Glioblastoma is an aggressive form of brain cancer and poses a challenge in treatment due to its profound heterogeneity and capacity for extensive infiltration into the brain parenchyma. Research has shown glioblastomas near the ependyma have poorer survival rates. Therefore, our aim was to identify distinct molecular features of glioblastoma invading the ependyma of lateral ventricles and neural stem cell region and the survival prognosis of these patients. A retrospective review of 170 patients with a new histologically confirmed diagnosis of glioblastoma between 2018 and 2019. Patients were excluded if they were less than 18-years-old, did not have a histological diagnosis, or had missing data. Overall survival (OS) data was analysed. Statistical analysis included Kaplan-Meier survival curves, log rank tests and Cox regression. A total of 170 patients were included (mean age 61 ± 11.3 years; 54% male). Tumours contacted the ependyma in 69 patients and did not in 101. The most common tumour locations were temporal (31%), frontal (29%), and parietal (21%) lobes. Preoperatively, 65% had a performance status of 0-1. Biopsy alone was performed in 19%, subtotal resection (STR) in 48%, and gross total resection (GTR) in 32%; GTR was more common in non-ependyma contacting tumours (40% vs. 20%). MGMT promoter was unmethylated in 64% of patients. Mean overall survival was significantly lower in patients with ependymal contact compared with non-contacting tumours (11.9 vs. 17.4 months, p = 0.004). On multivariable analysis, ependymal contact remained independently associated with poorer survival. No significant association was found between MGMT status and ependymal contact or tumour epicentre distance. Overall, our study reinforces the prognostic relevance of glioblastoma contact with the ependymal and subventricular zones. Tumours involving these regions were associated with poorer overall survival.

Long-term preservation of genetic lines is crucial for animal research. Current methods heavily rely on cryopreservation of haploid sperm, whereas diploid germline stem cells (GSCs) may provide a superior alternative. Here, using zebrafish, we establish an integrated platform combining in vivo expansion of GSCs (eGSCs), cryopreservation and direct genetic recovery. The eGSCs are amplified in cyp11a2-/- mutants and exhibit a total number of about 300-fold higher than conventional GSCs after cryopreservation and thawing. The eGSCs achieve a success rate of 82% after transplantation, compared with less than 1% for conventional GSCs. Furthermore, we develop the GSC-deficient nanos2-/- mutant as an ideal host for eGSC transplantation, enabling efficient production of both sperm and oocytes within a single maturation period. This allows rapid recovery of maternal-zygotic mutants directly in the F1 generation. Overall, our strategy provides a robust platform for preserving and restoring zebrafish homozygous mutants, opening new opportunities for zebrafish research.

Genome editing has progressed from a laboratory capability for targeted DNA manipulation to a clinically relevant strategy for correcting, silencing, or regulating genes implicated in human disease. In this Review, we synthesize the mechanisms, capabilities, and constraints of the principal programmable platforms-zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR-Cas systems-and highlight how base editors, prime editors, and epigenetic editors expand the range of achievable outcomes beyond double-strand break-dependent repair to precise nucleotide substitutions, small insertions/deletions, and transcriptional modulation. We compare genome-editing cargo formats, including plasmid DNA, viral-vector DNA, mRNA, guide RNA, and ribonucleoprotein complexes, together with the delivery modalities used to transport them, including AAV, adenoviral and herpesviral vectors, lipid nanoparticles (LNPs), electroporation, and virus-like particles. We then consolidate key biomedical applications enabled by these technologies, spanning endogenous gene tagging, high-throughput functional variant screening, molecular recording, and the generation of genetically faithful disease models. Across oncology, respiratory, hematologic, cardiovascular, metabolic, neurodegenerative, viral, ocular, and immune disorders, genome editing is advancing both ex vivo and in vivo interventions, including engineered cellular immunotherapies, hematopoietic stem and progenitor cell editing for hemoglobinopathies, and emerging liver-directed programs for lipid and coagulation targets. Finally, we discuss priorities for broad clinical implementation: improving editing fidelity and PAM flexibility, increasing performance in non-dividing cells, enabling tissue-selective delivery to difficult organs (for example, lung and central nervous system), and addressing manufacturing scalability, long-term monitoring, and equitable global access.

