Sporadic form of Alzheimer's disease (sAD), remains a complex neurodegenerative disorder with limited translational models. Brain organoids derived from human induced pluripotent stem cells (hiPSCs) have emerged as promising tools to recapitulate aspects of human brain development and pathology. Recent advances have introduced vascularized, immune-competent organoids capable of modeling hallmark features of sAD, including amyloid-β accumulation, tau pathology, and neuroinflammation. New strategies to enhance organoid maturation, cellular diversity, and aging phenotypes are pushing the boundaries of disease modeling. This review highlights cutting-edge developments in brain organoid systems for studying sAD, addresses key limitations, and outlines future directions to improve their translational relevance for therapy and mechanistic insights.

Functional persistence of chimeric antigen receptor T cells (CAR T cells) is limited by conventional CAR T cell manufacturing using anti-CD3/CD28 (αCD3/28) stimulation, which generates terminally differentiated and shorter-lived CAR T cells. We demonstrated that HCW9206, a unique protein scaffold linking interleukin-7 (IL-7), an IL-15/IL-15 receptor α (IL-15Rα) complex, and IL-21, generates CAR T cells without requiring αCD3/28 activation, which are highly enriched in long-lived T memory stem cells (TSCM cells) (>50%) and display potent activity across distinct disease models, HIV-1 or B cell leukemia. In a humanized mouse HIV infection model, HCW9206-generated anti-HIV duoCAR T cells suppressed viremia more effectively than αCD3/28-generated anti-HIV duoCAR T cells. In a xenograft leukemia mouse model, a recall proliferative response and complete clearance of leukemia rechallenge were displayed by HCW9206-generated but not by αCD3/28-generated anti-CD19 CAR T cells. HCW9206, a first-in-class cytokine scaffold-based platform, enables production of more potent CAR T cell-based immunotherapies by generating a CAR T cell population, which is highly functional and also markedly enriched for long-lived TSCM cells. This strategy is broadly applicable to increase persistence and functionality of CAR T cells, enhancing their efficacy for treating infectious disease and cancer.

To access the earliest stages of murine T cell development, we adapted a serum-free expansion system for hematopoietic stem and progenitor-like cells. These cells undergo normal T cell differentiation in vivo and in vitro, verified by gene expression trajectories from single-cell RNA sequencing. Their absolute differentiation speed is slower than that of fresh progenitors but is modulated with Flt3L priming. Leveraging this system, the earliest T cell lineage-initiating events in response to Notch signaling included chromatin opening and transcriptional activation of the TCR-Cβ locus. Acute CRISPR knockouts revealed that some transcription factors normally inherited from bone marrow progenitors impede early T cell development, with differing effects on proliferation. Among 23 factors tested, Lmo2 knockout greatly accelerated the onset of germline TCR-Cβ locus transcription and expression of Tcf7, Gata3, and Runx1 and their targets. Thus, normal endogenous expression of this progenitor- and leukemia-associated factor markedly restrains T cell program initiation.

While the cytosolic localization of cGAS is critical for cells to initiate immune responses and protect cells from viral infections, the activity of cGAS in the nucleus is inhibited to prevent autoimmune responses triggered by self-DNA. Therefore, the dynamically regulated distribution of cGAS in the cytosol and nucleus ensures its precise role in maintaining immune homeostasis. However, the molecular mechanism governing this spatial distribution of cGAS remains unclear. Here, we identify MSH6 as a regulator promoting cGAS nuclear localization by enhancing its association with importin-α proteins, consequently reducing cGAS condensation and activity. We further show that MSH6 attenuates antitumor immunity and that its deficiency in tumor cells leads to an effective tumor eradication by heat-inactivated modified vaccinia virus Ankara. Collectively, our results not only provide insights into understanding how cGAS activity is regulated but also suggest a therapeutic potential for treating MSH6-mutated tumors through the cGAS-mediated signaling pathway.

