CD90 (THY1) is a cell surface glycoprotein that plays a crucial role in the occurrence and development of various malignant tumors. It affects tumor cell proliferation, metastasis, angiogenesis, stem cell characteristics, and energy metabolism. CD90 is an important biomarker for many malignant tumors and is closely related to tumor prognosis. However, the role and specific mechanisms of CD90 in glutamine metabolism in gastric cancer (GC) cells remain incompletely understood. In this study, we report that CD90 affects glutamine metabolism in GC cells through SLC1A5, ultimately promoting GC progression. Mechanistically, CD90 promotes the binding of YY1 to the SLC1A5 promoter, enhances the transcriptional activity of the SLC1A5 promoter, and increases its transcriptional level. This, in turn, affects SLC1A5-mediated glutamine metabolism and ferroptosis in GC cells, ultimately facilitating GC progression. Our results suggest that CD90 plays an important role in regulating glutamine metabolism in GC cells, providing important experimental evidence for elucidating the pathogenesis of GC.

Isocitrate dehydrogenase 1 (IDH1) mutations confer distinct biological properties to gliomas, including the reshaping of the tumor immune microenvironment. While T cell dysfunction in glioblastoma has been extensively characterized, the role of innate lymphoid cells (ILCs)-critical regulators of tissue homeostasis and early immune responses- remains poorly understood.

Alternative lengthening of telomeres (ALT) is a telomere maintenance mechanism deployed in embryonic stem cells and cancer cells. High levels of the heterochromatin mark H3 lysine 9 trimethylation (H3K9me3) at telomeres are critical for ALT, but how this is achieved remains unclear. Telomeric association of orphan nuclear receptors (NRs)-such as COUP-TF1, COUP-TF2, TR2, and TR4-has been shown previously to facilitate ALT activation. Here, we show that orphan NRs regulate telomeric H3K9me3 through TRIM28, a corepressor of ZNF transcription factors, to promote ALT. We report that H3K9me3 is induced by telomeric association of orphan NRs in cultured human fibroblast and ALT cancer cell lines. Moreover, TRIM28 is required for the orphan-NR-induced H3K9 methylation and ALT phenotypes. Importantly, physical interaction of TRIM28 with orphan NRs induces telomeric localization of TRIM28. A TRIM28 variant defective in orphan-NR interaction fails to localize to telomeres and is unable to promote H3K9me3 and ALT phenotypes. These findings indicate that telomeric orphan NRs recruit TRIM28 for telomeric H3K9me3 and ALT activation, emphasizing the role of chromatin structure in ALT activation.

Parkinson's disease (PD), characterized by α-Synuclein aggregation and dopaminergic neuronal loss, has no current cure. Autophagy is critical for α-Synuclein clearance, yet its real-time dynamics remain challenging to assess in human-relevant systems. Here, we used live-cell imaging to assess autophagy within human neuronal cultures and midbrain organoids (hMOs) derived from induced pluripotent stem cells (iPSCs) of PD patients carrying a triplication of the α-Synuclein gene (3xSNCA). Using the LC3-Rosella dual-fluorescent reporter, we quantified autolysosomes dynamics in real time. In 3xSNCA neuronal cultures, we detected early autophagy defects. In 3xSNCA hMOs, reduced autolysosome area, increased total and phosphorylated α-Synuclein (pS129), and decreased electrophysiological activity were observed at 50 days of differentiation (DoD). By 70 DoD, autophagy impairment became more pronounced, overlapping with dopaminergic neuron dysfunction. These findings support the use of human iPSCs-derived models to study autophagy dysfunction in PD and demonstrate a temporal correlation between impaired autophagy, α-Synuclein pathology and neuronal degeneration.

The epigenetic silencing or remarkably diminished expression of STING in cancer cells, along with the structural and functional impairment of the endoplasmic reticulum (ER) and Golgi apparatus, represents a unique mechanism of tumor immune escape and poses an important challenge for STING-targeted therapies. Here, we develop a cell membrane-derived nanodisc system (ND-cGAMP-HP; HP, heparin), which is capable of presenting activated STING proteins in their native form by means of cell membrane-directed display and biological self-assembly techniques. It can directly introduce activated STING protein to tumor cells and circumvent the translocation process between the ER and Golgi apparatus, selectively activating the IFN-I signaling pathway without initiating the inflammation-related pathway NF-κB. ND-cGAMP-HP triggers potent cellular immune responses and remodels the tumor immune microenvironment. Moreover, it augments immune memory by promoting the differentiation of TCF1+ stem cell-like T cells. We thus manifest a strategy based on STING therapy that does not depend on the ER and Golgi apparatus pathways to activate the IFN-I pathway, for cancer immunotherapy.

