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Lei, Y.

Publications and source records attributed to Lei, Y..

At least 19 recordsLinked to original sources

Characterizing bilateral M1 excitability changes after learning a novel spatiotemporal coordination pattern

Bimanual coordination is essential for many skilled everyday actions and requires maintaining specific spatiotemporal relationships between the two hands according to task demands. However, the contribution of the primary motor cortices (M1s) to learning such relationships is only partially understood. Conventional bimanual tasks necessarily engage both M1s in controlling the moving hands, making learning-related changes difficult to distinguish from those associated with movement execution. To reduce this effector-related confound, we used a unimanual and bimanual rhythmic coordination paradigm in which participants learned the same novel 90-degree relative-phase relationship by coordinating either hand with a virtual partner and compared these conditions with conventional bimanual practice. In three experiments, performance accuracy and stability improved during practice and improved further after a 6-h retention interval with comparable time-dependent changes across unimanual and bimanual trained groups. In Experiment 2, motor-evoked potentials measured before and after training revealed an overall increase in corticospinal excitability across both M1s under unimanual and bimanual training. In Experiment 3, bidirectional dual-site paired-pulse stimulation revealed effector-dependent changes in interhemispheric inhibition, in which bimanual practice produced no reliable directional modulation, whereas unimanual practice produced asymmetric changes that depended on the trained hand. Neither corticospinal excitability nor interhemispheric inhibition showed a consistent relationship with behavioral improvement. Together, these findings demonstrate comparable behavioral learning of a novel spatiotemporal coordination relationship across bimanual and unimanual contexts, while showing that behavioral improvement is not straightforwardly reflected in M1 neurophysiological changes. Causal intervention studies are therefore needed to determine whether and when M1 contributes directly to acquisition of novel coordination relationships.

neuroscience↗

Tumor cell-intrinsic mtDNA instability drives cGAS-dependent temporal remodeling of the melanoma immune microenvironment

Mitochondrial DNA (mtDNA) instability is an emerging feature of cancer, yet its biological effects on the tumor microenvironment (TME) and tumor progression remain incompletely defined. Using melanocyte-specific conditional silencing of mitochondrial transcription factor A (TFAM) in an autochthonous BrafV600E/Pten-/- melanoma model, we demonstrate that tumor cell-intrinsic mtDNA instability accelerates melanoma growth and metastatic dissemination in a cGAS-dependent manner. Transcriptomic profiling of early and late tumor stages revealed that mtDNA instability drives temporal remodeling of the tumor microenvironment (TME), characterized by an early type I interferon (IFN)-rich inflammatory state that licenses immunoregulatory programs as tumors progress. Tumor-associated macrophages emerged as the principal immune cell type reshaped by tumor cell-intrinsic mtDNA stress signaling, with early enrichment of inflammatory states and a shift toward efferocytosis-associated, tissue-adaptive states. The early mtDNA instability-associated macrophage signatures mapped to human melanoma states associated with favorable survival and therapy response, whereas the late signatures mapped to states associated with poor prognosis and therapy resistance across independent cohorts. Anti-PD-1 treatment delayed progression of mtDNA-instable tumors, consistent with a therapeutic sensitivity window associated with the early IFN-rich immune state. These findings establish tumor cell-intrinsic mtDNA instability as a driver of temporal immune-stromal remodeling that promotes melanoma progression and shapes therapeutic response.

