Biology subjects

Xu, Y.

Publications and source records attributed to Xu, Y..

At least 253 records · Page 14Linked to original sources

bioRxiv2025

Decoding Lymphangioleiomyomatosis (LAM) Niche Environment via Integrative Analysis of Single Cell Multiomics and Spatial Transcriptomics

Lymphangioleiomyomatosis (LAM) is a rare, progressive lung disease characterized by cystic destruction and metastatic growth of smooth muscle-like cells. Despite advances in understanding its genetic basis, the cellular heterogeneity, regulatory mechanisms, and microenvironmental interactions driving LAM progression remain poorly defined. In this study, we employed an integrative multi-omics approach combining single-cell RNA sequencing (scRNA-seq), single-nucleus ATAC sequencing (snATAC-seq), and high-resolution spatial transcriptomics (Visium, Visium HD, and Xenium) to decode the LAM niche in its native environment. We identified two spatially and functionally distinct LAM subtypes: LAMCORE1 and LAMCORE2. LAMCORE1 cells exhibited a uterine smooth muscle-like phenotype, expressing associated markers (ACTA2, MYH11) and were enriched in MTORC1 signaling and myogenic pathways, supporting a uterine origin. In contrast, LAMCORE2 cells displayed fibroblast-like features, with upregulated extracellular matrix (ECM) remodeling genes (COL1A1, MMP11) and epithelial-to-mesenchymal transition (EMT) pathways, suggesting a role in niche formation. Pseudotime and regulon analyses revealed dynamic transitions between these subtypes, driven by distinct transcriptional networks (e.g., HOX/PBX in LAMCORE1, TWIST/ZEB in LAMCORE2). The presence of the two distinct LAM subtypes was further validated by RNAscope and immunofluorescence microscopy. We identified LAM-associated fibroblasts (LAFs) as activated stromal cells expressing canonical markers (FAP, S100A4, VIM, IGFBP7, SPARC) and localized within LAM lesions. Subpopulations of LAFs, LAF-seed (proximal to LAMCORE1) and LAF-niche (surrounding LAM niches), exhibited unique functional profiles, including ECM deposition, TGF-{beta} signaling, and myofibroblast activation. Regulatory network analysis pinpointed EGR1 as a central hub governing LAF phenotype. Our comprehensive spatial profiling revealed niche structures dominated by LAMCORE1 cells and surrounded by lymphatic endothelial cells (LECs), LAFs, scattered LAMCORE2 cells, macrophages, and reprogrammed alveolar epithelial cells (AT2). ECM remodeling and aberrant organization of cable-like structures (-smooth muscle actin+) of the LAM niches were further validated by second harmonic generation microscopy. These findings provide a high-resolution blueprint of LAM pathogenesis, highlighting the interplay between uterine-derived LAMCORE cells, activated fibroblasts, and the remodeled lung microenvironment. They significantly enhance our understanding of the LAM niche microenvironment and offer insights into potential therapeutic targets and strategies for managing this complex disease.

genomics↗

bioRxiv2025

Choreography of human mitochondrial leaderless mRNA translation initiation

It remains unclear how human mitochondrial ribosomal subunits assemble into an elongation-competent 55S particle on mRNAs devoid of 5 leader sequences. Here, we reconstituted and directly tracked human mitochondrial translation initiation using real-time single-molecule fluorescence spectroscopy. Corroborated with cryo-EM structural analysis, we show that the initiation factor mtIF2 and initiator fMet-tRNAMet are loaded to the 28S subunit to drive mRNAs binding via 5 start codon recognition. This enables sequential loading of the two ribosomal subunits onto the leaderless mRNA to initiate. In parallel, a preassembled 55S monosome can also be loaded with mtIF2 and fMet-tRNAMet to initiate on the mRNA. Both initiation pathways yield active complexes to enter translation elongation, which is gated by mtIF2. The monosome loading pathway can initiate promiscuously with non-formylated Met-tRNAMet, thus its usage may under tight regulation in cells, e.g. by mtIF3. Our work provides a dynamic framework for the distinct human mitochondrial translation initiation.

