FASEB bioAdvances最新文献

Sepsis remains a leading cause of pediatric morbidity and mortality, yet its molecular underpinnings are poorly understood. Here, we performed mass spectrometry-based plasma proteomics and cytokine profiling in pediatric sepsis patients at the acute phase (AP) and recovery phase (RP), alongside preoperative surgical controls. In AP vs. control, we identified 41 differentially abundant (DA) proteins, including acute-phase reactants and complement factors, with persistent but attenuated expression in RP. Pathway analysis revealed sustained enrichment in inflammatory and complement activation processes during both AP and RP, with partial restoration of immune surveillance and vascular homeostasis in recovery. Machine learning highlighted complement components (C9, C1R) and LRG1 as candidate AP biomarkers, and S100A9 as an RP-associated marker. Comparative analysis with adult sepsis proteomes uncovered age-specific complement activation patterns: adults displayed higher classical pathway activity, whereas pediatric patients exhibited enhanced alternative pathway activity. Cytokine profiling confirmed sustained immune activation and endothelial perturbation across sepsis phases. We also compared the sepsis cohort with the sterile inflammation (SI) cohort, which revealed distinct adaptive immune enrichment in sepsis while innate immune predominance in SI, enabling the identification of potential sepsis-specific protein signatures. Together, these findings delineate the dynamic immune and vascular proteomic landscape of pediatric sepsis, reveal biomarkers distinguishing sepsis from sterile inflammation, and highlight age-related complement pathway differences with potential therapeutic implications. Trial Registration: ClinicalTrials.gov: NCT04103268, NCT04299828.

Intestinal paracellular permeability was analyzed ex vivo by incubation of tissue segments at 0°C with the fluorescent dyes FM1-43FX (FM) or TRITC-dextran 3 kDa lysine-fixable (TD3L) and confocal microscopy in (i) healthy mice and (ii) mice submitted to chronic stress or lipid diets. In the small intestine of healthy mice, FM staining was restricted to the apical surface of enterocytes but fully penetrated around Goblet cells, enteroendocrine cells, tuft cells, and apoptotic cells. The same cell types were similarly labeled in the colon when located on the tissue surface but not within the crypts. TD3L exhibited a comparable labeling pattern but also showed moderate staining of the basolateral surface of enterocytes at the tips of small intestinal villi, and also substantial penetration around colonic epithelial cells at the surface or top of crypts. The study reveals patterns of permeability likely corresponding to the "leak" pathway of paracellular transport through the intestinal epithelium, because transcellular endocytosis is blocked at 0°C. This pathway is found around specific cell populations involved in the luminal detection of food, antigens, microbes, or their secretions. These trigger immune, neural, and tissue responses that maintain intestinal homeostasis. Chronic stress induced by glucocorticoid exposure increased FITC-dextran 4 kDa permeability in vivo. Using FM, increased paracellular permeability was also detected ex vivo and selectively localized in the colon of stressed mice. A single oral administration of a lipid-rich food also increased ex vivo permeability around jejunal enterocytes. Pathophysiological increases in paracellular permeability are therefore detectable using the FM methodology.

Breast cancer (BC) is one of the most common cancers in women around the world, and utilizing a combined approach is a crucial strategy. Induction of cuproptosis in tumor cells is a novel antitumor approach, though its standalone efficacy remains unclear. In this study, we prepared a novel liposome loaded with the photosensitizer indocyanine Green (ICG) and the cuproptosis inducer elesclomol-Cu (ES-Cu) to examine the synergistic effects of photodynamic-cuproptosis treatment on BC. The cuproptosis inducer ES-Cu and the photosensitizer ICG were encapsulated in nanoliposomes with a membrane hydration approach and then validated in vitro and in vivo. JC-1, MDA, GSH, and other cuproptosis-related indicators were used to confirm the ability of PDT to enhance ES-Cu-induced cuproptosis in MCF-7 breast cancer cells. For confirming the cytotoxic impact of PDT in conjunction with the cuproptosis inducer, tests for CCK-8 and cell death staining were performed. The drugs were administered to animals via tail vein injection to observe their tumor inhibition effects in vivo. Their safety was assessed by monitoring changes in body weight. The average particle size of liposomes loaded with ES-Cu and ICG was 208.3 ± 1.07 nm, exhibiting a consistent nanospherical morphology. ICG produced cytotoxic reactive oxygen species (ROS) that enhanced ES-Cu-induced cell cupping under NIR laser irradiation. The therapeutic effect of the synergistic treatment combining PDT and cuproptosis was validated in both in vitro and in vivo experiments. This investigation proved that PDT markedly augments the ES-Cu-induced cuproptosis in breast cancer cells, demonstrating a synergistic therapeutic effect. This synergistic effect presents a novel therapy approach for BC with substantial practical application potential.

J. A. Zegarra-Valdivia, M. Z. Khan, J. Fernandes, et al., “Interoceptive Information of Physical Vigor Through Circulating Insulin-Like Growth Factor 1,” FASEB BioAdvances 8, no. 1 (2026): e70084, https://doi.org/10.1096/fba.2025-00226.

