The FASEB Journal最新文献

Porcine epidemic diarrhea virus (PEDV) causes severe intestinal injury and economic losses in the pig industry. Arginine (Arg) promotes barrier repair and immune regulation, but its effects on PEDV-induced damage are unclear. This study aimed to investigate whether Arg could alleviate intestinal injury in PEDV-infected piglets. Thirty-two 7-day-old piglets were randomly divided into four groups: Control, Arg, PEDV, and PEDV+Arg. Piglets in the Arg and PEDV+Arg groups were orally administered 400 mg/kg BW Arg from Day 5 to Day 11. On Day 11, PEDV-infected piglets were orally challenged with the virus at a dosage of 1 × 10^5.5 TCID₅₀ per individual. On Day 14, tissue samples were collected after slaughter. PEDV infection markedly reduced villus height (VH) in the duodenum, jejunum, and ileum, increased crypt depth (CD), decreased plasma D-xylose concentration, and elevated diamine oxidase (DAO) activity, whereas Arg supplementation restored VH, decreased CD in the duodenum and jejunum, increased D-xylose, and reduced DAO activity. Arg also enhanced villin, occludin, and claudin-1 expression, indicating improved barrier integrity. Interestingly, Arg could promote PEDV replication in the small intestine. However, Arg reduced IL-1β, REG3G, and iNOS expression, while upregulating antiviral genes IFITM3, MX1, and DHX58 in the jejunum. Transcriptomic and proteomic analyses indicated that Arg administration may enhance interferon signaling cascades through the RIG-I-like receptor signaling pathway. In conclusion, our results show that Arg exhibits multifaceted effects upon PEDV infection. Although it increases PEDV replication, Arg could enhance intestinal barrier function, attenuate intestinal inflammatory responses, and alleviate PEDV-induced intestinal injury in piglets.

Inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), is an autoimmune disease with a strong genetic component. Although genome-wide association studies (GWAS) have identified multiple risk loci, the underlying causal genes and biological mechanisms remain predominantly elusive. GWAS data for IBD, UC, and CD were obtained from the FinnGen R10 dataset and integrated with expression quantitative trait locus (eQTL) data from the Genotype-Tissue Expression Project (GTEx) v8 to perform cross-tissue transcriptome-wide association studies (TWAS). Single-tissue TWAS validation was conducted using Functional Summary-based Imputation (FUSION). Multi-marker Analysis of GenoMic Annotation (MAGMA), Mendelian randomization (MR), colocalization, and summary data-based MR (SMR) analyses were applied to identify candidate susceptibility genes. GeneMANIA analysis was performed to explore functional implications. The same analytical framework was used on independent GWAS datasets to assess consistency. 6 susceptibility genes were identified for IBD, 1 for UC, and 2 for CD. These findings were supported by cross- and single-tissue TWAS, MAGMA, MR, colocalization, and SMR analyses. In independent datasets, partial gene-level concordance and substantial consistent pathway-level signals were observed. GeneMANIA analyses suggested that susceptibility genes in IBD, UC, and CD were mainly associated with 3′,5′-cyclic-nucleotide phosphodiesterase activity, transition metal ion transmembrane transporter activity, and CENP-A–containing chromatin organization, respectively. By integrating cross-tissue TWAS with multi-tier validation strategies, this study provides new insights into the genetic architecture of IBD. Further functional studies are necessary to elucidate the mechanisms by which these candidate genes contribute to IBD pathogenesis.

