S-palmitoylation, a reversible post-translational modification, regulates critical biological processes and represents a promising therapeutic target for diverse human diseases. Over the past decade, human α/β hydrolase domain (ABHD) proteins have emerged as key regulators of the dynamic S-palmitoylation/depalmitoylation cycle, yet a systematic overview of their roles in this modification remains lacking. This review provides a comprehensive summary of the functions of emerging ABHD proteins—including ABHD17A/B/C, ABHD16A, ABHD10, ABHD7, and ABHD8—in the dynamics of S-palmitoylation and depalmitoylation cycle, along with their molecular mechanisms, structural features, and substrate specificities. Associations between dysregulated ABHD protein-mediated S-palmitoylation and various diseases, including cancers, neurological disorders, degenerative diseases, viral infectious diseases, metabolic diseases, and reproductive disorders are discussed. Predictive criteria for uncharacterized ABHD proteins and prospective targeted therapeutic strategies are also proposed. This review provides mechanistic insights into emerging ABHD proteins in S-palmitoylation/depalmitoylation, and lays a foundation for developing novel diagnostic markers and precision therapies for S-palmitoylation-related diseases.
Millimeter-wave directed energy (mmWave DE) is increasingly used in telecommunications and military applications, yet its biological effects remain unclear. Here, we characterized the spatially dependent cellular and molecular impacts of 34.14 GHz mmWave DE exposure on mouse neuroblastoma (N2A) cells. N2A cells were exposed to mmWave DE for 48 h, generating Center (direct exposure), Penumbra (peripheral exposure), and Control groups. Morphology was assessed by phase-contrast microscopy and F-actin staining; viability and membrane integrity were evaluated by MTT, LDH, and DAPI/PI assays. Cytoskeletal protein degradation (αII-Spectrin, Vimentin, Nestin) was examined by Western blot and immunocytochemistry. Label-free LC-MS/MS proteomics, Gene Ontology (GO), Ingenuity Pathway Analysis (IPA), and PathwayStudio were used to define global molecular responses. Center-exposed cells showed severe morphological disruption, reduced viability (p < 0.0001), increased LDH release, and marked membrane permeabilization, consistent with dominant necrotic injury. Western blotting revealed enhanced proteolysis of αII-Spectrin, Vimentin, and Nestin. Proteomic profiling identified 181 statistically significant dysregulated proteins in the Center and 34 in the Penumbra (adjusted p < 0.05); of these, 12 and 14 proteins, respectively, also met the high-effect threshold of |log₂FC| ≥ 1. These alterations converged on cytoskeletal organization, mitochondrial stress, RNA processing, and proteostasis. mmWave DE exposure induces a spatially graded injury response, ranging from adaptive stress signaling in the Penumbra to proteostasis collapse, cytoskeletal failure, and membrane injury in the Center. These findings support further mechanistic evaluation of mmWave bioeffects and their potential relevance to neuroinjury-related pathways.
Selenium is a nonmetal element incorporated into a small subset of proteins as selenocysteine, the 21st amino acid. In humans, the so-called selenoproteins represent a unique and physiologically important family of proteins, some of which are essential to life, primarily through the maintenance of redox balance. Other selenoprotein functions include defense against various forms of oxidative stress, thyroid hormone metabolism, muscular homeostasis and integrity, as well as immune regulation, inflammatory responses, and protection against oxidative stress-mediated cell deaths. Increasing evidence shows that inhibiting global selenoprotein expression or targeting individual selenoproteins, such as glutathione peroxidase 4 and thioredoxin reductase 1, can greatly affect cellular redox status in different tissues and organs and trigger cell death through apoptosis, pyroptosis, ferroptosis, or even cuproptosis. This is particularly true in hematopoietic cells, where oxidative stress and reactive oxygen species production may profoundly alter differentiation of stem and lineage-committed progenitor cells and thus contribute to the progression and treatment resistance of hematopoietic malignancies. Thus, inhibition of selenoprotein synthesis or activity by genetic or pharmacological approaches has been shown to trigger apoptosis and/or ferroptosis, leading to the elimination of leukemic cells. Of note, glutathione peroxidases 1 and 4, SELENOM, SELENOO, and SELENOW are overexpressed in some subtypes of acute myeloid leukemia, often conferring a pejorative prognosis. Thus, targeting selenoproteins, either alone or in combination with conventional or targeted therapies, might represent an attractive strategy for the management of patients suffering leukemia.