biochemistry↗

bioRxiv2025

Genomic Touchstone: Benchmarking Genomic Language Models in the Context of the Central Dogma

The emergence of genomic language models (gLMs) has revolutionized the analysis of genomic sequences, enabling robust capture of biologically meaningful patterns from DNA sequences for an improved understanding of human genome-wide regulatory programs, variant pathogenicity and therapeutic discovery. Given that DNA serves as the foundational blueprint within the central dogma, the ultimate evaluation of a gLM is its ability to generalize across this entire biological cascade. However, existing evaluations lack this holistic and consistent framework, leaving researchers uncertain about which model best translates sequence understanding into downstream biological prediction. Here we present Genomic Touchstone, a comprehensive benchmark designed to evaluate gLMs across 36 diverse tasks and 88 datasets structured along the central dogmas modalities of DNA, RNA, and protein, encompassing 5.34 billion base pairs of genomic sequences. We evaluate 34 representative models encompassing transformers and convolutional neural networks, as well as emerging efficient architectures such as Hyena and Mamba. Our analysis yield four key insights. First, gLMs achieve comparable or superior performance on RNA and protein tasks compared to models pretrained on these molecules. Second, transformer-based models continue to lead in overall performance, yet efficient sequence models show promising task-specific capabilities and deserve further exploration. Third, the scaling behavior of gLMs remains incompletely understood. While longer input sequences and more diverse pretraining data consistently improve performance, increases in model size do not always translate into better results. Fourth, pretraining strategies, including the choice of training objectives and the composition of pretraining corpora, exert substantial influence on downstream generalization across different genomic contexts. Genomic Touchstone establishes a unified evaluation framework tailored to human genomics. By spanning multiple molecular modalities and biological tasks, it offers a valuable foundation for guiding future gLM design and understanding model generalizability in complex biological contexts.

bioinformatics↗

bioRxiv2025

Rictor CRISPR Gene Editing by Lipid Nanoparticle Delivery Stimulates Anti-Tumor Immunity in Breast Cancer Liver Metastasis Model

Triple-negative breast cancer (TNBC) is a highly aggressive subtype, accounting for 10-15% of breast cancer cases in the United States. Liver metastases, common in advanced TNBC, are linked to especially poor outcomes, with a 5-year survival rate of just 11%. Although immune checkpoint inhibitors (ICIs) targeting PD-1 or PD-L1 show promise, durable responses in TNBC remain uncommon. This is largely due to a profoundly immunosuppressive tumor microenvironment (TME), driven by tumor-associated myeloid cells. Tumor-associated macrophages (TAMs) and neutrophils (TANs) polarize into immunosuppressive M2 and N2 phenotypes, respectively, suppressing T cell activity through cytokines, ROS, and checkpoint ligands such as VISTA. Myeloid-derived suppressor cells (MDSCs) further inhibit immunity by depleting nutrients and inducing regulatory T cells. As a result, despite its immunogenic features, TNBC remains resistant to immunotherapy due to persistent myeloid-mediated suppression. Here, we developed ionizable lipid nanoparticles (iLNPs) engineered to deliver the CRISPR-Cas12a ribonuclease complex targeting Rictor, a critical component of mTORC2, for in vivo reprograming of myeloid cells. The intravenous (IV) injection of CRISPR Rictor-targeting iLNP (CR-Ric-LNP) showed efficient uptake by circulating myeloid cells and accumulation into the breast cancer liver metastases. Notably, Rictor gene editing triggered pro-inflammatory activation of myeloid cells in the TME, enhancing antitumor responses. Single-cell RNA sequencing revealed that Rictor silencing treated samples showed induced rapid remodeling of the TME, with a significant reduction in immunosuppressive macrophages within 24 hours of treatment. Concurrently, cytotoxic T-cell populations exhibited increased interferon-gamma (Ifng) production, driving the emergence of specific myeloid clusters that were responsive to Interferon signaling, particularly in macrophages and neutrophils. A shift from an immunosuppressive to an inflammatory TME was further evidenced by an elevated Cxcl10/Spp1 ratio in myeloid cells. CR-Ric-LNP treatment also enhanced T-cell activation, reducing exhausted T cells and regulatory T cells (Tregs) while expanding natural killer (NK) cells, naive CD4+, and CD8+ T cells. These changes correlated with a decreased proportion of tumor cells and proliferating cells, ultimately leading to a significant survival benefit in a 4T1 breast cancer liver metastasis model. Our findings demonstrate that myeloid-targeted Rictor silencing reprograms the TME, promoting antitumor immunity and improving therapeutic outcomes. O_FIG O_LINKSMALLFIG WIDTH=199 HEIGHT=200 SRC="FIGDIR/small/661471v1_ufig1.gif" ALT="Figure 1"> View larger version (109K): org.highwire.dtl.DTLVardef@10a5e7aorg.highwire.dtl.DTLVardef@1c0c005org.highwire.dtl.DTLVardef@17ec6b5org.highwire.dtl.DTLVardef@27c507_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2025

