The blood-brain barrier (BBB) comprised of the brain capillary endothelial cells (BCECs), with its tight junctions (TJ), transporters and receptors, regulates the passage of solutes, such as nutrients, metabolites, and xenobiotics, including drugs. In Alzheimer's disease (AD), characterised by the accumulation of amyloid-β peptide (Aβ) and the formation of hyperphosphorylated tau aggregates, a compromised BBB integrity was reported. There is a lack of knowledge about the effects of tau pathology on BBB function in AD. Advances in developing BBB models using human induced pluripotent stem cell (hiPSC)-derived BCECs have opened a new avenue for investigating AD-related changes in BBB functional integrity. Here, we characterised the BBB model derived from hiPSCs generated from an AD patient with a tau-related mutation (STBCi 062-A) versus the one based on a healthy person's cells (UKKi 011-A) in terms of mimicking AD-related changes in paracellular permeability, TJs, transporters, receptors and other proteins playing a role in BBB integrity. The STBCi 062-A-derived BCECs showed lower TEER values and increased permeability associated with downregulation of proteins regulating TJ organization and BBB integrity, as compared to UKKi 011-A-derived BCECs. We revealed AD-relevant increase in protein expression of efflux transporter BCRP and amino acid transporter ASCT1, as well as transferrin receptor protein 1 in the STBCi 062-A-derived BCECs compared to UKKi 011-A-derived BCECs. The developed AD-patient-hiPSC-derived BCEC model possesses several important characteristics that recapitulate changes in BBB integrity in AD and can serve as a robust tool for developing AD treatments.

Diabetic cataract is characterized by early lens opacification. This study identifies a pathogenic axis that involves the N6-methyladenosine (m6A) demethylase fat mass and obesity-associated protein (FTO) and the mitochondria-enriched long non-coding RNA (lncRNA) RMRP. Transcriptomic analysis of lens epithelial cells (LECs) under high-glucose stress revealed that lncRMRP was the most significantly downregulated lncRNA, while FTO was specifically upregulated. High glucose stimulated FTO to bind to RMRP, removed its m6A methylation, and expedited its degradation. Knockdown of RMRP led to mitochondrial DNA depletion, loss of respiratory chain subunits I, III, and V, bioenergetic failure, and structural damage. Notably, inhibition of FTO restored RMRP levels and rescued mitochondrial function under high-glucose conditions. In mice, overexpression of FTO in the anterior chamber induced lens opacification and mitochondrial defects, both of which were alleviated by co-expressing RMRP. In a mouse model of streptozotocin-induced diabetes, intracameral AAV-RMRP delivery restored RMRP expression specifically in LECs, suppressed cataract formation, alleviated cellular energy deficits, and restored mitochondrial DNA copy numbers and key mitochondrial biogenesis regulators. This work uncovers a new mechanism in diabetic cataracts: chronic hyperglycemia upregulates FTO, which degrades RMRP-causing mitochondrial dysfunction and lens opacity. Critically, rescuing RMRP function in a physiologically relevant diabetic model validates it as a promising therapeutic target for diabetic cataracts.

Alginate, a natural polysaccharide [(1 → 4)-linked β-D-mannuronate and α-L-guluronate], is commonly used in hydrogel form in the biomedical field, including wound healing, drug delivery, and tissue engineering applications. The type and density of cross-linking are key parameters determining the physicochemical and mechanical properties of the hydrogel. However, most of the crosslinking reagents used are generally toxic, requiring extensive modifications and limiting their applicability. Alginate readily forms ionic hydrogels through coordination with divalent cations such as Ca2+ and Ba2+; however, these physically crosslinked networks exhibit poor mechanical integrity under aqueous and in-vivo conditions. Consequently, there is a critical demand for cross-linking agents that are biocompatible, non-toxic, hydrolytically stable, rapidly gelling, and cost-effective to overcome these limitations. In this study, alginate-based 3D hydrogel scaffolds were prepared in a completely green synthetic way using caffeine-catalyzed citric acid-poly(ethylene glycol) (PEG)/poly(propylene glycol) (PPG) as the cross-linker system. The thermal stability, swelling, rheology as well as compressive strength of the hydrogels were investigated and optimized. The developed hydrogel scaffolds containing PPG withstood compressive deformation in both dry and swollen states without any damage and recovered their initial shape after unloading, indicating that the hydrogels possess a certain shape recovery property. In-vitro biological studies with human adipose stem cells revealed that both scaffold types were cytocompatible and showed good hemocompatibility. Overall, the green synthesis of biocompatible and mechanically resilient alginate-CA-PEG scaffolds highlights their potential as versatile biomaterials for prospective medical applications, particularly in soft tissue engineering.

