a common (CYP3A4 rs2242480) and two rare alleles (*3 and *22). Another three markers [rs16969968 in CHRNA5, rs11881222 in IFNL3 (IL28B), and SLCO1B1*14] were found to be highly distributed among the participants (MAF = 0.35, 0.30, and 0.14, respectively). Conversely, three rare markers: CYP2A6*2, NAT2*14, and rs115545701 in CFTR, were identified at low-frequency levels (MAF = 0.021, 0.011, 0.005, respectively). Statistically significant differences in allele frequencies were observed for eight variants between Saudi and African populations, five variants compared to East Asians, and two variants compared to Europeans.

CONCLUSION: This study provides novel insights into the distribution of clinically relevant PGx variants in the Saudi population. The findings have implications for personalizing treatments for various conditions, including rheumatoid arthritis, cystic fibrosis, and hepatitis C. These data contribute to the development of population-specific PGx testing panels and treatment guidelines.

PMID:40376268 | PMC:PMC12078325 | DOI:10.3389/fphar.2025.1559399

Nat Nanotechnol. 2025 May 15. doi: 10.1038/s41565-025-01903-6. Online ahead of print.

ABSTRACT

Liver macrophages capture circulating nanoparticles and reduce their delivery to target organs. Serum proteins adsorb to the nanoparticle surface after administration. However, the adsorbed serum proteins and their cognate cell receptors for removing nanoparticles from the bloodstream have not been linked. Here we use a multi-omics strategy to identify the adsorbed serum proteins binding to specific liver macrophage receptors. We discovered six absorbed serum proteins that bind to two liver macrophage receptors. Nanoparticle physicochemical properties can affect the degree of the six serum proteins adsorbing to the surface, the probability of binding to cell receptors and whether the liver removes the nanoparticle from circulation. Identifying the six adsorbed proteins allowed us to engineer decoy nanoparticles that prime the liver to take up fewer therapeutic nanoparticles, enabling more nanoparticles for targeting extrahepatic tissues. Elucidating the molecular interactions governing the nanoparticle journey in vivo will enable us to control nanoparticle delivery to diseased tissues.

PMID:40374797 | DOI:10.1038/s41565-025-01903-6

An Pediatr (Engl Ed). 2025 May 14:503857. doi: 10.1016/j.anpede.2025.503857. Online ahead of print.

ABSTRACT

Cystic fibrosis is a severe genetic disease caused by variants in the CFTR gene. Although it is a multisystem disease, respiratory involvement is the main cause of morbidity and mortality. Cystic fibrosis transmembrane conductance regulator modulator (CFTRm) therapies have advanced the treatment of this disease by improving function of this protein. Ivacaftor, the first CFTRm, has been found to significantly improve lung function and quality of life in patients with certain gating variants. However, only a small percentage of patients in Spain are eligible for this treatment. Combinations of correctors and potentiators, such as lumacaftor-ivacaftor or tezacaftor-ivacaftor, have been developed for treatment of patients with the most frequent variant (F508del), although with limited benefits. Triple therapy with elexacaftor-tezacaftor-ivacaftor has been found to significantly improve respiratory, gastrointestinal and nutritional outcomes as well as quality of life, thus changing the management of CF in eligible patients. The impact of triple therapy is also reflected in an increase in life expectancy and a decrease in mortality and lung transplantation. As regards hepatic and pancreatic involvement, while CFTR modulators have exhibited promising effects, further research is required. The use of CFTR modulators has also shifted nutritional status trends in patients with CF, reducing the risk of undernutrition but increasing the risk of obesity. The use of these drugs for treatment of less frequent variants and for potential antenatal treatment is currently being investigated. Despite these advances, there is still a subset of patients who are ineligible for treatment with modulators or highly effective therapy.

PMID:40374426 | DOI:10.1016/j.anpede.2025.503857

Diabetes Care. 2025 May 15:dc250044. doi: 10.2337/dc25-0044. Online ahead of print.

ABSTRACT

OBJECTIVE: A number of disease-modifying therapies have been introduced for people with cystic fibrosis (CF) over the past two decades. The cumulative effects of this changing landscape on CF-related diabetes (CFRD) are unclear. We examined trends in CFRD epidemiology over time using data from the U.S. Cystic Fibrosis Foundation Patient Registry (CFFPR).

