Frontiers in Cell and Developmental Biology最新文献

Backgrounds: Protein crotonylation is a novel post-translational modification implicated in tumorigenesis and progression. Its putative roles and mechanisms in lung adenocarcinoma (LUAD), however, remain incompletely elucidated.

Methods: We analyzed the expression of crotonylation-related genes (CRGs) in LUAD samples and identified differentially expressed genes for gene set variation analysis (GSVA). Using GSVA scores as phenotypic traits, weighted gene co-expression network analysis (WGCNA) was applied to identify key module genes. A putative prognostic model was subsequently constructed via Lasso-Cox regression. Functional enrichment, gene mutation analysis, and immune infiltration analyses were conducted to compare high- and low-risk groups. Furthermore, cellular experiments were performed to validate the putative role of the hub gene FAM83A and its regulation of key glycolytic enzymes PKM2 and LDHA.

Results: We established a putative crotonylation-related prognostic model for LUAD, which effectively stratified patients into high- and low-risk groups with significantly different overall survival. Functional analysis suggested putative disparities in metabolic pathways between the two groups. Mutation landscape analysis revealed distinct genomic variation patterns, while immune infiltration assessment indicated a putative immune-evasion phenotype in high-risk patients. Cellular assays demonstrated that FAM83A enhances lung cancer cell proliferation, putatively through promoting glycolysis. Our findings establish that FAM83A integrates histone H3K27 crotonylation signaling to drive transcriptional reprogramming of glycolytic metabolism, specifically upregulating key enzymes PKM2 and LDHA.

Conclusion: This study proposes a putative prognostic model based on CRGs for LUAD outcome prediction. The hub gene FAM83A may facilitate lung cancer cell growth by regulating glycolysis via PKM2 and LDHA, offering a novel theoretical foundation and a potential target for prognostic assessment and targeted therapy in LUAD patient.

Background: Exosome-hydrogel complexes have great potential in regenerative medicine, being able to combine biological signals with structural support. But overall, the knowledge structure and translational connections between academic discoveries and patent deployment are not clear.

Methods: A dual-source analysis framework was established to analyze academic papers and patents, illustrating the landscape of exosome-hydrogel research from 2016 to 2025. An interdisciplinary knowledge graph was constructed using topic modeling, entity-relation extraction, and evidence-ranking methods to quantify temporal trends, thematic differences, and translational gaps.

Results: The core components include mesenchymal stem cell-derived exosomes and hydrogels based on gelatin methacrylate (GelMA) or collagen, which form a well-established research foundation. Academic research focuses on osteogenesis, and recent progress mentions angiogenesis and immune regulation. The research application has strong temperature dependence, and patent activities lag behind academic publications. Several high-evidence yet unpatented propositions, such as "hydrogel-encapsulated exosomes" and "exosome-enhanced angiogenesis," represent potential innovation opportunities.

Conclusion: This study employs a data-driven framework to connect scientific research with transformation. The integration of semantic models and cross - source evidence reflects the evaluation logic of exosome - hydrogel research, and provides support for future research in the field of regenerative biomaterials and the priority of patent strategies.

In mammals, the fusion of sperm and oocyte gives rise to a totipotent zygote, which undergoes a series of cleavage divisions and differentiation events. During this process, the embryo transitions from totipotency to pluripotency, accompanied by extensive epigenetic reprogramming. With continuous innovation of low-input multi-omics technology and other methods, the relationship between epigenetic remodeling and embryonic development has been gradually revealed. This review synthesizes recent advances in our understanding of the dynamics of epigenetic reprogramming during early embryogenesis in mice and humans. It covers the remodeling of DNA methylation, histone modifications, chromatin accessibility, and three-dimensional chromatin architecture, with a particular focus on the dynamic features of histone modifications. The patterns of common histone modifications such as methylation, acetylation, and ubiquitination are elaborated. Furthermore, the review outlines both the emerging roles of metabolism-associated modifications such as crotonylation and lactylation in genomic targeting and transcriptional regulation, and the dynamic patterns of histone variant incorporation.