Chimeric antigen receptor (CAR)-T cell therapy has transformed treatment of relapsed/refractory DLBCL, yet resistance driven by regulatory T cells (Tregs) limits its efficacy. Here we identify Timosaponin AIII (TAIII), a clinical-stage natural product, as an effective modulator of CAR-T function that depletes CAR-Tregs while enhancing effector activity. Mechanistically, TAIII acts as an allosteric A2AR inhibitor by competing with cholesterol, suppressing CREB-dependent FoxP3 transcription and disrupting the A2AR-Treg axis. Ablation of A2AR or Tregs in vitro and in vivo abolishes TAIII activity, confirming specificity. Furthermore, TAIII reduces intratumoral Tregs, increases CD8⁺ T cells infiltration, and potentiates PD-1 blockade in solid tumor models. Importantly, TAIII promotes central memory T-cell formation and enhances CAR-T cytotoxic cytokine secretion. Combining or pretreating CAR-T cells with TAIII markedly improves antitumor efficacy and prevents late relapse across preclinical models. These findings establish TAIII as a combinatorial strategy to deplete CAR-Tregs, enhance CAR-T activity, and extend therapeutic durability.

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold tremendous promise for in vitro modeling to assess native myocardial function and disease mechanisms, as well as testing drug safety and efficacy. However, current hiPSC-CMs are functionally immature, resembling in vivo CMs of fetal or neonatal developmental states. The use of targeted culture media and organoid formats have been identified as potential high-yield contributors to improve CM maturation. This study presents an hiPSC-CM maturation medium formulation, designed using a differential evolutionary approach targeting metabolic functionality for iterative optimization. Relative to existing high-performing reference formulations, our medium significantly matured morphology, Ca2+ handling, electrophysiology, and metabolism, which was further validated by multi-omic screening, for cells in either pure or co-cultured microtissue formats. Together, these findings not only provide a reliable workflow for highly functional hiPSC-CMs for downstream use, but also demonstrate the power of high-dimensional optimization processes in evoking advanced biological function in vitro.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive and often fatal cancer with limited treatment options. Small nucleolar RNA host gene 10 (SNHG10) has emerged as a key regulator in the progression and metastasis of human cancers. However, the potential of SNHG10 in PDAC tumorigenesis, gemcitabine resistance, and the underlying molecular mechanisms remains poorly understood. Our data analysis revealed significant upregulation of the SNHG10 transcript in 179 PDAC cases compared with 171 normal pancreatic specimens, with a positive association with clinical stages of PDAC. Further, we confirmed the significant overexpression of the SNHG10 transcript in several PDAC cell lines compared to normal pancreatic cells. Our results revealed that the downregulation of SNHG10 significantly decreased the cellular proliferation, clonogenic ability, cell migration, and the epithelial to mesenchymal transition, leading to the induction of cell cycle arrest and apoptosis of PDAC cells. Mechanistically, the downregulation of SNHG10 significantly inhibited the expression of vimentin, N-cadherin, survivin, CDK4, CDK6, cyclin B1, cyclin D1, aurora kinase A, and aurora kinase B, with an increased expression of E-cadherin and p21. The bioinformatics analysis, RNA Immunoprecipitation, and qRT-PCR results showed the physical interaction among SNHG10, miR-150-5p, and VEGF-A, which are the integral parts of the ternary complex in PDAC cell lines. Interestingly, silencing of SNHG10 led to the significant induction of miR-150-5p, which repressed the expression of VEGF-A in PDAC cells. Moreover, the miR-150-5p rescued the VEGF-A expression in PDAC cells even during the silencing of SNHG10. Interestingly, the downregulation of SNHG10 enhanced gemcitabine sensitivity in gemcitabine-resistant PDAC cells. The depletion of SNHG10 in the PDAC xenograft model significantly reduced tumor growth, volume, and weight. Importantly, downregulation of SNHG10 suppressed the phosphorylation of EGFR, AKT, ERK1/2, mTOR, and c-MET signaling pathways in both in vitro and xenograft models of PDAC. Our study unveils the oncogenic potential of SNHG10 in tumorigenesis through modulation of the EGFR/AKT/ERK/mTOR, and miR-150-5p/VEGF-A axis, as well as gemcitabine resistance of PDAC as a prospective therapeutic strategy.The graphical abstract represented the potential role of SNHG10 and its regulated molecular mechanisms in the tumorigenesis and gemcitabine resistance in PDAC. Silencing of SNHG10 decreases cell survival, proliferation, clonogenicity, EMT tumor growth through the EGFR/AKT/ERK/mTOR axis, and restores the expression of miR-150-5p, which eventually downregulates VEGF-A. SNHG10 downregulation enhanced the gemcitabine sensitivity in gemcitabine-resistant PDAC cells.

