Acute kidney injury lacks definitive therapeutic options and clear molecular mechanisms. The proximal tubule demands high energy primarily derived from mitochondrial fatty acid beta-oxidation. ADP-ribosylation factor guanine nucleotide exchange factor 3 (Arfgef3) plays a role in tumor-associated diseases, but its function in nephrotoxicity remains unclear. We hypothesized that this exchange factor contributes to cisplatin-induced AKI by disrupting peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α)-mediated mitochondrial function and fatty acid oxidation. We utilized global knockout mice and human kidney 2 cells to investigate this exchange factor in cisplatin-induced injury in vivo and in vitro. Renal function was evaluated using biochemical markers and histopathological scores. Mitochondrial integrity and respiratory capacity were measured via transmission electron microscopy and Seahorse analysis. Intracellular lipid accumulation, key fatty acid oxidation enzymes, and physical interactions with PGC-1α were evaluated alongside small-molecule agonist rescue experiments. Arfgef3 was upregulated in renal tubular epithelial cells after cisplatin exposure. Genetic Arfgef3 knockout in mice significantly attenuated cisplatin-induced elevations in serum creatinine, blood urea nitrogen, tubular necrosis, inflammation, and cellular apoptosis. Furthermore, loss of this protein preserved mitochondrial ultrastructure, restored cellular adenosine triphosphate production, and reversed the suppression of key fatty acid oxidation enzymes. Mechanistically, co-immunoprecipitation confirmed a direct interaction between Arfgef3 and PGC-1α. Activating the coactivator with the agonist ZLN005 substantially rescued the aggravated mitochondrial damage and lipid accumulation caused by Arfgef3 overexpression. Arfgef3 promotes cisplatin-induced AKI through PGC-1α-mediated mitochondrial dysfunction and impaired fatty acid oxidation. Targeting this novel signaling pathway may represent a promising therapeutic strategy for preventing or treating cisplatin-associated nephrotoxicity.

Lupus nephritis (LN) is a critical complication of systemic lupus erythematosus (SLE) and a leading cause of mortality in those affected. Abnormal energy metabolism of macrophages is closely associated with LN pathogenesis. Adenosine diphosphate (ADP) is known to regulate macrophage proinflammatory function; however, its specific role in SLE progression remains unexplored. We employed a spontaneous SLE mouse model and an integrated transcriptomic and metabolomic approach to investigate the effect of ADP on SLE disease progression and macrophage function. Our study revealed that ADP stimulates the AMPK signaling pathway, curtails macrophage senescence, and diminishes the release of senescence-associated secretory phenotype factors, ultimately alleviating SLE progression. Mechanistically, ADP promotes KLF10 expression, which binds to Sirtuin1 to facilitate deacetylation at the promoter region of Cdkn1a in macrophages. This process leads to reduced p21 expression and macrophage senescence. Collectively, the findings uncover a novel regulatory role of ADP in the pathogenesis of SLE and macrophage senescence, indicating its potential as a target for treatment.

Cuproptosis is a type of cell death that depends on the energy production process inside cells, and it is important in cancer treatment. This study aimed to identify cuproptosis-associated genes in triple-negative breast cancer (TNBC) and reveal regulatory pathways. We applied weighted gene co-expression network analysis to screen gene expression data from TNBC patients. Cuproptosis-associated genes were identified at the intersection of the co-expression modules and the cuproptosis gene dataset. G protein-coupled receptor 68 (GPR68) was identified as a cuproptosis-associated gene that is highly expressed in TNBC cells. Knockdown of GPR68 inhibited TNBC cell growth, migration, and invasion. The knockdown of GPR68 enhanced cuproptosis in TNBC cells when stimulated with ES-Cu, leading to an accumulation of cellular copper. This effect was reversed by the copper chelator tetrathiomolybdate. Correlation analysis verified that GPR68 regulates glutaminase (GLS) expression in TNBC cells. GLS knockdown reversed the promoted role of GPR68 in TNBC progression and the inhibitory role of GPR68 in TNBC cell cuproptosis. We further investigated the role of GPR68 in TNBC through pathway enrichment analysis, which revealed significant enrichment in the mitogen-activated protein kinase (MAPK) signaling pathway. GPR68 overexpression increased the levels of ERK phosphorylation. The ERK signaling pathway inhibitor, Trametinib, blocked the effects of GPR68 on TNBC progression and cuproptosis. GPR68 knockdown hindered TNBC development and induced cell cuproptosis by regulating the MEK/ERK signaling pathway, which is linked to GLS. This study provides new insights into developing cuproptosis targets for TNBC treatment.