immunology↗

Redox activation of ATG5 licenses autophagy upon nutrient restriction

Dietary restriction (DR) protects against metabolic disease, extends lifespan, and is associated with remodeling of tissue reactive oxygen species (ROS). ROS control biological adaptation through reversible oxidation of protein cysteines, yet the targets of DR-initiated redox signaling are unknown. Here we generate OxiDR, a tissue-resolved atlas of the cysteine redox proteome that quantifies oxidation state under DR. Rather than oxidizing the proteome broadly, DR selectively targets a high-amplitude set of cysteines in a tissue-specific manner, allowing systematic classification of biological processes subject to DR-mediated redox regulation. Among the cysteines most highly oxidized upon DR is Cys19 of the core autophagy protein ATG5. We show oxidation of Cys19 is required for ATG5-mediated autophagosome formation and for autophagy triggered by nutrient restriction in human cells and mice. Reversible oxidation of this cysteine promotes ATG5 binding to ATG10, thus forming the ATG5-ATG12 conjugate that lipidates LC3B/ATG8 and matures the autophagosome. In mice, loss of this redox switch prevents effective initiation of autophagy upon nutrient restriction, resulting in gross tissue pathology and rapid onset of mortality. The autophagic response to nutrient restriction is thus gated by oxidation of a single cysteine.

cell biology↗

A quantitative redox proteome of the human muscle response to exercise

Reactive oxygen species (ROS) regulate protein function through reversible cysteine oxidation. In human skeletal muscle, exercise-induced ROS initiates adaptations such as mitochondrial biogenesis, increased insulin sensitivity, and hypertrophy. However, specific protein targets of ROS regulation during exercise remain unclear owing to longstanding challenges in analyzing redox proteomes in vivo. We applied cysteine derivatization and multiplexed proteomics to map muscle protein cysteine oxidation in humans during exercise. The OxiMuscle dataset quantifies reversible modifications across 9,177 unique cysteine sites on 2,782 proteins, comprising 17,492 individual cysteine site measurements in young men undergoing three types of exercise, providing the first comprehensive, site-resolved and quantitative analysis of the exercise-regulated redox cysteine proteome in humans. We systematically define cysteine oxidation targets regulated by at least one form of exercise, many of which reside in proteins with established roles in muscle physiology. Among these sites is a redox-regulated cysteine on the autophagy receptor protein p62. We demonstrate that reversible oxidation of this cysteine regulates p62-mediated autophagy upon myotube contraction and mouse muscle adaptation to exercise in vivo. Together, these results define a redox-driven mechanism linking exercise-induced autophagy to muscle adaptation. More broadly, our findings offer a comprehensive resource on redox-signaling networks in human muscle, accessible at http://oximuscle-alb-1899330623.us-east-1.elb.amazonaws.com/.

cell biology↗

Atypical Chemokine Receptor DARC/ACKR1 Is Expressed Selectively by Neurons with Regional Heterogeneity and Protects against Neuroinflammation and Cognitive Impairment

Atypical chemokine receptors (ACKRs) regulate inflammatory responses by scavenging and sequestering chemokines. The Duffy antigen receptor for chemokines (DARC), also known as ACKR1, binds multiple inflammatory chemokines and regulates inflammation in peripheral tissues. However, the distribution and functional role of DARC/ACKR1 in the brain remain to be defined. Here, we show that DARC/ACKR1 is widely expressed throughout the mouse brain, with marked regional heterogeneity. DARC/ACKR1 is predominantly expressed by neurons and is not detected in microglia or astrocytes. Importantly, this neuronal expression pattern was also observed in the human brain. Furthermore, DARC/ACKR1 deficiency resulted in enhanced neuroinflammation, characterized by increased microglial activation and astrogliosis across multiple brain regions, particularly the hippocampus, and was accompanied by cognitive deficits. Together, these findings identify DARC/ACKR1 as a previously unrecognized neuron-specific atypical chemokine receptor that plays an important role in restraining neuroinflammation and preserving cognitive function.

neuroscience↗

Cysteine reduction rather than methionine restriction promotes immunometabolic fitness to confer survival benefit in aging

Elevated sulfur-containing amino acids (SAA; methionine and cystine) are linked to human mortality. While methionine restriction (MR) extends lifespan in animals, it is often ignored that traditional longevity-promoting MR diets also lack cystine. When dietary cystine is eliminated, host redirects substrates from methionine cycle and generates cysteine via cystathionine {gamma}-lyase (CTH) in transsulfuration pathway, leaving it unclear which SAA controls aging. Here, we show that selective cysteine restriction in Caenorhabditis elegans enhanced lifespan and stress survival independently of methionine. Notably, cysteine-free diets (methionine-replete or restricted) in Cth-deficient mice induced pro-metabolic effects, whereas MR with normal cysteine was ineffective. Sustained 70% cysteine reduction in aged Cth-/- mice reprogrammed the immunometabolic axis, conferring healthspan benefits. Thus, lowering cysteine while keeping the methionine pool intact enhances healthy lifespan.