This study aims at implementing a 3D cell culture model of Alzheimer's disease (AD). To that end we engineered human induced pluripotent stem cell (iPSC)-derived neural stem cells to conditionally overexpress FAD mutant APP and PSEN1 variants. After differentiation in 3D basement membrane matrices, cultures exhibited increased Aβ42 and Aβ40 levels and a highly pathogenic shift of the Aβ42/40 ratio. Typical AD phenotypes such as amyloid deposition and tau pathology were observed alongside impaired mitochondrial integrity and neuronal damage. Pathophenotypes were ameliorated by γ-secretase inhibition, confirming amyloid toxicity as main driver of AD pathology. iPSC-derived microglia added to the cultures engulfed Aβ and apoptotic cells, underscoring the modularity of this experimental system. We expect our model to provide a useful tool for assessing the impact of amyloid reduction on downstream AD pathologies such as mitochondrial dysfunction, neuroinflammation, and neurodegeneration, in particular in light of recent progress in the development and use of amyloid-targeting drugs.

Neurons exhibit region-specific identities corresponding to functional distinctions across different brain areas. Region-restricted neural stem cells (NSCs) have previously been generated from pluripotent stem cells; however, maintaining their regional identity over extended passages remains challenging. Here, we report the generation of hindbrain-like induced NSCs (Hb-LiNSCs) with upregulated hindbrain-specific markers and downregulated forebrain, midbrain, and spinal cord markers under xeno-free and basic fibroblast growth factor (bFGF)-free conditions using three chemicals-CHIR99021 (at a high concentration), a potent activator of the Wnt pathway; A-83-01, a potent inhibitor of the TGF-β/Activin/Nodal pathway; and LDN193189, a potent inhibitor of the bone morphogenetic protein pathway. Hb-LiNSCs maintained their chromosomal integrity, multipotency, and differentiation capacity even after long-term culture for more than 60 weeks. This approach enhances our understanding of neurodevelopmental and neurodegenerative processes in the hindbrain region and paves the way for developing targeted cell-based therapy as well as disease modeling for drug discovery.

We developed HyPer-3D (hydrogen peroxide-NaN3-DMSO), a chemical treatment that addresses persistent challenges in 3D imaging of cleared tissues. Imaging across stem cell, zebrafish, and murine model systems demonstrated that HyPer-3D quenches tissue autofluorescence, increases signal-to-background ratios (SBRs) by 30×, potentiates optical clearing capacity by 6×, increases detection of poorly recognized antigens by 4.5×, and enables a 5× increase in fluorescent reporter detection, which are conversely often diminished in 3D imaging. These improvements enhanced multiplex interrogation of highly autofluorescent and fibrous organs, including previously undocumented nephron segments, cardiac conduction system elements, and diffuse spermatogonial stem cells. Optimization for human tissues, which remain among the most difficult to 3D imaging due to autofluorescence and structural density far exceeding those of commonly used animal tissues, yielded a 5× increase in SBRs and enabled multiplex imaging with cellular resolution. Collectively, HyPer-3D provides a robust approach for high-contrast 3D imaging of basic and clinical specimens.