Oral squamous cell carcinoma is an aggressive malignancy driven by aberrant signaling pathways, with Wnt signaling acting as a central regulator of proliferation, tumor-microenvironment interactions, and oncogenic growth. Here, we focus on Frizzled-4, a Wnt receptor with context-dependent functions in epithelial cancers. We show that pharmacological inhibition of Frizzled-4 delays cell cycle progression, and reduces proliferative capacity. These effects are accompanied by suppressed metabolism of retinoic acid, suggesting that Frizzled-4 inhibition stabilizes intracellular retinoic acid levels, which ultimately contributes to the observed cell cycle slowdown. Blocking retinoic acid signaling reverses the cell cycle effects of Frizzled-4 inhibition, confirming that retinoic acid-dependent regulation operates downstream of Frizzled-4. By restraining uncontrolled proliferation and attenuating oncogenic signaling, Frizzled-4 emerges as a key molecular node in oral squamous cell carcinoma, highlighting its potential as a therapeutic target to counteract Wnt-driven tumor growth.

Background: Lymph node metastasis is a critical event in the progression of EGFR wildtype lung adenocarcinoma (LUAD), a subtype lacking effective targeted therapies and associated with poor prognosis. The specific myeloid cell subsets and molecular mechanisms driving LNM in this context remain poorly understood.

Methods: To elucidate the cellular drivers of LNM, we performed an integrated analysis of multi-cohort single-cell RNA sequencing data from EGFR-wildtype LUAD patients and bulk transcriptomic data from The Cancer Genome Atlas (TCGA). Pathological lymph node status from the bulk RNA-seq cohort was mapped to the single-cell transcriptomes using the UCell algorithm. Myeloid cell heterogeneity was analyzed via sub-clustering, and the transcription factor regulon activity was inferred using pySCENIC. The functional role of the key regulator RELB was validated using siRNA-mediated knockdown in neutrophil-tumor cell co-culture systems, assessed by qPCR, proliferation, clonogenic, migration, and invasion assays. A RELB signature score was constructed based on its target genes and validated in independent cohorts for prognostic, immunotherapeutic, and drug sensitivity assessment.

Results: Myeloid cells were significantly enriched in the LNM group. The process of sub-clustering identified a pro-metastatic subset of ELANE-positive neutrophils that was specifically expanded in LNM samples, linked to advanced N stage, higher clinical stage, and shorter overall survival. RELB was identified as a central regulator of ELANE + Neu through transcription factor analysis, showing significantly heightened regulon activity in LNM. Using a neutrophil-tumor cell co-culture system, we found that RELB knockdown in neutrophils attenuated inflammatory signaling in tumor cells and subsequently reduced their proliferative, clonogenic, migratory, and invasive capacities. A high score of the RELB signature was a strong predictor of poor prognosis and was associated with pro-tumorigenic pathways, and the creation of an immunosuppressive microenvironment. Additionally, RELB scores were associated with lower tumor mutational burden, poorer response to immunotherapy, different drug sensitivity patterns, and increased expression of several antibody-drug conjugate targets.

Conclusion: The study highlights a pro-metastatic ELANE + neutrophil subpopulation, with RELB acting as its primary transcriptional regulator. The RELB signature may serve as a biomarker for prognosis and treatment response prediction, indicating a potential target for precision treatment in EGFR wildtype LUAD.

Retinal ganglion cells (RGCs) play a pivotal part transmitting visual data to the brain. Yet, damaged RGCs are unable to maintain and regrow axons and connectivity, as in the common blinding disease glaucoma. Thus, the idea of rescuing and replacing damaged RGCs holds immense therapeutic potential. In recent years pluripotent stem cells cultured in both 2D and 3D (retinal organoid) environments have generated RGCs from healthy- and patient-derived cells. These models can be used to study normal retinal physiology and compare it to the diseased retina. Although the effects of glaucomatous injuries on RGCs have been well-studied in animal models, much less is known about similar mechanisms in the human retina. Further, using in vitro-derived RGCs as a tool for cell characterization and replacement is still in its infancy. In particular, many distinct RGC subtypes have been described, and it remains unclear how well this diversity is reflected in the various differentiation protocols, or their functional roles in human health and disease. In this review we summarize the currently described subtypes of human RGCs and their markers and discuss recent evidence for subtype-specific vulnerabilities to injury and disease. Finally, we synthesize the limited evidence for subtype differentiation in human stem cell culture approaches. Increased understanding of this human RGC diversity will provide new tools to enrich for selective subtypes and ultimately fill key translational gaps in human glaucoma research.