Head and neck squamous cell carcinoma (HNSCC) remains a leading cause of cancer mortality worldwide, with treatment resistance posing a major clinical challenge. The tumor microenvironment, particularly the three-dimensional (3D) extracellular matrix architecture, plays a critical role in driving therapy resistance, yet the underlying molecular mechanisms remain incompletely understood. Here we investigate how the 3D collagen microenvironment orchestrates chemoresistance through NF-κB/STAT3 signaling and partial epithelial-mesenchymal transition (EMT) in HNSCC. Using 3D type I collagen culture systems, both FaDu and CAL27 HNSCC cells form spheroidal structures with cytoskeletal reorganization. Compared to 2D monolayers, 3D collagen culture activated NF-κB signaling, upregulated stemness marker ALDH1A1 and drug resistance proteins ABCG2/Bcl2, and significantly increased IC50 values for paclitaxel and cisplatin in FaDu and CAL27 cells. NF-κB inhibitor PDTC reversed these phenotypes, reducing resistance marker expression and restoring chemosensitivity. In FaDu xenografts, PDTC combined with paclitaxel exerted enhanced antitumor effects, achieving greatest tumor growth inhibition. Mechanistically, we identified a signaling hierarchy with NF-κB upstream of STAT3; PDTC suppressed both pathways, while Stat3 inhibitor Stattic only affected STAT3 downstream effectors. Clinical relevance was confirmed by immunohistochemistry of human HNSCC specimens, showing spatial enrichment of p65/p-p65 at the tumor-stroma interface specifically in lymph node-positive cases, with p-p65 H-scores significantly higher in metastatic patients (p < 0.05). Importantly, 3D collagen induced a partial EMT state with concurrent E-cadherin and N-cadherin upregulation, downregulation of ZO-1 and upregulation of Slug. Using a 14-gene p-EMT score in chemoresistant patient data, RELA and STAT3 strongly correlated with this hybrid score (RELA: R = 0.75; STAT3: R = 0.82) and activated mesenchymal genes while preserving epithelial identity. These findings demonstrate that the 3D collagen microenvironment activates an NF-κB-Stat3 signaling axis that drives cancer stemness, p-EMT, and chemoresistance in HNSCC, highlighting NF-κB inhibition as a promising therapeutic strategy to overcome microenvironment-mediated resistance.

Gestational diabetes mellitus (GDM) is a common metabolic disorder during pregnancy. Metformin (MET) has emerged as a promising alternative to insulin, but its mechanism of action in the placenta remains incompletely understood. In this study, we aimed to examine whether MET counteracts high-glucose-induced ferroptosis by regulating activating transcription factor 2 (ATF2) and to delineate the underlying molecular mechanism in GDM. We established in vitro GDM cell models using HTR-8/SVneo trophoblast cells and an in vivo STZ-induced hyperglycemic rat model, followed by treatments including ATF2 lentiviral manipulation and MET administration. We evaluated cell viability, ferroptosis markers (ROS, MDA, GSH, iron deposition), and PI3K/Akt signaling pathway activity. Our results demonstrated that in GDM patients and high-glucose-challenged HTR-8/SVneo cells, MET promoted cell viability by suppressing ATF2, reactivating PI3K/Akt signaling, enhancing nuclear Nrf2 translocation, and upregulating GPX4, while concurrently lowering ROS and MDA levels, elevating GSH, and reducing iron deposition. These protective effects were reversed by ATF2 overexpression or pathway inhibitors. In STZ-induced hyperglycemic rats, MET decreased placental ATF2 expression, restored the PI3K/Akt-Nrf2-GPX4 axis, alleviated ferroptosis, and reduced fetal weight. Collectively, our findings indicate that MET suppresses ATF2, activates PI3K/Akt, drives Nrf2 nuclear import, upregulates GPX4, and curbs high-glucose-induced trophoblast ferroptosis, offering new mechanistic insight and therapeutic avenues for GDM.