RESEARCH DESIGN AND METHODS: CFFPR data from 2003 to 2018 were queried to determine annual screening, incidence, and prevalence rates of CFRD. Individuals with incident CFRD were compared with individuals without CFRD. Survival analyses were performed to estimate the cumulative hazard of CFRD given predictors of interest over the 15 years of study. Data were also grouped into three time periods (2003-2008, 2009-2013, and 2014-2018) to investigate whether the hazard of developing CFRD varied over time.

RESULTS: CFRD screening rates increased from 2003 to 2018, particularly in 10- to 18-year-olds. Although screening rates increased in adults, overall rates remained low. In 10- to 18-year-olds, the incidence of CFRD was stable over time, while incident cases in adults steadily decreased, approaching incident rates in adolescents. Despite this, the prevalence of CFRD has gradually increased in adults, likely reflecting increased longevity. Age, female sex, Black race, severe mutation class, liver disease, poorer lung function, pancreatic insufficiency, enteric feeds, and low and high BMI were all risk factors associated with CFRD.

CONCLUSIONS: Findings support the need for the development of tailored CFRD screening algorithms and increased subspecialists to care for a growing population of adults with CF and CF-associated comorbidities.

PMID:40372381 | DOI:10.2337/dc25-0044

Antimicrob Agents Chemother. 2025 May 15:e0003025. doi: 10.1128/aac.00030-25. Online ahead of print.

ABSTRACT

Patients with cystic fibrosis have dysfunctional cystic fibrosis transmembrane conductance regulator (CFTR), and this predisposes them to nontuberculous mycobacteria (NTM), including Mycobacterium abscessus (MAB), infection. We found that one of the CFTR modulators, ivacaftor, kills MAB in a concentration-dependent manner, with killing efficacy comparable to amikacin and imipenem, drugs in guideline-based regimens. Using clinical isolates of MAB, amikacin 1/4× MIC concentration combined with ivacaftor killed 2.67 log10 CFU/mL MAB.

PMID:40372084 | DOI:10.1128/aac.00030-25

Otolaryngol Pol. 2025 Mar 19;79(3):1-8. doi: 10.5604/01.3001.0055.0503.

ABSTRACT

<b>Introduction:</b> In the pathophysiology of chronic upper respiratory tract inflammation, an important role is attributed to the disturbances of the patient's microbiome in terms of diversity and functioning, to the decreased abundance of commensal bacteria and the increase of pathogenic bacteria. In recent years, there has been growing scientific interest in the role of probiotics - administered both locally and orally - in the management of various diseases, particularly inflammatory conditions such as chronic rhinosinusitis. <br><br><b>Aim:</b> To assess the use of nasal rinsing with probiotics in patients with rhinitis and rhinosinusitis (primary and secondary). <br><br><b>Material and methods:</b> A total of 51 patients (31 women and 20 men) were included in the study, including 24 patients with granulomatosis with polyangiitis during immunosuppressive therapy (12 women and 12 men) and 27 patients (19 women and 8 men) with rhinitis (chronic rhinosinusitis with polyps, chronic rhinosinusitis without nasal polyps, atrophic rhinitis with nasal septum perforation, and allergic rhinitis). Exclusion criteria were: cystic fibrosis, primary ciliary dyskinesia, pregnancy, severe lung, heart, kidney disease, use of oral probiotics, use of intranasal probiotics in the last 6 months, sinus surgery in the last 6 months, lack of consent to participate in the study, antibiotic therapy in the last 2 months. Patients were scheduled to undergo nasal rinsing with a probiotic solution, with the following parameters assessed before and after the procedure: SNOT-22 scores and the severity of nasal lesions according to the Lund-Kennedy scale. In the group of patients with rhinitis, the ENS-6 questionnaire was also conducted and symptoms assessed on the VAS scale (visual analogue scale): nasal discharge, nasal obstruction, facial pain, impaired sense of smell, nasal irritation, nasal itching, and severity of crusting. <br><br><b>Results:</b> The study showed that nasal rinsing with a probiotic solution is well tolerated and does not cause any adverse effects. In both groups, a reduction in symptoms was observed based on the SNOT-22 questionnaire (p = 0.002 in GPA, ns - in rhinitis/ rhinosinusitis). According to the Lund-Kennedy scale, the reduction in the intensity of changes in both groups was statistically significant. In addition, patients with primary rhinitis or rhinosinusitis also experienced a reduction in nasal mucosa irritation and crusting intranasal (p<0.05). <br><br><b>Conclusions:</b> Probiotic nasal rinsing appears to have a beneficial effect on the condition of the nasal mucosa in patients with both primary and secondary (GPA-related) rhinosinusitis and is generally well tolerated.

