Journal of Cachexia, Sarcopenia and Muscle

Introduction. Skeletal muscle loss in sarcopenia and neuromuscular disorders remains a major unmet medical need. AAV9-delivered follistatin (FST), a myostatin/activin antagonist, induces muscle hypertrophy; however, fibre growth without adequate vascular adaptation may limit therapeutic efficacy. We evaluated whether co-administration of a VEGF-A165 plasmid enhances the hypertrophic and angiogenic effects of intramuscular AAV-FST gene transfer in C57BL/6 mice. Methods. Thirty-six C57BL/6 mice (18 males, 18 females) were assigned to PBS vehicle (n=10), AAV-FST (1 x 10^11 vg; n=10), VEGF plasmid (100 ug; n=6), or combination treatment (VEGF plus AAV-FST; n=10). The contralateral hindlimb served as an internal control. Endpoints at Day 115 included hindlimb muscle mass ratio (R/L), transgene expression, FST protein levels, muscle fibre morphometry, capillary density, and safety assessments. Results. Combination therapy produced the highest R/L ratio (1.176 +/- 0.091; p=0.004; d=2.04), whereas AAV-FST alone showed a borderline effect (R/L=1.113; p=0.050). Compared with AAV-FST monotherapy, combination treatment increased muscle FST mRNA approximately 2.1-fold, protein levels approximately 2.0-fold, and the muscle-to-liver expression ratio 2.6-fold. It also induced larger muscle fibres and doubled CD31+ vessel counts versus AAV-FST alone, indicating simultaneous hypertrophy and angiogenesis. No adverse haematological, biochemical, or histopathological findings were observed. Discussion. Combined AAV-FST and VEGF therapy enhanced local muscle hypertrophy, increased capillary density, and improved the muscle-to-liver transgene expression profile compared with AAV-FST monotherapy. The regimen was well tolerated and supports further evaluation of angiogenic preconditioning as a strategy to improve muscle-directed gene therapy for muscle-wasting disorders.

Skeletal muscle aging is characterized by the deterioration of muscle function, which can lead to negative quality-of-life outcomes including frailty and sarcopenia. While understanding the mechanisms of this process is increasingly important as the global population ages, previous molecular studies of skeletal muscle aging have been limited by statistical power and cell type resolution. In this study, we analyzed single-nucleus gene expression and chromatin accessibility data from 287 human skeletal muscle samples from individuals aged 20-79 years to explore sex- and cell type- specific aging effects. Across 467,126 nuclei from 13 cell types, we identify 384 age-associated genes and 4,061 age-associated chromatin regions. These age-associated molecular features are enriched for functional pathways, including metabolic processes, cell-to-cell communication, and senescence Kyoto Encyclopedia of Genes and Genomes KEGG terms. Age-associated closing chromatin was more common across fiber types and sexes than opening chromatin, and was enriched in active enhancer regions while depleted for active transcription start sites. We observe enrichment for specific transcription factor motifs in closing chromatin, including those of glucocorticoid and androgen receptors, both of which play a key role in the maintenance of healthy skeletal muscle. Together, these findings identify an age-associated regulatory shift, largely invisible in matched transcriptomic data, characterized by closing chromatin which reduces accessibility to hormone receptor binding sites and enhancer regions in the muscle fiber epigenome.

Skeletal muscle stem cells (MuSCs) give rise to a fusogenic cell population that provide new myonuclei to muscle fibers. Myonuclear functional heterogeneity has recently become appreciated, but the terminal identity of MuSC-Derived myonuclei remains undefined. We performed single-nucleus RNA-sequencing of myonuclei in Adult and Aged muscle to define MuSC-Derived and resident myonuclear responses to mechanical overload (MOV), which induces a hypertrophic stimulus. We found a MuSC-dependent induction of a youthful transcriptional signature in resident myonuclei after MOV in Aged muscle. Age determined terminal transcriptional states of MuSC-Derived myonuclei toward MTJ in Adult, NMJ in Aged, and muscle spindles in both ages. Microtubule-remodeling genes, Macf1, Map1b, and Nav3, along with the transcription factor Runx1, identified this post-fusion specialization with greater expression of these genes in Adult than in Aged MuSC-Derived myonuclei. In-silico transcription factor KO screen identified Runx1 as a regulator of post-fusion specialization and Esrrg as a driver of spindle (intrafusal) MuSC-Derived myonuclear maturation. By defining the age-associated fate of MuSC fusion to muscle fibers, we provide potential targets for modulating muscle plasticity.

