De novo lipogenesis (DNL) operates as a core metabolic pathway in cellular lipid homeostasis, yet its central effector enzyme fatty acid synthase (FASN) undergoes aberrant overexpression and hyperactivation in tumors and metabolic disorders. Herein, we systematically outline the versatile functions of FASN in tumor lipogenic metabolism and summarize recent advances in its covalent regulatory modifications. We highlight nucleic acids methylation modifications, acetylation, ubiquitination, glycosylation, ubiquitin-like modification, palmitoylation and phosphorylation modifications as an integrated network that tune FASN stability, catalytic activity, and subcellular localization. These modifications exhibit extensive crosstalk and disease-specific heterogeneity, offering potential molecular targets for therapeutic intervention in cancer and metabolic diseases.
Syndecans (SDCs) 1–4 are a family of transmembrane heparan sulfate proteoglycans (HSPGs) that regulate cell–cell communication, adhesion, extracellular matrix organization, and signaling pathways involved in tumor biology. In prostate cancer (PCa), accumulating evidence suggests that SDCs contribute to tumor progression, therapeutic resistance, and interactions within the tumor microenvironment. However, their specific, stage-dependent roles remain incompletely understood. This review provides an integrated synthesis of current experimental and clinical evidence on SDC1–SDC4 in PCa, complemented by exploratory analyses of publicly available transcriptomic, genomic, and proteomic datasets. In contrast, copy-number alteration (CNA) strata dichotomized by the mean for SDC1, SDC2, and SDC4 showed differences in progression-free interval. Specific CNA subclasses and relationships between CNA values and SDC mRNA or protein abundance could not be determined. Proteomic pseudotime analysis further suggested that SDC4 expression increases during PCa progression, supporting its potential involvement in advanced disease. We discuss the regulation and modulation of SDCs by androgen deprivation therapy (ADT), enzymatic shedding, integrin-mediated signaling, extracellular matrix interactions, lipid signaling pathways, and microRNA networks. In particular, SDC1–microRNA interactions may influence PCa cell proliferation, cellular senescence, epithelial–mesenchymal transition (EMT), and intracellular signaling pathways. Overall, this review highlights SDCs as context-dependent regulators of PCa biology with potential relevance as biomarkers or therapeutic targets. However, clinical translation will require independent validation, standardized assays, compartment-resolved analyses, and mechanistic confirmation.
Metastatic progression of prostate cancer involves complex remodelling of signalling networks that cannot be fully explained by changes in individual pathway activity. In this study, we performed a systems-level analysis of signalling module interactions using transcriptomic data from primary prostate tumours in the TCGA-PRAD cohort (n = 550) and metastatic tumours from the SU2C cohort (n = 118). The framework integrates thirteen metastasis-associated signalling modules encompassing oncogenic, inflammatory, stress-response, angiogenic and mechanotransduction pathways. Module activity scores were computed for key cancer-related signalling pathways and used to construct module co-occurrence networks. While differences in median pathway activity between cohorts were modest, metastatic tumours exhibited substantial rewiring of pathway coordination. Notably, this rewiring reflects changes in pathway coordination rather than absolute activity levels, supporting a network-centric view of established metastatic disease. Network analysis identified the CXCR4 signalling axis as a central hub showing increased connectivity with inflammatory and growth-factor pathways, including NF-κB, MAPK, and hypoxia modules. Unsupervised clustering of pathway activity profiles revealed two distinct signalling states characterized by differential androgen receptor signalling and activation of metastasis-associated pathways. These signalling states were significantly associated with PTEN and TP53 alterations. A logistic regression classifier based on module activity achieved moderate discrimination between primary and metastatic tumours (AUC = 0.685), indicating that coordinated pathway activity captures biologically relevant features of disease progression. Overall, our findings highlight the importance of network-level approaches for understanding metastatic prostate cancer and identify CXCR4-centred signalling architecture as a key component of metastatic signalling rewiring.