PMID:40371957 | DOI:10.5604/01.3001.0055.0503

Int Forum Allergy Rhinol. 2025 May 15:e23591. doi: 10.1002/alr.23591. Online ahead of print.

NO ABSTRACT

PMID:40371719 | DOI:10.1002/alr.23591

Development. 2025 May 15:dev.204277. doi: 10.1242/dev.204277. Online ahead of print.

ABSTRACT

The small intestine is well known for its nutrient-absorbing enterocytes; yet equally critical for homeostasis is a diverse set of secretory cells, all presumed to originate from the same intestinal stem cell. Despite their major roles in intestinal function and health, understanding how the full spectrum of secretory cell types arises remains a longstanding challenge, largely due to their comparative rarity. Here, we investigate the specification of a rare population of small intestinal epithelial cells found in rats and humans but not mice: CFTR High Expressers (CHEs). Using pseudotime trajectory analysis of single-cell RNA-seq data from rat jejunum, we provide evidence that CHEs are specified along the secretory lineage and appear to employ a second wave of Notch-based signaling to distinguish themselves from other secretory cells. We validate the transcription factors directing these cells from crypt progenitors and demonstrate that Notch signaling is necessary to induce CHE fate in vivo and in vitro. Our findings suggest that Notch reactivation along the secretory lineage specifies CHEs, which may help regulate luminal pH and have direct relevance in cystic fibrosis pathophysiology.

PMID:40371707 | DOI:10.1242/dev.204277

JAMA Netw Open. 2025 May 1;8(5):e2510298. doi: 10.1001/jamanetworkopen.2025.10298.

ABSTRACT

IMPORTANCE: Patients with dementia have considerable supportive care needs. Specialist palliative care may be beneficial, but it is unclear which patients are most appropriate for referral and when they should be referred.

OBJECTIVE: To identify a set of consensus referral criteria for specialist palliative care for patients with dementia.

DESIGN, SETTING, AND PARTICIPANTS: In this survey study using 3 rounds of Delphi surveys, an international, multidisciplinary panel of clinicians from 5 continents with expertise in the integration of dementia and palliative care were asked to rate 83 putative referral criteria (generated from a previous systematic review and steering committee discussion). Specialist palliative care was defined as an interdisciplinary team consisting of practitioners with advanced knowledge and skills in palliative medicine offering consultative services for specialist-level palliative care in (nonhospice) inpatient, outpatient, community, and home-based settings.

MAIN OUTCOMES AND MEASURES: Consensus was defined a priori as at least 70% agreement among experts. A criterion was coded as major if the experts advocated that meeting 1 criterion alone was satisfactory to justify a referral. Data were summarized using descriptive statistics.

RESULTS: Of the 63 invited and eligible panelists, the response rate was 58 (92.1%) in round 1, 58 (92.1%) in round 2, and 60 (95.2%) in round 3. Of the 58 panelists who provided demographic data in round 1, most were aged 40 to 49 years (28 of 58 [48.3%]), and 29 panelists (50%) each were men and women. Panelists achieved consensus on 15 major and 42 minor criteria for specialist palliative care referral. The 15 major criteria were grouped under 5 categories, including dementia type (eg, rapidly progressive dementia), symptom distress (eg, severe physical symptoms), psychosocial factors or decision-making (eg, request for hastened death, assisted suicide, or euthanasia), comorbidities or complications (eg, ≥2 episodes of aspiration pneumonia in the past 12 months); and hospital use (eg, ≥2 hospitalizations within the past 3 months).

CONCLUSIONS AND RELEVANCE: In this Delphi survey study, international experts reached consensus on a range of criteria for referral to specialist palliative care. With testing and validation, these criteria may be used to standardize specialist palliative care access for patients with dementia across various care settings.

PMID:40366652 | PMC:PMC12079294 | DOI:10.1001/jamanetworkopen.2025.10298

Inn Med (Heidelb). 2025 May 14. doi: 10.1007/s00108-025-01912-6. Online ahead of print.