Immune checkpoint inhibitors (ICIs) are the standard of care for advanced non-small-cell lung cancer (NSCLC) without targetable drivers, but resistance remains a challenge. Although the antidiabetic drug metformin has shown immunomodulatory potential, clinical evidence regarding its survival benefit in patients treated with ICIs is conflicting, and the molecular mechanisms driving potential synergy remain undefined. We conducted a multicentre, retrospective cohort study of patients with advanced NSCLC and type 2 diabetes treated with PD-1 or PD-L1 inhibitors at six tertiary hospitals in China. The primary outcome was durable clinical benefit (progression-free survival of 6 months or longer). Mechanistic studies used two in vivo models, a PBMC-humanized H1299 xenograft and an LLC syngeneic immunocompetent model, with the LLC tumours analysed by single-cell RNA sequencing (scRNA-seq). Lysine 2-hydroxyisobutyrylation (Khib) was quantified, HDAC3 and HSP90AA1 were modulated and quantitative proteomics mapped Khib sites. TBK1-IRF3 signalling and T-cell-mediated cytotoxicity were assessed. In total, 89 eligible patients were analysed. Concomitant metformin use (n = 54) was independently associated with improved durable clinical benefit [43 (80
Kidney disease progression is increasingly understood as a failure of cellular stress adaptation, in which renal cells cannot fully restore homeostasis after metabolic, hemodynamic, toxic, or inflammatory injury. Although mitochondrial dysfunction, proteostasis collapse, metabolic rewiring, regulated cell death, and immune activation have been widely studied as therapeutic targets, RNA homeostasis is emerging as a measurable layer of renal stress biology with selected pharmacological entry points. RNA homeostasis encompasses the coordinated regulation of RNA biogenesis, processing, chemical modification, localization, translation, decay, and quality control. These processes influence whether injured kidney cells regain adaptive function or progress toward maladaptive repair, senescence, inflammation, and fibrosis. Recent studies have identified transfer RNA (tRNA)-derived small RNAs, RNA autophagy, N6-methyladenosine RNA methylation, RNA-binding proteins, stress granules, and extracellular RNA signaling as functional regulators of kidney injury and repair. These pathways are increasingly accessible to mechanistic interrogation and, in selected preclinical settings, to modulation by small-molecule enzyme inhibitors, RNA mimics, antisense oligonucleotides, siRNAs, mRNA-based approaches, RNA editing, and kidney-targeted delivery systems. In this review, we synthesize RNA homeostasis as an emerging framework for understanding and potentially targeting kidney disease, while distinguishing mechanisms with causal preclinical support from more speculative therapeutic opportunities. We focus on targetable RNA-homeostatic mechanisms in acute kidney injury, diabetic kidney disease, podocyte injury, and renal fibrosis, and highlight strategies for translating RNA quality-control pathways into biomarkers, pharmacodynamic readouts and candidate disease-modifying interventions.
Liver organoids (LOs) are derived from pluripotent stem cells, adult stem cells (ASCs), or primary hepatic tissues using three-dimensional (3D) culture systems. These models self-assemble in vitro and recapitulate key hepatic structures and functions, thereby creating complementary platforms for liver development studies, as well as research on disease pathogenesis, drug metabolism/toxicity, and translational applications. The continuous improvement in engineering technologies has permitted LOs and liver-on-a-chip platforms to be applied to a variety of liver diseases such as fatty liver disease, inherited liver disorders, drug-induced liver injury (DILI), fibrotic liver disease, and hepatocellular carcinoma. Nevertheless, the widening deployment of LOs is impeded by numerous source-specific limitations. Although pluripotent stem cell-derived LOs are often immature, fetal-like, and lineage-biased, tissue-derived LOs continue to be hampered by limited sample availability, donor-to-donor variability, expansion-related phenotypic drift, and challenges in their standardization. Incomplete recapitulation of the native microenvironment, as well as high cross-batch variability, continues to hinder clinical translation in both systems.