Agropyron cristatum genome provides a new insight for wheat improvement with wild relatives

Exploring novel genetic variation for target traits is crucial for advancing wheat breeding and genetic improvement. Since 1990, we have been transferring genes from Agropyron cristatum, a promising wild relative genetic resource featuring robust spike morphology and stress tolerance, into common wheat by distant hybridization. To efficiently harness A. cristatum genes, we de novo assembled a 25.62-Gb high-quality genome of an autotetraploid accession Z559 with 28 chromosomes, and uncovered its specific genome features and an extensive repertoire of genes associated with yield and resistance to abiotic and biotic stresses. We systematically characterized A. cristatum genome introgressions by resequencing 431 wheat-A. cristatum derivatives and cloned the gene AcGNS1, which functions in the regulation of grain number per spike in wheat. Subsequently, we developed an optimized genome-selection model for wheat-A. cristatum breeding derivatives, enabling the rapid selection of new potential varieties. These findings demonstrate a genomic breeding strategy for leveraging novel genetic resources of wild relatives and advancing wheat improvement.

genomics↗

bioRxiv2025

UniCure: A Foundation Model for Predicting Personalized Cancer Therapy Response

Predicting drug efficacy across diverse patient contexts remains a major challenge in oncology, as models trained on cancer cell lines often fail to capture patient-specific biology. Emerging biological foundation models and patient-derived technologies offer a promising solution. Here, we present UniCure, the first pre-trained foundation model integrating both omics and chemical foundation models (UCE and Uni-mol) to predict transcriptomic responses to drugs across diverse cellular and tissue contexts, enabling personalized cancer therapy and drug prioritization at the individual level. Rather than encoder/decoder used in traditional models, UniCure utilizes parameter-efficient fine-tuning (PEFT) techniques for optimizing the training process, a novel FlexPert module for modeling flexible drug-cell interactions, and a Maximum Mean Discrepancy (MMD) loss for learning unpaired data. Trained on over 1.8 million perturbation RNA-seq profiles over 22,000 compounds, 166 cell types, and 24 tissues, UniCure achieves high accuracy in predicting both dose-dependent responses and drug combination effects, demonstrating strong generalization across bulk and single-cell transcriptomic data. In particular, fine-tuning on our patient-derived tumor-like clusters and real-world data of 800 profiles enables UniCure to generate individualized therapeutic predictions on patients tissue samples. In addition, UniCure enables patient stratification based on the predicted drug responses, providing a new way for subtyping patients. UniCures drug prioritization was validated across over 1000 patients from pan-cancer cohorts and supported by experiments of candidate therapeutics. By enabling the potential to screen millions of compounds per patient at scale, UniCure represents a biologically grounded tool that could advance personalized precision oncology and accelerate drug discovery.

