Human pluripotent stem cells (hPSCs) present considerable potential for regenerative medicine; however, the standardization and large-scale production of these cells are hindered by an incomplete understanding of the molecular mechanisms governing self-renewal. The long non-coding RNA ESRG is integral to maintaining hPSC self-renewal, with its depletion leading to reduced levels of nucleophosmin 1 (NPM1) protein, thereby compromising the self-renewal capacity of hPSCs. Mechanistically, ESRG physically interacts with NPM1, and a reduction in ESRG/NPM1 expression results in elevated bone morphogenetic protein 4 (BMP4) levels and decreased polypyrimidine tract-binding protein 1 (PTBP1) levels, which in turn destabilize TGF-β1 mRNA and attenuate TGF-β signaling activity. Notably, treatment with the TGF-β agonist SRI-011381 partially rescues the self-renewal defects caused by ESRG or NPM1 knockdown. Collectively, our findings elucidate that the ESRG-NPM1-BMP4-PTBP1 axis governs hPSC self-renewal by post-transcriptionally regulating TGF-β1 mRNA stability and sustaining TGF-β signaling. This study enhances the understanding of the molecular regulatory network underlying hPSC self-renewal maintenance and validates ESRG as a promising target for improving the in vitro expansion of hPSCs.

CRISPR technology has profoundly transformed both genome editing and the next generation of nucleic acid detection. In this study, we characterized a Cas12a ortholog, Gs12-9 (EsoCas12a), which recognizes a broad PAM motif (NYYN, where Y denotes C or T) and maintains robust cis- and trans-cleavage activity across a wide temperature range (16–60 ℃), outperforming established Cas12a variants such as LbCas12a. Leveraging Gs12-9, we established a rapid, highly sensitive, and temperature-tolerant nucleic acid detection platform that enables precise genotyping without stringent thermal control. By integrating recombinase polymerase amplification (RPA) with CRISPR/Gs12-9-mediated detection, we accurately discriminated myostatin (MSTN) knockout pigs from wild-type individuals, achieving a detection limit of 40 copies per reaction with 100% accuracy. While the MSTN knockout (KO) pig served as a proof-of-concept model in this work, the versatility of Gs12-9 supports its broader application in CRISPR-based diagnostics, including pathogen screening and genetic variant detection.

Diffuse large B-cell lymphoma (DLBCL) is an aggressive subtype of non-Hodgkin lymphoma. Fatty acid synthase (FASN) is a key lipogenic enzyme implicated in tumor progression, but its regulation in DLBCL is poorly understood. The function of FASN in DLBCL was explored using database analysis, clinical sample analysis, and cellular phenotyping experiments. Candidate m6A sites on FASN mRNA were predicted using the SRAMP database. Bioinformatics analyses and experimental techniques were conducted to elucidate the role of methyltransferase-like 3 (METTL3) on cell phenotypes and its regulation of FASN. The findings were further confirmed in mice models and clinical sample analyses. FASN was highly expressed in DLBCL and positively correlated with poor prognosis. FASN knockdown inhibited the malignant phenotypes in DLBCL cells by suppressing the PI3K/AKT and MAPK/ERK signaling pathways, as well as promoting endoplasmic reticulum (ER) stress. The analysis of FASN mRNA revealed the presence of m6A modification sites, and a positive correlation was identified between METTL3 and FASN. The impact of METTL3 knockdown on the malignant phenotypes of DLBCL cells was consistent with the effects induced by FASN knockdown, whereas METTL3 overexpression reversed the effects of FASN knockdown. Mechanistically, METTL3 stabilized FASN expression by mediating m6A modification of FASN mRNA, thereby facilitating DLBCL progression. METTL3 stabilizes FASN expression through m6A modification, thereby facilitating the DLBCL progression by activating the PI3K/AKT and MAPK/ERK signaling pathways and inhibiting the ER stress response pathway. The METTL3/FASN axis represents a potential therapeutic target, especially in METTL3/FASN-high DLBCL subgroups.

0 . 1 1 8 6 / s 1 3 5 7 8 -0 2 5 -0 1 5 0 7 -6In this article [1], Fig. 3C, the H&E staining for the WT/ NCD group was incorrect, and we have made the necessary corrections.Although the incorrect image also depicts H&E staining for the WT/NCD group, it originates from a different cohort of animals.This mix-up may have occurred due to the identical ear tags of the mice used in both sets of images.For completeness and transparency, the incorrect and correct versions are displayed below.The original article has been corrected.

Most neural stem cells (NSCs) maintain self-renewal to ensure proper development of central nervous system (CNS) until a specific time and undergo terminal differentiation after neurogenesis. However, the regulation of the NSC maintenance during brain development is not well understood. Our study reveals that casein kinase 2 (CK2) is essential for maintaining NB self-renewal in late larval stages in Drosophila. Deficiency of CK2 results in increased nuclear localization of the differentiation promoting factor Prospero (Pros), and consequently a significant reduction in NB number. Mechanistically, Pros is phosphorylated by CK2 and sequestered in the cytoplasm, which prevents premature differentiation of the NB in late larval stages. While in pupal stages, the developmentally declined expression of CK2 allows NBs to undergo terminal differentiation. Overall, our study uncovers a critical role of CK2 as a time switch on NB transition from self-renewal to differentiation, highlighting a novel mechanism of the temporal regulation of neurogenesis by post-translational modifications (PTMs).