Inadequate autophagy promotes smoking-induced cellular senescence involved in chronic obstructive pulmonary disease (COPD) pathogenesis. Transcription factor EB (TFEB) is a master regulator of the autophagy-lysosome axis. For the first time, we investigated the therapeutic potential of pemafibrate, a putative TFEB inducer. COPD lung epithelial cells showed reduced TFEB expression. Pemafibrate enhanced autophagy/mitophagy flux and restored lysosomal acidification observed during cigarette smoke (CS) extract exposure in human bronchial epithelial cells, resulting in reduced cellular senescence. TFEB knockdown demonstrated involvement of pemafibrate-induced TFEB in these effects. Pemafibrate induced TFEB expression, mitigated alveolar enlargement and airflow obstruction, and attenuated the CS-induced increase in static lung compliance in a long-term CS-exposed mouse model. It reduced the CS exposure-induced cellular senescence, possibly through autophagy/mitophagy, as suggested by bulk RNA sequencing of mouse lungs. A retrospective cohort study showed that patients given pemafibrate displayed attenuated FEV1.0 decline compared with those given bezafibrate or fenofibrate. In conclusion, pemafibrate is a promising therapeutic agent for COPD, potentially exerting its effects through the regulation of the TFEB-autophagy/mitophagy-lysosome axis.

Heat shock protein family B member 8 (HSPB8) is a chaperone involved in the chaperone-assisted selective autophagy (CASA) complex. HSPB8 in conjunction with cochaperone BAG3, promotes autophagy-mediated removal of misfolded proteins associated with various neurodegenerative diseases. Mutations in HSPB8, previously associated with Charcot Marie Tooth disease type 2L, have recently been linked to an autosomal dominant rimmed vacuolar myopathy (MFM13), and is considered a multisystem proteinopathy. Patients have distal and proximal limb girdle myopathy with muscle biopsy showing fatty replacement, endomysial fibrosis, and rimmed vacuoles leading to muscle atrophy and early demise. We have demonstrated reduced expression of HSPB8, altered autophagy and TDP-43 accumulation in patient fibroblasts. Using CRISPR technology, we generated a knock-in Hspb8 mouse model of the c.515dupC hot spot frameshift variant to study disease pathology. Overexpressed murine Hspb8 frameshift mutant (c.515dupC, fs) displays insolubility and aggregation propensity in Murine Neuroblastoma X Spinal Cord 34 (NSC-34) cells. Mutant Hspb8 mice developed late-onset muscle weakness beginning at 15 months. Muscle biochemical analyses revealed reduced HSPB8 levels, increased TDP-43, and altered autophagy markers, partially recapitulating the human phenotype. Fiber type analysis, neuromuscular junction integrity, and motor neurons show mild myopathy without neurodegeneration. Given the lack of available treatments, we evaluated trehalose, a natural disaccharide that induces HSPB8 and enhances autophagy. Administration of 2% trehalose in drinking water improves motor performance, restores HSPB8 expression, and ameliorates autophagic and TDP-43 pathology in mutant mice. These findings support the value of our preclinical models for translational studies, and autophagy enhancement as a potential therapeutic strategy for HSPB8-related myopathy.