Although cellular senescence contributes to tissue dysfunction in inflammatory bowel disease (IBD), the heterogeneity of senescence at the single-cell resolution remains poorly defined. In this study, we systematically characterized the senescence landscape within the IBD microenvironment, with particular emphasis on fibroblast subpopulations and their systemic consequences. Single-cell RNA sequencing (scRNA-seq) dataset was applied to profile senescence-associated feature heterogeneity. Fibroblasts were classified into high-senescence (HS-Fibro) and low-senescence (LS-Fibro) groups according to senescence scores. HS-Fibro-specific gene signatures were identified and functionally annotated. Intercellular communication networks were constructed using CellChat to delineate the HS-Fibro interactome atlas, with emphasis on identifying cellular targets responsive to senescence-associated secretory phenotype (SASP) signals. Integrated microarray datasets were employed to elucidate the systemic impact of HS-Fibro in IBD. Through the integration of differential expression analysis, Weighted Gene Co-expression Network Analysis (WGCNA), and an ensemble of 113 machine learning algorithms, seven hub genes implicated in IBD progression were systematically identified and prioritized. In the IBD microenvironment, fibroblasts exhibited enhanced senescence activity. HS-Fibro displayed potent pro-inflammatory properties and enhanced communicative capacity. IBD patients with high HS-Fibro infiltration showed sustained immune-inflammatory activity. Systematic bioinformatic analysis and immunohistochemistry validation identified ADAMTS2 as a robust diagnostic indicator for IBD, with substantially elevated expression in patients relative to controls. The human colon fibroblast cell line CCD-18Co was treated with H2O2 to establish a cellular senescence model, demonstrating marked upregulation of ADAMTS2, SASP markers, and fibrosis-related genes. Moreover, siRNA-mediated knockdown of ADAMTS2 resulted in obvious attenuation of these SASP and fibrosis-associated genes.

Hypertrophic cardiomyopathy (HCM) is frequently linked to sarcomeric gene mutations, yet many familial cases remain unexplained by known pathogenic genes. Transcriptional coactivator TAZ serves as the core effector of the Hippo signaling pathway that regulates cardiomyocyte proliferation and myocardial hypertrophy, and its PDZ-binding motif is critical for maintaining the normal function of TAZ. Deletion of this motif triggers TAZ dysfunction and subsequently induces HCM; however, the underlying molecular mechanism remains to be further elucidated. PDZ-binding motif-deleted TAZ (TAZ-ΔPDZ) was identified as the pathogenic mutation in HCM. Functional experiments revealed that TAZ-ΔPDZ exacerbated phenylephrine (PE)-induced cardiomyocyte hypertrophy, manifested as enlarged cardiomyocyte surface area, upregulated expression of hypertrophy marker proteins (ANP/BNP/β-MHC), and intracellular calcium overload. Mechanistically, TAZ-ΔPDZ blocked the transcriptional activation of mitofusin 2 (MFN2) by T-box transcription factor 5 (TBX5), thereby triggering calcium overload and aggravating myocardial hypertrophy. Further in vivo animal experiments verified that TBX5 overexpression reduced heart size, alleviated myocardial collagen deposition and inflammatory cell infiltration, and decreased intracellular calcium concentration to exert cardioprotective effects; however, TAZ-ΔPDZ markedly reversed such protective effects mediated by TBX5 overexpression. TAZ-ΔPDZ impairs TBX5 transcriptional function, reduces MFN2 expression, triggers cardiomyocyte calcium overload, and ultimately causes cardiac hypertrophy. This study is expected to provide novel insights for the prevention, control, and screening of pathogenic genes underlying hereditary HCM.