Activation of macrophages with pro-inflammatory stimuli leads to the generation and secretion of large amounts of reactive oxygen species (ROS), which cause DNA double-strand breaks (DSBs). Here, the role of DNA polymerase μ (Polμ), a component of the non-homologous end-joining (NHEJ) DNA repair pathway of DSBs, in macrophage functional activity during inflammation is evaluated. Polμ is expressed in macrophages upon pro-inflammatory stimuli depending on ROS. Under pro-inflammatory stimuli, Polμ-deficient macrophages exhibit reduced proliferation and enhanced apoptosis, a phenotype associated with deficient DSB repair. In a mouse model of macrophage-dependent muscular regeneration, Polμ deficiency impaired tissue repair associated with macrophage survival. In models of inflammation and infection, Polμ is required for macrophages to subsist at inflammatory foci and mount an effective inflammatory response. These findings show that Polμ is crucial for macrophages to block the unwanted effects of ROS during inflammatory responses.

Herpes simplex encephalitis (HSE) is a life-threatening disease of the central nervous system caused by herpes simplex virus type 1 (HSV-1). Despite being a standard treatment, antiviral acyclovir and its derivatives often face limitations in clinical application due to their side effects and viral drug resistance. Inspired by viral entry through recognition of nectin-1 on the host cell surface, we engineered enucleated mesenchymal stem cells with high nectin-1 expression (eMSCs) to serve as "decoys" for capturing HSV-1. We found that eMSCs competitively captured the virus in the presence of neurons while inhibiting its replication and spread by removing the nucleus in advance. Interestingly, due to the absence of nuclei, eMSCs capturing the virus trigger macrophage efferocytosis through intrinsic apoptosis after approximately 60 h, thereby accelerating existing viral clearance. This is a property lacking in current antiviral drugs, including ACV. In summary, this strategy significantly improved the quality of life of HSE mice.

ObjectiveThis study aimed to evaluate the clinical and biological outcomes of a one-step regenerative treatment using bone marrow aspirate concentrate (BMAC) on a hyaluronic acid scaffold combined with intraarticular platelet-rich plasma (PRP) for treatment of full-thickness cartilage lesions in the knee.Design/MethodsA total of 165 patients (mean age 26.4 years) with full thickness cartilage defects were included in the study. In a single-step procedure, bone marrow was aspirated from the iliac crest and centrifuged to obtain BMAC concentrate. PRP was prepared from whole blood. The BMAC concentrate was seeded onto a hyaluronic acid scaffold and implanted into the debrided cartilage defect and PRP was added intraarticularly. Clinical outcomes were assessed using the Knee Injury and Osteoarthritis Outcome Score (KOOS) and a semiquantitative grading scale (Excellent, Good, Fair) at one-year follow-up. All patients underwent MRI preoperatively and at one year follow-up, with cartilage repair evaluated using the MOCART-2.0 scoring system.ResultsSignificant improvements were observed in all KOOS subscales at one year. Semiquantitative grading revealed excellent outcomes in 50% of patients, good in 28%, and poor in 22%. MRI analysis showed an average MOCART-2.0 score of 71, with 36% of patients achieving excellent cartilage repair (MOCART 2.0 >80).ConclusionThe combined use of BMAC on a hyaluronic acid scaffold and intraarticular PRP resulted in improved subjective clinical outcomes and demonstrated favourable biological cartilage healing responses at one year postoperatively. This one-step cartilage repair technique appears to be a clinically valid option for treatment of full-thickness cartilage lesions in the knee.