ABSTRACT

Adolescence represents a period of significant transformation for individuals with chronic conditions such as cystic fibrosis (CF) and asthma. Due to substantial progress in highly effective cystic fibrosis transmembrane conductance regulator (CFTR) modulator therapy, nearly everyone with CF now reaches adulthood, necessitating ongoing interdisciplinary medical care. Conversely, approximately 70% of children with asthma experience a resolution of symptoms as they reach adulthood. Factors contributing to the persistence of asthma into adulthood include the presence of severe asthma, diminished lung function in childhood and allergic comorbidities. Especially for young adults with severe asthma, continuous pneumological care including uninterrupted access to biologics, must be provided. During the transition process, considerations regarding career choices are pertinent for both patient groups. Additionally, issues related to family planning and prenatal diagnostics become particularly relevant for young adults with CF and should be addressed during the transition process. In both patient groups, an early and structured initiation of the transition process is essential. The use of checklists and transition plans can facilitate the transfer of critical information and ensure a seamless transition. Joint pediatric and internal medicine consultations foster trust and ensure medical quality. Ultimately, a successful transition should enable individuals with CF as well as those with asthma to assume responsibility for their condition and treatment, recognize clinical deterioration, and seek timely assistance.

PMID:40369362 | DOI:10.1007/s00108-025-01912-6

Eur Respir Rev. 2025 May 14;34(176):240220. doi: 10.1183/16000617.0220-2024. Print 2025 Apr.

ABSTRACT

INTRODUCTION: The Montreal Cystic Fibrosis Related Diabetes Screening Cohort (MCFC) was established in 2004 to study the prevalence, risk factors and management of cystic fibrosis-related diabetes, a significant extrapulmonary complication of cystic fibrosis with an increasing prevalence due to improved cystic fibrosis survival rates. The aims of this review are to highlight the key insights gained from monitoring the MCFC over 20 years, and to discuss the challenges and advantages of establishing such a cohort in a rare disease like cystic fibrosis.

METHODS: Adult people with cystic fibrosis were recruited from 2004 onward in Montreal, Canada, excluding those already diagnosed with cystic fibrosis-related diabetes. Clinical and biological results (including oral glucose tolerance tests) were recorded regularly.

RESULTS: Findings from the MCFC contributed to a better understanding of cystic fibrosis-related diabetes pathophysiology (in particular, the joint roles of reduced insulin secretion and added insulin resistance) and its relationship with lung function. Over the years, we observed a shift towards overweight and obesity among cystic fibrosis patients, along with improved lung function. This could be due to improved cystic fibrosis care and to the introduction of cystic fibrosis transmembrane conductance regulator modulators. We were also able to validate new, simplified screening modalities and management strategies (e.g. physical activity) for cystic fibrosis-related diabetes.

CONCLUSION: The MCFC has contributed to the understanding of cystic fibrosis-related diabetes and informed best practice guidelines. Future research will focus on how cystic fibrosis transmembrane conductance regulator modulators influence glycaemic control and cardiometabolic health in people with cystic fibrosis.

PMID:40368427 | DOI:10.1183/16000617.0220-2024

Eur Respir Rev. 2025 May 14;34(176):240261. doi: 10.1183/16000617.0261-2024. Print 2025 Apr.

ABSTRACT

The increasing life expectancy of people with cystic fibrosis (pwCF), largely driven by advancements in early diagnosis, multidisciplinary care and the recent introduction of cystic fibrosis transmembrane conductance regulator (CFTR) modulator therapies, is likely to herald a shift in the focus of care toward managing the complexities of ageing. This review highlights key challenges and research priorities for addressing the health needs of an ageing CF population. A growing body of evidence underscores the heightened risks of cancers, cardiovascular diseases and changing nutritional and metabolic profiles as pwCF age. CFTR modulators have improved clinical outcomes, but their effects on inflammation, immunity and long-term disease trajectories remain incompletely understood. Nutritional management, particularly the implications of obesity and body composition, poses new challenges, as does the potential accelerated ageing of immune and pulmonary systems in CF. Emerging issues such as menopause in females with CF, lifetime antimicrobial resistance and the interplay between chronic inflammation and ageing further complicate the care landscape. The review emphasises the urgent need for multidisciplinary research programmes that integrate clinical, patient and community perspectives. Leveraging established CF registries, clinical trial networks and collaborations with ageing research frameworks is critical to addressing these challenges. Ultimately, the goal is to ensure that pwCF not only live longer but also experience improved quality of life and holistic wellbeing as they realise the full benefits of therapeutic advances.