Post-translational modifications (PTMs) of proteins represent central regulatory mechanisms within cells, enabling rapid and targeted modulation of protein functions. PTMs can directly influence protein activity, stability, or protein-protein interactions, while histone modifications also play a crucial role in gene expression regulation. To date, over 400 types of PTMs have been identified, with new types continually emerging. Among these, lysine lactylation (Kla), crotonylation (Kcr), and lysine 2-hydroxyisobutyrylation (Khib) have recently been recognized as novel PTMs with critical roles in plant growth, development, and stress responses. Unlike previous reviews that broadly summarize plant PTMs, this article provides a focused synthesis on these three newly characterized acylations, highlighting their distinct biochemical features, regulatory mechanisms, and functional relevance in plant growth, development, and stress adaptation. We summarize recent proteomic and functional studies that uncover (i) the role of histone and non-histone lactylation in metabolic reprogramming and stress resilience; (ii) the contribution of crotonylation to transcriptional regulation, enzyme activity, and abiotic stress tolerance; and (iii) the emerging function of 2-hydroxyisobutyrylation in photosynthesis, stem development, and plant pathogen interactions. Furthermore, we discuss cross-talk among Khib, Kcr, and Kac (lysine acetylation), revealing a coordinated acylation network that fine-tunes chromatin dynamics and metabolic homeostasis. By integrating findings across multiple species including rice, wheat, maize, pepper, and fungi this review proposes a comparative and mechanistic framework for understanding how these acylations bridge cellular metabolism with epigenetic and physiological regulation.

We report on the generation of human induced pluripotent stem cell (iPSC) lines, BIHi005-A-86 and BIHi005-A-87, carrying the KIT D816V mutation associated with Indolent Systemic Mastocytosis (ISM). To overcome the confounding genetic backgrounds of existing leukemic models, we introduced this gain-of-function mutation into the healthy BIHi005-A line using CRISPR/Cas9 editing. The resulting clones were validated via whole-genome sequencing (WGS) to confirm specific on-target editing and lack of predicted or disease-relevant off-target effects, while maintaining genomic stability. Together with the parental line, this resource provides an isogenic controlled platform for investigating KIT D816V-driven pathogenesis.

Oral mucosal ageing represents a fundamental reprogramming of the tissue microenvironment, a dynamic process that underlies the functional decline and heightened disease susceptibility observed in the elderly. This review synthesises current evidence to reconceptualise oral mucosal ageing as an active reprogramming of the tissue microenvironment, delineating the interplay between structural, molecular, and immunological changes, and exploring how these alterations drive functional decline and increase susceptibility to age-related oral diseases. Through a comprehensive analysis of experimental and clinical studies from human and animal models, we demonstrate that the ageing process fundamentally transforms the oral mucosa. Key findings include structural changes such as epithelial atrophy, extracellular matrix remodelling, and salivary gland degeneration, driven molecularly by genomic instability, accumulation of proinflammatory senescent cells, stem cell exhaustion, and dysregulated stress responses. These are compounded by an immunological state of 'inflammaging' and functional decline in innate and adaptive immunity, further exacerbated by shifts in the oral microbiome. Collectively, these deficits lead to impaired regeneration, diminished sensory function, and reduced salivary secretion, creating a permissive landscape for chronic oral diseases. In conclusion, oral mucosal ageing is a dynamic process of microenvironmental reprogramming driven by cellular senescence, immunosenescence, and structural decay. This actively underpins the heightened vulnerability to oral disease in the elderly, providing a mechanistic foundation for developing targeted interventions to preserve oral health in ageing populations.

Magnesium alloys have significant advantages as biodegradable orthopedic implants. However, their rapid degradation in physiological environments and lack of efficient osteointegration severely limit clinical applications. To effectively regulate the degradation rate of ZK60 magnesium alloy and enhance its bone defect regeneration ability, this study employed micro-arc oxidation (MAO) and hydrothermal treatment to create a microporous coating, followed by a layered double hydroxide (LDH) coating to seal the micropores and enhance corrosion resistance. Meanwhile, polydopamine (PDA) modification and BMP2 loading were applied to construct a functional composite coating with both corrosion resistance and osteogenic activity. The corrosion resistance and bone defect regeneration performance were systematically evaluated in vitro and in vivo. Results showed that the LDH coating exhibited a typical layered structure and effectively sealed the MAO coating micropores, forming a dense protective layer. Electrochemical results showed that compared with pristine ZK60, the corrosion current density of BMP2@PDA-LDH/ZK60 decreased from 6.18 × 10-5 A/cm² to 8.15 × 10-6 A/cm², indicating the coating significantly retarded the corrosion rate. In vitro and in vivo biological assessments demonstrated that the coating effectively promoted rat bone marrow mesenchymal stem cells (BMSCs) adhesion and bone defect regeneration; specifically, the bone mineral density (BMD) value of the BMP2@PDA-LDH/ZK60 group was 454.25 ± 31.41 mg/cm³ , which was 1.44 times higher than that of the LDH/ZK60 group.This study provides a novel strategy for biodegradable magnesium alloys with excellent corrosion resistance and osteogenic potential.