IntroductionPeripheral nerve ageing leads to profound proteomic remodelling, with shifts in metabolic and inflammatory signalling pathways resembling changes that occur during nerve degeneration and regeneration following injury. Moreover, aged nerves exhibit impaired degeneration and regeneration, contributing to age-related peripheral neuropathies that show sex-specific differences in prevalence. However, it remains unclear whether these alterations arise from intrinsic nerve changes or an altered systemic environment. Therefore, we investigated the impact of sex on age-related proteome changes and nerve-intrinsic proteomic responses in young and aged male and female nerves using an ex vivo degeneration model. MethodsMass spectrometry-based proteomics were performed on young and old nerves from male and female animals, as well as on contralateral nerves after seven days of ex vivo nerve degeneration. A comparative bioinformatic analysis was then used to identify changes during ageing and ex vivo nerve degeneration that were independent of sex, as well as changes that were sex-specific. ResultsEx vivo nerve degeneration induced extensive proteome remodelling in mouse sciatic nerves that was largely independent of age and sex. Principal component and clustering analyses clearly separated intact from degenerated nerves, while revealing only subtle age- and sex-related effects, with more pronounced ageing-associated changes in males. Approximately 20% of age-regulated proteins and 7-10% of degeneration-regulated proteins exhibited sex-specific expression patterns. Degeneration was characterised by increased abundance of lysosomal and repair-associated proteins alongside reduced myelin and axonal proteins, consistent with active tissue remodelling. In aged nerves, impaired protein clearance and partial pre-activation of degeneration-associated pathways suggested altered injury responses. Comparative analyses demonstrated positive correlations of protein abundance changes between ex vivo and in vivo degeneration datasets, although the temporal dynamics were altered in aged nerves. Pathway enrichment analyses identified coordinated regulation of metabolic, RNA-processing and vesicular transport pathways, while ageing was associated with enhanced immune signalling and reduced lipid metabolism. Sex-specific analyses revealed stronger inflammatory signatures in males, whereas females exhibited enrichment of metabolic pathways, including folate biosynthesis. ConclusionThese findings reveal distinct sex-specific molecular features of peripheral nerve ageing, characterised by enhanced inflammatory signalling in males and metabolic adaptations that may confer resilience in females. Our datasets provide a comprehensive molecular resource of sex-dependent changes in peripheral nerve ageing and nerve-intrinsic injury responses, offering a foundation for identifying therapeutic strategies to promote healthy peripheral nerve ageing. Plain English summaryAge-related peripheral neuropathies are common disorders that can cause pain, numbness, weakness and reduced mobility, affecting millions of people worldwide. They become more common from around the age of 50 and affect men and women differently. These conditions are thought to result from age-related changes in the structure and function of peripheral nerves, which reduce their ability to repair themselves after injury. In this study, we used advanced protein analysis (proteomics) to investigate how ageing affects peripheral nerves in male and female mice. We also used an ex vivo model, in which nerves are studied outside the body, to examine how age and sex influence the molecular changes that occur during nerve degeneration. We found that degeneration caused widespread changes in the proteins present in the sciatic nerve in both young and old mice. Most of these changes were similar in males and females, but some important differences emerged. Male nerves showed stronger signs of inflammation, whereas female nerves showed increased activity of metabolic pathways, including those involved in folate metabolism. Ageing nerves also appeared less able to remove damaged material and showed signs of activating degeneration-related processes even before injury. Overall, the ex vivo model reproduced many of the molecular changes seen after nerve injury in living animals, although it did not fully capture the inflammatory response, suggesting that signals from the rest of the body, including factors carried in the blood, also contribute to nerve degeneration. HighlightsO_LIEx vivo nerve degeneration caused major protein changes in young and old mouse sciatic nerves. C_LIO_LIMost degeneration-related protein changes were shared between males and females. C_LIO_LIAgeing altered the nerve proteome, with stronger ageing-related shifts in males. C_LIO_LIMale nerves showed stronger inflammatory and immune-related signatures. C_LIO_LIFemale nerves showed enrichment of metabolic pathways, including folate biosynthesis, and ex vivo degeneration did not fully reproduce the inflammatory response seen after injury in vivo. C_LI

Background Therapy resistance in pancreatic ductal adenocarcinoma (PDAC) is facilitated by the desmoplastic tumor microenvironment (TME) orchestrated by cancer associated fibroblasts (CAFs). Upon activation, pancreatic stellate cells (PSCs) deplete their intracellular retinoic acid (RA)-containing lipid droplets and secrete stromal remodeling proteins like pentraxin 3 (PTX3), leading to cancer progression. Preclinical evidence indicates that all-trans RA (ATRA) reprograms the TME, while circulating vitamin A and PTX3 were proposed as biomarkers for ATRA response in PDAC. To support further clinical development of RA-based therapies in PDAC, we studied the effects of ATRA on CAFs and patient-derived organoids (PDO) and evaluated the clinical relevance of these biomarkers in PDAC patients. Methods We employed viability assays in human and murine organoid mono- and co-culture models to explore the efficacy of adding ATRA to gemcitabine (GEM). In parallel, we conducted a prospective observational study and assessed vitamin A and PTX3 as response biomarkers in peripheral blood collected before first treatment and at cycles 2 and 4 of treatment among patients with advanced PDAC receiving GEM with or without nab-paclitaxel (NAB-P). Results In PDO monocultures, a significant additive effect of ATRA in combination with GEM on viability was observed in 5 (41%) of 12 PDOs and this effect was numerically more frequent in organoids from patients who had clinically responded to GEM. In human and murine 3D PDO+PSC/CAF co-cultures, ATRA demonstrated an additional direct impact on the viability of stromal cells. Clinically, among 18 patients with PDAC treated with GEM+/-NAB-P, patients with no treatment response (n=10) showed an increase in PTX3 and concomitant decrease in vitamin A levels under therapy. In contrast, response was associated with stable vitamin A levels and a trend towards lower PTX3 levels during chemotherapy. Conclusions Our preclinical data support the repurposing of ATRA, an agent with favorable toxicity profile, to potentiate the efficacy of GEM in PDAC treatment. Complementing these results, our clinical data suggest vitamin A and PTX3 as promising response biomarkers in PDAC treatment, not restricted to ATRA containing regimens.