Endothelial-to-mesenchymal transition (EndMT) plays a pivotal role in fibroblast proliferation and cardiac fibrosis during diabetic cardiomyopathy (DCM) development, yet its underlying regulatory mechanisms remain unknown. The objective of this study is to investigate the role of general control non-repressed 5 protein (GCN5) and its mediated protein lactylation in regulating EndMT during DCM progression. We combined in vitro (high glucose (HG)-treated HUVECs) and in vivo (high-fat diet (HFD)/streptozotocin (STZ)-induced DCM mice) models with endothelial-specific GCN5-knockout mice. Pathological changes and fibrosis in cardiac tissue were assessed using HE staining and Masson staining, respectively. Cell migration was determined using wound healing and Transwell assays. Co-IP assay was employed to detect YY1 lactylation, as well as the interaction between YY1 and GCN5. ChIP and dual-luciferase reporter assays were employed to analyze the binding relationship between YY1 and the Twist1 promoter. GCN5 was upregulated in diabetic hearts and endothelial cells, and its knockdown reduced Twist1-dependent EndMT in HG-treated HUVECs. Mechanistically, GCN5 catalyzed YY1 lactylation at K203, which enhanced YY1 phosphorylation at Thr398, activating Twist1 transcription. GCN5 knockdown reversed the promoting effect of wildtype YY1 overexpression on HG-induced EndMT but didn't affect the effect of mutant YY1 overexpression on EndMT. Endothelial-specific GCN5 deletion reversed HFD/STZ-induced cardiac dysfunction and fibrosis in mice by inhibiting Twist1-dependent EndMT. GCN5 enhanced Twist1 transcription by promoting lactylation of YY1 at K203 and its phosphorylation at Thr398, thereby increasing EndMT in DCM. This GCN5-YY1-Twist1 axis may offer novel therapeutic targets for DCM management.

Pathological neovascularization drives blinding retinal diseases, including diabetic retinopathy, retinopathy of prematurity, and neovascular age-related macular degeneration. Endoplasmic reticulum (ER) stress and unfolded protein response (UPR) signaling are key mechanisms underlying neovascularization. Mice lacking the UPR regulator Activating Transcription Factor 6 (ATF6) show normal retinal vasculature yet are resistant to pathologic neovascularization, suggesting that ATF6 inhibition may mitigate retinal damage and vision loss from aberrant retinal angiogenesis. We tested Ceapin-A7, a selective ATF6 inhibitor, in Vldlr[-/-] mice, a genetic model of retinal neovascularization. Intravitreal Ceapin-A7 reduced ectopic vascular projections into the outer retina and significantly increased scotopic b-wave amplitude, while a-wave and photopic responses showed no significant difference between groups. Single-nucleus RNA sequencing revealed suppressed ATF6/UPR gene programs and angiogenic pathways in retinal endothelial cells and reduced Müller cell gliosis. These findings demonstrate that pharmacologic ATF6 inhibition mitigates retinal neovascularization in Vldlr[-/-] mice and identifies ATF6/UPR as a therapeutic target for neovascular retinal diseases.

The schematic workflow illustrates next-generation sequencing of lung-derived mesenchymal stromal cells (MSCs) from IPF patients and controls. Rather than broad epigenetic reprogramming, IPF-MSCs exhibit a remarkably restricted down-regulation of only three critical microRNAs: hsa-miR-373-3p, hsa-miR-486-5p, and hsa-miR-449a. This selective dysregulation leads to the targeted upregulation of key profibrotic genes (SMAD4, BCL2, TGF-β, CXCL12, VEGF), driving TGF-β cascade activation, altered autophagy, and profibrotic and proangiogenic signaling.

Flavin cofactors (FMN and FAD) are essential mediators of mitochondrial respiration, antioxidant defense, and redox signaling in metabolically active tissues. The neural retina exhibits one of the highest energetic and redox demands in the body, yet mechanisms governing local flavin homeostasis remain poorly defined. Retbindin (RTBDN), a retina-specific riboflavin-binding protein, has previously been shown to regulate total retinal flavin levels and photoreceptor survival. Here, we demonstrate that RTBDN abundance and subcellular localization are dynamically modulated by developmental stage, riboflavin availability, and the light-dark cycle. RTBDN expression emerges during outer segment maturation, peaks in early adulthood, and declines with aging in parallel with retinal flavin content. Fasting reduces retinal flavin levels without altering total RTBDN protein abundance but induces redistribution of RTBDN from the outer segment-RPE interface toward cone-associated domains. Riboflavin repletion restores its localization. Similarly, RTBDN redistributes around cones during the dark phase and returns to outer segment tips at light onset, independent of changes in total protein levels. These findings identify RTBDN as a spatially dynamic regulator of flavin-dependent homeostasis that adapts to nutrient flux and circadian metabolic demand. This adaptive redistribution may represent a protective mechanism to preserve cone function under fluctuating energetic conditions and suggests that disruption of flavin homeostasis may contribute to cone degeneration.