Aims/hypothesisMetabolic dysfunction–associated steatotic liver disease (MASLD) is a leading cause of chronic liver disease globally and is closely associated with type 2 diabetes (T2D) and obesity. Glucagon-like peptide-1 receptor agonists (GLP-1RAs) offer partial therapeutic benefits in MASLD, but their efficacy is often limited in advanced insulin-resistant states, highlighting the need for adjunctive strategies to improve treatment responsiveness.MethodsWe investigated the metabolic and mechanistic effects of combining semaglutide (GLP-1RA) with Akkermansia muciniphila 11 (Akk11) in db/db mice, a model of T2D-associated MASLD. Mice were treated with semaglutide alone or in combination with Akk11. Metabolic parameters, hepatic and adipose tissue morphology, gut microbiota composition, transcriptomic profiles, and inflammatory signaling were evaluated.ResultsGLP-1RA monotherapy elicited only modest improvements in glycemia and hepatic lipid accumulation, with effects waning over time. However, Akk11 supplementation significantly enhanced semaglutide therapeutic effects, leading to reduced subcutaneous and visceral fat, improved liver histology, and decreased serum triglycerides. Mechanistically, combination treatment suppressed fatty acid synthesis, promoted mitochondrial function, and remodeled the gut microbiota. Notably, both GLP-1RA and AKK attenuated intestinal pyroptosis pathways, and the combination further suppressed pro-inflammatory markers in the liver and intestine.Conclusions/interpretationOur findings highlight a synergistic interaction between GLP-1RA and Akk11, likely mediated through microbiota remodeling and parallel improvements in lipid metabolism across intestinal, hepatic, and adipose compartments. This study supports the development of microbiota-guided strategies to enhance therapies for MASLD and other metabolic diseases.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13578-025-01525-4.

BackgroundThe aggregation of α-Synuclein (αS) into amyloid fibrils and their deposition in intraneuronal Lewy bodies are hallmark features of Parkinson's disease (PD) and other synucleinopathies. Among the molecular players implicated in αS toxicity, the cellular prion protein (PrPC) has emerged as a potential modulator of αS-neuron interactions.ResultsUsing confocal microscopy, colocalization analysis and both siRNA-induced PrPC silencing and antibody-based blockade, we investigated the contribution of PrPC to αS-induced neurotoxicity in human iPSC-derived dopaminergic neurons, primary rat cortical neurons and human SH-SY5Y neuroblastoma cells. We show that PrPC facilitated the early recruitment of αS prefibrillar type B* oligomers (OB*) and short fibrils (SF) to neuronal membranes, enhancing αS-induced Ca2+ influx and membrane permeabilization. However, PrPC levels remained unchanged following prolonged exposure with OB* and SF, suggesting no feedback modulation of PrPC expression. While PrPC blockade partially inhibited the release of toxic soluble oligomers from αS fibrils, downstream cell death was only marginally reduced, indicating a limited contribution of PrPC to the final neurotoxic outcome. By contrast, extracellular Ca2+ emerged as a major driver of αS toxicity, directly promoting the membrane recruitment, internalization and cytotoxic effects of αS aggregates.ConclusionsCollectively, our findings indicate that while PrPC facilitates early events in αS aggregate interaction with neurons, the sustained neurotoxicity induced by αS prefibrillar oligomers and fibrils is predominantly mediated by extracellular Ca2+. This promotes aggregate-membrane interactions, membrane permeabilization, and intracellular Ca2+ dyshomeostasis, thereby establishing a vicious cycle of neuronal dysfunction and death.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13578-025-01479-7.

Embryonic stem cells (ESCs) possess remarkable pluripotency and self-renewal capacity, making them a cornerstone of regenerative medicine. A defining yet underappreciated feature of ESCs is their attenuated innate immune response, characterized by suppressed expression of pattern recognition receptors, cytokines, and interferons. While this immune hyporesponsiveness helps preserve pluripotency and rapid proliferation during early embryogenesis, it also presents both challenges and opportunities for clinical application. This review summarizes recent findings on the molecular mechanisms underlying the innate immune deficiency of ESCs, including underdeveloped pathogen–sensing machinery and deficiencies in downstream signaling. We further explore the biological significance of this immune state in embryonic development and its implications for stem cell-based therapies. Specifically, we discuss how immune silence can mitigate inflammation during transplantation and be harnessed to improve mRNA-based differentiation and cancer vaccine strategies. Lastly, we highlight the challenges in balancing immune activation with stemness and outline future directions for optimizing ESCs-based therapeutics. A better understanding of ESC immunobiology will enable safer and more effective applications in regenerative medicine, oncology, and immunotherapy.

BackgroundIncreased pulmonary blood flow (IncPBF), one of the most important features of many children with congenital heart diseases, is well-known as a prerequisite for the induction of pulmonary arterial hypertension. However, due to the lack of neonatal mouse models of IncPBF, it remains largely unknown how IncPBF affects postnatal lung development.Methods and resultsA neonatal mouse model of IncPBF was created via abdominal aorta and inferior vena cava fistula microsurgery at postnatal day 7 (P7) and verified by abdominal ultrasound and cardiac ultrasound. Hematoxylin–eosin staining demonstrated that at P14, the number of alveoli was significantly reduced in the IncPBF group compared with the sham group. Immunostaining further confirmed the results, showing that the markers of alveoli type 1 (AT1), alveoli type 2 (AT2), and endothelial cells were significantly reduced in the IncPBF group compared with the sham group. Moreover, RNA-sequencing analysis demonstrated a substantial difference of gene expression profile between IncPBF and sham lungs, and many gene ontology terms or reactome enrichment that are associated with normal alveolar development and pulmonary function, such as angiogenesis, cell migration, and lipid metabolism, were downregulated. Mechanistically, suppression of Mfap5-positive myofibroblasts or Shh-Gli1 signaling could ameliorate IncPBF-induced alveolar hypoplasia.ConclusionsIncPBF led to alveolar dysplasia during the early developmental stage, and a neonatal mouse model of IncPBF was successfully created. This study introduced a platform for understanding IncPBF-associated pediatric diseases.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13578-025-01502-x.