Mechanical stimulation plays a crucial role in odontoblast differentiation. However, the underlying molecular mechanisms remain unclear. We have previously shown that hydrostatic pressure (HP) applied to stem cells from human exfoliated deciduous teeth (SHED) promotes odontoblast differentiation by translocating RUNX2 and increasing WNT16 expression through PIEZO1 signaling. In this study, we further explored the downstream signaling cascade linking PIEZO1 activation and odontoblast differentiation. HP stimulation increased the expression of odontoblast differentiation markers PANX3 and DSPP, as shown by qPCR, and enhanced Alizarin Red staining-results significantly suppressed by siRNA targeting either PIEZO1 or WNT16. RT-PCR analysis revealed that, among the two known human WNT16 isoforms, only WNT16b was expressed in SHED. qPCR demonstrated that HP-induced WNT16 expression was reduced by siPIEZO1 and further decreased by siRUNX2. Promoter analysis identified four RUNX2-binding sites within the upstream region of WNT16. A luciferase reporter assay using plasmids containing the WNT16 promoter showed that RUNX2 overexpression in HEK293 cells significantly increased luciferase activity. Moreover, HaloChIP assays with a HaloTag-RUNX2 expression vector confirmed RUNX2's binding to the WNT16 promoter. These findings suggest that PIEZO1-mediated mechanical stress promotes odontoblast differentiation through the RUNX2-dependent transcriptional activation of WNT16.

Inflammatory bowel disease (IBD) is secondary to an abnormal immune response to the microbiota. To study this, models of host-microbe interactions that represent mucosal bacterial communities and inter-patient diversity are required. Human intestinal organoids (HIOs) are an established model to investigate epithelial responses. Here, we describe a technique of culturing bacteria directly from the sites of inflammation in IBD, while simultaneously sampling host tissue. We generated HIOs from a cohort of newly diagnosed paediatric IBD patients, without confounding treatments or comorbidities, and explored their response to site-specific bacteria. A unique biobank of matched HIOs and cultured mucosa-attached bacteria was established from 27 paediatric patients. Transcriptional profiling revealed differential gene expression between control and IBD-derived organoids. We used microinjection to introduce bacteria to the apical surface of the epithelium, to determine the effect of bacteria on host epithelial cells. We measured survival and growth of bacteria within the HIOs and tested several related bacterial isolates for their impact on the epithelium. An isolate from a control patient stimulated inflammatory signalling pathways but this was not observed in response to a closely related isolate originating from an IBD patient. This study demonstrates the feasibility of isolating bacteria and generating organoids from the same biopsy tissue, to explore personalised host-microbe interactions. The microinjections, while labour-intensive, demonstrate that closely related bacteria can induce very different epithelial responses, with downstream implications for immune response. This highlights the importance of understanding host-microbe interactions in a strain- and site-specific manner and developing techniques for personalised microbiome-based therapeutics.

Bladder cancer is a common urogenital malignancy that remains difficult to treat, particularly due to therapeutic resistance, such as resistance to cisplatin, in which cancer stem cells (CSCs) play a central role. This study investigates the combination of 5-aminolevulinic acid-mediated photodynamic therapy (5-ALA-PDT) and berbamine as a potential multimodal treatment strategy using the bladder cancer cell lines RT112 and J82, their cisplatin-resistant variants, and generated CSC-like cells. Berbamine is a natural plant compound and was confirmed in this study to have anticancer properties by inhibiting cell migration and invasion, and by inducing apoptosis. This study also showed that berbamine enhances the accumulation of protoporphyrin IX (PpIX), the photosensitizer induced by 5-ALA. 5-ALA-PDT destroys cancer cells by stimulating PpIX via 635 nm red laser light to produce reactive oxygen species (ROS). This was found to happen in all tested cell lines, whereas berbamine could modulate the cell destruction in a concentration-dependent manner and was influenced by the specific biological characteristics of the tested cell variants. CSCs showed the strongest response to the combination therapy approach, suggesting that they may represent more vulnerable cell variants to the tested treatment. Cisplatin-resistant cell lines could also be treated successfully with 5-ALA-PDT, whereas berbamine could enhance its efficacy in the cisplatin-resistant J82 LTT. These findings suggest that the combination treatment of 5-ALA-PDT and berbamine may serve as a promising approach to overcome therapeutic resistance in bladder cancer, particularly in cisplatin-resistant and CSC-enriched tumour types.