Neuroblastomas are heterogeneous pediatric tumors of the sympathetic nervous system for which treatments are still limited. Fundamental and applied approaches have been enabled thanks to the generation of patient-derived tumoroids (PDT), ex vivo 3D structures used as avatars of the original tumor. We generated neuroblastoma PDT and quantified the spatial distribution of CD133+ cancer stem cells using immunohistochemistry. We observed that those cells tend to aggregate in the PDT. In order to better understand the set of rules needed for generating such structures, we implemented a multiscale agent-based neuroblastoma tumoroid model. Model rules specify single cell's fate based on their intracellular content, which dynamically evolves according to a stochastic gene regulatory network. The state of this network can be modulated by cell-to-cell signaling through neighbor cell fate decisions and, possibly, spatial location. We first observed that in the absence of any spatial rules for inter-cellular interactions, no spatial structure emerged. The addition of simple rules (signaling by cell-to-cell contact or differential cell adhesion) only marginally improved the quantitative agreement to the experimental dataset. In sharp contrast, the addition of short-range pro-stem cell diffusive signaling among stem cells produced very realistic 3D PDT-like structures. This works highlights the power of our multiscale approach to discard too simplistic rules and to propose a minimal set of hypotheses required to reproduce qualitatively and quantitatively experimentally observed spatial structures. In the case of neuroblastoma-derived PDT, short-range spatial diffusion of stem-to-stem cell signaling proved to play a key role in successfully reconstructing the spatial structure.

There is currently no biomarker to predict the maximum morphine dose (MMD) for newborns experiencing withdrawal from chronic prenatal opioid exposure (POE). This is due, in part, to a lack of understanding about how the developing brain is altered by chronic opioid exposure and withdrawal on a molecular level. We previously developed a human induced pluripotent stem cell-derived model of POE and withdrawal to examine the impact on neural progenitor cell fates. Here, we leveraged our model to investigate the role of two microRNAs implicated in both neural stem cell differentiation and opioid signaling: miR-149 and miR-23b. Further, we asked if these microRNAs were related to the need for morphine treatment and MMD in the saliva of infants with POE. Levels of miR-149 (One-way ANOVA, F = 34.18, p < 0.0001), but not miR-23b (One-way ANOVA, p = 0.14), were significantly decreased in human neural progenitors after chronic morphine exposure (Tukey's, adj. p = 0.004), and decreased further in those that underwent withdrawal compared to vehicle exposed controls (Tukey's, adj. p < 0.0001). The relevance of miR-149 to neonates experiencing withdrawal after POE was confirmed by decreased salivary levels of miR-149 compared to levels in healthy infants 24-96 hours after birth (n = 56, 28 unexposed and 28 infants with POE) (Mann-Whitney U, p < 0.0001). Stratifying infants with POE by need for pharmacotherapy revealed a further decrease in levels of miR-149 in infants that required treatment (One-way ANOVA, p < 0.0001). In a hierarchical linear regression model utilizing infant demographic factors, addition of miR-149 levels in neonatal saliva improved performance for predicting the MMD necessary for symptom control (R = 0.673, p = 0.002). These results indicate the potential relevance of miR-149 levels in infants with prenatal opioid exposure. Validation in larger cohorts is necessary.

Human induced pluripotent stem cell (hiPSC)-derived microglia (hiMG) constitute a powerful platform to study human microglial biology in health and disease. Here, we present a protocol for microglial differentiation and efficient gene delivery to these cells using adeno-associated virus (AAV). We describe steps for differentiation, viral transduction, and validation using flow cytometry and fluorescence imaging. Finally, to demonstrate the utility of this approach, we outline procedures to record and analyze calcium activity in hiMG using the genetically encoded sensor GCaMP8s. For complete details on the use and execution of this protocol, please refer to Fruhwürth et al.1 and Heiss et al.2.