The epithelial-to-mesenchymal transition (EMT) is a widely studied cell state change, yet differences in model design and measurement approaches limit comparison across studies. Addressing this challenge requires experimental model systems and analysis frameworks that support standardization across contexts. Here, we show that human induced pluripotent stem (hiPS) cells in defined cell culture geometries, two-dimensional colonies and three-dimensional lumenoids, enable multimodal measurements of EMT dynamics within a single experimental platform. Using fixed-cell and live-cell image-based assays, we quantify changes in cell migration, EMT-related molecular markers, cell-cell junction organization and interactions with the basement membrane, a specialized form of the extracellular matrix, during EMT induced in hiPS cells. We identify cell culture geometry-dependent differences in the timing of migration onset and show that basement membrane integrity can be quantitatively linked to these differences. Together, these results establish an imaging-based framework for analysis of cell state transitions and provide accessible datasets and tools.

Adipose-derived mesenchymal stem cells (ADMSCs) possess modulatory functions in adipose tissue, which could help combat the development of obesity. It has been reported that the functions of ADMSCs are impaired by obese microenvironment. However, the mechanisms are not clearly understood. The study investigates the role of leptin on immunoinhibitory functions of ADMSCs in obesity, and primarily explored its possible mechanisms. ADMSCs were isolated and identified by morphological observation, flow cytometry and differentiation potential. The lipopolysaccharides (LPS)-stimulated macrophages were co-cultured with ADMSCs pretreated with or without leptin. The expression of macrophage-associated markers was detected by flow cytometry. The secretion of inflammatory cytokines was evaluated by ELISA. Autophagy and MAPK signaling related proteins were detected by western blotting. Furthermore, the expression of tumor necrosis factor-alpha-stimulated protein 6 (TSG-6) was detected by western blotting, qRT-PCR, and ELISA. ADMSCs belonged to CD73+ CD90+ CD105+/ CD14- CD34- CD45- HLA-DR- cells, and capable differentiation to adipogenic, osteogenic and chondrogenic cells. The immunosuppressive effect of ADMSCs on macrophage activation were restrained by leptin mediated the expression of TSG-6. Inhibition of leptin induced autophagy by Atg5 knockdown increased the expression of TSG-6, and promoted ADMSCs immunosuppression for macrophage activation. Furthermore, leptin induced autophagy and decreased the expression of TSG-6 in ADMSCs was dependent on the activation of p38 MAPK pathway. In conclusion, our study confirmed that leptin-induced autophagy regulated the immunomodulatory potential of ADMSCs through downregulating the expression of TSG-6 via the activation of p38 pathway. These findings reveal a novel mechanism and provide a potential molecular target for understanding the immunomodulatory dysfunction of ADMSCs in obesity.

Relapsed/refractory acute myeloid leukemia (AML) in children remains challenging due to leukemia stem cells (LSCs) and poor immunogenicity. This study aimed to construct a novel immunoconjugate combining BCG heat shock protein 70 (HSP70) with an anti-CD123 monoclonal antibody (MAb) and evaluate its targeted anti-leukemic efficacy and mechanisms.

Microglia are the brain's resident immune cells, essential for homeostasis and implicated in common neurodegenerative diseases like Alzheimer's and Parkinson's disease (PD), where their early activation and sustained inflammatory mediator release contribute to neuronal loss. However, their role in rare disorders is unclear. β-propeller protein-associated neurodegeneration (BPAN), caused by WDR45 mutations, shares key features with PD, including iron accumulation and dopaminergic neuron loss, but the impact of microglia and mutant WDR45 in BPAN pathophysiology remains unexplored. To address this, we established the first induced pluripotent stem stell (iPSC)-derived microglia model from BPAN patients. Parallel targeted transcriptomic and secretomic profiling revealed a shift from a homeostatic microglial toward a stress-adapted and transcriptionally reprogrammed state characterized by selective remodeling of immune signaling pathways and dysregulation of autophagy and cellular stress responses. Complementary secretomic analysis identified reduced secretion of lysosomal enzymes alongside increased shedding of immune-associated surface proteins, indicating altered lysosomal trafficking and remodeling of microglial immune signaling. These findings identify a distinct microglial phenotype in BPAN and implicate microglial dysfunction as a potential contributor to disease mechanisms, highlighting new avenues for therapeutic strategies targeting neuroimmune pathways.