Skeletal muscle, a primary site of insulin-mediated glucose uptake, plays a central role in the pathogenesis of type 2 diabetes. It is therefore critical to understand the disease-associated alterations in skeletal muscle and identify the underlying drivers of this dysregulation. Here, we characterize type 2 diabetes associated transcriptional dysregulation using 301 skeletal muscle biopsies from living donors with and without diabetes. Using weighted gene co-expression network analysis, we identify 56 distinct gene modules, which we further characterize using single-nucleus RNA-seq-derived cell type signatures and pathway enrichment analysis. We identify numerous cell type-associated dysregulated pathways in skeletal muscle tissue from individuals with diabetes, including muscle fiber-associated mitochondrial function and mRNA splicing and processing; endothelial vascularization and phospholipase D signaling; and macrophage- and T-cell-associated inflammation. Through analysis of module hub genes and transcription factor regulatory network analysis, we further identify candidate driver genes of this dysregulation including ATP5L, ATF2, SIRT1, and THRAP3 in muscle fibers; JAM2 and CLEC14A in endothelial cells; and F13A1 and IRF8 in immune cells. Finally, we integrate our co-expression networks with single-nucleus ATAC-seq data to identify proximal and distal genomic regulatory elements and identify context-specific enrichment for type 2 diabetes and related trait GWAS signals in muscle fiber and endothelial modules. Together, our results reveal dysregulation in pathways in muscle tissue from individuals with diabetes, identify candidate drivers, and connect the genomic drivers of this dysregulation across type 2 diabetes and related metabolic traits.

Background: Pancreatic ductal adenocarcinoma (PDAC) has remained one of the most serious malignancies and is still a leading cause of cancer-related deaths worldwide. Great attention has been paid to the relationship between diabetes mellitus and PDAC. The aim of our study was to analyse the mutual association of PDAC and diabetes mellitus in the entire population of the Czech Republic within a 5-year period. Methods: The incidence of PDAC in 2018-2022 was estimated based on the individual population data from the Czech National Cancer Registry. Another data source, the National Registry of Reimbursed Health Services that collects data from health insurance companies, was used to identify individuals recently diagnosed with diabetes mellitus. For the purpose of this study, diagnosis of new-onset of diabetes mellitus was defined as the time of the first prescription of any antidiabetic drug or another related health care service. Subsequently, patients diagnosed with PDAC in 2022 were followed retrospectively to see if they had been diagnosed with diabetes mellitus in the last five years before diagnosis of pancreatic cancer. Results: In 2022, 2,189 patients aged 60 years or older were diagnosed with pancreatic cancer. New-onset diabetes was observed in 17.4% within five years prior to diagnosis, with the highest occurrence (12.4%) within the last three years. Among patients aged 60-74 years, the respective proportions were 14.4% within three years and 5.7% four to five years prior to pancreatic cancer diagnosis. Conclusion: The incidence of pancreatic cancer in the Czech Republic is among the highest in Europe. One-fifth of PDAC cases are diagnosed following new-onset diabetes mellitus in patients over sixty. Unintended significant weight loss combined with new-onset diabetes thus must not be overlooked, as these can be early signs of PDAC. An individualised diagnostic work-up should follow without any unnecessary delay.

BCL2L13 is a mitochondrial BCL2 family protein linked to mitophagy and ceramide metabolism, but its role in NSCLC metastatic plasticity remains unclear. Human lung cancer Tissue Microarray and matched patient specimens showed subtype and site dependent BCL2L13 expression, with higher cytoplasmic granular staining in primary NSCLC and reduced, heterogeneous staining in lymph node metastases, most evident in adenocarcinoma and squamous cell carcinoma. Because Epithelial mesenchymal transition and anoikis resistance are central requirements for metastatic dissemination, this primary to node attenuation provided the rationale to test BCL2L13 knockdown and overexpression in metastasis relevant NSCLC models. In A549 and LLC cell lines. TGF beta 1 induced coordinated mitophagy and EMT with mitochondrial enrichment of BCL2L13. BCL2L13 knockdown impaired TGF beta 1 and carbonyl cyanide m chlorophenyl hydrazone associated mitophagy, reducing LC3 beta mitochondria colocalization, TOMM20, LAMP1 overlap and mitochondrial LC3 II, p62, TOMM20 turnover; BNIP3 and NIX redistribution did not compensate. BCL2L13 loss enhanced EMT marker switching and migration, whereas overexpression partially opposed these changes. During detachment, BCL2L13 knockdown reduced anoikis associated apoptosis despite preserved mitochondrial recruitment of BAX, BAK, BNIP3,NIX, altered BID processing, non parallel caspase activity and shifted FAK phosphorylation. Pharmacological autophagy modulation did not reverse this anoikis phenotype. Lipidomics identified adhesion state dependent ceramide synthases CerS2, CerS6 linked sphingolipid remodeling: BCL2L13 knockdown increased C24 linked sphingolipid species in attached cells but reduced C16, C24 ceramide related profiles during anoikis. These findings identify BCL2L13 downregulation as a metastasis associated mitochondrial-lipid state that limits mitophagic quality control while favoring EMT and detachment survival in NSCLC adenocarcinoma.