Traumatic brain injury (TBI) is a major global health issue leading to high mortality and disability. Activated astrocytes are one of the pivotal driving factors in the neuroinflammatory cascade following TBI. This study aims to investigate the role of esketamine on TBI and the underlying mechanism. Mice received a mouse weight-drop cortical impact or sham surgery and TBI mice were treated with either vehicle or esketamine at 2 h post-injury for 7 consecutive days. The modified Neurological Severity Scoring system, Rotarod test, Open Field test and Novel Object Recognition test were used to assess the neurological function after TBI. And cortical tissues surrounding focal trauma were obtained for Nissl staining, immunofluorescence, ELISA assay and western blotting. In vitro, astrocytes were induced with LPS, followed by the addition of esketamine to the culture medium. After a 24 h exposure, the astrocytes were collected for CCK-8 assay, qRT-PCR, western blotting, immunofluorescence and Co-IP analysis. Esketamine dramatically improved the neurological outcome of mice and reduced neuronal cell death (P < 0.05) and neuroinflammation after TBI. Its anti-inflammatory benefits stem from its ability to suppress astrocyte activation (P < 0.05), inhibit pro-inflammatory A1 astrocyte differentiation (P < 0.01), and promote the formation of protective A2 astrocytes (P < 0.01). Esketamine exerts its effects by inhibiting the METTL5/c-Myc/PD-L1 signaling pathway. Esketamine can effectively alleviate activated astrocytes and promote the polarization of activated astrocytes toward A2 following TBI by inhibiting the METTL5/c-Myc/PD-L1, demonstrating significant anti-inflammatory and neuroprotective effects.

Biocompatibility between bone cells and biomred for the process of osteogenesis. The aim of this study was to assess the effect of Photobiomodulation (PBM) in vitro on dental pulp stem cells (DPSCs) treated with Bio-Oss® (BO) bone substitute xenograft.

Myocardial ischemia-reperfusion injury remains a major unresolved challenge in cardiovascular medicine. Although timely restoration of blood flow is essential to limit ischemic damage, reperfusion triggers a complex network of maladaptive biological responses, including oxidative stress, calcium overload, mitochondrial dysfunction, metabolic impairment, and sterile inflammation. These processes converge on cardiomyocyte death, adverse ventricular remodeling, and long-term functional deterioration. Mesenchymal stem cells have been widely investigated as cardioprotective agents; however, accumulating evidence indicates that their beneficial effects are predominantly mediated by paracrine mechanisms. Among these, extracellular vesicles released by mesenchymal stem cells have emerged as key biological effectors. Experimental studies demonstrate that mesenchymal stem cell-derived extracellular vesicles modulate multiple signaling pathways involved in ischemia-reperfusion injury, including activation of the phosphoinositide 3-kinase (PI3K) and protein kinase B (PKB) axis, regulation of signal transducer and activator of transcription 3 (STAT3) signaling in a cell-specific manner, suppression of nuclear factor kappa B (NF-κB)-driven inflammatory responses, and stabilization of hypoxia-inducible factor-1α (HIF-1α)-dependent adaptive programs. At the subcellular level, these vesicles preserve mitochondrial structure and function, support energy metabolism, regulate mitophagy, and limit oxidative damage. Their molecular cargo, comprising regulatory microRNAs, metabolic enzymes, and stress-response proteins, enables coordinated modulation of survival, inflammatory, and reparative pathways rather than single-target effects. This review synthesizes current experimental evidence on the mechanistic basis of mesenchymal stem cell-derived extracellular vesicle-mediated cardioprotection and discusses their potential as cell-free, mechanism-based therapeutic strategies to limit myocardial ischemia-reperfusion injury.

T-cell exhaustion is a state of functional decline in T cells, driven by chronic antigen exposure and inhibitory signals within the tumor microenvironment. Exhausted CD8+ T cells (Tex) derive from precursor exhausted T cells (Tpex), a self-renewing population responsible for the proliferative burst in anti-PD-1 therapies. Exhausted T cells are exposed to adenosine in the tumor, yet the role of the CD73/adenosine axis and Tpex/Tex differentiation remains unclear. Using an in vitro model for CD8+ T-cell exhaustion, we found that CD73 expression increased during Tpex formation and that its expression was negatively correlated with Tex generation. CD73-deficient OT-I cells (OT-I/CD73KO) showed impaired activation and reduced progression to Tex. Moreover, we demonstrated that CD73 promotes transcriptional expression of the adenosine receptor A2A (A2AR). RNA-seq analysis of exhausted OT-I/CD73KO cells revealed a more stem-like transcriptomic profile and enrichment in genes associated with the immune response compared to their OT-I counterparts. In vivo, the cotransfer of naïve OT-I/CD73KO and OT-I antigen-specific CD8⁺ T cells into tumor-bearing mice resulted in increased Tpex frequency and numbers among OT-I/CD73KO cells in tumors. Conversely, in vitro exhaustion in the presence of A2AR agonists decreased Tex frequency and activation/exhaustion markers, while increasing CD73 and CD62L, which are markers associated with stemness. Supporting this, A2AR blockade with SYN115 in tumor-bearing mice reduced Tpex markers and increased Tex differentiation. Altogether, our data suggest that CD73 promotes Tpex-to-Tex differentiation, whereas adenosine A2AR signaling supports Tpex maintenance in the tumor microenvironment.

