The Management of Myelomeningocele Study (MOMS) established that prenatal repair of myelomeningocele (MMC) improved the rate of ventriculoperitoneal shunt insertion for the treatment of hydrocephalus, compared to postnatal repair. While there was mild improvement of motor function, most children were unable to ambulate independently. Therefore, the ongoing phase 1/2a clinical trial Cellular Therapy for In Utero Repair of Myelomeningocele (CuRe trial) aims to further improve distal neurological function using placenta-derived mesenchymal stem/stromal cells seeded on an extracellular matrix (PMSC-ECM) to augment open fetal repair of MMC. Here, the authors present a video of the neurosurgical repair using the PMSC-ECM. The video can be found here: https://stream.cadmore.media/r10.3171/2026.1.FOCVID25229.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen threatening the global swine industry, with existing vaccines failing to provide broad protection due to the high viral genetic variability and immune evasion capacity. Retinoic acid (RA), a vitamin A metabolite known for immunomodulatory functions, has been reported to enhance antiviral innate immunity and suppress excessive inflammation. However, whether RA affects PRRSV infection remains unclear.

Glioma-associated microglia/macrophages (GAMs) have traditionally been described as immunosuppressive. However, within their highly heterogeneous repertoire, pro-phagocytic and cytotoxic subsets with anti-tumoral properties exist. Although macrophages (MACs) can exhibit tumor-suppressive functions, their anti-glioma properties largely remain elusive. To identify anti-glioma myeloid cell effectors, we performed directionally concatenated ligand-receptor (L-R) interactome analyses from dendritic cells (DCs) and microglia (MG) to lymphoid (CD4+T, Tregs, CD8+T, NK, and NKT) cells identified by our recent single-cell transcriptomics interrogation of tumor-associated CD45+ leukocytes from tumor brains of eighteen isocitrate dehydrogenase (IDH)-stratified glioma patients. Within DC-specific and MG-specific interactomes, we identified LGALS9, which encodes Galectin-9, as a key mediator of cell-cell interactions in IDH-wild type (IDH-wt) gliomas. Spectral cytometry, immunohistochemistry, and Western blotting analyses confirmed the abundant expression of Galectin-9 in glioma-associated MG, but not in tumor cells. Furthermore, differential gene enrichment analyses revealed transcripts associated with cellular adhesion (coronin 1A, integrin) and phagocytosis (FcγR, phospholipase D, Rab family proteins, etc.) pathways that were significantly upregulated in Galectin-9+ compared to Galectin-9- MG and MACs. Using an ex-vivo primary human microglia (pMG) and patient-derived glioma stem cells (GSCs) co-culture system, we evaluated the functional role of Galectin-9. Confocal imaging analyses of co-cultured pMG with GSCs revealed that siRNA-mediated knockdown or antibody-based neutralization of Galectin-9 significantly impaired pMG-GSC adhesion. In addition to reduced adhesion, phagocytosis of GSCs was dramatically attenuated across all Galectin-9 silenced or neutralized pMG. Altogether, our study underscores the unappreciated non-canonical role of Galectin-9 in MG as a regulator of glioma cell adhesion and phagocytosis that can be harnessed for glioblastoma immunotherapy.

Glaucoma, the leading cause of irreversible blindness worldwide, is characterized by the progressive loss of retinal ganglion cells (RGCs) and their axons. While elevated intraocular pressure (IOP) is a core risk factor, the pathogenesis of normal-tension glaucoma (NTG) remains unclear, as static IOP is within the normal range. Based on circadian fluctuations of IOP and cerebrospinal fluid pressure (CSFP), and the pressure-dependent function of the ocular glymphatic system, we propose the "dynamic trans-lamina cribrosa pressure difference (TLCPD) imbalance" hypothesis. This hypothesis posits that optic nerve damage may stem from abnormal pulse synchrony between IOP and CSFP (phase mismatch, amplitude mismatch, or abnormal frequency) rather than static TLCPD elevation alone, pending further validation. Dynamic imbalance induces RGC injury through dual mechanisms: mechanical stress on the lamina cribrosa (collagen fiber rupture, astrocyte activation) and metabolic dysfunction (ocular glymphatic clearance impairment, toxic waste accumulation), which ultimately converge on the activation of the programmed axonal degeneration (PAD) pathway-a conserved final common effector of RGC axon loss. Phase mismatch is the core pathological pattern in NTG. In contrast, high-tension primary open-angle glaucoma (POAG) is characterized mainly by amplitude mismatch and abnormal frequency, with potential coexistence and mutual influence of these mechanisms. Verifiable clinical (24-h IOP-CSFP synchronous monitoring) and animal experiments are proposed. This hypothesis may help explain unresolved clinical phenomena, provides novel diagnostic markers (a transient peak in TLCPD) and therapeutic strategies (CSFP regulation, glymphatic function enhancement, modulation of the PAD pathway), and opens new avenues for personalized glaucoma management.