Inflammation accompanies a wide range of pathological conditions and negatively impacts the male reproductive function by interfering with steroidogenesis and spermatogenesis. Steroidogenesis is sustained by Leydig cells (LCs), but it can be impaired by local or systemic inflammatory events known to alter the expression and function of aquaporins (AQPs). In addition, glycerol acts as an osmolyte and a metabolic substrate that supports cellular homeostasis and contributes to the proper function of testicular cells. To investigate the effects of inflammation on aquaglyceroporins (AQGPs) and glycerol transport, Mus musculus LCs (TM3) were exposed to bacterial lipopolysaccharide (LPS) for 24hours to induce an inflammatory-like state; this promoted a decrease in glycerol permeability, assessed by stopped-flow light scattering, and increased expression of the Aqp9 transcript. Furthermore, inflammatory stimulation was associated with impaired mitochondrial respiration activity, reflected by decreased oxygen consumption rates linked to ATP production (OCR), as well as reduced expression of the steroidogenic genes Star and Cyp17a1 and lower testosterone production. Collectively, these findings demonstrate that inflammation is associated with alterations in Aqp9 expression, glycerol permeability, mitochondrial respiratory parameters, and steroidogenic activity in TM3 mLCs. Though the mechanisms linking glycerol homeostasis, cellular energy metabolism, and hormone production under inflammatory conditions warrant further investigation, this work provides first-time evidence of a correlation between inflammation-induced metabolic and hormonal disruptions and impairment of glycerol transport in TM3 LCs.
The processes by which an individual develops as a male (testis) or female (ovary) are remarkably diverse across taxa. While sex is determined by genetic factors in some species, it can be determined by environmental factors in others. Despite these discrepancies, common features are however observed. For species as distant as drosophila and humans, the ovary or testis arises from a bipotential gonad. Thus, each embryo has the potential to develop as a male or female. Given the growing evidence that global change can affect the sexual fate and fertility of individuals across taxa, there is a critical need to understand the mechanisms involved in sex differentiation and maintenance. In the present work, we followed the transcription and methylation levels of genes known, or suspected, to be involved in sex differentiation throughout zebrafish development. Early sex differentiation was marked by sex-specific transcription patterns of genes encoding for methyl-sensitive transcription factors of the zbtb family and for sox9a and zar1, establishing common features with mammals. In contrast, our results suggest that sex markers such as cyp19a1a, dmrt1, foxl2a or amh are mainly involved in sex stabilization and maintenance. DNA methylation appeared to be at the interface, playing a role in sex differentiation and maintenance by progressively constraining plasticity and stabilizing the transcription of genes under study. Our results also highlighted significant changes in the transcription level of genes involved in prostaglandins production during the ovary-to-testis transition, suggesting a potential role of these mediators in DNA methylation reprogramming and sex differentiation.
BACKGROUND:Fibroblast-like synoviocytes (FLSs) are key effector cells in osteoarthritis (OA), contributing to joint degeneration through inflammatory and mechanobiological mechanisms. Triamcinolone acetonide (TA) is widely used as an intra-articular therapy for OA; however, its effects on synoviocyte mechanobiology remain poorly defined. This study investigated TA-induced changes in FLS viability, cytoskeletal organization, biomechanical properties, mechanically evoked Ca²⁺ signaling, and mechanotransduction-related gene expression. METHODS:Primary FLSs isolated from the synovial tissue of patients with osteoarthritis (N = 3) were treated with increasing concentrations of TA. Cell morphology and viability were assessed by cytooskelton laelling andflow cytometry. Cellular stiffness was measured by atomic force microscopy (AFM). Mechanically evoked Ca²⁺ responses were assessed by combining AFM-based single-cell stimulation with live-cell calcium imaging. Mechanotransduction-associated transcripts were quantified by qPCR, while PIEZO1, PIEZO2 and TRPV4 protein expression was evaluated by immunolabelling and ELISA. RESULTS:TA induced concentration-dependent alterations in FLS morphology and viability, with marked cytotoxicity above 0.35 mM. At sub-cytotoxic levels, TA significantly increased cellular stiffness at 0.2 mM (p = 0.0219), while 0.35 mM showed a non-significant increase. Mechanically evoked Ca²⁺ signaling was enhanced following TA treatment, particularly at lower concentrations, with notable inter-donor variability. Immunolabelling revealed PIEZO1, PIEZO2 and TRPV4 distribution across all conditions, while quantitative ELISA showed increased protein expression following TA exposure despite only modest changes in the transcript levels. CONCLUSIONS:TA exerts concentration-dependent effects on OA-derived FLSs, modulating cytoskeletal organization, biomechanical properties, mechanically evoked Ca²⁺ responses and mechanosensitive ion-channel regulation. These findings suggest that TA modulates FLSs mechanobiology beyond its established anti-inflammatory effects.