In the original paper, the name of one of the authors was incorrect. It was incorrectly stated as “M. Zahid Kahn.” The author's correct name is “M. Zahid Khan.”

Additionally, in the original paper, Supplementary Figures: S1B, S2B, S2D, S3, S4B, S4D, S4E, S5A, S5B, S5C, S5E, and S6D contained blank panels. The correct graphical data that were peer reviewed have been returned to the supplementary figures.

We apologize for these errors.

Dysfunction of the circadian clock has been implicated in the pathogenesis of various diseases, including metabolic disorders, inflammatory conditions, and cancer. While the significance of circadian rhythm in diabetic nephropathy is gaining attention, the specific alterations in circadian profiles in diabetic nephropathy remain unexplored. In the present study, we performed RNA sequencing on renal cortex samples collected every 4 h across the day from both control and diabetic mice. The rhythmicity of genes was identified using the JTK_CYCLE algorithm for each group. Genes that lost, acquired, or sustained rhythmicity in diabetic mice were denoted the circadian dysregulation gene set. Subsequent bioinformatic analyses focused on this gene set to investigate the circadian reprogramming in diabetic nephropathy. We observed significant circadian disruption in the kidney of diabetic mice, marked by both the gain and loss of rhythmicity, along with alterations in the phase and relative amplitude of genes that retained rhythmic expressions. Circadian disturbances, such as phase shifts and alterations in relative amplitude or mesor, were also noted in core clock genes. Furthermore, genes that lost rhythmicity in diabetic nephropathy were predominantly associated with protein homeostasis and glycolipid metabolism, whereas those that gained rhythmicity were mainly linked to gene regulation, fatty acid metabolism, and protein transport. The genes in the circadian dysregulation gene set that exhibit differential expression at least at one Zeitgeber time were most prominently enriched in the lipid metabolic process. WGCNA and correlation analysis revealed co-expression networks involving core clock genes and PPAR signaling pathway with renal triglyceride levels. Our study reveals substantial circadian disruption in diabetic nephropathy, with significant impacts on protein homeostasis and glycolipid metabolism. Furthermore, our findings highlight the potential influence of circadian system dysregulation on the disorder of fatty acid metabolism in diabetic nephropathy.

The National Institutes of Health (NIH) has launched a major initiative to expand human-based New Approach Methodologies (NAMs) in biomedical research and reduce reliance on animal models. While NAMs offer powerful complementary tools, animal-based research remains indispensable in musculoskeletal science for understanding complex cellular and systemic processes, disease onset and progression, and developing effective therapies. Foundational knowledge of embryonic development, disease mechanisms, tissue regeneration, gene function, and systemic pharmacology has emerged from animal models and will continue to do so. This review underscores the essential role of animal models in five key areas of musculoskeletal biology: osteoporosis, osteoarthritis, bone fracture repair and regeneration, bone cancer, and Inherited Skeletal Disorders (ISDs). We also examine NAMs including organoids, engineered scaffolds, organ-on-chip platforms, and Artificial Intelligence (AI)/computational modeling, highlighting their strengths in mechanistic and high-throughput studies but also their limitations in replicating in vivo structural, physiological, biomechanical, and systemic complexity. Animal models remain the gold standard for exploring disease mechanisms, testing preclinical therapeutic and diagnostic efficacy and safety, and translating discoveries into clinical practice. Rather than replacing animal research, NAMs should be integrated as complementary approaches to advance understanding and innovation. Curtailing animal research would jeopardize medical progress and hinder life-saving interventions for humans and animals alike. This review aims to inform the public and policymakers on the continued necessity of ethically conducted animal research as a cornerstone of musculoskeletal health.

Peripheral arterial disease progression to critical limb ischemia remains a significant burden in the aged, necessitating revascularization. Therapies to enhance neovascularization, such as angiogenesis or arteriogenesis, may provide an option for patients not suitable for surgery. The thrombospondins (TSPs) are secreted matricellular glycoproteins, with TSP-1, TSP-2, and TSP-5 implicated in neovascularization. Currently, there is little data regarding the effects of TSP-2, TSP-5, sex, and aging on angiogenesis and arteriogenesis. In vitro, the effect of TSP-2 or TSP-5 on early or late passage endothelial cell (EC) tubule formation or disruption was assessed. In vivo, angiogenesis, and arteriogenesis in male and female, young (14–16 weeks) and old (105–110 weeks) wild-type, TSP-2 and TSP-5 knockout mice were compared. To assess the effect of sex hormones, neovascularization was assessed in ovariectomized young female mice. After 14 days, angiogenesis and arteriogenesis were quantified using immunohistochemistry. Laser Doppler was used to confirm > 50% decrease in blood flow. In vitro: TSP-2 inhibited angiogenesis in early passage ECs and disrupted tubules in late passage ECs. TSP-5 prevented early passage tubule disruption. Late passage cells showed greater disruption than early passage. In vivo: TSP-2 was anti-angiogenic in young mice and anti-arteriogenic in old mice; TSP-5 had no effect on angiogenesis but largely negatively impacted arteriogenesis. Females showed reduced angiogenesis/arteriogenesis versus males, aging blunted both responses, and TSP-2/TSP-5 knockouts altered sex- and age-specific ischemic responses. In conclusion, we broadly elucidate the differential effects of TSP-2, TSP-5, sex and aging on neovascularization after inducing limb ischemia.