HIV infection is accompanied by chronic inflammation-related co-morbidities, even when viral replication is suppressed by therapy. This persistent inflammatory state suggests that long-lived immune cell lineages may acquire stable pro-inflammatory programming. Here, we investigate whether inflammatory programming can be imprinted within hematopoietic lineages, following the exposure of mice and bone marrow-derived macrophages (BMDMs) to extracellular vesicles (EVs) carrying Nef, a key inflammatory factor of HIV. Multi-omics profiling shows that hematopoietic cells exposed to Nef-EVs undergo epigenetic remodeling and reprogramming of energy and lipid metabolism characteristic of trained innate immunity. The inflammatory phenotype in BMDMs is partially reversed by inhibition of glycolysis, a key metabolic driver of trained immunity. We demonstrate that following competitive bone marrow transplantation, hematopoiesis in mice receiving bone marrow from Nef-EV-treated donors displays a sustained bias toward myelopoiesis, and BMDMs retain enhanced inflammatory potential. These findings demonstrate that Nef-EVs can imprint a lasting inflammatory memory, mechanistically similar to trained immunity, in hematopoietic cells. This memory persists beyond the initial exposure and may contribute to chronic inflammation in people with HIV.

Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease with limited therapeutic options. Stem cell-based tissue engineering, particularly utilizing human periodontal ligament stem cells (hPDLSCs), represents a promising approach for cartilage regeneration. However, we have previously demonstrated that chronic inflammation and hypoxic stress in the TMJOA microenvironment markedly accelerate cellular senescence in hPDLSCs, severely impairing their regenerative potential. Here, we identify the YTHDC1-m⁶A-GADD45B axis as a critical regulator of senescence and chondrogenic differentiation in hPDLSCs. We show that YTHDC1, an m⁶A reader protein, is downregulated under inflammatory and senescent conditions. Functional studies reveal that YTHDC1 overexpression attenuates senescence and enhances chondrogenesis, whereas its knockdown exacerbates senescence and suppresses differentiation. Mechanistically, YTHDC1 recognizes m⁶A modifications on GADD45B mRNA and promotes its decay, leading to inhibition of the p53/p21 signaling pathway. Mutation of the m⁶A site in GADD45B abolishes the regulatory effects of YTHDC1. In rats with TMJOA, transplantation of YTHDC1-overexpressing hPDLSCs ameliorated disease phenotypes, an effect reversed by co-expression of wild-type GADD45B. Our findings reveal a novel epitranscriptomic mechanism that regulates hPDLSCs senescence and subsequently affects chondrogenic differentiation, and highlight the therapeutic potential of targeting the YTHDC1-GADD45B-p53/p21 axis.

In pulpitis treatment, vital pulp therapy (VPT) aims to preserve pulp vitality and promote dentin regeneration. However, conventional pulp-capping materials usually lack reparative dentin-inducing activity and fail to regulate the local immune microenvironment effectively. In this work, valproic acid (VPA)-loaded zeolitic imidazolate framework-8 (ZIF-8) nanocomposites (VPA@ ZIF-8 NPs) were designed to achieve high VPA loading capacity, excellent encapsulation efficiency, and sustained and pH-responsive VPA release kinetics. Using a VPA-equivalent (VPA-eq) dosing strategy, we systematically evaluated its biocompatibility, antimicrobial properties, immunomodulatory effects, and dentin-regenerative potential across multiple methods. In vitro studies showed that when treated with VPA@ZIF-8 NPs, stem cells from human exfoliated deciduous teeth (SHEDs) exhibited enhanced proliferation, migration, and mineralization ability, and oral opportunistic pathogens Veillonella parvula and Streptococcus mutans showed suppressed growth and biofilm formation activities. Further, macrophage polarization assays revealed that VPA@ZIF-8 NPs promoted the M2-like polarization in THP-1 cells, indicating an anti-inflammatory function. Notably, the BMP2/SMAD1/RUNX2 pathway was activated by VPA@ZIF-8 NPs in the SHEDs. In vivo, local VPA@ZIF-8 NP application recapitulated the macrophage M2-like polarization and BMP2/SMAD1/RUNX2 pathway activation phenotype and contributed to marked reparative dentin bridge formation in a pulp exposure rat model. In summary, the multifunctional VPA@ZIF-8 NPs integrate biosafety, antibacterial, immunomodulatory, and dentin-regenerative-inducing capabilities, providing a promising and integrated strategy for vital pulp therapy against pulpitis.