Endometrial-derived hydrogels are rich in extracellular matrix proteins as well as bioactive molecules that promote cell adhesion, proliferation, and angiogenesis.

Elevated interleukin-1 beta (IL-1β) in orchitis triggers immune-mediated inflammation, disrupting spermatogenesis and impairing reproductive function.

Despite the advent of potent tyrosine kinase inhibitors (TKIs), resistance and disease persistence remain significant clinical challenges in chronic myeloid leukemia (CML). This perspective aims to synthesize concepts derived from recent advances in single-cell and multi-omics analyses, which have revealed profound heterogeneity among leukemic stem cells (LSCs). These findings augment traditional models that focus solely on clonal selection and resistance-conferring mutations. We discuss how LSCs, like normal hematopoietic stem cells (HSCs), exist in a spectrum of transcriptionally and epigenetically defined cell states, each governed by distinct gene regulatory networks (GRNs) that confer unique lineage biases and responses to therapy. Incorporating recent insights from single-cell analysis, our perspective highlights evidence for a conserved chronic phase (CP) LSC state characterized by lineage skewing, altered metabolic and environmental responsiveness, and epigenetic dysregulation, features that are likely to be underpinned by specific GRN configurations that collectively contribute to intrinsic TKI resistance. We explore how both intrinsic factors (such as germline polymorphisms and lineage bias) and extrinsic cues (including microenvironmental signals, immune interactions, and hypoxia) are likely to modulate GRN activity and LSC states, thereby affecting apoptotic thresholds, primary resistance, and the potential for treatment-free remission (TFR). Emerging data support the concept of GRNdefined LSC states at diagnosis that are predictive of TKI responses. Furthermore, multiple studies suggest that blast crisis (BC) converges on a common high-risk transcriptomic and GRN state that is agnostic to mutational diversity, and driven by polycomb and DNA methylation-dependent epigenetic reprogramming. Given that BCR::ABL1-independent mechanisms, regulated at the level of GRNs, may contribute to resistance and LSC persistence, these observations support placing greater emphasis in CML management on addressing GRN-defined cell-state vulnerabilities, with the goal of lowering the risk of blast crisis in high-risk patients and improving control of therapy-resistant CP LSCs.

Mobilized hematopoietic stem and progenitor cells (HSPCs) are essential for transplantationbased therapies, including curative gene therapies for sickle cell disease (SCD). While granulocyte colony-stimulating factor (G-CSF, filgrastim) remains the standard mobilization agent, many patients respond inadequately, and it can trigger life-threatening vaso-occlusive crises in SCD. The CXCR4 antagonist AMD3100 (plerixafor) is routinely combined with G-CSF for non- SCD settings but is ineffective as a single agent in SCD, underscoring the urgent need for alternative strategies. We previously identified 27-hydroxycholesterol (27HC) as a physiological inducer of HSPC mobilization during pregnancy. Here, we show that exogenous 27HC enhances AMD3100-induced HSPC mobilization in mice, either alone or with G-CSF. Because 27HC is metabolized by the enzyme Cyp7b1, we tested whether pharmacological Cyp7b1 inhibition could mimic this effect. Treatment with clotrimazole, an antifungal and Cyp7b1 inhibitor, significantly enhanced AMD3100-induced HSPC mobilization in wild-type, SCD, and humanized mice. Importantly, intravenous administration of voriconazole, a clinically approved systemic antifungal with Cyp7b1-binding activity, similarly augmented AMD3100-induced HSPC mobilization in wildtype and SCD mice without altering steady-state hematopoiesis. These findings establish Cyp7b1-inhibiting azoles as novel and clinically relevant enhancers of HSPC mobilization, particularly for SCD patients who cannot safely receive G-CSF but require robust HSPC yields for gene therapy.