Patients with high-risk (HR) leukemia remain at substantial risk of early relapse, treatment-related toxicity, and poor survival, underscoring the need for effective relapse prevention therapies. To our knowledge, this first-in-human, disease burden-guided study evaluated the feasibility, safety, and efficacy of donor-derived allogeneic interleukin-15-activated cytokine-induced killer cells (IL15-CIK) combining T-cell and natural killer cell properties for post-transplant disease control.

Both the levels and duration of gene expression play critical roles in many biological processes. Recent studies have further revealed that the dynamics of oscillatory versus sustained gene expression also provide essential regulatory information during cell proliferation and differentiation. Oscillatory expression, governed by intracellular negative feedback loops and intercellular coupling with appropriate delays, promotes the proliferation of stem cells, whereas sustained expression typically drives cells toward quiescence or differentiation. Over time, oscillations can result in the gradual upregulation or downregulation of downstream factors or shifts in phase relationships between distinct oscillators, thereby functioning as a timer for cell state transition. Moreover, oscillation frequency encodes critical cues for cell fate choice. Thus, oscillatory dynamics add an extra dimension to the informational landscape of gene expression. Here, we discuss recent advances in our understanding of how oscillatory gene expression is regulated and how it influences the proliferation and differentiation of stem cells.

Adult stem cells maintain tissue homeostasis, yet are themselves vulnerable to loss. One common mechanism to replace lost stem cells is dedifferentiation, in which progeny revert to stem cell identity. It is a paradox how stem cells and progeny retain the same stem cell potential while exhibiting distinct current identities of self-renewal, differentiation, and dedifferentiation. Here, we show that the Drosophila male germline lineage solves this paradox via two parallel and complementary mechanisms to separate potential and identity. First, differentiating progeny maintain stem cell potency by inheriting perdurant stem cell mRNAs without actively transcribing them. Second, two known niche signals (Bmp and Jak-Stat) activate distinct sets of targets, defining three identities (self-renewal, differentiation, and dedifferentiation) based on the combination of their on/off states. Together, this study reveals how a pool of dedifferentiation-competent progeny is maintained to regenerate stem cells as needed without resulting in stem cell overproduction and resolves the puzzle of why most stem cell systems require multiple independent niche signals.

Osteogenesis depends on the self-renewal and differentiation of mesenchymal stem cells (MSCs). Emerging research underscores the regulatory functions of RNA methylation on bone homeostasis. Here, we show PCIF1, the N6,2'-O-dimethyladenosine (m6Am) methyltransferase, is essential for maintaining bone mass and promoting osteogenic differentiation of MSCs. Multiple complementary analyses-including GWAS, TWAS, and single-cell transcriptomics-collectively point to PCIF1 as a regulator of human bone mineral traits and early-stage mesenchymal differentiation. Global or MSC-specific Pcif1 deletion elicits osteoporotic pathology in mice, although myeloid cell-specific Pcif1 knockout does not induce femur bone alterations. Mechanistically, Pcif1 knockout decreases m6Am signals of Wnt-related genes (Wnt11, Fzd4, and Fgfr2) and accelerates mRNA degradation. This down-regulates active β-Catenin protein, and thus impairs osteogenic function of MSCs. Additionally, the WNT agonist attenuates the osteoporosis-like phenotype induced by Pcif1 deletion. These findings highlight the crucial role of PCIF1-mediated m6Am modification in regulating osteogenesis and suggest potential therapeutic implications for bone disorders.

Without rapid revascularization, ischemic heart disease leads to irreversible loss of cardiomyocytes, scar formation, and decreased cardiac function. The ability to model hypoxia-reoxygenation injuries and evaluate potential therapeutic interventions in human tissues is an unmet need. Human cardiac organoids (hCOs) are an emerging model system, but beginning work with hCOs can be challenging due to their small size, requirements for suspension culture, and variability in differentiation efficiency. This protocol offers a user-friendly guide for generating and maintaining human iPSC-derived self-assembling hCOs; methods to process and histologically analyze hCOs; and an approach to injure hCOs with hypoxia-reoxygenation that mimics the fibrosis and apoptosis seen with human ischemia/reperfusion. This protocol provides guidance for using hCOs as a complement to animal models of ischemia-reperfusion injury and could be used to identify and test factors that may boost human myocardial recovery.