Suppressor of cytokine signaling 2 (SOCS2) is a classical feedback inhibitor of JAK-STAT signaling, but its role in porcine adipose-derived mesenchymal stem cells (PADSCs) and fat deposition remains unclear. Here, we investigated the function and mechanism of SOCS2 in PADSC proliferation and adipogenic differentiation. SOCS2 overexpression reduced colony number, slowed CCK-8 growth curves, and induced G0/G1 arrest, whereas SOCS2 knockdown enhanced proliferation and promoted the G1/S transition. In parallel, SOCS2 overexpression decreased lipid droplet accumulation and downregulated adipogenic markers, while SOCS2 silencing produced the opposite effects, indicating that SOCS2 negatively regulates both PADSC expansion and adipocyte formation. Transcriptomic analysis showed that SOCS2 mainly influenced pathways related to extracellular matrix organization, cell cycle, and cytokine/inflammatory signaling. At the protein level, SOCS2 silencing increased total JAK2 and STAT3, whereas SOCS2 overexpression reduced JAK2 abundance and STAT3 activation, accompanied by concordant changes in Cyclin D1, CDK2, and PCNA. Collectively, these results suggest that SOCS2 exerts dual negative effects on PADSC proliferation and adipogenic differentiation, accompanied by changes in JAK2-STAT3 signaling. These findings provide functional evidence supporting SOCS2 as a candidate gene related to porcine fat deposition and provide a basis for further genetic validation in pig breeding populations.

Pulmonary fibrosis, particularly idiopathic pulmonary fibrosis, is a progressive and fatal interstitial lung disease with limited curative therapies. This narrative review provides a concise overview of emerging molecular targets and novel therapeutic strategies for pulmonary fibrosis, including mesenchymal stem cell therapy and nanomaterial-based inhalation systems, focusing on their mechanisms, preclinical evidence, and clinical potential. We highlight critical challenges in translating preclinical discoveries into clinical benefit, such as animal model limitations and patient heterogeneity, and outline future research priorities. Notably, this review is not a systematic review; it does not include formal meta-analysis or a comprehensive literature search following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, which may have introduced selection bias. Thus, findings should be interpreted with appropriate caution.

Endochondral ossification is essential for the development of appendicular bones, physiological bone remodelling and fracture healing. Recent studies have identified mesenchymal stromal cell-derived FABP5+ septoclasts (SCs) as key mediators for the growth and repair of long bones, particularly in cartilage matrix degradation and growth plate remodelling via the secretion of matrix metalloproteinases. Our previous study has shown that soluble epoxide hydrolase (sEH) inhibitor, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU), promotes long bone growth and bone repair by enhancing H-type vessel-coupled osteogenesis. However, whether TPPU treatment regulates SC activity, thereby promoting long-bone growth and fracture healing, remains unclear. Here, our in vitro and in vivo results showed that TPPU treatment promoted long-bone growth in newborn mice and regulated the hypertrophic layer in the growth plate, with a reduced ratio of hypertrophic cartilage (HC) to proliferative cartilage (PC) width. Furthermore, TPPU treatment enhanced SC activity, as evidenced by elevated expression of MMP9 and FABP5 in the metaphysis near the growth plate. Simultaneously, TPPU induced FABP5+ SC-like cells to degrade chondrocytes in co-cultured human umbilical vein endothelial cells (HUVECs) and human dental pulp stem cells (hDPSCs). Mechanistically, TPPU enhanced the crosstalk of co-cultured HUVECs and hDPSCs to activate the NOTCH signalling pathway in hDPSCs by upregulating HIF-1α expression in HUVECs. Furthermore, TPPU enhanced fracture healing by inducing more FABP5+ SCs and MMP9 secretion at the fracture site. Collectively, these findings highlight sEH as a promising therapeutic target that regulates endochondral ossification through inducing SC activity, offering new opportunities for bone development and repair.

Chemotherapy-induced myelosuppression is a common and severe complication in clinical oncology treatment. Chemotherapeutic agents trigger this condition by damaging bone marrow mesenchymal stem cells (BMSCs) and disrupting hematopoietic microenvironment homeostasis. Danggui Jixueteng Decoction (DJD), a classical blood-tonifying and blood-activating formula, has been confirmed to alleviate post-chemotherapy myelosuppression, but its specific molecular mechanisms in protecting BMSCs remain unclear.