physiology↗

The Capillary endothelial cell-governed niche sustains angiogenesis in proliferating infantile hemangioma via multilayer stromal support

Infantile hemangioma (IH), the most common benign vascular tumor in infants, is characterized by the aberrant proliferation of hemangioma endothelial cells (HemECs) and excessive angiogenesis. However, it is unclear how the tumor microenvironment sustains these active angiogenic signals. We performed single-cell RNA sequencing and spatial transcriptomic to profile 30 samples from proliferating IH, involuting IH, and normal skin. The results indicated that HemECs comprised distinct subpopulations. Among them, capillary endothelial cells (Cap.ECs) had an immature phenotype and functioned as the core drivers of IH. The endothelial-to-mesenchymal transition endothelial cells (endoMT-ECs) secreted collagen and engaged in CD44-mediated crosstalk with Cap.ECs to construct the angiogenesis-active niche in the tumor microenvironment (TME) of proliferating IH. Blockade of CD44-mediated intercellular communication effectively suppresses the angiogenesis of HemECs and promotes tumor regression. Beyond the endothelial layer, pericytes tightly enveloped Cap.ECs, with SERPINE2+ subtypes sustaining their angiogenic activity via ANGPT signaling. Pseudotime trajectory analysis revealed that the mesenchymal stem cells (MSCs)-enriched niche serves as a progenitor pool for Cap.ECs in proliferating IH. Our findings thus revealed that IH progression relies on CD44-mediated endoMT-EC-Cap.EC crosstalk and on both pericyte and MSC niches to sustain angiogenesis. HIGHLIGHTSO_LICapillary endothelial cells drive infantile hemangioma progression, with pericytes tightly ensheathing them. C_LIO_LICD44-mediated capillary and endoMT-derived endothelial cells crosstalk construct the angiogenic-active niche in the tumor microenvironment of proliferating infantile hemangioma. C_LIO_LIThe MSC-enriched niche provides a progenitor pool for the capillary endothelial cells in proliferating infantile hemangioma. C_LI

cancer biology↗

Design and Validation of a Hybrid Stimulus fMRI Paradigm for Simultaneous Retinotopy and Mapping of Reading-Selective Visual Cortex

Reading selectively recruits specialized cortical patches within the left ventral occipitotemporal cortex (vOTC), such as the visual word form area (VWFA). Under the framework of the interactive account of word recognition, these neural responses emerge from dynamic, bidirectional loops where top-down linguistic predictions continuously modulate and constrain bottom-up sensory visual inputs. Consequently, identifying and isolating these top-down cognitive signals fundamentally requires a precise characterization of the underlying low-level sensory baseline. This dense-sampling fMRI study (11 right-handed adults, 10 sessions each) systematically addresses the spatial and functional architecture of the reading network using population receptive field (pRF) modeling and functional localizers (fLoc). First, we conduct a rigorous group- and subject-level replication of stimulus-dependent pRF eccentricity shifts (words and false fonts versus checkers) observed in previous research. To overcome known barriers to individual parameter stability, we systematically manipulate and evaluate stimulus Frequency, bar Width, and element Size (FWS) across eight sessions per subject to isolate the precise factors driving test-retest reliability. Second, we design and validate a novel Word-Center (WC) paradigm acquired across one/two sessions per participant. This hybrid paradigm was designed to decouple moving spatial mapping carriers (checkerboard bars) from central reading processes (word or false-font streams in the central fixation location). Functional data were denoised with NORDIC, preprocessed via fMRIPrep, and projected to the cortical surface for analysis via Nilearn GLMs to model word-responsive regions and mrVista to obtain pRF estimates. Individual-level spatial maps replicated previous results in approximately 70% of subjects. The lack of replication in the remaining subjects, whether driven by methodological factors or inherent individual differences, highlights ongoing baseline mapping challenges. Optimizing this mapping is sensitive to stimulus carrier type and specific FWS configurations. Furthermore, the hybrid WC paradigm successfully demonstrates stimulus equivalence, concurrently yielding robust reading-selective functional localization and reliable retinotopic estimates. By breaking the spatial-lexical confound of traditional mapping protocols, this paradigm provides an innovative framework to separate bottom-up sensory sweeps from top-down central linguistic processing. This vision-centric framework serves as a methodological baseline toward a comprehensive, multi-signal understanding of the human reading hierarchy.