Crosstalk between parenchymal and nonparenchymal cells of the liver mediates the activation of hepatic stellate cells (HSCs). Activated HSCs transform into myofibroblasts and produce the extracellular matrix (ECM), and massive ECM accumulation can affect liver function and eventually lead to liver fibrosis. Ovarian tumor domain protein 1 (OTUD1) is a deubiquitinases (DUB) that controls diseases progression. However, the physiological functions and underlying mechanisms of action of OTUD1 in liver fibrosis have not been investigated. In this study, bile duct ligation (BDL) and carbon tetrachloride (CCl4) injection were used to investigate the role of OTUD1 in liver fibrosis. The human liver cell line, LO-2, was co-cultured with human HSCs LX-2 to investigate the role of OTUD1 in HSCs activation and its related mechanisms. Our results show that OTUD1 was upregulated in fibrotic liver tissues. Additionally, OTUD1 knockout improved BDL- and CCl4-induced liver injury and fibrosis in vivo. In vitro, OTUD1 overexpression induced LX-2 activation by promoting TGF-β1 secretion in LO-2 cells. Meanwhile, we found that OTUD1 deubiquitinated and stabilized HSP90β. Our results suggest that OTUD1 in hepatocytes affect TGF-β1 secretion by stabilizing HSP90β protein level, thereby promoting the activation of LX-2 cells. We identified HSP90β as a target of OTUD1 and propose OTUD1 as a potential therapeutic target for liver fibrosis.
Erlotinib, an epidermal growth factor receptor (EGFR) small molecule inhibitor approved by the Food and Drug Administration (FDA) for the treatment of pancreatic cancer, faces significant drug resistance challenges. To address this, we investigated the potential of agrimoniin, a key active compound in the traditional Chinese medicine Agrimonia pilosa, which is known for its potent anticancer activity. This study aims to explore the combined effect of these two compounds in pancreatic cancer. We sought to verify the combined inhibitory effect of pancreatic cancer by agrimoniin and erlotinib in vitro and in vivo using various assays, including subcutaneous xenograft experiments and western blot. Single-cell RNA sequencing, bioinformatics, western blot, and lactic acid assays were performed to study how agrimoniin and erlotinib affect the tumor microenvironment and aerobic glycolysis. Finally, using signal pathway inhibitors/agonists, lactic acid assays, and Western blot, we aimed to reveal two positive feedback loops between signal pathways and lactic acid production. Agrimoniin combines with erlotinib to inhibit proliferation of pancreatic cancer cells. Together, they effectively suppress signal interactions between tumor cells, stromal cells, and macrophages within subcutaneous tumors. Simultaneously, they jointly inhibit aerobic glycolysis and lactic acid production in pancreatic cancer. Notably, this study proposes two potentially novel positive feedback loops between the PI3K/AKT/HIF-1α/HK2, MEK/ERK/HIF-1α/HK2 signaling pathways and lactic acid production. The combination of agrimoniin and erlotinib suppresses signal interactions between tumor cells, macrophages, and stromal cells in the tumor microenvironment, reprograms aerobic glycolysis, disrupts two positive feedback loops, thereby cooperatively inhibiting the proliferation of pancreatic cancer cells. This study provides a theoretical basis for agrimoniin as a sensitizer or resistance-reversing agent for erlotinib, and offers new insights and potential targets for pancreatic cancer drug development.
Transcription factor ETV4 is a member of the E26 transformation-specific (ETS) family and has recently emerged as a pivotal regulator in various diseases, functioning as a dynamic transcriptional integrator that links extracellular signaling cues to chromatin remodeling and transcriptional reprogramming. Accumulating evidence has expanded its functional spectrum far beyond conventional growth control, revealing critical roles in ferroptosis suppression, DNA damage, metabolic reprogramming, stemness maintenance, immune evasion, and tumor microenvironment regulation. In this review, we present a conceptual synthesis of current advances in ETV4 biology, emphasizing emerging mechanistic insights that redefine its role across cancer and nonmalignant disease contexts. We further highlight emerging insights into its potential involvement in phase separation and cancer neuroscience, proposing that ETV4 may serve as a molecular bridge integrating neural, immune, and metabolic cues within malignant contexts. By situating these recent developments within the broader landscape of cancer research, we outline unresolved challenges and offer new perspectives for the precision targeting of “undruggable” transcription factors, aiming to facilitate the translation of ETV4 from basic discovery to clinical application.