Skeletal muscle dynamically adapts to physiological stimuli such as exercise through coordinated molecular and structural remodeling processes. Circulating microRNAs (miRNAs) represent promising non-invasive biomarkers of exercise responsiveness and skeletal muscle physiological states; however, most analytical frameworks rely solely on annotated miRNAs and overlook novel candidates. Here, we present nf-sarcopipe, a modular Nextflow pipeline that integrates de novo and reference-guided miRNA discovery with transcriptomic analysis and regulatory network reconstruction. The pipeline is organized into three complementary modules: 1) Preprocessing, 2) miRNA Discovery, and 3) Target Prediction & mRNA Integration. Using publicly available datasets from active and sedentary young women, the pipeline identified reproducible miRNA signatures and prioritized a small set of structurally supported, high-confidence de novo candidates. Previously reported exercise-associated miRNAs compiled from the literature were additionally incorporated for comparative candidate evaluation. Although the available datasets were derived from different tissues, confounding-aware analyses enabled the identification of coherent transcriptional signatures associated with exercise responsiveness. Integrative miRNA-mRNA analysis uncovered consistent regulatory interactions linking circulating miRNAs--both novel and known--to pathways involved in immune response, extracellular matrix remodeling, autophagy, and skeletal muscle adaptation. Together, these results establish nf-sarcopipe as a robust and scalable framework for complementary miRNA discovery and for investigating regulatory mechanisms associated with exercise-induced skeletal muscle adaptation.

Abstract Background: Intraductal papillary mucinous neoplasms (IPMNs) are recognized as precursor lesions to pancreatic ductal adenocarcinoma (PDAC). However, the molecular programs underlying progression from low-grade dysplasia to advanced disease remain incompletely characterized. Herein, we performed an integrated plasma and tissue-proteomic analyses coupled with spatial and single-cell transcriptomics to identify biologically coherent remodeling programs reflected in circulation that distinguish IPMN by dysplasia grade and invasive disease. Methods: Using the O-link proximity extension assay platform, a panel of 1,104 proteins were quantified in plasma samples collected from patients with low-grade (LG) IPMN (n=30), high-grade (HG) IPMN with or without associated PDAC (IPMN/PDAC; n=40) and PDAC without IPMN (n=8). Predictive performance of individual biomarkers were assessed; likelihood ratio testing was performed to identify protein biomarkers that were complementarity with CA19-9 for risk of malignancy of IPMN. Findings were intersected with available spatial (N= 13) and single-cell (N= 6) transcriptomic datasets of IPMN tissues as well as mass spectrometry-based proteomic profiles of an independent set of resected human IPMN tissues (N= 9). Results: A total of 28, 43, and 35 circulating proteins were found to be differential in HG, IPMN/PDAC, and HG + IPMN/PDAC cases compared to LG IPMN. Among differential proteins were known PDAC-associated markers CEACAM5, CTRC, and REG3A as well as several biomarkers reflecting cytoskeletal and extracellular matrix remodeling and inflammatory processes. Focusing on cytoskeletal and ECM-related proteins and using likelihood ratio testing, an OR rule considering CA19-9, BGN, and ITGB1BP1 achieved overall sensitivity of 48.7% for HG + IPMN/PDAC, including 38.1% sensitivity for HG IPMN, at an overall specificity of 90%, which was improved compared to that of CA19-9 alone (overall sensitivity of 28.2%; McNemar Exact test 1-sided p-value: 0.011). Integrated proteomic and spatial transcriptomic datasets of IPMN tissues revealed coordinated alterations cytoskeletal and ECM remodeling and elevated matrix stiffness as prominent features associated with IPMN/PDAC, which paralleled concordant increases in BGN and ITGB1BP1. Cell-type of origin analyses based on spatial and single-cell data further revealed fibroblasts and myeloid cells as primary contributors to expression levels of BGN whereas ITGB1BP1 was primarily expressed in neoplastic epithelium. Conclusion: Advanced IPMN dysplasia and invasive disease are characterized by coordinated tissue remodeling programs that are systemically reflected in circulating proteomic profiles. Blood-based biomarkers identified through our study, such as BGN and ITB1BP1, have potential to improve upon CA19-9 for risk stratification of IPMN to better guide clinical management.