neuroscience↗

Accumulation of Lipid Droplets in Microglia following Neonatal Brain Hypoxia-Ischemia

BackgroundHypoxic-ischemic encephalopathy (HIE) is a major cause of neonatal mortality and neurodevelopmental impairments. Following brain hypoxia-ischemia (HI), microglia face substantial metabolic stress; and upon phagocytosis, they become overloaded with lipids derived from engulfed dead neurons and myelin debris. It is unclear how microglia respond to and process the lipid cargo, and whether lipid accumulation may affect microglia function following neonatal HI. MethodsThe postnatal day 10 mice were subjected to HI using the Vannucci model. Lipid droplets (LD) were assessed by histology and immunofluorescent staining. Single-nucleus RNA sequencing (snRNA-seq) was performed using brain tissue from HI-injured and sham-operated mice at 72 hours after HI. LD-accumulating microglia (LDAM) were identified by a specific LD marker gene perilipin 2 (Plin2). Differential gene expression was analyzed between Plin2-positive and Plin2-negative microglia after HI. Human HIE brain sections were also examined for LD accumulation. The dynamic changes of PLIN2-expressing microglia and infiltrating monocyte-derived macrophages (MDM) at 24 hours, 72 hours and 7 days after HI were compared using flow cytometry. In addition, mouse BV2 microglia were subjected to oxygen-glucose deprivation (OGD) to study phagocytosis and cytokine expression. ResultsLipid droplets accumulated primarily in microglia after HI in neonatal mice and in human HIE brain. LD were not found in astrocytes or neurons. Plin2-expressing LDAM emerged as new microglia clusters after HI. Compared with microglia without LD, LDAM showed a distinct transcriptional profile with upregulation of genes linked to microglial activation, enhanced cholesterol and lipid processing, and a shift towards phagocytic and pro-inflammatory state. Blocking LD biogenesis reduced elevated phagocytosis and IL-1{beta} expression in BV2 cells following OGD. ConclusionOur study revealed that microglia accumulate lipid droplets as part of their metabolic responses to HI in the neonatal brain. Microglial lipid droplet formation is associated with a pro-inflammatory phenotype at early stage after HI, and increased phagocytosis in vitro. The lipid metabolic changes may regulate microglial function and influence HI outcomes.

neuroscience↗

Designing Fidelity of CRISPR-Cas Endonucleases by Kinetic Insights

Finding high-fidelity CRISPR-Cas variants is critical for both the precision of in vitro DNA detection and the safety of in vivo gene-editing therapeutics. However, the large size of the Cas enzyme and the distinct selection criteria between its natural evolution and clinical practice lead to extensive experimental trials and limited success rate for de novo design, directed evolution, protein language model (PLM)-based filtering. Here, we present a PLM-assisted physics-driven approach that utilizes atomistic molecular dynamics simulations and automated path searching to efficiently obtain the complete kinetic insights, including the transition state structures, for the conformational changes of Cas before DNA cleavage. We show that these kinetic insights can pinpoint a few fidelity-diminishing protein residues during the early stage of target recognition, and have led to SpyCas9 and FnCas12a variants with ultra-high fidelity surpassing previously reported counterparts at minimal cost of a few wet-lab trials.

biophysics↗

Neuropeptide Y4 receptor activation delays autoimmune diabetes by reprogramming β-cell stress and immune tolerance