bioinformatics↗

bioRxiv2025

A bottom-up approach to understanding ultrasound mediated blood-brain barrier opening using long and short pulses

Ultrasound-mediated blood-brain barrier (BBB) opening is promising for non-invasive, localized, and reversible drug delivery to the brain. However, the underlying mechanisms remain unclear, particularly at microscopic level in real time. Recently developed short-pulse ultrasound enhances safety by reducing side effects compared to conventional long-pulse ultrasound. Using vessel-mimicking microchannels cultured with brain endothelial monolayers, we directly observed bubble dynamics and cellular bioeffects under flow conditions. Intriguingly, cyclic jetting during stable cavitation occurs at long-pulse mode, accompanied by a few localized cell detachments and extensive sonoporation. In contrast, short-pulse ultrasound induced milder, uniform bubble dynamics, reversible sonoporation and calcium signaling, promoting safer BBB modulation. In vivo two-photon imaging and histology in mice confirmed these findings, showing that long-pulse ultrasound enabled higher drug delivery efficiency but caused localized endothelial damage, while short-pulse ultrasound facilitated uniform delivery and faster BBB recovery. The differential bubble dynamics and cellular responses correlated well between in vitro and in vivo models. This study establishes a cross-scale framework for real-time analysis of ultrasound-mediated BBB opening, revealing key biophysical factors governing safety and efficacy. The findings provide guidance for the optimization of ultrasound protocols for vascular drug delivery, with potential applications in treating brain tumors, neurodegenerative diseases, and other disorders.

biophysics↗

bioRxiv2025

Mapping genetic modifiers of epimutation rates reveals a punctuated-equilibrium model of CG methylome evolution

Spontaneous epimutations--stochastic changes in cytosine methylation--can persist across generations in plants and are thought to contribute to phenotypic variation. Although epimutations are increasingly studied for their potential long-term effects, it remains unclear why their accumulation varies across genotypes. Here, we tracked DNA methylation across ten generations in [~]400 mutation accumulation lineages derived from [~]70 Arabidopsis Ler x Cvi recombinant inbred lines. Treating epimutation rates as quantitative molecular traits, we mapped a major QTL to a Cvi-derived deletion near VIM2 and VIM4, two genes involved in CG methylation (mCG) maintenance. We show that this deletion rapidly reduces genome-wide methylation to a lower steady-state and compromises mCG maintenance fidelity across generations, resulting in a [~]1.5-fold increase in epimutation rates. Genotypes with elevated rates exhibited accelerated epigenetic drift and phenotypic divergence. Our findings support a punctuated-equilibrium model of mCG evolution, in which sudden disruptions to methylation homeostasis can destabilize epigenetic inheritance over longer time-scales.

genetics↗

bioRxiv2025

Unbiased recording of clonal potency reveals species-specific regulation of mammalian intestine

The mammalian intestine regenerates rapidly after damage, yet the clonal dynamics and species-specific regulation of different populations remain poorly understood. Here we used synthetic or naturally occurring DNA alterations to reconstruct clonal histories of the mouse and human intestinal epithelium at single-cell resolution. In mice, we uncovered the clonal architecture of different cell types and their roles in regeneration, supporting a hierarchical regenerative response model. We identified a rare embryonic precursor population that persisted in the adult and was crucial for regeneration after irradiation. This population was marked by Tob2, which is required for nuclear transport of Ascl2. A parallel clonal analysis of 65 human colonic biopsies revealed secretory lineage bias and an age-associated decline in clonal diversity in the distal colon. Unlike highly proliferative murine Lgr5+ stem cells, human LGR5+ cells were found largely quiescent, revealing species-specific difference in clonal potency, and suggesting a distinct regulation of intestinal stemness.

genomics↗

bioRxiv2025

Locust cGAS-like receptors recognize derivatives of a Gypsy retrotransposon to synergize with RNAi against viral invasion