Reduced nicotinamide mononucleotide (NMNH) is a reduced NAD precursor with reported NAD- augmenting activity in preclinical models; however, controlled human data remain limited. This was a randomized, double-blind, placebo-controlled, parallel-group phase I trial evaluating oral NMNH-Ca in healthy adults aged 40-65 years. Eighty participants received placebo or NMNH-Ca 125, 250, or 500 mg once daily for 90 days. The primary objective was safety and tolerability. Whole-blood NAD was assessed as the key pharmacodynamic endpoint, including a 24-hour post-dose substudy, with biomarker-derived blood phenotypic age, treadmill-based six-minute walk distance, body mass index, and SF-36 domains analyzed as exploratory outcomes. NMNH-Ca was well tolerated at all doses, with no serious adverse events, treatment-related adverse events, or discontinuations. In the acute substudy, whole-blood NAD increased after single-dose NMNH-Ca, with peak mean concentrations at 12 hours. Over 90 days, NAD increased in a dose-related pattern; Day 90 mean changes from baseline were 2.33 {+/-} 18.53 M with placebo and 8.22 {+/-} 10.25, 15.85 {+/-} 11.16, and 39.90 {+/-} 14.11 M with NMNH-Ca 125, 250, and 500 mg, respectively. Exploratory analyses showed hypothesis-generating favorable signals in blood phenotypic age, treadmill-based six-minute walk distance, and health-related quality of life, most consistently at 500 mg. Oral NMNH-Ca was safe and pharmacodynamically active over 90 days, supporting larger and longer confirmatory trials with prespecified geroscience endpoints and tissue-relevant NAD metabolomics.

INTRODUCTION: Asthma has been associated with obesity in both adults and children. We investigated the association between longitudinal BMI trajectories and asthma at ages 8-9, 12-13, and 16-17 years in a UK cohort of White and South Asian children and adolescents. METHODS: We analysed data from the Leicester Respiratory Cohorts, population-based cohort studies that recruited 0-4-year-old children in 1990 (N = 1650) and 1998 (N = 8700) and followed them up until 2010. Outcome data on asthma came from postal questionnaires throughout infancy, childhood, and adolescence. BMI data came from birth records, well-child visits, questionnaires, and a clinical study visit. We used Group-based trajectory modelling to identify distinct BMI trajectories and used multivariable logistic regression to investigate the association with asthma. RESULTS: Out of 10,350 participants in the Leicester Respiratory Cohorts, we were able to model BMI trajectories for 5571 (54%); 1801 (17%) had information on asthma at 8-9 years of age, 1269 (12%) at 12-13 years, and 575 (6%) at 16-17 years. We identified five BMI trajectories: stable normal BMI (47%), persistent low BMI (30%), early overweight resolving (8%), childhood onset obesity (4%) adolescent onset overweight (11%). The persistent low BMI trajectory was associated with lower odds for asthma at age 8-9 years (0.56 [0.37-0.83]) and 12-13 years (0.38 [0.22-0.63]). Early overweight resolving was very similar to the reference stable normal BMI. The childhood onset obesity trajectory was associated with higher odds for asthma at 16-17 years (5.58 [1.35-24.40]). The adolescent onset overweight trajectory was not associated with asthma. When analysed separately, boys and girls and children of European and South Asian ancestry showed similar associations. CONCLUSION: This study strengthens the current understanding of obesity as a contributing factor in asthma development and highlights that early intervention and management of overweight and obesity in childhood, to achieve a normal BMI, may prevent secondary health impairments such as asthma.