Type 1 diabetes (T1D) involves immune-mediated destruction of pancreatic {beta}-cells, yet current disease-modifying therapies mainly target immunity without enhancing {beta}-cell resilience. We show selective neuropeptide Y4 receptor (Y4R) agonism protects {beta}-cells while reshaping islet immunity across T1D models. Multi-modal localisation using cell sorting, qPCR, RNAscope and fluorescent ligand competition demonstrated predominant Y4R expression and functional accessibility on mouse and human {beta}-cells. Selective Y4R agonism was non-toxic and did not impair islet network integrity, Ca{superscript 2} dynamics, glucose-stimulated insulin secretion or systemic glucose tolerance. Y4R activation conferred cytoprotection against inflammatory cytokines, streptozotocin, lipotoxicity and ER stress, reducing caspase-3/7 activation and {beta}-cell loss whilst sustaining insulin release and promoting proliferation in both mouse and human islets. Bulk RNA-seq revealed a coordinated {beta}-cell resilience programme characterised by reinforced identity and insulin processing, KEAP1-NFE2L2-driven antioxidative and proteostatic activation, and suppression of EIF2 signalling and associated biosynthetic and ER stress pathways. Concurrently, Y4R agonism dampened pathogenic chemokine and cytokine networks, including CXCL10, CCL3/4/7 and IL-6, while preserving IL-2 and Foxp3 signals, thereby limiting CD8 T cell, CD4 T cell and macrophage chemotaxis toward cytokine-stressed islets. Reduced immune-cell recruitment was conserved in a fully human immune-islet system, where Y4R activation significantly attenuated IL-2-activated human PBMC migration and invasion toward cytokine-stressed human islets. In a stringent NY8.3 CD8 T cell adoptive-transfer model, systemic Y4R agonism significantly delayed diabetes onset. These data position Y4R as a {beta}-cell-centric therapeutic target coupling intrinsic resilience with local immune modulation, offering a complementary approach for {beta}-cell preservation in T1D and islet replacement therapies. Graphical abstractThe selective Y4 receptor agonist K22 binds {beta}-cell-enriched NPY4R in mouse and human islets, activates a {beta}-cell resilience programme that preserves insulin secretion under inflammatory and metabolic stress, and simultaneously dampens islet chemokine output, thereby limiting innate and adaptive immune-cell recruitment and delaying autoimmune diabetes onset. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=119 SRC="FIGDIR/small/736290v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@7efd48org.highwire.dtl.DTLVardef@1268860org.highwire.dtl.DTLVardef@c38a81org.highwire.dtl.DTLVardef@1ba3e6c_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

Alginate-Coated Collagen Hydrogel Tubes for Scalable Cell and Biotherapeutic Particle Manufacturing

Efficient, scalable, and cost-effective production of mammalian cells and biotherapeutic particles remains a major challenge for both research and clinical applications. Conventional 2D and 3D culture systems suffer from low volumetric yields, poor scalability, and high costs. Previously, we developed collagen hydrogel tube microbioreactors (ColTubes) that support high-density, high-viability cell culture by preventing excessive cell aggregation and minimizing hydrodynamic stress. However, ColTubes exhibit adhesion to culture vessels and to each other, and leaked cells frequently attach to outer tube surfaces -- behaviors that would limit scalability. Here we introduce AlgColTubes: collagen hydrogel tubes coated with a thin, ionically crosslinked alginate layer to overcome these limitations. Scanning electron microscopy confirms that alginate penetrates the collagen wall and forms a stable interpenetrating hydrogel network, whose depth can be tuned by coating concentration and duration. The alginate coating remains structurally intact under static and dynamic culture conditions without impairing nutrient transport or cell growth. AlgColTubes eliminate tube-tube and tube-vessel adhesion, and prevent exogenous cell attachment to the outer surface, while maintaining cell viability and proliferation comparable to uncoated ColTubes. Their unique architecture -- an adhesive collagen interior and non-adhesive alginate exterior -- further enables a truncated-tube format for continuous release of biotherapeutic particles through open tube ends. We demonstrate that lentivirus is released from truncated AlgColTubes in a segment length-dependent manner, reaching ~100% release efficiency at 1-mm segment lengths. AlgColTubes provide a scalable, cost-effective platform for high-yield cell and particle manufacturing, with broad potential across basic research, translational studies, and industrial bioprocessing.