Mechanotransduction, the process by which cells convert mechanical stimuli into biochemical signals, serves as a fundamental biological mechanism driving tissue adaptation and repair in orthopedic rehabilitation. The present review explores how mechanical forces regulate cellular behavior in bone, cartilage, tendon and ligament healing, emphasizing their critical role in optimizing regenerative outcomes. Specialized mechanosensors, including integrins, ion channels and primary cilia, detect physical cues such as compression, tension and shear stress, activating downstream pathways that direct stem cell differentiation, matrix synthesis and tissue remodeling. The extracellular matrix functions not only as a structural scaffold but also as a dynamic mediator of mechanical signaling, influencing cellular responses to therapeutic loading. Clinically, mechanotherapy strategies, including controlled weight‑bearing, eccentric exercises and devices providing dynamic compression, are designed to exploit these principles, promoting anabolic activity while preventing catabolic damage. Advances in biomechanically optimized scaffolds, bioreactor systems and technologies (such as low‑intensity pulsed ultrasound) further demonstrate how targeted mechanical conditioning enhances tissue‑engineered constructs and accelerates functional recovery. However, challenges remain in defining optimal loading parameters across diverse tissues and individual patients. Future directions should prioritize personalized rehabilitation protocols informed by real‑time biomechanical monitoring and genetic profiling, alongside biomaterials that can adapt to in vivo mechanical cues. The integration of mechanobiology with regenerative medicine is paving the way for a new era in orthopedic rehabilitation. This evolution promises more precise, effective and biologically driven interventions that harness the innate mechanoresponsive capacity of the body to restore function.

Obesity and its metabolic complications represent a major and growing public health burden, for which effective therapeutic strategies remain limited. Although super-enhancers are recognized as key regulators of cell identity and adipogenesis, the roles of super-enhancer- microRNAs in obesity and adipocyte dysfunction remain insufficiently characterized. Here, we identify miR-1260b as a super-enhancer-associated miRNA that influences human adipogenesis. Analysis of adipose-related datasets from SEdb 2.0 revealed that MIR1260B is consistently linked to a clustered SE region across multiple human adipose tissues. Integrated ATAC-seq and Hi-C analyses demonstrated progressive chromatin opening and dynamic enhancer-promoter interactions between this SE and the MIR1260B promoter during adipogenic differentiation. Functionally, miR-1260b overexpression markedly inhibited adipocyte differentiation of human adipose-derived stem cells. Quantitative proteomic profiling revealed that miR-1260b suppresses adipogenic and lipogenic programmes while activating lipid catabolic pathways. Notably, the MIR1260B-associated SE overlaps with a previously reported diabetes-associated SNP, and external clinical datasets indicate reduced miR-1260b levels in umbilical cord serum from children at increased risk of obesity. Together, these findings suggest that miR-1260b may function as a super-enhancer-associated regulator that inhibits adipogenesis and indicate that SE-informed multi-omics approaches can aid in identifying miRNA regulators relevant to metabolic disorders of obesity.