Gallbladder cancer (GBC) is characterized by a bile acids (BAs)-rich microenvironment that imposes significant endoplasmic reticulum (ER) stress. While the transcription factor AP-2α (TFAP2A) is upregulated in GBC, its functional role in this specific stress context remains unknown. This study investigates how TFAP2A enables GBC cell adaptation and survival under BAs stress. TFAP2A expression was analyzed in the tissues of patients with GBC via RNA sequencing, immunohistochemistry, and western blot. In vitro functional assays and patient derived organoid models were used to assess TFAP2A’s role in GBC. Interaction partners were identified through astral data-independent acquisition (DIA) proteomics and confirmed by coimmunoprecipitation. Protein stability was evaluated using cycloheximide chase and ubiquitination assays. In vivo validation was performed using cell-derived and patient derived xenograft models. Golgi integrity and ER stress markers were analyzed by immunofluorescence and western blot. TFAP2A was highly expressed in GBC tissues and shows cytoplasmic accumulation correlating with BA levels. Under BAs stress, TFAP2A translocated to the cytoplasm, where it interacted with the Golgi protein GOLIM4 and stabilized it by inhibiting its ubiquitin-mediated degradation. This TFAP2A/GOLIM4 maintained Golgi structure and alleviated ER stress, as evidenced by downregulation of proapoptotic CHOP and upregulation of GRP78/BiP. Consequently, it promoted GBC cell survival, proliferation, and tumor growth. Knockdown of either TFAP2A or GOLIM4 suppressed malignant phenotypes and increased ER stress-induced apoptosis, both in vitro and in vivo. This study identified a novel, nontranscriptional function of TFAP2A in response to BAs stress. By stabilizing GOLIM4 and enhancing secretory capacity, the TFAP2A/GOLIM4 axis represented a UPR-independent adaptive pathway that confers a survival advantage to GBC cells. Targeting this interaction may offer a new therapeutic strategy for this aggressive malignancy.
The adenosine deaminases acting on RNA (ADAR) family of enzymes (ADAR1 and ADAR2) catalyze adenosine-to-inosine (A-to-I) RNA editing. This post-transcriptional change is remarkably prevalent in the central nervous system (CNS). ADAR-mediated editing is critical for proper brain development, synaptic plasticity, and immunological homeostasis in the central nervous system (CNS) via recoding neurotransmitter receptors and ion channels. Conversely, a wide range of CNS disorders, such as neurodegenerative diseases (Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis), neuropsychiatric conditions (schizophrenia, bipolar disorder, and major depression), cerebrovascular diseases, and gliomas, are now linked to dysregulation of ADAR activity, whether through loss-of-function mutations, altered expression, or mislocalization. To address the main question of whether altered RNA editing is a fundamental driver of pathogenesis, a compensatory response, or a context-dependent modulator, this review critically synthesizes existing evidence. The basic processes of ADAR enzymes and their regulation throughout neurodevelopment are first described. Next, we thoroughly assess the unique molecular fingerprints of ADAR dysregulation across several CNS disorders, emphasizing recurring themes such as Alu RNA hypo-editing, induction of innate immunity, and GRIA2 editing, which can cause excitotoxicity. Lastly, we examine new treatment approaches that use or reinstate ADAR activity, such as small-molecule modulators and site-directed RNA editing tools (leveraging endogenous ADAR for programmable editing of RNA [LEAPER], clustered ADAR-recruiting guide RNAs (gRNAs) for effective RNA editing [CLUSTER], and mimicking inverted repeats to recruit ADARs using engineered oligoribonucleotides [MIRROR]). We summarize by reviewing key obstacles to clinical translation, including crossing the blood–brain barrier, the risks of off-target editing, and the challenges of achieving spatiotemporal accuracy. We also list important open topics for further investigation.