Prostate cancer commonly remains refractory to checkpoint blockade, consistent with an immune-cold microenvironment and tumor-intrinsic defects in interferon signaling. We report that loss of the basic helix-loop-helix transcription factor TCF3 defines a tumor-intrinsic mechanism that simultaneously drives aggressive progression and immune evasion. TCF3 expression is reduced in advanced and metastatic prostate cancer across multiple cohorts and in a Pten-driven model. In androgen-independent prostate cancer cells and organoids, TCF3 depletion enhances proliferation, clonogenicity, migration, and EMT-associated programs, while overexpression of TCF3 isoforms suppresses these malignant phenotypes. Transcriptomic profiling further shows that TCF3 loss profoundly downregulates interferon-α/γ response programs. Mechanistically, TCF3 promotes transcription of STAT2, preserving basal and inducible interferon signaling and downstream ISG expression; TCF3-deficient tumors exhibit reduced immune infiltration and impaired CD8+ T cell effector function in vivo. Notably, therapeutic innate immune activation partially restores interferon responses, and poly(I:C) plus PD-1 blockade reinvigorates cytotoxic T cell activity and markedly suppresses tumor growth. These data support TCF3 as a biomarker of interferon competence and a rational basis for selecting patients for innate agonist-checkpoint combinations.
Background Oral leukoplakia (OLK) is the most prevalent and extensively studied oral potential malignant disorder (OPMD), with a high risk of progression to malignancy. Ferroptosis has been implicated as a key process in the progression of OLK, and iron metabolism significantly influences the ferroptosis process. DIRAS2, a member of the GTPase family within RAS proteins, is associated with ferroptosis, however, how DIRAS2 affects iron metabolism remains unknown. Methods Immunohistochemical staining was used to assess the expression of SLC7A11 and LCN2 in human normal oral mucosa and OLK tissues. Erastin-induced Leuk-1 cells were employed to investigate ferroptosis through CCK8 assays, MDA and Fe²⁺ measurements, and alterations in mitochondrial membrane potential and morphology. qRT-PCR and western blotting were performed to detect the expression of iron metabolism-related molecules, additionally, LCN2 inhibitor and small interfering RNAs were employed in this study. Dual-luciferase assays, co-ip, CHX chase assay and Duolink assay were conducted to elucidate the interactions among DIRAS2, HIF1α, and LCN2. The findings were validated in 4-Nitroquinoline-1-oxide (4NQO) -induced mouse OLK models too. Results SLC7A11 and LCN2 expression was upregulated in OLK specimens compared to normal oral mucosa tissues. DIRAS2 knockdown in Erastin-induced Leuk-1 cells exacerbated mitochondrial morphology alteration, increased MDA and Fe²⁺ levels, and further upregulated LCN2 and NCOA4 expression while decreasing FTH1 protein expression. Knockdown of LCN2 expression by inhibitor or small interfering RNAs decreased Fe²⁺ levels and related protein expression in Leuk-1 cells. Compared with normal tongue tissues, the expression of ACSL4, SLC7A11, and the LCN2-NCOA4/FTH1 pathway was elevated in 4NQO-induced mouse tongue tissues. Conditional knockout of DIRAS2 resulted in decreased expression of SLC7A11 and FTH1, while increased expression of ACSL4, LCN2, and NCOA4. Furthermore, DIRAS2 regulated the expression of LCN2 via HIF1α mediation. Conclusions DIRAS2 may regulate iron metabolism in ferroptosis during the progression of OLK.