Gastrointestinal dysfunction often precedes motor symptoms in Parkinson's disease (PD), suggesting the enteric nervous system (ENS) is central to early pathogenesis. How α-synuclein contributes to ENS dysfunction, and how inflammation modulates this, remains unclear. Here we show that Tumor Necrosis Factor alpha enhances α-synuclein accumulation in induced pluripotent stem cell-derived enteric neurons and glia, and impairs the malate-aspartate shuttle, a key pathway for mitochondrial energy production. This drives a metabolic shift toward glutamine oxidation in patient cells. This metabolic impairment reduces overall mitochondrial function, which is partially rescued by the neuroprotective compound Chicago-Sky-Blue 6B. Furthermore, transcriptomic and histological analyses of human gut tissue from inflammatory bowel disease patients reveal that inflammation-associated metabolic suppression and α-synuclein upregulation occur beyond PD, representing general hallmarks of intestinal inflammation. These findings highlight a conserved metabolic vulnerability in the ENS and establish patient-derived enteric lineages as a robust platform to model inflammatory ENS pathology.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with limited treatment options, where neuroinflammation and gut microbiota dysbiosis are emerging as interconnected therapeutic targets. This study evaluated the therapeutic potential of non-gene-edited human chemically induced pluripotent stem cell-derived neural stem cells (hCiPSC-NSCs) in a prenatal valproic acid (VPA)-induced rat model of ASD, using a dual-pathway administration strategy (intravenous systemic delivery combined with an intracerebroventricular boost). The treatment significantly ameliorated core ASD-like behaviors, including improved sociability (increased stranger interaction time, P < 0.0001), reduced repetitive behaviors (decreased marble-burying, P < 0.0001; and self-grooming, P < 0.05), and enhanced spatial memory (shorter escape latency in the Morris water maze, P < 0.01). At the mechanistic level, hCiPSC-NSCs attenuated neuroinflammation (suppressed IL-1β, IL-6, and TNF-α; elevated IL-10, all P < 0.0001), reduced oxidative stress (restored GSH and SOD, decreased MDA and NO), diminished microglial activation in the hippocampus and cortex, and restored synaptic ultrastructure by replenishing synaptic vesicles. Furthermore, 16S rRNA sequencing revealed a rebalancing of the gut microbiota, characterized by a reduced Firmicutes/Bacteroidota ratio, enrichment of beneficial taxa like Bacteroidota and Alloprevotella, suppression of pathobionts such as Desulfovibrionales, and partial restoration of microbial diversity. These findings demonstrate that non-gene-edited hCiPSC-NSCs can simultaneously address neural pathophysiology and gut ecosystem disruption in ASD, highlighting their potential as a gut-brain axis-targeting therapy for neurodevelopmental disorders.

Habitat destruction, changing climate and other anthropogenic impacts have resulted in the recorded extinctions of hundreds of species, with many more undocumented extinctions being likely to have occurred. Approaches to conserving threatened species include protection or improvement of habitat, fenced conservation reserves, species translocations and reintroductions, elimination of environmental toxins, breeding programs in reserves or captivity, and genetic rescue and management. The latter includes storage of gametes, stem cells or embryos, to both conserve species and maintain or expand their genetic diversity. Many of these approaches require a basic knowledge of the reproductive biology of the species of interest. Such knowledge is difficult to achieve because of the astonishing diversity of species-specific reproductive strategies that have evolved. Unfortunately, for many species we simply do not have that knowledge. This report summarises key discussions from a workshop titled Reproductive Biology Research Needed for Saving our Wildlife held in Melbourne, Australia, and attended by stakeholders from zoos, wildlife organisations, universities, museums and government organisations. The workshop prioritised aspects of reproductive biology knowledge needed, how this knowledge might be obtained, and how it should be deployed. Using examples of planned and successful conservation strategies for individual species, the workshop participants considered environmental challenges, managing introduced species, captive breeding programs, challenges for assisted reproductive technologies, de-extinction science in conservation efforts, examination of reproductive steroid hormones across species, endocrine disruption, and cryopreservation of genomic diversity to assist the management of wild and captive populations. The workshop highlighted the magnitude of the issues involved and identified reproductive approaches to be used to direct future conservation efforts for saving threatened species.