PMID:40368426 | DOI:10.1183/16000617.0261-2024

J Allergy Clin Immunol Pract. 2025 May 12:S2213-2198(25)00437-4. doi: 10.1016/j.jaip.2025.05.013. Online ahead of print.

ABSTRACT

The pathophysiology of atopic dermatitis (AD) involves cutaneous inflammation, predominantly mediated by innate immunity and T cells, with Immunoglobulin E (IgE) playing a marginal role in most patients. Over previous decades, however, there has been an ongoing debate regarding the relevance of IgE-mediated allergy testing in AD patients. Patients with AD have a defective skin barrier that facilitates a high inflammatory response to environmental antigens, placing them at greater risk for food allergies. Nevertheless, because these patients often produce very high levels of IgE, the positive predictive value of skin prick tests and specific IgE measurements is low; such tests should be performed only when there is a concordant immediate hypersensitivity reaction (i.e. urticaria or angioedema) rather than eczema. In recent years, numerous studies have emphasized the importance of maintaining oral exposure to foods in order to prevent the development or progression of food allergies in atopic patients. While acknowledging that food allergens may contribute to AD in certain cases, it is critical that patients understand the risk of developing IgE-mediated food allergies if they exclude allergenic foods from their diet. Ultimately, controlling AD while retaining these foods in the diet should be the goal for all patients.

PMID:40368248 | DOI:10.1016/j.jaip.2025.05.013

Einstein (Sao Paulo). 2025 May 12;23:eAO1312. doi: 10.31744/einstein_journal/2025AO1312. eCollection 2025.

ABSTRACT

BACKGROUND: Santos et al. analyzed the clinical characteristics and pulmonary function of children with cystic fibrosis infected with SARS-CoV-2. Infected children showed higher rates of dyspnea, coughing, hospitalization, and pulmonary exacerbations. Despite a temporary decline in pulmonary function, the recovery rates matched those of the uninfected children during follow-up. ■ SARS-CoV-2 infection leads to mild-to-moderate disease in children with cystic fibrosis. ■ No worsening of cystic fibrosis was observed months after infection.

OBJECTIVE: This study aimed to evaluate the clinical manifestations of SARS-CoV-2 in children and adolescents with cystic fibrosis.

METHODS: This was a case-control analysis of clinical variables and pulmonary function test results in 43 children with cystic fibrosis, 17 (39.5%) of whom tested positive for SARS-CoV-2.

RESULTS: The infected children exhibited a higher frequency of dyspnea and cough and a greater need for hospitalization. One infected child died. Pulmonary exacerbations were more frequent among the infected children. Additional data indicated a subsequent reduction in pulmonary function in the infected children, although this was not significantly different from that in the uninfected children.

CONCLUSION: Children with cystic fibrosis who contracted SARS-CoV-2 experienced mild to moderate symptoms and required hospitalization but generally had high recovery rates.

PMID:40367008 | DOI:10.31744/einstein_journal/2025AO1312

J Man Manip Ther. 2025 May 14:1-7. doi: 10.1080/10669817.2025.2505096. Online ahead of print.

ABSTRACT

BACKGROUND: Cystic fibrosis (CF) is a severe hereditary disease that affects multiple organ systems. Among these, the musculoskeletal system is an under-explored area. This interview study aimed to explore experiences of musculoskeletal symptoms and of manual therapies as complementary care in this context.

METHODS: Semi-structured interviews were used to collect data from ten respondents. The data were subsequently analyzed through content analysis with an inductive approach in accordance with the method of Elo and Kyngäs.

RESULTS: The analysis resulted in three main categories; 1) Living with CF involves musculoskeletal health challenges, 2) Manual therapies impact daily life for people with CF, and 3) People with CF aspire for broader and more collaborative respiratory care.

CONCLUSION: The respondents described musculoskeletal symptoms in and around the thoracic cage. They experienced symptom relief and increased body awareness following manual therapy interventions (MTI) and recommended that these methods be offered as complementary care to enhance quality of life.

PMID:40366667 | DOI:10.1080/10669817.2025.2505096

mBio. 2025 May 14;16(5):e0133824. doi: 10.1128/mbio.01338-24. Epub 2025 Apr 9.