The environmental pollutant benzo(a)pyrene (B(a)P), a representative polycyclic aromatic hydrocarbon (PAH), is a recognized carcinogen, and chronic pulmonary inflammation is closely associated with lung carcinogenesis. Although alveolar type 2 (AT2) cells are the origin of lung adenocarcinoma, the genetic and functional changes in AT2 cells and the mechanisms involved in inflammation-related lung tumorigenesis have not been elucidated. Here, C57BL/6J mice are exposed to B(a)P and the inflammatory irritant lipopolysaccharide (LPS) to establish a model of inflammation-related lung tumorigenesis. Single-cell RNA sequencing is performed on lung tissues. DNA mutations in AT2 cells are analyzed via whole-exome sequencing. The protein expression of AT2 cells in lung cancer tissue is determined by immunofluorescence staining. The results reveal that LPS promotes B(a)P-induced lung tumorigenesis; in the whole lungs of B(a)P/LPS, a decreased proportion, altered differentiation trajectory, and increased gene mutation number in AT2 cells are observed. Additionally, in B(a)P/LPS-treated lung cancer tissue, the levels of γ-H2AX DNA damage and the proliferation marker Ki67 in AT2 cells are increased, whereas the levels of differentiation markers are decreased. Single-cell RNA transcriptomics reveals that the autophagy-related genes Foxo3 and Ppp2r5, which are enriched in the PI3K-Akt pathway, and the autophagy-related genes in AT2 cells in lung cancer are decreased in the B(a)P/LPS group. Thus, chronic inflammation promotes DNA damage, gene mutation and dysfunction in AT2 cells, and decreased autophagy in AT2 cells may be an important mechanism for inflammation-related lung tumorigenesis.

Interleukin 16 (IL16) is a pleiotropic cytokine that does not belong to any interleukin family and functions as a CD4+ T-cell chemokine, a regulator of T-cell activation, and an inhibitor of human immunodeficiency virus replication in acquired immunodeficiency syndrome. Studies in the hemato-oncological disease multiple myeloma have demonstrated that IL16 plays an important role in promoting tumor cell growth and is associated with tumor burden and prognosis. However, the role of IL16 in acute myeloid leukemia (AML) remains largely unexplored.

Allogeneic hematopoietic stem cell transplantation can achieve durable HIV remission when donor cells exhibit reduced or absent C-C chemokine receptor type 5 (CCR5) function, but CCR5Δ32/Δ32 donors are rare and cord blood banks (CBBs) do not routinely screen inventories for CCR5-deficient units. We evaluated a scalable flow cytometry-based phenotyping strategy to identify cord blood units (CBUs) with intrinsically reduced CCR5 surface expression and to determine whether low-expression phenotypes enrich for CCR5Δ32 and other CCR5 variants.

The thymic medulla is essential for establishing central tolerance, orchestrating the development of a diverse yet self-tolerant T cell repertoire, and preventing autoimmunity. This process is primarily mediated through interactions between developing thymocytes and antigen-presenting cells, including thymic epithelial cells (TECs) and dendritic cells (DCs), with additional regulatory contributions from endothelial cells, mesenchymal cells, and macrophages. Despite its critical role, the complexity of late-stage thymocyte development and the dynamics of their medullary residency remain incompletely understood. Recent advances in single-cell, epigenomic, and transcriptomic technologies have begun to reveal previously unappreciated layers of cellular and molecular heterogeneity within the thymic medulla throughout life. In this review, we explore how the medulla shapes the fate of both conventional and non-conventional T cells, examine the diversity of thymocyte populations it supports, and discuss how this specialized microenvironment adapts during aging and regeneration.

In mammalian genomes, at least several thousand copies of transposable elements (TEs) may function as enhancers or promoters that regulate gene expression, cellular processes, and development. However, it is still largely unknown how many TEs have been co-opted into regulatory processes and under which cellular situations they are functional. In particular, few studies have addressed how TE functions change during cell differentiation.

Embryonic hematopoiesis involves successive waves of progenitors from distinct anatomical sites, but the origins and contributions of early hematopoietic stem and progenitor cells (HSPCs) remain incompletely defined. Here we use genetic fate mapping in mice to temporally label hemogenic endothelium (HE) subsets and track their progeny. We show that a wave of fetal-restricted HSPCs arises from HE in the vitelline and umbilical arteries between embryonic days 8.5 and 9.5, preceding the emergence of definitive hematopoietic stem cells. Lineage tracing, single-cell transcriptomic analyses and functional assays revealed that these progenitors are transient and distinct from erythro-myeloid progenitors, contribute extensively to fetal lympho-myelopoiesis but decline postnatally. Our findings reveal a previously unrecognized early HE wave as a key source of fetal-restricted HSPCs, refining the spatial-temporal understanding of layered hematopoiesis and informing developmental origins of blood cell diversity.