Cellular plasticity is a fundamental driver of tumor heterogeneity, cancer stemness, immune evasion, therapeutic resistance, and disease progression. In malignancies such as breast cancer and glioblastoma, tumor cells undergo reversible phenotypic transitions between proliferative, stem-like, invasive, and drug-tolerant states in response to intrinsic regulatory programs and extrinsic signals from the tumor microenvironment. These adaptive dynamics are governed by complex interactions among signaling pathways, transcriptional networks, chromatin remodeling, DNA methylation, histone modifications, non-coding RNAs, and immune-mediated microenvironmental cues. Such epigenetic instability enables stochastic and therapy-induced shifts between alternative cellular states, thereby contributing to tumor evolution, metastasis, resistance to targeted therapies, and variable responses to immunotherapy. Understanding the mechanisms that govern epigenetic plasticity remains a central challenge in cancer biology. Recent advances in CRISPR/dCas9-based epigenome editing have provided powerful experimental tools for investigating the functional consequences of locus-specific chromatin modifications without altering the underlying DNA sequence. Catalytically inactive Cas9 (dCas9) fused to epigenetic effector domains, including DNMT3A, TET1, KRAB, and p300, enables targeted modulation of gene expression programs implicated in cell-state transitions, lineage specification, and adaptive resistance. These technologies offer a versatile platform for interrogating causal relationships between chromatin states and cellular phenotypes and for modeling mechanisms of tumor adaptation. This review examines the molecular basis of epigenetic plasticity in cancer, evaluates current CRISPR-based epigenome editing strategies, and discusses their application in studying tumor heterogeneity, microenvironment-driven adaptation, immune escape, and therapy resistance. This study highlights emerging opportunities and persistent challenges associated with epigenome editing, including delivery barriers, durability of epigenetic modifications, context-dependent biological responses, and translational limitations. Collectively, these approaches provide valuable experimental frameworks for dissecting the regulatory logic of cancer cell plasticity while informing future therapeutic development.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by alpha-synuclein (α-syn) aggregates termed Lewy bodies. To model PD pathology in vitro, preformed fibrils of α-syn (PFFs), which can be taken up by cells, provide a seed that drives misfolding and aggregation of endogenous α-syn, with new aggregates amplifying this process. External application of PFFs to dopaminergic neurons (DNs) increases aggregate formation, marked by α-syn phosphorylation at serine 129 (pS129-syn), a pathological PD marker. Building on this, we developed an automated synuclein seeding assay to quantify new α-syn aggregates in iPSC-derived DNs. Using pS129-syn as a readout, we show that PFFs elicit a time- and dose-dependent increase in pS129-syn aggregates. Our automated seeding assay further revealed that aggregate formation depends on endogenous α-syn levels. Treatment with PFFs produced a greater increase in pS129-syn aggregates in iPSC DNs derived from a PD patient with a triplication in the SNCA gene, which encodes the α-syn protein and which elevates total α-syn levels, relative to DNs from an isogenic iPSC line from the same individual, in which the SNCA gene mutation had been corrected by CRISPR/Cas9. In contrast, no pS129-syn signal was detected in neurons in which all copies of the SNCA gene had been knocked out (KO). This proof-of-principle automated high-content imaging workflow for synuclein seeding has been validated using isogenic cell lines with defined SNCA copy number variants and it offers a platform for assessing compounds and therapeutics that may impede α-syn aggregate formation.

Cell therapy with adipose-derived mesenchymal stem cells (ASCs) is a promising biological augmentation strategy for rotator cuff repair; however, before clinical translation, regulatory frameworks mandate rigorous preclinical characterization of safety. We report the complete preclinical program evaluating both the safety of an allogeneic ASC-based cell therapy medicinal product (CTMP) and its efficacy in a chronic rotator cuff tear model.

Introduction and Objectives Stem cell therapies have shown potential in treating liver cirrhosis and acute-on-chronic liver failure by promoting liver regeneration. However, their efficacy and safety remain uncertain due to inconsistent results. This systematic review and meta-analysis aimed to evaluate efficacy and safety of stem cell therapies in patients with liver cirrhosis and acute-on-chronic liver failure, compared to standard of care or placebo. Materials and Methods We conducted a systematic review and meta-analysis following PRISMA guidelines. Randomized controlled trials (RCTs) comparing stem cell therapies to standard medical care or placebo for patient important outcomes were included. Risk of bias was assessed using Cochrane RoB 2 tool, and quality of evidence was evaluated with GRADE framework. Results A total of 19 studies were included. Meta-analysis of 15 RCTs (n = 925) provided very low-certainty evidence regarding effects of stem cell therapy on overall mortality. The pooled risk ratio (RR) for mortality was 0.63 (95% CI 0.43 to 0.91; I² = 60%). Ten RCTs (n = 250) assessed effect of stem cell therapy on Model for End-Stage Liver Disease (MELD) scores. The pooled mean difference (MD) was -1.22 (95% CI -2.25 to -0.19; I² = 51%). The certainty in evidence was rated down for risk of bias and imprecision for most outcomes. Conclusions This systematic review and meta-analysis provides very low-certainty evidence regarding efficacy of stem cell therapies for liver cirrhosis and acute-on-chronic liver failure. Further high-quality research is necessary to clarify role of stem cells in treating liver diseases and to ensure safety and efficacy.