The hepatitis E virus (HEV) is a leading cause of acute hepatitis worldwide. Although most infections are self-limiting, zoonotic genotypes can persist in immunocompromised individuals. Transmitted via the fecal-oral route, HEV has been suggested to directly infect the intestinal epithelium, a tissue with high regenerative capacity. Here, we demonstrate that HEV predominantly infects proliferative transit-amplifying and intestinal stem cells within the crypts of human pluripotent stem cell-derived intestinal organoids (hIOs). Supporting this, we detected HEV RNA in the intestinal crypts of an HEV-infected patient. We further found that HEV infection spreads through cell division and is maintained in hIOs for more than 40 days, contrasting with acute hepatitis A virus, whose infections are rapidly cleared from hIOs. Given the self-renewal ability and metabolic constraints of proliferative intestinal progenitor cells, our findings suggest that intestinal crypts could serve as reservoirs for chronic HEV infection and highlight the intestinal crypt as a primary target for viral infection in the gastrointestinal tract.

Dengue virus infection is a major global health problem with clinical outcomes ranging from mild febrile illness to severe disease. Although a reduction in platelet counts is a common feature of dengue, the mechanisms underlying the presence of viral antigens in circulating platelets remain incompletely understood. In this study, we investigated whether infection of megakaryocytic precursors contributes to the generation of platelets carrying dengue virus proteins. Using the human megakaryocytic precursor cell lines K562 and MEG-01 cell lines as in vitro models, we show that reference strains and clinical isolates of dengue virus infect megakaryocytic precursors and modulate their differentiation, as evidenced by increased expression of the differentiation marker CD41 and the production of platelet-like particles. Flow cytometry analysis demonstrated that PLPs released from infected precursors contained precursor-derived PLP-associated dengue virus envelope protein. Correlation analyses revealed significant associations between the extent of precursor infection and both differentiation markers and PLP-associated E protein levels at early time points. Consistent with these findings, dengue virus E protein was detected in platelets from dengue patients, while no statistically significant differences were observed between clinical severity groups, a trend toward higher proportions of E+ platelets were observed in patients with dengue with warning signs or severe dengue. Spearman correlation analyses further supported an association between in vitro precursor infection and the generation of E-positive PLPs. In contrast, platelets from healthy donors incubated ex vivo with dengue virus did not acquire E protein, indicating that direct uptake or infection of circulating platelets is inefficient. Together, these results support a model in which infection of megakaryocytic precursors contributes to the generation of platelets carrying dengue virus antigens, suggesting that platelet-associated E protein may reflect bone marrow involvement during dengue infection.

Transition from a pluripotent to a differentiated cell state is accompanied by significant changes in genome organization. Activity dependent neuroprotective protein (ADNP) is a chromatin regulator with critical roles in neurodevelopment and limits the genomic occupancy of CTCF, a master architectural protein in genome organization, in embryonic stem cells. However, ADNP localization, function, and relationship with CTCF in differentiated neural lineages are not well studied. Here we develop a dual degron model which allows us to acutely deplete ADNP in neural progenitor cells (NPCs). We find that ADNP depletion does not impact NPC survival in the short term, but results in a genome organization switch, which favors the formation of short-range chromatin looping interactions coinciding with CTCF accumulation. Furthermore, ADNP localizes to active gene promoters in NPCs that are unoccupied by CTCF, where it prevents over-expression of genes that are activated upon neurodifferentiation and represses those involved in commitment to other lineages. Our findings uncover CTCF-dependent as well as CTCF-independent regulatory mechanisms of ADNP in NPC-specific chromatin organization and gene expression programs that may underlie its essential function in neurodevelopment.