The co-option of transposable elements (TEs) as immune sentinels represents an evolutionarily conserved strategy across metazoans, yet the molecular mechanisms linking retrotransposon reactivation to antiviral defense remain enigmatic. Here, we identify LmGypsy, a long terminal repeat (LTR) retrotransposon in Locusta migratoria, as a critical mediator of biphasic antiviral immunity against Acrididae reovirus (ARV). ARV infection triggers selective de-repression of LmGypsy, which orchestrates dual antiviral pathways: (1) its encoded reverse transcriptase synthesizes viral DNA (vDNA) from ARV RNA, fueling RNA interference (RNAi)-mediated viral RNA degradation through Dicer-2-dependent vsiRNA biogenesis; (2) LmGypsy-derived nucleic acids activate cGAS-like receptors (LmcGASs) and induce immune responses. Strikingly, vDNA persists in infected locusts from 24 h post-infection until host death, suggesting a role in sustaining antiviral activity akin to immune memory mechanisms observed in Diptera. This study provides the first evidence that insect cGAS homologs function as retrotransposon sensors, mirroring mammalian cGAS-STING recognition of endogenous retroviral elements. Our findings redefine retrotransposons as central hubs in arthropod antiviral immunity, bridging RNAi and nucleic acid-sensing pathways to establish a coordinated defense network. These results illuminate conserved principles of TE-immune crosstalk and highlight LmGypsy as a paradigm for understanding host-transposon coevolution in antiviral contexts.

immunology↗

bioRxiv2025

Towards Unmanned Proteomics Data Generation: A Fully Automated Sample-to-Data System for Proteomic Experiments

The {pi}-HuB project requires proteomic measurements of millions of low-input samples that are challenging with current technology. Here, we introduce a pilot {pi}-HuB data factory that features a fully automated sample-to-data system to enable high-throughput digitization of human samples. Our platform, as a proof-of-concept towards unmanned proteomics, was demonstrated to show longitudinal robustness and applied for spatial proteomics and the profiling of plasma and cell lines, thereby serving as the workhorse to drive {pi}-HuB.

biochemistry↗

bioRxiv2025

Controllable Gap Junctions by Vitamin B12 and Light

Gap junctions, mediate rapid signal transduction between contiguous cells, which are indispensable for multicellular organisms to coordinate cellular activities across numerous physiological processes. However, precise control of gap junctions remains elusive. Herein, we present CarGAP, a single-component chemo-optogenetic tool that utilizes the C-terminal adenosylcobalamin (AdoB12) binding domain of a photoreceptor protein (i.e., CarHC) to achieve reversible control over both vertebrate and invertebrate gap junctions with spatiotemporal precision. The vertebrate CarGAP (i.e., Cx-CarGAP), created by genetically fusing connexins with CarHC in mammalian cells, can efficiently block the gap junction channels through AdoB12-induced protein oligomerization, and subsequently reinstate them via green light-induced protein disassembly. We further introduced the CarGAP system (i.e., Inx-CarGAP) to the Drosophila ovary, enabling reversible control over the heterotypic gap junctions formed by innexin2 (Inx2) and innexin4 (Inx4, also known as zero population growth, Zpg), thereby uncovering the roles of gap junctions in stem cell-niche interactions. This study illustrates CarGAP as a generalizable chemo-optogenetic tool for interrogating the functions of gap junctions in various biological contexts.

bioengineering↗

bioRxiv2025

Kre6-Dependent β-1,6-glucan Biosynthesis Only Occurs in the Conidium of Aspergillus fumigatus