Obesity-related changes in adipose tissue alter local secretory profiles and influence interactions with other surrounding cells, including cancer cells. The continued delivery of nutrients shifts the metabolism of fat cells, making them act as a lipid reservoir. Typically, research on obesity mechanisms relies on animal models, which, however, do not allow analysis of metabolic processes at the cellular level. Hence, there is a need to develop a reliable in vitro model of human obesity that can help explore changes in the function of obese adipocytes compared to their lean counterparts. This work presents a protocol for the differentiation of human adipocytes from their precursors (adipose-derived stem cells) using a mixture of insulin, isobutyl-1-methylxanthine, dexamethasone, and indomethacin. Subsequently, differentiated adipocytes were incubated with a set of fatty acids, including linoleate, palmitate, and oleate, to promote lipid accumulation. As a result, a valuable model mimicking the hypertrophic cells under obesity conditions was achieved, which can be used for further studies.

Chimeric antigen receptor (CAR) T-cell therapy has been a breakthrough in many hematological malignancies but success in relapsed/refractory (R/R) acute myeloid leukemia (AML) has been limited due to underwhelming response rates and high on-target/off-tumor toxicity. This Phase 1 dose-escalation trial evaluated the safety and efficacy of KITE-222, an autologous CAR T-cell therapy that recognizes C-type lectin-like molecule 1 (CLL-1) predominantly expressed on myeloid cells but absent on normal hematopoietic stem cells and other tissues.

Bone maintains a dynamic and stable state through the orchestration of osteoclasts and osteoblasts. Osteoblasts are derived mainly from mesenchymal stem cell (MSC) and are responsible for bone formation. The inhibition of osteoblast proliferation and differentiation is involved in many diseases, including osteoporosis, osteoarthritis, infected bone defects, and inflammatory aseptic loosening of implants. Given the currently limited treatment options, exploring new methods to promote bone formation is an important focus significant for orthopedists. Platelet-rich plasma (PRP), an autologous substance that is rich in various growth factors, is widely used in regenerative medicine. However, the effect of PRP on inflammatory bone destruction remains unclear. We investigated the effects of PRP on the viability of MSC, and the impact of different concentrations of PRP (1% and 3%, respectively) on cell death, proliferation, and differentiation. Furthermore, we tested the therapeutic effect of different concentrations of PRP (1% and 3%) on LPS-induced inflammatory bone destruction in vivo. PRP enhanced the cellular activity of MSC and promoted osteogenesis. A higher concentration of PRP (3%) primarily reduced the death and increased the proliferation of in LPS-treated MSC via the PI3K/AKT pathway, while a lower concentration of PRP (1%) promoted MSC differentiation into osteoblasts through the MAPK pathway. Consistent with in vitro experiments, we validated the protective effect of PRP against LPS-induced bone loss by increasing bone formation in vivo. These results suggest that different concentrations of PRP can ameliorate LPS-induced inflammatory bone loss through distinct mechanisms.

We have previously defined the constitutive photomorphogenic protein 1 (COP1) gene as a therapeutic target in hepatocellular carcinoma (HCC). A recent study demonstrated that COP1 induces non-alcoholic fatty liver disease (NAFLD), a precursor to HCC, in normal hepatocytes and that reducing COP1 expression significantly improves high-fat diet-induced hepatic steatosis. Thus, in this study, we investigated if the function of COP1 was associated with HCC metabolism and evolution. Silencing of COP1 expression by a target siRNA significantly suppressed long-term colony formation in Huh7, HepG2, Huh1, and PLC/PRF/5 HCC cell lines. RNA sequencing of COP1-silenced Huh7 and HepG2 cells revealed the same directional regulation of 24 (14 up- and 10 down-regulated) genes. Notable molecular alterations were upregulation of AKR1D1 and downregulation of TMEM65, which involves negative regulation of lipid metabolism and promotion of metastasis, respectively. Correlation analysis using GEPIA2 supported inverse relationship between COP1 and AKR1D1 expression and positive relationship between COP1 and TMEM65 expression in HCC clinical samples. Targeting of COP1 reduced fat accumulation and metastatic potential in both HCC parental cells and CD133+ liver cancer stem cells. Overexpression of COP1 reversed the phenotypic changes. Collectively, our findings indicate that the COP1 is functionally correlated with HCC lipid metabolism and stemness.