Mitochondrial function is essential for skeletal muscle health, and its disruption leads to atrophy and functional decline. This study examines the impact of denervation on skeletal muscle mitochondria in polymerase gamma (PolG)^(+/mut) mice, which accumulate mitochondrial DNA (mtDNA) mutations due to a partial deficiency in polymerase gamma proofreading. Using a 14-day denervation protocol, we assessed muscle mass, mtDNA copy number, oxidative stress and mitochondrial dynamics in wild-type (WT) and PolG^(+/mut) mice. Our findings reveal that while denervation significantly reduced muscle wet weight and mitochondrial enzyme activity, no genotype-specific differences in muscle atrophy were observed. However, PolG^(+/mut) mice displayed more disorganized mitochondrial cristae and elevated oxidative stress markers, indicating greater mitochondrial vulnerability. Despite these changes, the lack of significant differences in mitochondrial proteins and gene expression between genotypes may reflect an adaptive antioxidant response, including increased catalase expression, although the compensatory nature of this response cannot be conclusively determined. These results suggest that oxidative stress–related responses are involved in mitochondrial adaptations during denervation-induced muscle atrophy. The increased expression of antioxidant enzymes, such as catalase, in PolG^(+/mut) mice suggests that antioxidant mechanisms are activated in response to increased oxidative stress. These findings underscore the importance of controlling oxidative stress for maintaining muscle health.

The cellular and molecular complexity of acne pathogenesis has hindered progress toward effective targeted therapies. While keratinocytes are known to influence skin inflammation, their precise transcriptional programs and regulatory circuitry in acne remain unclear. We developed an integrative computational framework that combines single-cell RNA sequencing (scRNA-seq), gene co-expression network analysis (WGCNA), and two complementary machine learning algorithms (SVM-RFE, LASSO) to identify disease-relevant biomarkers. We mapped acne lesion cellular composition, reconstructed keratinocyte differentiation trajectories, and integrated miRNA–transcription factor–drug interaction networks to link molecular signatures to potential interventions. We uncovered marked keratinocyte heterogeneity and enriched late-stage pro-inflammatory states in acne lesions, accompanied by increased macrophage/monocyte and T cell infiltration. Six keratinocyte-associated biomarkers (PYGL, C10orf99, C12orf75, S100A2, PI3, CARD18) were identified, achieving high diagnostic accuracy (AUC > 0.85). Functional enrichment connected these genes to cytokine and chemokine signaling, while regulatory analysis revealed upstream modulators (hsa-let-7b-5p, FOXC1). Drug–gene network mapping suggested repurposing potential for cyclosporin A and valproic acid. In conclusion, our study delineates a keratinocyte-centered molecular signature that shapes acne pathogenesis and provides potential therapeutic biomarkers.

Gamma-aminobutyric acid (GABA) is the primary inhibitory neurotransmitter in the central nervous system (CNS). Many aspects of GABAergic neurotransmission, including the densities of GABAergic neurons, the synthesis of GABA and its interaction with the respective receptors, are believed to be altered during aging, contributing to increased neuronal excitability seen in multiple neurodegenerative conditions, such as dementias, Alzheimer's disease, and traumatic brain injury (TBI). Oral administration of a nuclear fraction extract of the bovine thymus gland (thymus nuclear fraction—TNF) to rats was recently reported to improve their functional recovery from controlled cortical impact (CCI)—an animal model of TBI. Given that individual thymic peptides and mixed thymus fractions were also found to have broad neuroprotective effects and anti-neuroinflammatory activity, we sought to investigate the impact of TNF on GABAergic neurotransmission in the aging mouse brain. Using biochemical investigation, electrophysiological recordings, obtained using electroencephalography (EEG), and power spectral density analysis, we evaluated GABAergic protein expression and cortical neuronal activity in aged control mice and in mice supplemented with a low dose (LD) or a high dose of TNF for 14 weeks. We uncovered increased expression of two isoforms of glutamic acid decarboxylase, GAD65 and GAD67, and increased levels of β2/β3 subunits of GABA_A receptor in the brains of TNF-supplemented mice compared to the control group, suggesting possible enhancement of inhibitory neurotransmission. Decreased neuronal excitability, evidenced by reduced EEG amplitudes, power spectral densities, and peak amplitudes of high-frequency cortical oscillations, further confirmed a dose-dependent attenuation of neuronal excitability by TNF. Our results suggest that TNF supplementation may have the potential to mitigate age-related alterations in GABAergic neurotransmission, thereby modulating neuronal excitability.