Introduction: Precise and longitudinal monitoring of Neural Stem Cell (NSC) differentiation is pivotal for advancing regenerative medicine. However, traditional identification methods rely on invasive immunochemical staining, which terminates cell viability and precludes real-time analysis.

Methods: To address these limitations, we propose CG-RecNet, a specialized deep learning framework for accurate, label-free classification of NSC differentiation lineages-specifically neurons, astrocytes, and oligodendrocytes-directly from brightfield imaging flow cytometry (IFC) data. The architecture integrates a LinAngular Cross-Channel Attention (LinAngular-XCA) Fusion Module to capture global morphological dependencies and a Gated Convolutional Neural Network (GatedCNN) Block to suppress background noise.

Results: Validation on rat embryonic NSCs indicates that CG-RecNet achieves an overall accuracy of 96.40% and a macro-average AUC of 0.9979, representing a 1.82% improvement over established baselines. Notably, the model achieved high precision in identifying the minority oligodendrocyte lineage without synthetic oversampling.

Discussion: Grad-CAM analysis indicates that the model's attention aligns with biologically relevant hallmarks, such as neurite outgrowth and soma texture. CG-RecNet provides a reliable, non-invasive, and qualitatively interpretable tool for neural stem cell research.

Introduction: Conventional T-flasks (T-25) filled to capacity are frequently employed to minimize shear stress arising from fluid motion during ground-based microgravity simulations using a clinostat or random positioning machine (RPM). However, this approach can introduce confounding factors, such as hypoxia and CO₂ accumulation that affect cell metabolism and function. Therefore, in vitro platform simulating microgravity is crucial to distinguish true gravity-dependent responses from culture artifacts. Here, we proposed an innovative engineered culture system (F-25) with a growth area of 25 cm² primarily designed for full-filled and clinostat experiments.

Methods: We assessed the effects of static and rotational (i.e., microgravity) full-filled cultures including conventional T-25 and eighteen customized F-25 with different medium depths, gas exchange areas and membrane types on mitochondrial function of intact C₂C₁₂ myoblasts by high resolution respirometry.

Results: After 24 h, conventional T-25 flasks, full-filled to height of H₀ (2.25 cm) and with a hydrophobic-type gas exchange area of A₀ (0.2 cm²) showed intact cellular respiration (ICR) and maximal uncoupled respiration (ET) rates that were more than twice those measured in partially-filled controls, whose values (40 ± 3 for ICR and 60 ± 4 for ET pmol O₂ s^-1 10^-6 cells^-1) remained unchanged between time zero and 24 h. At each medium depth (¹/₃H₀, ²/₃H₀, and H₀) increasing the gas exchange area from (6A₀, 12A₀, and 18A₀) led to a progressive decrease in ICR and ET rates reaching control values. The best optimized F-25 flask configuration, combining reduced medium depth (¹/₃H₀) with an enhanced hydrophilic gas exchange membrane of 18A₀, maintained ICR and ET rates similar to partially-filled controls. The F-25 flask was further tested to assess mitochondrial function under simulated Mars, Moon, and space gravity conditions following 24 h of exposure. Under different extraterrestrial gravity conditions, ICR and ET rates were again twice than those of partially-filled controls but remained unchanged in optimized F-25 flask.

Discussion: The latter one provides a reproducible and relevant baseline, avoiding confounding factors related to O₂ delivery for clinostat-based simulations. The F-25 flask setup, which allows controlled oxygenation and minimized hydrostatic artifacts, offers a versatile platform not only for space biology, but also for hypoxia studies, 3D culture systems, and tissue engineering applications requiring a defined O₂ microenvironment.

[This corrects the article DOI: 10.3389/fcell.2025.1755318.].