Paroxysmal kinesigenic dyskinesia (PKD) is a genetically heterogeneous movement disorder primarily associated with PRRT2 variants. Recently, TMEM151A and KCNJ10 have emerged as additional PKD-associated genes. However, genotype–phenotype correlations remain poorly defined. In this study, we retrospectively analyzed 41 PKD patients from a single center in Southeastern China. All patients underwent comprehensive clinical evaluation and whole-exome sequencing (WES), with variant classification based on ACMG guidelines. Additionally, we conducted a literature review of PKD cohorts published since 2021 to compare the clinical characteristics of patients carrying PRRT2, TMEM151A, KCNJ10 variants, and those without identified mutations. A genetic diagnosis was achieved in 19/41 patients (46.3%), with PRRT2 being the most frequent. We identified five novel variants, including two in KCNJ10, two in TMEM151A, and one in PNKD. Compared to other groups, PRRT2-positive patients had the earliest onset and highest treatment response. TMEM151A-positive patients tended to exhibit more frequent attacks and a lower response to carbamazepine. KCNJ10-positive patients presented with later onset and ultra-brief attacks. Genetically negative cases displayed distinct features, including fewer auras and more unilateral, ultra-brief episodes, yet responded well to carbamazepine. PKD exhibits significant genotype-dependent clinical heterogeneity. Novel variants in TMEM151A and KCNJ10 expand the mutational spectrum and suggest emerging genotype-specific phenotypic trends. Systematic genetic and phenotypic profiling may guide more precise diagnosis and management of PKD.

As health awareness grows, more people are turning to weight loss—yet excessive slimming or dietary imbalance is quietly triggering a wave of fertility problems. Researches show that low BMI or excessive exercise can negatively impact fertility, but its specific cellular mechanism remain unclear. In this study, we investigated how adipose tissue influences ovarian function under energy-restricted conditions by establishing a peri-ovarian adipose tissue (POAT) removal mouse model. We showed that POAT removal resulted in a significant decrease in litter size, impaired follicle development, and an increased apoptosis rate in granulosa cells (GCs). Additionally, the lipid droplets (LDs) content in the ovaries and GCs was significantly reduced. We also observed that Rab14 expression was upregulated in GCs after POAT removal. Functional experiments further showed that Rab14 overexpression significantly enhanced lipophagy activity, reduced the number and volume of LDs, and inhibited the expression of lipid droplet protein PLIN2. Meanwhile, Rab14 overexpression inhibited the phosphorylation level of AKT and induced the apoptosis of granulosa cells, suggesting that Rab14 may activate the cell death pathway mediated by lipophagy. Taken together, POAT may support ovarian function by maintaining lipid homeostasis within GCs. The role of Rab14 in the regulation of lipophagy may link local adipose-derived signaling with GCs survival and female fertility.

Acute respiratory distress syndrome (ARDS) involves impaired macrophage function in clearing apoptotic cells. The link between clinical hyperlactatemia in ARDS patients and poor outcomes prompted this study on the immunometabolic role of lactate in disease progression. In an LPS-induced ARDS mouse model, mice received either exogenous lactate or a lactate dehydrogenase inhibitor. Inflammatory cell infiltration was evaluated through flow cytometry and histological analysis with hematoxylin and eosin staining. Lactate signaling was confirmed in GPR81-deficient mice. In vitro, lactate metabolism during efferocytosis was studied using primary Alveolar Macrophages (AMs). Lactate accumulation, neutrophil infiltration, and elevated inflammatory factors were observed in this ARDS model. External lactate delayed inflammation resolution and worsened lung injury. GPR81^−/− mice exhibited reduced neutrophil infiltration and better outcomes. Macrophages produced substantial amounts of lactate during efferocytosis in vitro, concurrent with upregulated expression of the glucose transporter Glut1, the lactate transporter MCT1, and the lactate receptor GPR81. Pharmacological inhibition using an LDH inhibitor, an MCT1 antagonist, or extra lactate significantly impaired efferocytic capacity. Efferocytosis triggered Myc upregulation in vitro, which was suppressed by exogenous lactate. Genetic ablation of GPR81 elevated both MCT1 and Myc expression. Silencing Myc via siRNA significantly impaired efferocytosis in vitro. These findings indicate that the activation of GPR81 by lactate delays the resolution of inflammation in acute lung injury. This effect may be attributed to the suppression of alveolar macrophage efferocytosis, which subsequently impairs the clearance of apoptotic cells and exacerbates lung injury.