Cancer stem cells (CSCs) constitute a subpopulation of malignant cells with self-renewal and differentiation capabilities that drive tumorigenicity, metabolic adaptability, immune evasion, and therapeutic resistance, key factors contributing to metastasis and poor clinical outcomes. While genetic drivers of tumorigenesis are well-characterized, the epigenetic machinery sustaining the CSC state remains complex. Long noncoding RNAs (lncRNAs) represent a vast yet poorly understood class of regulatory molecules that influence gene expression at epigenetic, transcriptional, and post-transcriptional levels. Emerging evidence indicates that lncRNAs play a crucial role in shaping tumor cell plasticity and stemness-associated phenotypes. In this mini-review, we summarize current findings on how lncRNAs regulate CSC biology. We categorize their mechanisms of action, ranging from chromatin remodeling to the modulation of mRNA and protein stability. Furthermore, we discuss how the advent of high-resolution omics, including bulk tissue, single-cell, and spatial transcriptomics studies, is revolutionizing the identification CSC-associated lncRNAs and contributing to the development of clinically relevant biomarkers. Finally, we explore advanced methodologies for manipulating lncRNA expression, assessing the challenges and opportunities of lncRNA-directed therapeutics as a novel strategy to dismantle tumor plasticity and overcome drug resistance.

Brain microvascular endothelial cells (BMECs), which constitute the blood-brain barrier (BBB), are essential for maintaining central nervous system homeostasis. Like BMECs, multipotent mesenchymal stem cells (MSCs) originate from the mesodermal lineage. Thus, MSCs may serve as a direct and efficient cellular source for BMEC-like differentiation. Notably, differentiation of human induced pluripotent stem cells (hiPSCs) into BMECs typically involves a 2-step protocol: inducing mesodermal commitment followed by endothelial specification. In contrast, direct differentiation from MSCs could bypass the initial mesodermal induction step, offering a streamlined alternative. This study tested a novel strategy for differentiating MSCs into brain-like endothelial cells (BLECs), circumventing the conventional mesodermal induction step.

Peripheral blood samples are widely used in HLA genotyping due to their easy accessibility and the high-quality DNA from nucleated leukocytes. However, in cases of disease relapse requiring a second transplantation, clinicians encounter significant challenges in performing HLA genotyping and interpreting complex results from patients who have undergone haploidentical hematopoietic stem cell transplantation (haplo-HSCT). Furthermore, systematic studies investigating the impact of haplo-HSCT on recipients' HLA genotypes across different tissues remain scarce. Therefore, this study aims to analyze HLA genotypes in various tissues of the recipient after haplo-HSCT.

The identification of stem-like CD8+ T cells, also termed progenitor or precursor of exhausted T cells (TPEX), has reshaped our understanding of durable antitumor immunity. These cells exhibit progenitor-like properties, including self-renewal capacity and multilineage differentiation potential, giving rise to both effector-like and terminally exhausted CD8+ T cell subsets. Accordingly, the abundance of stem-like CD8+ T cells correlate strongly with improved clinical outcomes in patients receiving immune checkpoint inhibitors, adoptive cell therapy, or cancer vaccines across multiple tumor types. This review synthesizes recent advances in TPEX cells biology, highlighting interconnected research pillars, including: specialized niche microenvironments that sustain stemness of TPEX cells through coordinated chemokine signaling and antigen-presenting cell interactions; core molecular circuitry that dynamically balances self-renewal versus effector differentiation via transcription factors and cytokines; and therapeutic reprogramming strategies that harness TPEX cells as the primary driver of immunotherapy efficacy. Further, we explore strategies to augment the functionality of TPEX cells through niche modulation, stem-like CAR-T engineering, and combinatorial approaches, highlighting the trend that targeting TPEX cells thus emerge as a transformative future strategy to overcome immunotherapy resistance and achieve a durable response.