Claudin-low breast cancers (BCs), representing approximately 1.5-14% of all BCs, are characterized by high aggressiveness, enriched cancer stem cell (CSC) population, and poor prognosis. Despite the established role of Notch signaling in mammary gland development and BC progression, its status in claudin-low BCs remains inadequately explored. In this study, Notch pathway activation was evaluated in BC cell lines and 107 patient samples. Claudin-low subtype exhibited elevated Notch activity. Notch1 was observed to be the predominant receptor in the above subtype, whereas Notch3 was predominant in the claudin-high cancers. Notch1 and HES1 expression showed a significant inverse correlation with claudins 3, 4, and 7, and were positively associated with aggressive tumor features including high Ki67 index, higher grade, and increased metastasis. Immunohistochemical analysis further confirmed a correlation between nuclear Notch1 (N1ICD) expression and claudin-low status, supporting its potential as a biomarker for identification of aggressive BC. Combined treatment with celecoxib (10 µM) and doxorubicin (1 µM) in claudin-low cells not only significantly inhibited Notch signaling and claudin expression, but also suppressed viability, proliferation, migration, and BCSC populations. Since Notch signalling may be an essential factor in these latter events, our findings suggest that Notch1/N1ICD can serve as therapeutic targets for the better management of claudin-low BCs. However, validation of the same requires detailed functional studies involving modulation of each type of Notch receptor or other players involved in Notch signaling using more robust approaches.

Microtubules scaffold cells, supporting signaling and cargo transport. They assemble from GTP-tubulin, which hydrolyzes to GDP-tubulin during polymerization. GTP-microtubule lattices are stable; GDP lattices depolymerize rapidly. In vitro, hydrolysis triggers lattice compaction. Lattice spacing regulates motors and microtubule-associated proteins; however, the conformation of tubulin in microtubules in cells is unknown. Here, we present the atomic-resolution cryo-electron microscopy structure of human microtubules in situ, in the axons of human cortical neurons derived from induced pluripotent stem cells (iPS cells). Our 2.7-Å-resolution reconstruction delineates bound water molecules and reveals that axonal microtubules adopt an expanded GTP-like lattice, despite being GDP bound. Using cryo-electron tomography and power spectrum analysis, we find that, unlike in axons, microtubules in undifferentiated iPS cells are compacted. Therefore, lattice expansion is part of neuronal differentiation. Our work provides molecular insights into neurogenesis and has implications for understanding microtubule stability and effector recruitment in neurons.

Lung cancer leads to a series of physiological abnormalities. The remodeling of extracellular matrix (especially elastin and collagen fibers) has been drawing increasing attention as it is suggested to be a hallmark of tumorigenesis. However, the interaction between these crucial matrix components, together with their relationship to mechanical changes, remains poorly understood. Here, we develop a quantitative multiphoton microscopy system to elucidate the relationship between tissue stiffening and elastin-collagen interplay in lung cancer. Based on label-free images of both fibers, we establish a metric termed resemblance metric (RM) to characterize their interaction by quantifying the similarity of their morpho-structural distributions. Specifically, RM is found to increase with lung tumorigenesis, and exhibits superior sensitivity in identifying human lung cancer through ex vivo quantitative imaging. Nanoindentation results suggest a strong correlation between tissue stiffness and inter-channel interaction, notably greater than that between stiffness and any individual morpho-structural feature of either fiber type. Finally, the translational potential of RM-based imaging is demonstrated through tumor boundary identification via in vivo imaging within a mouse model harboring human lung cancer.

Cell-type-specific promoters are used in gene therapy to restrict expression of the therapeutic payload. However, these promoters often have suboptimal strength, selectivity and size. Here, leveraging recent insights into the function of enhancers, we developed synthetic super-enhancers (SSEs) by assembling functionally validated enhancer fragments into multipart arrays. Focusing on the core SOX2-driven and SOX9-driven transcriptional regulatory network in glioblastoma stem cells (GSCs)1, we engineered SSEs with robust activity and high selectivity. Single-cell profiling, biochemical analyses and genome-binding data indicated that SSEs integrate neurodevelopmental and signalling-state transcription factors to trigger the formation of large multimeric complexes of transcription factors. Moreover, GSC-selective expression of a combination of cytotoxic (HSV-TK and ganciclovir) and immunomodulatory (IL-12) payloads, delivered using adeno-associated virus vectors, as a single treatment led to curative outcomes in a mouse model of aggressive glioblastoma. Notably, IL-12 induced an immunological memory that prevented tumour recurrence. The activity and selectivity of the adeno-associated virus and SSE were validated using primary human glioblastoma tissue and normal cortex samples. In summary, SSEs harness the unique core transcriptional programs that define the GSC phenotype and enable precision immune activation. This approach may have broader applications in other contexts when precise control of transgene expression in specific cell states is necessary.