PD-1+TCF1+ stem-like CD8 T cells are a progenitor population that provides a proliferative burst of effector CD8 T cells upon PD-1 blockade therapy, making them a key therapeutic target in antiviral and anticancer immunotherapy. Here, we show that IL-7 therapy preferentially expands these stem-like CD8 T cells, using NT-I7, a long-acting Fc-fused recombinant human IL-7 (efineptakin alfa). Gene profile analysis showed that a proliferating stem-like cluster induced by NT-I7 exhibited enrichment of genes related to lymphocyte migration. After NT-I7 treatment, proliferation of stem-like cells in the spleen was initiated within the white pulp, followed by egress into the red pulp. NT-I7 treatment led to an increase in stem-like CD8 T cells in circulation and peripheral tissues, contrasting with their resident property in lymphoid organs. These findings suggest NT-I7 as a promising strategy to expand and mobilize stem-like CD8 T cells to enhance antiviral and anticancer immunotherapies.

High recurrence rates, significant metastatic potential, and limited overall survival make triple-negative breast cancer (TNBC) the most challenging subtype among breast cancers. Previous studies have indicated that the downregulation of TGFB2-AS1 can enhance the stem-like properties of tumor cells, thereby promoting TNBC progression. Bioinformatics analysis has revealed the regulatory role of GATA6 in TGFB2-AS1 transcription, providing insights into the transcriptional regulation of TGFB2-AS1 by GATA6 and offering potential prognostic biomarkers and therapeutic strategies for TNBC.

Fanconi anemia (FA) is a rare inherited disorder characterized by genomic instability, bone marrow failure, and a markedly increased risk of developing acute myeloid leukemia (AML). The intrinsic hypersensitivity of FA cells to DNA-damaging agents renders conventional chemotherapy particularly toxic and often ineffective.

The prognosis of relapsed or refractory T-cell and NK-cell lymphoid proliferations and lymphomas (R/R T/NK-LPD/LYM) remains poor. Modified dendritic cell–cytokine-induced killer (DC–CIK) cells loaded with tumour stem cell (TSC) membrane microparticles (MMPs) can improve the targeted killing activity against tumours. However, the use of this treatment has not been reported for R/R T/NK-LPD/LYM. Herein, we report 3 cases. Case 1 involved an 8-year-old male who presented with bilateral cervical painless lymphadenopathy and a left elbow mass. Case 2 involved a 44-year-old man with recurrent nasal congestion and a runny nose. Case 3 involved a 49-year-old man whose bilateral cervical, axillary, and inguinal lymph nodes were enlarged. The biopsy results revealed T-lymphoblastic leukaemia/lymphoma, not otherwise specified (NOS); extranodal NK/T-cell lymphoma, nasal type; and nodal T follicular helper (TFH) cell lymphoma, angioimmunoblastic type (nTFHL-AI) for Patients 1, 2, and 3, respectively. All three patients ultimately underwent modified dendritic cell–cytokine-induced killer cell therapy loaded with tumour stem cell membrane microparticles following disease progression, relapse, and chemotherapy-associated complications after first-line therapy and multiple lines of salvage therapy. Following treatment with modified dendritic cell–cytokine-induced killer cells loaded with tumour stem cell membrane microparticles, two of these three patients achieved complete and durable remission, whereas the third patient achieved a partial response, with no treatment-related adverse events reported. At the most recent follow-up, two patients maintained stable disease (SD) with preserved quality of life, while one patient experienced progressive disease (PD) and died because of multiple organ dysfunction syndrome secondary to transplantation-associated colitis. To our knowledge, this is the first report of the use of modified dendritic cell–cytokine-induced killer cells loaded with tumour stem cell membrane microparticles for the treatment of R/R T/NK-LPD/LYM. Our findings warrant further evaluation of this novel targeted immunotherapy in future prospective clinical trials.

Cytokines mediating epithelial and immune cell interactions modulate mucosal healing-a process that goes awry with chronic inflammation as in inflammatory bowel disease. TNFSF13 is a cytokine important for B cell maturation and function, but roles for epithelial TNFSF13 and putative contribution to inflammatory bowel disease are poorly understood. We evaluated functional consequences of a novel monoallelic TNFSF13 variant using biopsies, tissue-derived colonoids and induced pluripotent stem cell (iPSC)-derived colon organoids. TNFSF13 variant colonoids exhibited a >50% reduction in secreted TNFSF13, increased epithelial proliferation, and reduced apoptosis, which was confirmed in iPSC-derived colon organoids. Single cell RNA-sequencing and flow cytometry suggested FAS as the predominant colonic epithelial receptor for TNFSF13, which was confirmed by co-immunoprecipitation and binding assays. Imaging mass cytometry revealed an increase in epithelial-associated B cells in TNFSF13 variant colon tissue sections. Finally, TNFSF13 variant colonoids co-cultured with memory B cells demonstrated a reduction in immunoglobulin-producing plasma cells compared to control colonoid cocultures. Our findings support a role for epithelial TNFSF13 as a regulator of colonic epithelial growth and epithelial crosstalk with B cells.