To examine placental histomorphological and ultrastructural features in normal monochorionic diamniotic (MCDA) and selective fetal growth restriction (sFGR) pregnancies and investigate high temperature requirement A4 (HTRA4)'s potential role in mediating pathology. Included normal MCDA, sFGR placentas of type I and type III. Placental terminal villi maturity and syncytiotrophoblast (STB) ultrastructural features were assessed by immunohistochemistry and transmission electron microscopy. RNA sequencing identified differentially expressed genes, validated by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. Functional studies using HTRA4-overexpressing BeWo cells. Type III sFGR-S placentas exhibited the most pronounced structural defects among the groups, including: diminished terminal villous maturity (manifested by reduced microvessel density and β-HCG with thickened villous syncytial membrane), elevated STB autolysosomes, and placental lipid droplet accumulation. HTRA4 was identified as a significantly upregulated gene in sFGR-S placentas, with mRNA and protein levels highest in type III, and fold change significantly greater than in type I or normal MCDA. Functional experiments confirmed that HTRA4 overexpression impaired syncytialization, disrupted autophagic flux, and altered lipid metabolism. This study reveals significant terminal villous hypoplasia and STB dysfunction in sFGR type III. HTRA4 upregulation mediates these abnormalities and reflects disease severity.

G-protein-coupled receptor kinase 5 (GRK5) plays a pivotal role in various pathological conditions, particularly in cancer. While our previous work identified elevated GRK5 expression in gliomas, its functional significance remains unclear. Using clinical glioma specimens and U251 cell models with GRK5 knockdown or overexpression, we investigated the oncogenic functions of GRK5 through Cell Counting Kit-8, flow cytometry, transwell, and wound healing assays. Underlying mechanisms were explored via Western blot, immunofluorescence, and transcriptomic analysis. GRK5 overexpression correlated with glioma malignancy and drove tumor progression by enhancing proliferation, migration, and invasion while suppressing apoptosis through Bax downregulation. This oncogenic activity partially involved β-arrestin-mediated desensitization of the G-protein-coupled receptor pathway. Crucially, GRK5 facilitated the translocation of HDAC7 from the nucleus to the cytoplasm. Cytosolic HDAC7 subsequently deacetylated cortactin, which in turn stabilized F-actin and enhanced the MSN/CD44 complex expression and stability, thereby driving the MES transition—a hallmark of aggressive glioblastoma. Inhibition of HDAC7 abolished the GRK5-induced upregulation of MSN and CD44. We unveil MSN as a non-canonical GRK5/HDAC7 axis that promotes glioma malignancy by modulating the cortactin/MSN/CD44 pathway. Targeting this axis potentially represents a promising therapeutic strategy against gliomas.

Neuropsychiatric symptoms (NPS), commonly concomitant with Alzheimer's disease (AD), substantially impair the quality of life and accelerate disease progression, yet reliable biomarkers for early identification of individuals at high NPS risk remain elusive. In this study, we leveraged the data from the Alzheimer's Disease Neuroimaging Initiative (ADNI), incorporating longitudinal data from 509 participants diagnosed with mild cognitive impairment (MCI) or mild AD at baseline and followed up for 1 and 2 years. The dataset included cerebrospinal fluid (CSF) proteomic profiles comprising 6361 proteins, along with comprehensive data on NPS diagnosis, cognitive function, and AD pathology. LASSO regression and recursive feature elimination were applied to identify NPS-related CSF proteins, followed by random forest modeling to predict NPS risk at baseline, 1 year, and 2 years. Incorporating selected CSF proteins significantly improved NPS prediction compared to the reference model, with AUCs increasing from 0.64 to 0.76 at baseline, 0.63 to 0.80 at 1 year, and 0.63 to 0.81 at 2 years. Notably, Cyclin-Dependent Kinase-Like 2 (CDKL2), Nidogen 2 (NID2), and Lin-7 Homolog B (LIN7B) were consistently associated with NPS across all time points. Among them, CDKL2 and NID2 were significantly associated with AD biomarkers and cognitive scores, and their expression changes were independently validated in cerebrospinal fluid from a mouse model, highlighting their potential as stable predictive biomarkers. Our findings highlight CSF proteomic signatures that robustly predict NPS progression in individuals with MCI and mild AD, offering a framework for early risk stratification and precision intervention in NPS.