Although dysregulation of iron metabolism is known to contribute to tumorigenesis, the upstream mechanisms driving iron overload in gastric cancer (GC) remain poorly understood. This research aims to elucidate the oncogenic role of eukaryotic translation initiation factor 3 subunit I (EIF3I) in GC and its impact on iron metabolism. EIF3I expression in GC was evaluated using clinical tissue samples and correlated with clinicopathological features. The functional impact of EIF3I on tumor progression was evaluated through both in vitro and in vivo assays. Molecular mechanisms were investigated using protein interaction studies, ubiquitination assays, and transcriptional analyses. A therapeutic peptide targeting EIF3I– was screened and tested in zebrafish and mice models. EIF3I was significantly overexpressed in GC and associated with aggressive tumor phenotypes. EIF3I depletion suppressed GC cell proliferation and tumor growth, while its overexpression enhanced malignancy. Mechanistically, EIF3I stabilized yes-associated protein 1 (YAP) by directly interacting with it and preventing its ubiquitination and degradation by STUB1. This stabilization promoted YAP nuclear liquid–liquid phase separation and transcriptional activity, leading to increased expression of transferrin receptor 1 (TFR1), and consequent intracellular iron accumulation. Targeting the EIF3I interface on YAP with a screened peptide restored STUB1-mediated YAP degradation and significantly inhibited tumor growth in vitro and in vivo. EIF3I promotes GC progression by stabilizing YAP and reprogramming iron metabolism through the EIF3I–YAP-iron axis. Targeting this interaction represents a promising therapeutic strategy for GC.
Senile osteoporosis (SOP) is driven largely by bone marrow mesenchymal stem cell (BMSC) senescence and mitochondrial dysfunction. S-glutathionylation is an important redox modification regulating mitochondrial homeostasis, yet its role in skeletal aging remains unclear. The objective of this study is to systematically investigate the role of Grx2 deficiency-mediated CD36 S-glutathionylation in BMSC senescence and SOP development. Grx2, p53, and p21 expression in human and mouse BMSCs were analyzed by immunohistochemistry (IHC), western blotting (WB), and polymerase chain reaction (PCR). Aging-related and ovariectomy-induced osteoporosis models were established in Grx2−/− mice for bone metabolism and senescence assessments. BMSC senescence, osteogenesis, and adipogenesis were evaluated by SA-β-Gal and WB, ALP/ARS staining, and ORO staining. Transcriptomic and S-glutathionylated proteomic analyses were performed to identify underlying mechanisms. Fatty acid uptake was quantified using two-color flow cytometry. DAG and MDA levels were measured to assess lipid overload and oxidative injury. Mitochondrial structure and function were evaluated by TEM, Mitotracker, qPCR, NAD+/NADH, ATP, ROS, JC-1, and Seahorse assays. The PI3K/AKT pathway was assessed by WB. Co-IP confirmed CD36 S-glutathionylation, and molecular docking predicted C272 as the key modification site. CD36-C272S mutation and Grx2 overexpression were applied to validate functional mechanisms in vitro and in vivo. Grx2 deficiency, in both male and ovariectomized female mice, accelerates bone loss, inhibits osteoblast formation without altering osteoclast function, and exacerbates BMSC senescence. Through integrated transcriptomic and S-glutathionylated proteomic analysis, we identified the fatty acid transporter CD36 as a critical downstream target of Grx2. Notably, Grx2 deficiency markedly increases the S-glutathionylation of CD36, which not only enhances its fatty acid uptake capacity, leading to the accumulation of toxic lipid metabolites and oxidative damage, but also impairs mitochondrial energy metabolism by inhibiting the PI3K/AKT signaling pathway. Overexpression of Grx2 or the C272S mutation in CD36, which disrupts its S-glutathionylation, can break this harmful cycle and inhibit BMSC senescence. Grx2 deficiency-mediated CD36 S-glutathionylation drives BMSC senescence and SOP, providing new insight into the redox regulatory mechanisms underlying skeletal aging.