Cellular senescence in osteogenic mesenchymal cells contributes to age-related bone loss. The bone marrow hosts myeloid cells, the precursors of immune cells, as well as mesenchymal cells, which give rise to osteoblasts and osteocytes. The senotype and senolytic response of bone marrow cells, particularly hematopoietic cells, in age-related bone loss is unclear. In this issue, Doolittle et al. showed that of all immune cells, myeloid cells had the strongest senescence profile, yet the relative level of senescence remained lower than that of mesenchymal stromal cells. Mesenchymal cells displayed a profound senotype, rendering them susceptible to senolytic clearance protecting against bone loss. By contrast, selective clearance of p16+ myeloid cells was not long-lasting and, hence, did not fully protect against age-related bone loss. These findings underscore the challenges of developing senolytic strategies for tissues with mixed senotypes, such as bone.

Cellular senescence plays a critical role in the pathogenesis and progression of osteoarthritis (OA), contributing to articular cartilage degradation, chronic inflammation, and joint function impairment. Mesenchymal stem cells/stromal cells (MSCs) and their derivatives have emerged as potential targets for novel therapeutic strategies against cell senescence in OA, as they exert anti-aging effects in repairing damaged cartilage through multiple mechanisms-including regulating age-related signaling pathways, reducing the secretion of pro-inflammatory cytokines, and improving mitochondrial function.

β-thalassemia is an inherited blood disorder characterized by chronic anemia, ineffective erythropoiesis, and in its most severe form, lifelong transfusion dependence. The standard of care for transfusion-dependent thalassemia (TDT) is regular red blood cell transfusions to relieve the anemia and suppress ineffective erythropoiesis and iron chelation therapy to mitigate morbidity and mortality related to iron overload. Allogeneic hematopoietic stem cell transplantation is a curative option but is only available to patients with an appropriate donor and carries risks of graft-versus-host disease and other transplant-related morbidity. In recent years, the therapeutic landscape for TDT has changed dramatically with the approval of two autologous gene therapies in the United States: betibeglogene autotemcel (beti-cel) and exagamglogene autotemcel (exa-cel). Clinical trials for both gene therapies have demonstrated high rates of sustained transfusion independence for both pediatric and adult age groups. However, despite these advances, challenges remain. Gene therapy requires myeloablative busulfan-based conditioning chemotherapy, which carries the risk of short- and long-term toxicities. Furthermore, centralized manufacturing and high treatment costs are likely to limit access to gene therapy. In this review, we discuss the available clinical trial and real-world data for beti-cel and exa-cel. We describe how gene therapy fits into the current treatment landscape and introduce areas of ongoing investigation to improve access to transformative therapy for TDT.

Myocardial infarction (MI), a lethal coronary artery disease primarily triggered by atherothrombosis or an imbalance in myocardial oxygen supply and demand, stands as a leading cause of mortality worldwide. Promoting angiogenesis is recognized as an effective therapeutic strategy for MI, a process highly dependent on the functional status of endothelial cells (ECs). Small extracellular vesicles (sEVs), which are membrane-bound vesicles secreted by cells and enriched with bioactive molecules including proteins, lipids, and RNAs, are ubiquitously present in the secretome of diverse cell types such as stem cells, immune cells, and cardiac cells. Studies have confirmed that sEVs can deliver specific "cargo" such as miRNAs and cytokines via paracrine or endocrine pathways, activating key downstream signaling cascades. This effectively promotes EC proliferation, migration, and tube formation, thereby enhancing angiogenic capacity and ultimately mitigating pathological cardiac remodeling while improving prognosis post-MI. This review focuses on sEVs derived from various cellular sources, systematically summarizing their roles in promoting angiogenesis and the latest research advances in regulating EC function, aiming to provide novel insights for the effective treatment of MI.