bioengineering↗

An arthropod-specific TMEM16 protein accelerates olfactory response termination in Drosophila

Rapid termination of odor responses is essential for accurate olfactory coding in insects, where odorant receptors function as ligand-gated ion channels rather than G protein-coupled receptors. However, the mechanisms that restore olfactory receptor neuron (ORN) excitability after stimulation remain poorly understood. Here, we identify DmTMEM16O (CG6938), an arthropod-specific member of the TMEM16 (Anoctamin) family, as a key regulator of ORN response termination in Drosophila melanogaster. DmTMEM16O is highly enriched in Orco-positive ORNs in both larval and adult olfactory organs. Loss of DmTMEM16O prolongs odor-evoked neuronal activity, increasing decay time constants and causing persistent depolarization. DmTMEM16O mutant ORNs fail to resolve repeated odor stimulation and show impaired temporal coding, accompanied by reduced behavioral responses to both attractive and aversive odorants. Cell-specific rescue demonstrates that DmTMEM16O acts within ORNs to accelerate response termination. Although DmTMEM16O does not exhibit detectable Ca2+-activated chloride channel activity in heterologous cells, our results support a model in which it increases membrane conductance during the decay phase of the response, thereby shortening the membrane time constant and promoting rapid repolarization. This function is consistent with a role for chloride influx in insect ORNs, in contrast to mammalian systems where TMEM16B-mediated chloride efflux amplifies depolarization. Together, our findings identify DmTMEM16O as a lineage-specific regulator of ORN dynamics that enables precise temporal coding in insect olfaction.

physiology↗

PARP1-mediated 5' flap dynamics facilitate Okazaki fragment maturation

Abstract/SummaryPrecise regulation of enzyme recruitment during Okazaki fragment maturation (OFM) is essential for faithful and efficient lagging-strand DNA synthesis. Emerging evidence suggests that PARP1 contributes to OFM yet its specific functions remain unclear. Here, we define context-dependent functions of PARP1 during OFM. Under physiological conditions, PARP1 co-localizes with PCNA in early S phase and restrains Pol {delta}-PCNA- mediated strand-displacement DNA synthesis, thereby preventing the formation of long 5' flaps, which is refractory to FEN1 cleavage. On the other hand, in LIG1-deficient cells, in which DNA nicks and unexpectedly long 5' flaps accumulate, PARP1 promotes the recruitment of LIG3 to catalyze OF ligation and DNA2 to facilitate long 5' flap processing. Collectively, our findings uncover previously unrecognized roles of PARP1 in regulating 5' flap dynamics to ensure efficient OFM and cell viability. HighlightsO_LIPARP1 plays context-dependent regulatory functions in Okazaki fragment maturation (OFM). C_LIO_LIPARP1 controls strand displacement DNA synthesis by the PCNA-Pol{delta} complex to dictate generation of short over long RNA-DNA flaps during canonic OFM. C_LIO_LIPARP1 senses unligated Okazaki fragments in DNA Ligase 1 deficient cells and suppresses unwanted conversion of DNA nicks into 5 flaps. C_LIO_LIProcessing of unligated nicks or flaps by DNA ligase 3 or DNA2, respectively in LIG1 deficient cells depends on PARP1. C_LIO_LIPARP1 inhibitors induce synthetic lethality with DNA ligase 1 or DNA2 inhibition. C_LI

molecular biology↗

Activation of DMH GABAergic neurons, but not local GABAergic AgRP neurons, attenuates chronic stress-induced POMC neuron hyperactivity