Despite extensive characterization of sperm structures, certain specialized subcellular structures are still not well understood. Among these, the posterior ring has been recognized for over half a century; yet, its molecular composition and biological role remain unknown. This research aims to define the molecular components and functional roles of the sperm posterior ring. 3D-rendered confocal microscopy was used to determine the localization and developmental dynamics of SPEM3 and TEX50 in mature spermatozoa and spermatids at different steps of spermiogenesis. Spem3 and Tex50 knockout mouse models were generated to explore their physiological functions. Sperm morphology and ultrastructure were investigated through immunofluorescence along with transmission and scanning electron microscopy, whereas sperm motility was evaluated using computer-assisted sperm analysis. Proteomics, coimmunoprecipitation, and immunoblotting were performed to identify SPEM3 and TEX50 as structurally interdependent interacting components of the posterior ring and to elucidate how their loss disrupts sperm architecture and fertility. We identified SPEM3 as a core component of the posterior ring located at the sperm head–tail linkage. The posterior ring arises from the marginal ring at the acroplaxome periphery during early spermiogenesis. Loss of Spem3 led to severe bending of the head–tail linkage, accompanied by a marked reduction in sperm motility, ultimately leading to male infertility. Furthermore, SPEM3 interacts and colocalizes with TEX50, and deficiency of either protein resulted in similar phenotypes, including disorganization of the posterior ring and postacrosomal region as well as impairment of the sperm connecting piece. These findings establish SPEM3 and TEX50 as critical posterior ring components essential for maintaining postacrosomal integrity and anchoring the sperm head to the tail. By resolving the long unknown molecular composition of the posterior ring, this study provides new mechanistic insight into sperm head–tail integrity and the pathogenesis of certain forms of male infertility.
Toll-like receptor 4 (TLR4) activation within the bone marrow (BM) microenvironment regulates proinflammatory cytokines and chemokines, which influence immune activity and bone remodeling. However, the immune cell factors mediating TLR4-dependent cross-talk in the bone/BM microenvironment remain insufficiently defined. We provide a transcriptomic analysis of significant cellular and immune changes in total BM cells of wild-type mice following lipopolysaccharide (LPS)-induced TLR4 activation in vivo compared with TLR4 knockout (KO) mice. To obtain direct evidence that interleukin (IL)-22 contributes to TLR4-driven bone destruction, we analyzed their bone phenotypes following LPS injection into IL-22 KO mice. In addition, co-culture of pre-osteoblasts (OBs) and BM cells was conducted to evaluate the effect of IL-22 in osteoclast (OC) formation in a pre-OB-dependent manner. Differentially expressed gene analysis from BM cells of wild-type (WT) mice revealed an increase in IL-22-producing cells, along with increased IL-22 production in a TLR4-dependent manner. Flow cytometry confirmed that LPS enhanced the number of CD4+IL-22+ cells in the BM and decreased in TLR4-deficient BM. LPS also enhanced the differentiation of CD4+IL-22+ T cells and promoted IL-22 induction in T cells. IL-22 KO mice showed minimal LPS-induced bone loss, whereas WT mice exhibited severe bone destruction with increased formation of tartrate-resistant acid phosphatase-positive OCs. Interestingly, while IL-22 did not directly affect OC differentiation, IL-22 enhanced OC differentiation only when BM cells were co-cultured with pre-OBs through the elevation of the receptor activator of nuclear factor-κB ligand (RANKL) in pre-OBs during osteogenesis. Mechanistically, IL-22 induced cyclooxygenase-2 in pre-OBs through extracellular signal-regulated kinase and p38 signaling, driving prostaglandin E2 production and subsequent RANKL induction. TLR4 activation enhanced IL-22 production in T cells, which stimulates RANKL production in pre-OBs to further induce OC formation. These results support that IL-22 acts as a TLR4-driven mediator to shift bone remodeling toward catabolic bone degeneration.