The structural role of {beta}-1,6-glucan has remained under-investigated in filamentous fungi compared to other fungal cell wall polymers, and previous studies have shown that the cell wall of the mycelium of A. fumigatus did not contain {beta}-1,6-glucans. In contrast, the current solid-state NMR investigations showed that the conidial cell wall contained a low amount of {beta}-1,6-glucan. ssNMR comparisons of the A. fumigatus and C. albicans {beta}-1,6-glucans showed they are structurally similar. Deletion of the KRE6 gene which is the only KRE gene in the A. fumigatus genome resulted in a mutant depleted of {beta}-1,6-glucan which has a growth phenotype similar to the parental strain. Even though it is not an essential polymer in A. fumigatus, {beta}-1,6-glucan play a role in cell wall organization since the kre6{Delta} mutant showed a higher sensitivity to Congo-red and Calcofluor white which are known to be general cell wall inhibitors. It is also another example of the significant structural differences seen between conidium and mycelium of filamentous fungi.

biochemistry↗

bioRxiv2025

Metabolic basis of the astrocyte-synapse interaction governs dopaminergic-motor connection

Perisynaptic astrocyte processes are constitutive attachments of synapses in the central nervous system. However, the molecular mechanisms that control perisynaptic astrocyte ensheathment and their implications in the wiring of neural circuits remain unclear. Here, we report that glycolysis controls astrocyte-synapse contact. In the Drosophila larval dopaminergic (DAergic) circuit, blocking astrocyte glycolysis stimulated perisynaptic ensheathment by attenuating astrocyte-to-DAergic neuron neuroligin 2-neurexin 1 signaling. As a result, the larvae executed more reorientation actions during locomotion. At the circuit level, behavioral alterations were found to arise from increased DAergic neuronal synaptogenesis and DAergic-motor connection. Our research uncovers an ancient metabolic basis that determines perisynaptic astrocyte ensheathment abundance through a conserved neuroligin-neurexin signaling pathway and demonstrates the role of astrocyte glycolysis in controlling DAergic-motor circuit assembly and function.

neuroscience↗

bioRxiv2025

CSFeatures improves identification of cell type-specific differential features in single-cell and spatial omics data

Recent advancements in single-cell and spatial omics have enabled the analysis of gene expression patterns in cells and tissues with unprecedented precision. Genes that exhibit significant variation across different cell types or states are typically closely linked to cellular functional states and disease processes. However, current differential analysis methods often fail to adequately account for cell type specificity when identifying differentially expressed genes. CSFeatures effectively addresses this issue by comprehensively considering the expression level, the smoothness of expression distribution, and the proportion of gene expression in the target cell population as well as in each of the other cell populations. Through comprehensive experiments on eight single-cell RNA-seq datasets, it was demonstrated that CSFeatures can effectively identify cell type-specific genes that are highly expressed in the target cell population while being lowly expressed in all other populations. Moreover, it was validated that the underlying principle of CSFeatures can be generalized to other omics data, including single-cell ATAC-seq, spatial transcriptomics, and spatial ATAC-seq data. Furthermore, the differential features identified by CSFeatures are enriched in pathways, functional categories, and regulatory regions that are highly relevant to the specific functional states of the target cell population, providing insights into its gene regulation.

bioinformatics↗

bioRxiv2025

SPLiCR-seq: A CRISPR-Based Screening Platform for RNA splicing Identifies Novel Regulators of IRE1-XBP1 Signaling Under ER Stress

RNA splicing is fundamental to cellular function, yet systematic investigation of its complex regulation has been limited by existing methods. Here, we present SPLiCR-seq (SPLicing regulator identification through CRISPR screening), a high-throughput CRISPR screening platform that enables direct measurement of RNA splicing outcomes for pooled genetic perturbations, overcoming limitations of traditional fluorescence-based approaches. Applying SPLiCR-seq to investigate XBP1 splicing during the unfolded protein response (UPR), we conducted targeted and genome-wide screens across diverse cellular contexts, revealing both common and cell-type specific regulators. Notably, we identified GADD34 (PPP1R15A) as a novel modulator of IRE1-XBP1 signaling, demonstrating that it directly interacts with IRE1 and functions independently of its canonical role in eIF2 dephosphorylation. Pharmacological inhibition of GADD34 using Sephin1 effectively suppressed XBP1 splicing and alleviated CAR-T cell exhaustion in an ex vivo model, leading to enhanced tumor-killing capacity across multiple cancer models. This work not only establishes a powerful new tool for systematically studying RNA splicing regulation but also uncovers a promising therapeutic strategy for improving CAR-T cell immunotherapy through modulation of the IRE1-XBP1 pathway.