Alternate day fasting (ADF) improves glucose metabolism, but the underlying mechanism remains unclear. In this study, we demonstrate that ADF upregulates HDAC5 in pancreatic β-cells, which in turn suppresses RASA3 expression, leading to subsequent suppression of β-cell proliferation, increase of insulin secretion, and improvement of glucose metabolism. Under both normal chow and high-fat dietary feedings, ADF reduced islet mass, enhanced insulin secretion induced by glucose, and improved glucose tolerance. ADF significantly increased HDAC5 expression in islets. HDAC5 deficiency abrogated the metabolic benefits of ADF, resulting in increased islet area, reduced insulin secretion, and impaired glucose tolerance. Transcriptomic sequencing identified RASA3 as a key downstream molecule. HDAC5 negatively regulated RASA3 in cultured MIN6 cells. Overexpression of RASA3 promoted β-cell proliferation and reversed the effects of HDAC5. Collectively, this study demonstrates that HDAC5 contributes to the effects of ADF in the improvement of glucose metabolism and provides an alternative strategy for drug targeting that mimics the effects of fasting.

Sertoli cells provide essential structural and nutritional support for germ cell development and play critical roles in spermatogenesis. Post-transcriptional regulation exerts important functions in Sertoli cells. Complex alternative splicing processes exist in Sertoli cells. This study focuses on splicing factor SRSF1 to investigate its function and mechanisms in Sertoli cell development. Sertoli cell-specific SRSF1 ablation from embryonic development arrests cell proliferation and increases apoptosis, thereby reducing Sertoli cell number, constricting the germ cell microenvironment, and causing progressive germ cell loss, ultimately resulting in male infertility. Mechanistically, SRSF1 regulates CDK pathway and Bax/Bcl2 pathway-related gene transcription and forms spliceosomal complexes with DDX5 and RBMX/RBMXL1 to modulate AS of Cidea and Rif1, thereby governing Sertoli cell proliferation, apoptosis, and DNA damage response. This study establishes SRSF1 as a multifunctional spliceosomal hub in Sertoli cells, refining our understanding of post-transcriptional regulation in spermatogenesis and providing potential therapeutic targets for male infertility.

Uranyl acetate (UA)-induced renal injury disrupts mitochondrial ultrastructure and homeostasis through oxidative stress-mediated pathways, with activating transcription factor 4 (ATF4) identified as a critical molecular switch. This study demonstrates ATF4 directly suppresses peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) transcription via promoter binding, as validated by chromatin immunoprecipitation and luciferase reporter assays. Integrated bioinformatics of renal injury datasets (GSE34351, GSE87024) revealed co-expression networks converging on mitochondrial regulators. In UA-exposed mice, ATF4 silencing rescued mitochondrial biogenesis by activating PGC-1α, restoring cristae architecture observed by transmission electron microscopy and upregulating biogenesis markers (NRF1, Tfam, CPS1). Adipose-derived stem cell exosomes (ADSC-exo) were isolated as biological mediators that attenuated renal uranium accumulation, serum oxidative stress, and histopathological damage. Mechanistically, ADSC-exo inhibited ATF4 to unleash PGC-1α-driven mitochondrial biogenesis, thereby reestablishing redox homeostasis. These findings support the ATF4/PGC-1α axis as an important regulatory pathway involved in mitochondrial impairment during uranium nephrotoxicity.

Podocyte injury is a primary driver of progression in diabetic kidney disease (DKD). Although single-cell RNA sequencing (scRNA-seq) is a powerful tool for identifying novel disease-associated genes, podocytes are frequently underrepresented in whole-kidney scRNA-seq datasets because they constitute only a minor fraction of renal cells. To overcome this limitation, we performed scRNA-seq on magnetically isolated glomeruli from diabetic db/db mice, generating a high-resolution, glomerulus-focused transcriptomic dataset. Diabetic glomeruli exhibited a marked reduction in podocyte abundance and extensive transcriptional remodeling. By integrating in vivo scRNA-seq data with in vitro analyses of cultured human podocytes exposed to diabetic stimuli-specifically palmitate-induced lipotoxicity-we identified four candidate genes (OPCML, ADM, NELL2, and DHRS3) associated with diabetic podocyte injury. Among these candidates, in vitro mechanistic validation demonstrated that knockdown of OPCML or ADM, as well as overexpression of DHRS3, significantly attenuated palmitate-induced apoptosis and preserved actin cytoskeletal integrity. These findings identify OPCML, ADM, and DHRS3 as novel mediators of podocyte injury and promising therapeutic targets for DKD.