Hypopharyngeal squamous cell carcinoma (HPSCC) is a rare, highly aggressive malignancy, with recurrence playing a pivotal role in treatment failure and cancer-related mortality. The tumor microenvironment (TME) characteristics and molecular mechanisms driving recurrence in HPSCC remain poorly defined. Primary HPSCC (P-HPSCC) samples (n = 6) and recurrent HPSCC (R-HPSCC) samples (n = 3) were analyzed. Single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics were employed to dissect the cellular composition and molecular characteristics of the TME. Key regulatory networks were identified through gene expression analysis, pathway enrichment, and intercellular communication profiling. Functional validation experiments were performed to confirm the biological relevance of the pathways identified. ScRNA-seq analysis revealed that malignant epithelial cells (maEpCs) in recurrent tumors exhibited increased stemness, epithelial–mesenchymal transition (EMT), and immune evasion programs. Mechanistically, IGF2BP2 was selectively upregulated in R-HPSCC and functioned as an m6A reader to stabilize SMAD3 mRNA, thereby activating transforming growth factor beta (TGF-β) signaling and facilitating malignant progression. Integrated spatial transcriptomics and functional assays demonstrated that cancer-associated fibroblasts in recurrent tumors promote progression via FN1-mediated signaling. Concurrently, the recurrent TME underwent myeloid remodeling toward an immunosuppressive state, characterized by increased infiltration of SPP1+ tumor-associated macrophages and further enrichment of LAMP3+ dendritic cells with impaired antigen-presenting capacity, alongside aggravated CD8+ T-cell exhaustion, marked by upregulation of CTLA4 as a potential immune escape driver. This study identifies recurrence-associated cellular and molecular features in HPSCC. Recurrent tumors exhibited maEpC reprogramming via IGF2BP2-mediated activation of TGF-β signaling, coupled with FN1-dependent stromal activation and immune evasion marked by myeloid immunosuppressive remodeling and CD8+ T-cell exhaustion, characterized by CTLA4 upregulation. Collectively, these findings reveal a pro-recurrent TME and suggest potential therapeutic targets in R-HPSCC.
Swine leukocyte antigen class I (SLA-I) molecules are swine orthologs of human MHC class I molecules and are encoded by three classical loci, SLA-1, SLA-2, and SLA-3. By engaging T-cell receptors (TCRs), these cell-surface proteins present antigenic peptides, whose binding specificity is largely dictated by the architecture of the peptide-binding groove (PBG) and its constituent pockets. Although early SLA-I research primarily emphasized molecular and functional features, recent advances in structural biology have yielded an increasing number of crystal structures of SLA-I–peptide complexes, providing critical insights into the principles of peptide presentation and T-cell recognition in pigs. In this review, we examine the structural features of SLA-I molecules, focusing on the PBG and binding pockets that accommodate peptide anchor residues. Structural studies revealed that the SLA-I PBG, while architecturally conserved, exhibits pronounced allelic polymorphism and plasticity, predominantly within its six binding pockets (A–F). A key insight is the profound functional impact of micropolymorphisms; for instance, single-residue variations in pockets such as D can dramatically alter the peptide-binding specificity and repertoire. Finally, we highlight current knowledge gaps and future research directions to facilitate the exploitation of SLA-I structural features for targeted vaccine design and immunotherapeutic development, thus addressing the challenges of porcine immune recognition.
Lipotoxicity-induced endothelial dysfunction impairs wound healing in obesity and type 2 diabetes, yet the underlying mechanisms remain elusive. While Angiogenin promotes endothelial function, its role under lipotoxic stress has been unknown. Here, we demonstrate for the first time that Angiogenin effectively protects against lipotoxicity-induced delayed skin repair. In a cellular lipotoxicity model induced by oleic and palmitic acids (OPA) using human umbilical vein endothelial cells (HUVECs), the levels of Angiogenin decreased in a time- and dose-dependent manner. Meanwhile, a similar reduction was also observed in the skin tissue of high-fat diet/streptozotocin (STZ)–induced diabetic mice and Apolipoprotein E (APOE)−/− mice. Restoring Angiogenin levels significantly enhanced endothelial proliferation, migration, and angiogenesis, rescuing OPA-induced impairment. Through RNA-seq and subsequent validations, we provide novel mechanistic insight into Angiogenin’s protective action: Angiogenin directly binds and stabilizes Caveolin-1 (Cav1) mRNA, leading to increased Cav1 expression. Critically, we show that the Angiogenin-mediated endothelial rescue is strictly dependent on this Cav1 upregulation. Furthermore, topical administration of Angiogenin peptide significantly accelerated wound closure and neovascularization in both APOE−/− and high-fat diet/STZ-induced diabetic mice. Collectively, our study unveils a novel Angiogenin–Cav1 axis in endothelial protection, positioning Angiogenin as a promising therapeutic candidate for lipotoxicity-induced impairment of skin wound healing.