Trained immunity, an innate immunological memory induced by epigenetic and metabolic reprogramming, has changed the paradigm of host defense and pathogenesis of chronic inflammatory disease. Unlike adaptive immunological memory, trained immunity is characterized by the ability of innate immune cells and their progenitors to respond more robustly or differently to subsequent stimulations and contributes to chronic inflammatory conditions. Emerging data suggests that this process might be essential in autoinflammatory and immune-mediated inflammatory illnesses by enhancing sterile inflammation, decreasing activation thresholds, and boosting disease chronicity. This narrative review summarizes the existing evidence relating trained immunity to monogenic and polygenic autoinflammatory diseases. The greatest evidence in monogenic disease is for mevalonate kinase deficiency, where dysregulated mevalonate metabolism directly overlaps with conventional trained immunity pathways. Moderate evidence exists for familial Mediterranean fever, cryopyrin-associated periodic syndromes, and tumor necrosis factor receptor-associated periodic syndrome. For other rare hereditary autoinflammatory diseases, data are still inadequate. There is convincing evidence for a role of trained immunity in polygenic disorders like gout, atherosclerosis, obesity-associated "metaflammation", and type 2 diabetes and increasing evidence in Behçet's disease, adult-onset Still's disease, psoriasis, hidradenitis suppurativa, inflammatory bowel disease, and related inflammatory spectrum disorders. A major conceptual finding is that autoinflammatory illnesses may be a dynamic interplay between hereditary susceptibility and dysfunctional innate immune memory, rather than isolated static inflammatory abnormalities. However, information gaps still exist in reprogramming at the progenitor level, disease-specific epigenetic markers, and the reversibility of trained states. Understanding these systems may allow the development of therapeutic techniques to de-train abnormal innate immunological memory and obtain resilience for diseases.
Among the intracellular sensors of innate immunity against infection, RIG-I-like receptors (RLRs) serve as cytosolic surveillance sensors that detect viral RNA species. However, the role of kinases in modulating RIG-I in a catalytically-independent manner is not well known. In this study, we report that protein kinase DYRK1B is a novel positive regulator of RIG-I-mediated antiviral innate immunity. Overexpression of DYRK1B markedly amplified RNA virus-induced IFN-I production, whereas CRISPR-mediated knockout of DYRK1B substantially attenuated these antiviral responses. Mechanistic investigation revealed that this regulatory role operates independently of DYRK1B's catalytic kinase function; instead, DYRK1B functions as an adaptor protein that connects TRIM25 with RIG-I, thereby promoting TRIM25-mediated K63-linked polyubiquitination of RIG-I, an essential modification required for RIG-I functional activation. Collectively, our findings reveal that DYRK1B enhances innate immunity against RNA viruses by strengthening the physical association of RIG-I with TRIM25, providing a more profound understanding of the mechanisms involved in antiviral immune regulation.
INTRODUCTION:The nasolacrimal duct (NLD) holds significant clinical importance in diseases represented by primary acquired NLD obstruction (PANDO). This study aims to culture human NLD epithelial cells using organoid culture systems, with the goal of establishing cell models that recapitulate the in vivo characteristics of the NLD in an in vitro setting. METHODS:Human NLD epithelial specimens were surgically obtained from PANDO patients during endoscopic dacryocystorhinostomy. Human NLD epithelial cells were isolated via enzymatic digestion and expanded using Expansion Medium. Air-liquid interface culture was performed to induce differentiation, evidenced by active ciliary beating within 28 days. Both apical-in and apical-out NLD organoids were successfully generated using Matrigel-based 3D culture systems and AggreWell™ 400 24-well plate. RESULTS:Histological analysis confirmed that the human NLD epithelium comprised a stratified columnar architecture with ciliated, goblet, and basal cells. Using an air-liquid interface system, primary epithelial cells were successfully differentiated into a polarized layer exhibiting native tissue characteristics. Furthermore, both apical-in and apical-out organoids were established, recapitulating key structural and cellular features of the native NLD epithelium. CONCLUSION:This NLD epithelial organoid system provides a versatile platform for studying NLD physiology, pathology, drug screening, and naso-ocular interactions, with potential for further expansion into broader research applications.
The spatiotemporal regulation of protein phosphorylation regulates cellular homeostasis, yet the phosphatase networks that actively dismantle oncogenic kinase signaling remain elusive. Here, we identify Mg2+-dependent protein phosphatase 1 (PPM1B) as a critical tumor suppressor in cervical carcinoma. Biochemically, PPM1B directly interacts with and dephosphorylates the myosin phosphatase (MP) regulatory subunit MYPT1 at its inhibitory Thr853 residue. We show that Rho-associated kinase (ROCK) activation by lysophosphatidic acid generates a high-affinity substrate profile, recruiting PPM1B to the Thr853-phosphorylated MYPT1. Crucially, this dephosphorylation drives the nuclear export of the MP holoenzyme, triggering a dual tumor-suppressive cascade. In the cytoplasm, reactivated MP dephosphorylates myosin light chain (MLC20) at Ser19 to prevent actomyosin-driven invasion. Concurrently, nuclear clearance of MYPT1 disrupts the PRMT5/histone H4 epigenetic axis, diminishing PRMT5 Thr80 phosphorylation and H4 symmetric dimethylation. Proteomic and phospho-kinase profiling indicates that PPM1B globally reprograms the malignant state, upregulating tumor suppressors (p53, Maspin) while attenuating epithelial-mesenchymal transition (EMT) drivers and survival networks (JNK, CREB). Finally, clinical biopsies of human squamous cell carcinoma demonstrate a strong correlation between reduced PPM1B expression and MYPT1 Thr853 hyperphosphorylation. Collectively, our findings establish PPM1B as a master catalytic switch that abrogates oncogenic signaling at both the cytoskeletal and epigenetic levels.
Ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, has emerged as a critical mechanism in tumor suppression and a promising target for cancer therapy. As a reversible and dynamic post-translational modification, phosphorylation orchestrated by protein kinases serves as a pivotal regulatory layer in the ferroptotic process. This review systematically delineates the roles of two major kinase families- mitogen-activated protein kinases (MAPKs) and Adenosine 5'-monophosphate-activated protein kinase (AMPK)-in the initiation, execution, and modulation of ferroptosis. We discuss how MAPK subfamilies, including extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38), and extracellular signal-regulated kinase (ERK5), regulate iron metabolism, lipid peroxidation, and antioxidant responses through context-dependent signaling cascades. In parallel, AMPK acts as an energy-sensing hub that influences ferroptosis via lipid metabolic reprogramming and the control of glutathione peroxidase 4 (GPX4) expression. We further explore the dual implications of ferroptosis in tumor biology, highlighting its role as a barrier to cancer progression and the adaptive resistance mechanisms evolved by tumor cells. Finally, we evaluate the therapeutic potential of targeting kinase-mediated ferroptosis, offering insights into combination strategies involving kinase inhibitors and ferroptosis inducers, and addressing the challenges and future directions for translating these insights into clinical oncology.
Glioblastoma is the most aggressive primary brain tumor in adults. Resistance of glioblastoma to the standard chemotherapy agent temozolomide (TMZ) correlates with the expression of the DNA repair enzyme O6-Methylguanine-DNA methyltransferase (MGMT). Additionally, tumor initiation and recurrence are associated with a subpopulation of glioblastoma stem cells (GSCs) which exhibit self-renewal properties, multipotency, and resistance to both chemotherapy and radiotherapy. To identify novel molecular biomarkers and therapeutic targets, we investigated the possible role of the oncoprotein Golgi phosphoprotein 3 (GOLPH3) in glioblastoma. GOLPH3 is overexpressed in many solid tumors and confers resistance to DNA-damaging chemotherapeutic agents. Its overexpression is also associated with poor prognosis in a variety of cancers, including glioblastoma. Here, we show that GOLPH3 knockdown in U87MG glioblastoma cells inhibits cell proliferation, promotes senescence and induces striking cellular and molecular changes toward a neuron-like phenotype. We analyzed the constitutive expression of GOLPH3, MGMT and mechanistic target of rapamycin (mTOR) signaling proteins in U87MG cells and in three GSCs representative of the three glioblastoma molecular subtypes. Our results suggest that these glioblastoma cell models may exhibit distinct responses to mTOR inhibition and TMZ. Finally, we show that GOLPH3 depletion reduces the activity of both mTOR complex 1 and 2 leading to decreased phosphorylation of their respective downstream targets S6K and Akt in both U87MG cells and the BT379 GSCs subtype. Our findings identify a potential therapeutic vulnerability that can be exploited to develop patient-tailored treatments in glioblastoma.
Epididymal epithelial cells are critical for sperm maturation. Although previous studies have determined the development of epididymal epithelial cells in rat epididymis, the postnatal developmental trajectories of distinct epithelial cell types in the mouse epididymis remain elusive. Here, we examined the developmental timeline of mouse epididymal epithelium differed from that of the rat by confocal immunostaining and single-cell RNA sequencing (scRNA-seq). In mice, principal and basal cells appeared as early as postnatal day 7 (PD7) across all segments, labeled by AQP9 and KRT5, in contrast to the later emergence in rats (e.g., basal cells at PD14 in rat cauda). ATP6V1A-positive clear cells were detected at PD28 in caput, corpus, and cauda but not in the initial segment, while narrow cells were observed in all segments beginning at PD14. These contrasted with the rat, in which clear cells appeared at PD14. Notably, scRNA-seq and immunostaining identified a previously unrecognized KRT5⁺/ATP6V1A⁺ narrow-like basal cell subpopulation, enriched in lysosomes and mainly localized to the caput and corpus. These findings revealed an earlier epithelial maturation in mice compared to rats, underscoring the importance of choosing animal models. The newly identified basal subpopulation may contribute to luminal acidification and homeostasis, offering new insights into epididymal physiology and reproductive biology.