What is the current landscape of randomized controlled trials (RCTs) evaluating stem cell-based therapies for women's reproductive diseases, and how effectively has preclinical research informed their clinical translation?

Nucleic acid-based therapeutics, which involve the manipulation of genetic materials to treat or prevent diseases, have gained considerable attention, leading to the approval of medicines such as COVID-19 vaccines, patisiran (Onpattro), and nusinersen (Spinraza). However, their clinical application is hindered by challenges such as nuclease degradation, poor biodistribution, limited cellular uptake, and inefficient endosomal escape. Extracellular vesicles (EVs), which are natural nanoscale drug delivery systems derived from various eukaryotic and prokaryotic cells, offer a safe, efficient, specifically targeted, and non-pathogenic method for nucleic acid delivery. In this review, we summarize the classical methods and the latest research advances in EV preparation and nucleic acid loading. Additionally, we review the primary administration routes for nucleic acid-loaded EVs, such as intravenous, local, oral, intranasal, and inhalation delivery. By addressing these aspects, this review aims to guide the optimal design and clinical application of nucleic acid-loaded EVs.

In vitro gametogenesis (IVG) is a technology that allows the creation of gametes from stem cells. Given that IVG makes possible the production of cross-sex gametes, IVG, if successful, can, among other uses, allow a single person to solo reproduce (i.e. have children without a sperm or egg donor). This would involve using IVG to produce one cross-sex gamete and fusing that gamete with a same-sex gamete which is either artificially or naturally derived from the same person. Because solo reproduction is a highly experimental, artificial and asexual method of reproduction, it might be comparable in many ways with reproductive cloning. One of the arguments that has been made against cloning and, by extension, solo reproduction is that it expresses and encourages vices like hubris and narcissism. In this paper, we argue that the vice argument is insufficient to justify a ban on solo reproduction. This is because the mere fact that an act is vicious does not give anyone any kind of enforceable claim on us to not act viciously.

Necroptosis is an inflammatory programmed cell death pathway linked to diverse physiological and pathological disorders, yet its role in Enterotoxigenic Escherichia coli (ETEC)-induced intestinal inflammation and mucosal injury remains poorly understood. This study aimed to elucidate the contribution of intestinal epithelial cell necroptosis to the development of intestinal inflammation and injury induced by ETEC infection in piglets. Following ETEC challenge in piglets, key proteins involved in necroptosis, including phosphorylated receptor-interacting protein kinase 3 (p-RIP3) and high-mobility group box 1 (HMGB1), were upregulated in jejunal crypt epithelial cells, which are primarily composed of Paneth cells and stem cells, in a time-dependent manner. In addition, ETEC challenge triggered time-dependent pyroptosis in jejunal lamina propria lymphocytes, a population that includes macrophages, as demonstrated by elevated levels of NLRP3, Caspase-1, GSDMD-N, and Cleaved -IL-1β (p17) proteins in lamina propria lymphocytes. Necroptosis of jejunal crypt epithelial cells occurred prior to pyroptosis of lamina propria lymphocytes, indicating that epithelial cell necroptosis may contribute to the induction of pyroptosis in lamina propria lymphocytes. ETEC challenge induced progressive TNF-α and IL-1β upregulation in plasma, jejunal crypt epithelial cells, and lamina propria lymphocytes of piglets. These changes coincided with intestinal injury and barrier loss, which were indicated by increased plasma i-FABP and decreased jejunal ZO-1 and Occludin. Notably, Nec-1 pretreatment mitigates ETEC-induced intestinal inflammation and tissue damage in piglets by inhibition of crypt epithelial cells necroptosis and the ensuing pyroptosis of lymphocytes. These results indicate that targeting upstream epithelial-cell necroptosis is an important strategy to attenuate inflammation and preserve barrier integrity.