It is generally known that kidney stone disease (KSD) is associated with alterations in urinary microbiome, but the roles of the urinary microbiome in KSD pathogenesis remain unclear. This study addressed the impact of Lactobacillus acidophilus (a commensal bacterium found in normal urine) on renal epithelial integrity, calcium oxalate monohydrate (COM) crystal-cell adhesion, expression of membrane receptors of COM crystals, and oxalate degradation under oxalate-induced stress (a known inducer of KSD). Inner medullary collecting duct cells (mIMCD-3) were cultured for 24 h under control or oxalate-induced (by 0.6 mM sodium oxalate; NaOx) conditions without or with L. acidophilus (at 1 × 103 colony-forming unit (CFU)/ml) co-incubation. NaOx reduced transepithelial resistance (TER) of the mIMCD-3 monolayer and downregulated ZO-1, a tight junction (TJ) protein. Additionally, NaOx enhanced the COM crystal-binding capability of mIMCD-3 cells by upregulating a COM crystal receptor, annexin A2, on cell membranes. Such harmful effects of NaOx were abolished when mIMCD-3 cells were co-cultured with L. acidophilus. Moreover, culturing L. acidophilus in artificial urine (AU) supplemented with NaOx for 24 h revealed that the oxalate level in AU decreased, suggesting the oxalate-degrading activity of the bacterium in an AU environment. L. acidophilus prevented oxalate-mediated renal epithelial barrier disruption and COM crystal adhesion to renal epithelial cells by preserving ZO-1 and annexin A2 expression at their basal levels, at least in part, via its oxalate-degrading property. Not applicable (This is not a clinical trial).
O-GlcNAcylation is a post-translational modification (PTM) uniquely catalyzed by O-GlcNAc transferase (OGT), which has been linked to tumorigenesis and neurodegeneration. However, its roles in mammalian spermatogenesis remain unexplored. This study aims to elucidate the functional mechanisms of OGT in spermatogenesis and male fertility. We employed immunoprecipitation-mass spectrometry (IP-MS) to identify candidate O-GlcNAcylated substrates of OGT in juvenile mouse testes. To explore the physiological roles of OGT and O-GlcNAcylation, we constructed a mouse model with postnatal germ cell-specific deletion of Ogt via Stra8-Cre. In addition, we performed integrated bulk and single-cell RNA sequencing analyses to investigate the potential mechanisms by which OGT and O-GlcNAcylation deficiency impairs spermatogenesis. The results showed stage-specific OGT enrichment and O-GlcNAcylation in mouse testicular spermatogonia and early spermatocytes. Furthermore, OGT was found to interact with and O-GlcNAcylate transcription factors (e.g., HCFC1) as well as splicing regulators (e.g., SRSF1 and SF3B3) in mouse testes. Postnatal germ cell-specific Ogt deletion impaired spermatogonial differentiation, disrupted meiotic initiation and progression, and induced apoptosis, ultimately leading to male infertility. Mechanistically, Bulk RNA sequencing (RNA-seq) analysis revealed that OGT deficiency dysregulated transcriptional and alternative splicing programs, affecting genes critical for the mitotic–meiotic transition (e.g., Ythdc2 and Rbm46) and meiotic progression (e.g., Stra8, Stag3, and Syce2) in the testes. Single-cell RNA sequencing further uncovered aberrant retention of mitotic transcripts (e.g., Ccna2 and Ccnb1) in spermatocytes and impaired mRNA metabolism during spermatogonial differentiation. In addition, OGT deficiency caused cytoplasmic mislocalization and reduced expression of core transcription factors and splicing regulators in spermatocytes. These findings establish that OGT and its mediated O-GlcNAcylation coordinate essential gene expression and mRNA metabolism during mitotic-to-meiotic transition and meiotic progression. Moreover, our study provides mechanistic insights into the pathogenesis of male infertility associated with O-GlcNAcylation dysregulation.