Nuclear pore complexes (NPCs), massive assemblies of approximately 30 distinct nucleoporins (NUPs), serve as the major gateways for nucleocytoplasmic transport. Recent research highlights that NUP aberrations-ranging from gene fusions and mutations to pathological protein accumulation-are increasingly implicated in the pathogenesis of a broad spectrum of human diseases. The underlying pathogenic mechanisms are highly multifactorial, encompassing the structural collapse of the NPC, transport network paralysis, epigenetic hijacking via liquid-liquid phase separation, and off-pore transcriptional dysregulation. This review synthesizes the multifaceted molecular etiology of NUP-associated pathologies. A deeper understanding of these diverse functions will shed light on the broader biological roles of NUPs and guide future research into targeted and personalized therapeutic strategies for NUP-associated disorders.
For decades, apoptosis has reigned supreme in cell death therapeutics, yet its clinical limitations in resistant cancers and degenerative diseases have unveiled the critical role of non-apoptotic regulated cell death (RCD) pathways, including ferroptosis, necroptosis, pyroptosis, and parthanatos. This review examines chromatin as a key regulatory layer influencing these pathways through dynamic histone modifications, DNA methylation, non-coding RNAs, and 3D genome architecture. We dissect how chromatin landscapes integrate metabolic, oxidative, and inflammatory signals in a cell-type- and lineage-dependent manner to steer cell fate, thereby enabling context-specific RCD activation or suppression. Emerging evidence suggests that epigenetic dysregulation can silence tumor-suppressive cell-death regulators such as GSDME and RIPK3 in some cancers and may contribute to neuronal susceptibility to parthanatos in specific neurodegenerative models. Therapeutically, the reversibility of epigenetic marks makes HDAC/DNMT inhibitors, BET-targeting agents, and CRISPR/dCas9-based editing attractive candidates for re-sensitizing selected preclinical models to RCD inducers; however, their clinical value will depend on improving tissue selectivity, minimizing toxicity, and demonstrating durable efficacy in heterogeneous patient tumors. Nanotechnology may improve delivery, but it does not fully overcome systemic exposure or targeting barriers. Emerging frontiers (single-cell epigenomics, phase-separated biomolecular condensates, and mitochondrial-nuclear crosstalk) may help identify candidate biomarkers and vulnerabilities, but these remain incompletely validated. By shifting from a genetic to a chromatin-centric paradigm and explicitly accounting for cell-type-specific chromatin states, this review highlights a promising framework for overcoming cell-death resistance, while recognizing that most pathway links, biomarkers, and delivery strategies still require robust validation in vivo and across patient cohorts before broad clinical translation.
Mutations in GATA1 that cause skipping of exon 2, which encodes the N terminus, are associated with the myeloid leukemia of Down syndrome and Diamond-Blackfan anemia (DBA). To elucidate the molecular function of this N-terminal region, we used single-cell RNA sequencing (scRNA-seq) on fetal liver cells from Gata1-mutant embryos that express only the short isoform of GATA1 (GATA1s) lacking the N terminus of full-length GATA1 (GATA1FL). scRNA-seq revealed defects in erythropoiesis and aberrant upregulation of glycolytic genes, including PKM, which encodes pyruvate kinase to catalyze the final and irreversible step of glycolysis. Using precision nuclear run-on sequencing and cleavage under targets and release using nuclease (CUT&RUN) after acute GATA1 deletion in erythroid cells, we identified PKM as a direct target of GATA1. Substitution of GATA1FL with GATA1s induced histone lactylation at the PKM promoter, increased pyruvate kinase M (PKM) expression and activity, and enhanced glycolytic flux in erythroid pro genitors, without affecting mitochondrial respiration. Importantly, PKM expression is also significantly elevated in patients with DBA with RPS19 mutations, which is associated with reduced levels of GATA1, further supporting a link between GATA1s-driven defective erythropoiesis and dysregulated glycolysis. Together, these findings reveal that GATA1 controls not only heme metabolism but also glycolytic reprogramming.