ABSTRACT

Strict and facultative anaerobic bacteria are widely associated with both acute and chronic airway diseases. However, their potential role(s) in disease pathophysiology remains poorly understood due to inherent limitations of existing laboratory models and conflicting oxygen demands between anaerobes and host cells. To address these limitations, here, we describe a dual oxic-anoxic culture (DOAC) approach that maintains an oxygen-limited microenvironment at the apical epithelial interface while host cells are oxygenated basolaterally. This platform enables epithelial-anaerobe co-culture for ~48 h, and we demonstrate its utility by evaluating reciprocal interactions between the oxygen-sensitive anaerobic bacterium, Fusobacterium nucleatum, and oxygen-demanding airway epithelial cells at the transcriptional level. Using bulk RNAseq, we demonstrate that epithelial colonization results in altered gene expression by F. nucleatum, highlighted by the differential expression of genes associated with virulence, ethanolamine and lysine metabolism, metal uptake, and other transport processes. We also combine DOAC with single-cell RNA sequencing to reveal a cell type-specific transcriptional response of the airway epithelium to F. nucleatum infection, including the increased expression of inflammatory marker genes and cancer-associated pathways. Together, these data illustrate the versatility of DOAC while revealing new insights into anaerobe-host interactions and their mechanistic contributions to airway disease pathophysiology.IMPORTANCEConflicting oxygen demands between anaerobes and host cells present a significant barrier to in vitro modeling of how these cell types interact. To this end, the significance of our dual oxic-anoxic culture (DOAC) approach lies in its ability to maintain anaerobe and epithelial viability during co-culture, paving the way for new insights into the role(s) of anaerobic microbiota in disease. We use DOAC to interrogate reciprocal interactions between the airway epithelium and Fusobacterium nucleatum-an anaerobic commensal with pathogenic potential. Given its link to a range of diseases, from localized infections to various cancers, these data showing how F. nucleatum bacterium re-shapes its metabolism and virulence upon epithelial colonization provide new mechanistic insight into F. nucleatum physiology and how the host responds. We use F. nucleatum as our model, but the DOAC platform motivates additional studies of the gut, lung, and oral cavity, where host-anaerobe interactions and the underlying mechanisms of pathogenesis are poorly understood.

PMID:40366160 | DOI:10.1128/mbio.01338-24

mSphere. 2025 May 14:e0013725. doi: 10.1128/msphere.00137-25. Online ahead of print.

ABSTRACT

Sickle cell disease (SCD) is a chronic blood disorder that disrupts multiple organ systems and can lead to severe morbidity. Persistent and acute symptoms caused by immune system dysregulation in individuals with SCD could contribute to disease either directly or indirectly via dysbiosis of commensal microbes and increased susceptibility to infection. Here, we explored the nasal and oral microbiomes of children with SCD (cwSCD) to uncover potential dysbiotic associations with the blood disorder. Microbiota collected from nasal and oral swabs of 40 cwSCD were compared to eight healthy siblings using shotgun metagenomic sequencing. Commensal taxa were present at similar levels in the nasal and oral microbiome of both groups. However, the nasal microbiomes of cwSCD contained a higher prevalence of Pseudomonadota species, including pathobionts such as Yersinia enterocolitica and Klebsiella pneumoniae. Furthermore, the oral microbiome of cwSCD displayed lower α-diversity and fewer commensal and pathobiont species compared to the healthy siblings. Thus, subtle but notable shifts seem to exist in the nasal and oral microbiomes of cwSCD, suggesting an interaction between SCD and the microbiome that may influence health outcomes.

IMPORTANCE: The oral and nasal cavities are susceptible to environmental exposures including pathogenic microbes. In individuals with systemic disorders, antibiotic exposure, changes to the immune system, or changes to organ function could influence the composition of the microbes at these sites and the overall health of individuals. Children with sickle cell disease (SCD) commonly experience respiratory infections, such as pneumonia or sinusitis, and may have increased susceptibility to infection because of disrupted microbiota at these body sites. We found that children with SCD (cwSCD) had more pathobiont bacteria in the nasal cavity and reduced bacterial diversity in the oral cavity compared to their healthy siblings. Defining when, why, and how these changes occur in cwSCD could help identify specific microbial signatures associated with susceptibility to infection or adverse outcomes, providing insights into personalized treatment strategies and preventive measures.

PMID:40366139 | DOI:10.1128/msphere.00137-25

Pediatr Pulmonol. 2025 May;60(5):e71124. doi: 10.1002/ppul.71124.

NO ABSTRACT

PMID:40365926 | DOI:10.1002/ppul.71124