Diabetic neuropathy is associated with substantial systemic disease burden, but short-term mortality risk among affected intensive care unit (ICU) patients remains difficult to characterize. We evaluated whether temporal information from the first 24 hours of ICU care improves post-landmark mortality prediction beyond severity scores and static clinical summaries. Patients aged > 18 years with diabetic neuropathy were identified in MIMIC-IV v3.1. A 24-hour landmark was used: only patients alive and still hospitalized at 24 hours were included, and the outcome was subsequent in-hospital death. The final cohort included 1,347 patients, including 83 deaths (6.16%). Data were divided into an 80% development set and a locked 20% test set. Feature selection, hyperparameter tuning, calibration, and threshold selection were restricted to development data. Logistic regression, random forest, and XGBoost were evaluated. Random forest had the highest development cross-validated PR-AUC and was selected for interpretation. On the locked test set, random forest achieved an AUROC of 0.851 (95% CI 0.765-0.924), PR-AUC of 0.339, and Brier score of 0.051; XGBoost and logistic regression achieved AUROCs of 0.847 and 0.806. In a post hoc strictly nested analysis, adding temporal predictors increased discrimination across all three algorithms; random-forest AUROC increased from 0.815 with severity and static predictors to 0.870 with the full temporal representation. First-day temporal information therefore showed additional prognostic value, but external validation is required before clinical use.

Liver metastases (LM) remain a major cause of death from different cancer types, in particular malignancies of the gastrointestinal tract. Liver metastases predict a poor response to immunotherapy due, among others, to the immunotolerant microenvironment (ME) of the liver and loss of local and systemic cytotoxic T cells. Thus, strategies that can reprogram the immune ME of the liver and restore cytotoxic T cell reactivity are being sought. We previously reported that estrogen signaling blockade impedes the growth of LM by reducing MDSC accumulation and monocyte/macrophage polarization. The aim of this study was to elucidate the underlying mechanism(s) and assess whether estrogen signaling in the myeloid lineage was driving the immunotolerant ME of LM. To this end, we generated mice with conditional myeloid cell-specific deletions of estrogen receptors (ER) or ER{beta} and analyzed in these mice the effect of ER loss on the liver immune ME and the outgrowth of LM. In mice with ER, but not with ER{beta} deletion, we observed a marked reduction in the growth of murine colon carcinoma MC-38 liver metastases as compared to their respective controls. Flow cytometry and immunohistochemistry revealed a decrease in macrophages that were polarized to the pro-tumorigenic M2-like phenotype and a concomitant increase in activated CD8+ T and NK cells relative to controls. Bulk RNAseq analysis performed on hepatic immune cells infiltrating the liver revealed changes in the expression of key cytokines/chemokines mediating immune cell recruitment, activation and polarization, including Ccl5 (upregulated) and Csf1 (downregulated). Taken together, the data suggest that ER signaling in myeloid-derived cells programs the immune landscape and contributes to an immunosuppressive and metastases-growth permissive ME in the liver.

Purpose Modified FOLFIRINOX (FFX) and gemcitabine plus nab-paclitaxel (GNP) are standard first-line treatments for metastatic pancreatic ductal adenocarcinoma (PDAC), but no validated biomarker guides treatment selection. We developed MULTIPL, a multimodal machine learning system, and established the PASS-01 Challenge to benchmark prognostic and predictive biomarkers. Patients and Methods MULTIPL was trained in the COMPASS study (N=268), integrating clinical, digitized histopathology, whole-genome, and RNA-seq data. MULTIPL, PurIST, hENT1 expression, and HRDetect were evaluated in the PASS-01 trial, a randomized phase II trial of FFX versus GNP (N=160), within the Challenge. The primary endpoint was differential treatment benefit measured by concordance-for-benefit for progression-free survival. Results MULTIPL had the highest concordance index for OS among individually evaluated biomarkers (0.595; 95% confidence interval [CI], 0.55-0.65) and separated high- versus low-risk patients (hazard ratio, 1.62; 95% CI, 1.13-2.33; P=0.009). Patients recommended for GNP by MULTIPL had significantly longer OS with GNP than with FFX (hazard ratio, 0.47; 95% CI, 0.28-0.82; P=0.007), whereas patients recommended for FFX had similar OS between treatments. Interpretability analysis of MULTIPL in COMPASS identified KDM6A alterations and SSTR1 expression as prognostic biomarkers, which were validated in PASS-01. However, none of the tested biomarkers significantly predicted differential treatment benefit in the PASS-01 Challenge. Conclusion MULTIPL demonstrated robust prognostic performance in external validation, identified a subgroup enriched for benefit from GNP, and enabled discovery and validation of prognostic biomarkers in metastatic PDAC. However, no biomarker met the primary endpoint for differential treatment benefit, underscoring the value of the PASS-01 Challenge.