Radioresistant thymic cells encompass minor subsets of lymphoid precursors of T cells (TLPs), innate lymphoid cells (ILCs), as well as stromal-epithelial and endothelial populations. This review focuses on radioresistant TLPs and their regenerative and functional roles in thymic regeneration following damaging influences, particularly irradiation, as well as their secretory product, referred to as thymocyte growth factor (THGF). Retrospective analysis of experimental data assumes that THGF-producing and THGF-responsive cells correspond to the earliest stage of thymocyte precursors, double-negative (DN) TLPs, of CD117-Thy-1+Sca-1+CD44+CD25-CD4-CD8- phenotype, and may be a target for thymic oncogenesis, when they are in the activated DN1→DN2 stage. Unique features of THGF-driven proliferation of these cells include a colchicine-resistant DNA synthesis and, presumably, the formation of a "daughter" cell pool within "mother" cell-like structures, as well as the formation of colony-cluster-like structures, which are presumably composed mainly of single activated mother DN1 and surrounding daughter TLPs progressing from DN2 to DN4 stage. This atypical proliferation mode may represent an evolutionarily conserved mechanism of "defended mitosis" and/or amitotic or endomitotic pathways division, protecting against radiation-induced injury and thus allowing the cell expansion. THGF, which is induced by γ-irradiation and appears essential for autocrine expansion of radioresistant TLPs, initiates a cascade that enables subsequent responsiveness to IL-7, SCF, IL-2, and additional cytokines. The presented analysis proposes the concept of intrathymic dormant stem cells, which become activated under extreme conditions, and insights into parallels between THGF-responsive cells and other radioresistant thymic populations, suggesting an integrated network of stromal and lymphoid elements that orchestrate thymic regeneration. Together, this review proposes a model in which THGF acts as a critical regulator of dormant intrathymic stem cells, enabling their activation, protected proliferation, and differentiation, and thereby contributing crucially to the lymphoid lineage of thymic regeneration after irradiation, in addition to the concept of the IL-22-dependent pathway of stromal-epithelial regeneration of intrathymic niches microenvironment.

Human pluripotent stem cells (hPSCs) offer a versatile platform for modeling human cardiac development and generating cardiomyocytes for research and translational applications. Cardiac differentiation protocols are well established and rely on the sequential activation and inhibition of WNT, BMP, and FGF signaling pathways to guide lineage progression. While these intracellular signaling events are well characterized, less attention has been given to the temporal behavior of extracellular components present at the cell surface during differentiation. Glypicans (GPCs) are a family of membrane-bound heparan sulfate proteoglycans within the glycocalyx that are known to interact with morphogens in multiple developmental contexts. In this study, we profiled the expression of GPC1-6 during a standard chemically defined cardiac differentiation protocol, in the absence of targeted interventions. Gene expression analysis across stages revealed distinct, stage-associated patterns: GPC3 and GPC6 were upregulated during the WNT activation phase; GPC4 was suppressed after WNT inhibition and maintained low during cardiac commitment. GPC2 and GPC5 expressions peaked during the formation of cardiac progenitors, and GPC1 expression increased following cardiac specification. These findings provide a temporal map of GPC expression coinciding with established differentiation stages, demonstrating that members of the glypican family are dynamically expressed during human cardiac differentiation. By documenting when specific glypicans are expressed during a commonly used differentiation workflow, this study offers a descriptive reference framework that may inform future mechanistic studies investigating how extracellular components intersect with canonical cardiac signaling pathways.

The xenotransplantation of human cells into porcine hosts holds immense potential in the fields of regenerative medicine and organ transplantation. However, the low survival rate of human-derived cells within porcine remains a critical bottleneck constraining the application of xenotransplantation. Whether porcine cells exert negative effect on human cell growth is not studied. Here, we established an in vitro direct co-culture model of human and porcine mesenchymal stem cells (hMSCs and pMSCs) to investigate the competitive relationship between human and porcine-derived cells. The results demonstrated that the proliferation capability of hMSCs in the co-culture system was significantly suppressed compared to those cultured in isolation. Moreover, an increasing number of pMSCs exhibited enhanced inhibition of hMSC proliferation. Notably, results from transwell assays and treatment with porcine-conditioned medium indicated that the inhibition of hMSCs by pMSCs was not mediated through soluble cytokines. To elucidate the underlying molecular mechanisms, RNA sequencing analysis was performed and the result revealed that direct co-culture significantly downregulated the expression of proliferation-related genes in hMSCs, including CYP1B1, SLC7A11, TFAP2C, and PSAT1. Concurrently, the co-culture paradigm disrupted endoplasmic reticulum function and multiple amino acid transport processes within hMSCs, while activating the NF-κB signaling pathway, thereby achieving negative regulation of hMSC proliferation. Collectively, our primary study characterized the competitive interactions between hMSCs and pMSCs and uncovered possible underlying mechanisms which provided new experimental foundations for improving human cell survival in porcine hosts to advance xenotransplantation application.