genetics↗

bioRxiv2025

Universal Dual-Target CAR-γδT Cells Targeting B7-H3 and IL13Rα2 for Glioblastoma Therapy

Glioblastoma (GBM) represents the most prevalent and aggressive primary malignant neoplasm in the adult central nervous system, exhibiting marked infiltrative growth patterns, inevitable recurrence, and dismal therapeutic outcomes with current treatment modalities. While CAR-T cell immunotherapy has demonstrated remarkable success in hematological malignancies, its clinical translation for GBM has been hampered by several fundamental limitations. A key factor among these is tumor-intrinsic heterogeneity, which drives antigen escape and therapeutic resistance. Furthermore, although autologous CAR-T approaches dominate current clinical investigations, they encounter substantial barriers including manufacturing variability, scalability constraints, and practical limitations for widespread clinical deployment. In contrast, allogeneic "off-the-shelf" CAR-T therapy holds greater potential for the future applications. {gamma}{delta} T cells are a particularly compelling candidate for universal CAR therapy, offering several advantages including innate MHC-unrestricted target recognition obviating the need for HLA matching, polyfunctional cytotoxic mechanisms capable of addressing heterogeneous tumor populations, and intrinsic tropism for solid tumors. However, translational implementation has been constrained by their physiological rarity, ex vivo expansion difficulties, and genetic modification inefficiencies. To address these challenges, we adopted a dual-pronged targeting strategy focusing on B7-H3 and IL13R2 - two surface antigens demonstrating preferential overexpression across GBM subtypes while maintaining limited distribution in normal tissues. Using phage display platform and function-based nanobody screening we identified high-affinity binders against both targets. Subsequent optimization of {gamma}{delta} T cell expansion protocols and lentiviral transduction parameters enabled the development of a bispecific, allogeneic CAR-{gamma}{delta} T cell platform. Our in vitro studies revealed that dual-target CAR-{gamma}{delta}T cells sustained proliferative capacity under GMP-compatible culture conditions, exhibited potent and specific cytotoxicity against antigen-positive glioma cells, and critically, they showed superior elimination of target-heterogeneous tumors compared to monospecific CAR-T constructs. These results establish a robust preclinical foundation for clinical translation and highlight the therapeutic potential of combinatorial antigen targeting coupled with allogeneic {gamma}{delta} T cell engineering to overcome the persistent challenges in GBM immunotherapy.

immunology↗

bioRxiv2025

Deregulated Myt3 translation predisposes islet β-cells to dysfunction under obesity-induced metabolic stress

In response to obesity-related metabolic stress, islet {beta}-cells adapt (or compensate) by increasing their secretory function and mass. Yet, for unknown reasons, this compensation is reversed in some individuals at some point to induce {beta}-cell failure and overt type 2 diabetes (T2D). We have previously shown that transcription factor Myt3 (St18) and its paralogs, Myt1 and Myt2, prevent {beta}-cell failure. Myt3 was induced at post-transcriptional levels by obesity-related stress in both mouse and human {beta} cells and its downregulation accompanied {beta}-cell dysfunction during T2D development. Single-nucleotide polymorphisms in MYT3 were associated with an increased risk of developing human diabetes. We now demonstrate that Myt3 translation is regulated by an upstream open-reading frame that overlaps with the main Myt3 open-reading frame in mice. Disrupting this overlap enhances Myt3 translation in mouse {beta} cells without metabolic stress but decreases it under high-fat-diet challenges. Consequently, this deregulation results in {beta}-cell dysfunction and glucose intolerance in mice, accompanied by compromised expression of several {beta}-cell function genes. These findings suggest that stress-induced Myt3 translation is part of the compensation mechanism that prevents {beta}-cell failure.

genetics↗