Background: Hemoglobin (Hb) elevation is the hallmark of high-altitude adaptation, yet its effect on surgical outcomes may depend on arterial oxygen saturation (SpO2)--previously uninvestigated. Objectives: To explore whether preoperative Hb effect on postoperative length of stay (LOS) after laparoscopic cholecystectomy (LC) reverses across SpO2 strata. Methods: Retrospective single-center cohort of 612 adults undergoing elective LC (2018-2023) at Qinghai Red Cross Hospital, Xining, China (2260 m). Exposure: preoperative Hb (82-233 g/L) and SpO2 (86%-99%), stratified as low (<93%), mid (93%-95%), high (>=96%). Primary analysis: multivariable linear regression with Hb x SpO2 interaction, adjusted for BMI, age, sex, season. Results: Among 612 patients (65.8% female; mean age 43.5 [11.9] years; mean Hb 151.4 [20.7] g/L; mean SpO2 94.6% [2.3%]), the Hb x SpO2 interaction was significant (beta = -0.0095; P = .009). Hb effect reversed: in SpO2 >=93%, each 1 g/L Hb prolonged LOS by 0.003 days (P = .079); in SpO2 <93%, each 1 g/L reduced LOS by 0.006 days. In mid-SpO2 stratum (n = 251), Hb >=180 g/L had longer LOS (1.88 vs 1.62 days; P = .001; d = 0.54). Five computational robustness analyses confirmed the interaction (leave-one-out: 100% P < .05 across 612 iterations). Conclusions: In this exploratory cohort, we observed an SpO2-dependent reversal of the Hb effect on postoperative LOS, designated the "Plateau Hemoglobin Paradox." Given single-center design and achieved power of 0.754, findings require replication. If replicated, this pattern may inform future perioperative risk stratification at high altitude.

Background Clinical assessments of Amyotrophic Lateral Sclerosis (ALS) are typically collected infrequently in clinic visits and may not fully capture domain-specific functional decline in daily life. Digital Health Technologies (DHTs) can support remote monitoring, but passive free-living measures often require prolonged wear time and may be influenced by non-motor factors. This study evaluated whether short, standardized, at-home lower limb exercises recorded with ankle-worn accelerometers provide objective and interpretable measures of lower limb disease progression in ALS. Methods We analyzed data from 349 participants with ALS enrolled in the decentralized ALS Research Collaborative Study. Participants completed repeated self-entry ALS Functional Rating Scale-Revised (ALSFRS-RSE) assessments and wore bilateral ankle accelerometers during monitoring periods between September 2014 and January 2023. During each period, participants performed brief seated knee flexion-extension exercises at home. A previously developed signal processing pipeline was used to derive four exercise metrics: count, duration, intensity, and similarity. We examined baseline correlations with ALSFRS-RSE total and subdomain scores, longitudinal change using linear mixed-effects models, associations with gross motor item scores, differences by anatomical site of disease onset, and comparisons with free-living gait metrics. Results At baseline, exercise-derived metrics, particularly intensity and similarity, showed the strongest associations with the gross motor subdomain. Longitudinally, duration increased while intensity and similarity decreased, consistent with progressive slowing, reduced movement vigor, and reduced movement consistency (all p < 0.001); count did not change significantly. Worsening responses to gross motor items related to turning in bed, walking, and stair climbing were consistently associated with fewer, slower, less vigorous, and less consistent lower limb repetitions. Baseline intensity and similarity were lower in participants with lower limb disease onset on the corresponding side. Exercise-derived intensity showed model fit comparable to the strongest free-living gait metrics, while requiring substantially less observation time. Conclusions Short at-home lower limb exercises recorded using ankle-worn accelerometers provide scalable, objective, and interpretable measures of amyotrophic lateral sclerosis-related functional decline. Movement quality metrics, particularly intensity and similarity, may complement passive free-living monitoring and support remote digital clinical outcome assessment in ALS research. Trial registration NCT06885918.

Biomarkers perturbed by exercise-mediated molecular mechanisms, in women with early-stage (stage I-III, non-metastatic) breast cancer are poorly defined, and especially in under-represented Asian cohorts. In this exploratory Breast Cancer Exercise Intervention (BREXINT) pilot study, 15 Asian women were randomized to a combined aerobic and resistance exercise intervention program (n=8) and a control group (n=7). Fasting blood sampling was performed at baseline, 8,16, and 24-week timepoints. Blood parameters were imputed, transformed and screened for intervention-specific variations using IQR-trimmed, paired Wilcoxon tests. Twenty-one blood parameters were found to meet a differential change rule (significance observed in 1 group but not the other). Exercise-associated signatures displayed hematological and cytokine remodeling at 16-weeks. Control-associated signatures include adipokine and renal markers at 16 and 24-weeks. Of note, exercise-driven decrease of IL-10 at 16-weeks (p=0.022) retained significance following linear mixed effects confirmation among screened candidates. IL-10-centred modulation is the most convergent exercise-associated blood derived signature but warrants further validation in larger exercise oncology trials.