Gliomas are the most common primary malignant tumors of the adult central nervous system, characterized by rapid growth, high recurrence rates, and limited response to standard treatments, with median survival under 15 months. The SIX transcription factor family has been implicated in tumor development, but the role and regulatory mechanism of SIX5 in glioblastoma (GBM) remain unclear. This study systematically investigates the biological function of SIX5 and its regulatory network in GBM. Differential expression and weighted gene co-expression network analyses of GSE4290 and GSE50161 datasets, combined with machine learning algorithms including LASSO, identified SIX5 as a core candidate gene. Functional enrichment analyses and evaluation using TCGA and UALCAN databases revealed that SIX5 is highly expressed in GBM and associated with poor prognosis. Single-cell RNA sequencing and spatial transcriptomics showed enrichment of SIX5 in the tumor core and in astrocyte-like and stem cell-like subsets at the invasion front. In vitro, U87 and U251 cells with lentivirus-mediated SIX5 knockdown or overexpression were assessed for proliferation, migration, invasion, apoptosis, and colony formation. SIX5 knockdown significantly inhibited proliferation, migration, invasion, epithelial-mesenchymal transition, and tumorigenicity, while promoting apoptosis. Mechanistically, KDM5C positively regulates SIX5, which directly binds the UBE2C promoter to activate its transcription, enhancing AKT/mTOR signaling and promoting aerobic glycolysis via upregulation of GLUT1, HK2, PGK1, and LDHA. Rescue experiments showed that UBE2C overexpression partially restored malignant phenotypes under SIX5 downregulation. In vivo xenograft studies confirmed that the KDM5C-SIX5-UBE2C axis drives GBM growth. In conclusion, SIX5 functions as a critical oncogenic driver in GBM, regulated by KDM5C and promoting tumor progression through UBE2C-mediated activation of AKT/mTOR signaling and glycolytic reprogramming. The KDM5C-SIX5-UBE2C regulatory axis represents a potential prognostic biomarker and therapeutic target in glioblastoma.

ARG1 catalyzes the conversion of L-arginine to L-ornithine, urea, polyamines, and L-proline, thereby balancing nitrogen detoxification with tissue-specific roles in proliferation and immunity. This review delineates the context-dependent functions of ARG1 across diverse cell types-including tumor cells, immune cells, endothelial cells, keratinocytes, and stem cells. In tumors, ARG1 drives immunosuppression and metabolic reprogramming but can paradoxically suppress tumorigenesis. Immune modulation via ARG1-polyamine crosstalk regulates T cell differentiation, macrophage polarization, and microbiota interactions, influencing infection and autoimmunity. Endothelial ARG1 exacerbates obesity-related vascular dysfunction, while keratinocyte ARG1 impacts wound healing and psoriasis. Emerging therapies-such as ARG1 inhibitors, engineered extracellular vesicles, and microbiome interventions-show preclinical promise in cancer, cardiovascular, and neurodegenerative diseases. By mapping ARG1's spatiotemporal metabolic networks, this work highlights its dual roles and positions ARG1 as a central player for precision medicine in complex pathologies.

A diverse range of innovative biological therapies is being developed to treat various human diseases. The safety assessment of these biologics is a critical factor determining clinical success. Enhanced humanised mouse models have the potential to revolutionise immunotoxicological profiling by refining procedures for effective in vivo safety assessments.

Mechanical forces act throughout the body across multiple scales, from organs and tissues to cells and molecules, playing a vital role in maintaining tissue integrity, regulating cellular functions and supporting physiological performance. Importantly, alterations in mechanical forces and properties can be hallmarks of tissue injury and disease, and can thus serve as valuable biomarkers for disease monitoring and diagnostics and can be harnessed to modulate biological processes for therapeutic benefit. This concept, termed mechanomedicine, offers an important strategy in disease diagnosis and therapy. In this Review, we first introduce biomechanics and mechanobiology as the underlying principles of mechanomedicine and outline the properties and measurements of key mechanical signatures in health and disease. We then explore the application of mechanomedicine across scales, from organ-level and tissue-level diagnostics to cellular and molecular mechanotherapeutics, including strategies for tissue regeneration and rehabilitation. Finally, we highlight challenges and opportunities in the clinical translation of mechanomedicine approaches, in particular with regards to the innovation of materials and devices, the manufacturing of cells and organoids, the definition and standardization of mechanical biomarkers, and the integration of artificial intelligence.