Identifying the neural circuits engaged and reshaped by chronic stress is critical for understanding how adaptive responses shift to maladaptive behaviors that contribute to stress-related disorders. Our previous work demonstrates that chronic unpredictable stress (CUS) induces a persistent increase in the firing activity of proopiomelanocortin (POMC) neurons in the arcuate nucleus (ARC). This hyperactivity is due, in part, to a reduction in GABAergic synaptic transmission onto POMC neurons, indicating a disruption in inhibitory control. However, the sources of GABAergic inputs responsible for this effect of chronic stress are unknown. Although AgRP neurons provide local GABAergic input onto POMC neurons and are suppressed by chronic stress, chemogenetic activation of AgRP neurons during stress exposure failed to reduce POMC neuron hyperactivity. GABAergic projections originating from the dorsomedial hypothalamus (DMH) represent another source of inhibitory input to POMC neurons. We found that CUS decreased the firing activity of DMH GABAergic neurons with sex differences, with females exhibiting greater vulnerability to stress-induced suppression. Chemogenetic activation of these neurons during chronic stress markedly attenuated POMC neuron hyperactivity in both sexes, indicating that DMH GABAergic neurons function as a critical upstream regulator of POMC neuron activity under chronic stress. These findings suggest that reduced inhibitory input from DMH GABAergic neurons, rather than local GABAergic AgRP neurons, drives POMC neuron hyperactivity. The weakening of the DMHGABA[->]ARCPOMC circuit activity may represent a novel mechanism underlying maladaptive stress responses and a potential therapeutic target for stress-related disorders.

neuroscience↗

Increases in BCL2L1 and ID1 dosage synergistically drive fate bias and competitive advantage in human pluripotent stem cells

BackgroundGains of chromosome 20q11.21 are among the most common culture-acquired abnormalities in human pluripotent stem cells (hPSC), conferring a well-defined survival advantage while altering differentiation capacity. However, it remains unclear whether this advantage persists during differentiation, how the aneuploidy alters ectodermal and retinal pigment epithelium (RPE) lineage specification, and which genes within the minimal amplicon drive these effects. MethodsWe used three isogenic human embryonic stem cell line pairs (wild-type and 20q11.21 gain) and assessed their behaviour in two neuroectoderm differentiation systems: directed neuroectoderm induction (dual SMAD inhibition) and long-term spontaneous RPE differentiation. Competitive dynamics were measured in mixed cultures, and lineage outcomes were analysed using immunostaining, gene expression profiling and single-cell RNA sequencing. To identify driver genes, we generated BCL2L1 and ID1 overexpression lines and tested their effects under both directed and spontaneous differentiation conditions. ResultsAcross all lines and conditions, 20q cells expanded from a minor fraction to dominate mixed cultures, indicating that their competitive advantage persists beyond the undifferentiated state. Despite this dominance, pure 20q cells failed to specify to neuroectoderm or RPE. Single-cell transcriptomics revealed consistent diversion toward non-neural ectodermal and extraembryonic fates. Mechanistically, overexpression of BCL2L1 and ID1 alone or in combination impaired neuroectoderm specification, while synergistic effect of both genes promoted non-neural ectodermal outcomes under directed differentiation conditions. In spontaneous differentiation, both genes could disrupt differentiation. ConclusionsThe 20q11.21 gain couples a persistent survival advantage with a disruption of neural and RPE lineage competence, redirecting cells toward alternative ectodermal and extraembryonic fates. These effects arise from the combined action of two dosage-sensitive genes BCL2L1 and ID1 within the amplicon, illustrating how regional gene dosage can reshape developmental signalling responses in hPSC.

developmental biology↗

Kindlin-2-Moesin interaction orchestrates sprouting angiogenesis via modulating endothelial membrane mechanics and VEGF signaling

Membrane mechanics play a crucial role in cellular signaling and fate determination, yet their impact on angiogenesis remains poorly understood. Here, we identify Kindlin-2 as a key regulator of sprouting angiogenesis via regulating endothelial membrane tension through its interaction with Moesin, a crucial linker protein between the cell membrane and the actin cortex. Mechanistically, Kindlin-2 binds to the N62 residue of Moesin, limiting its overactivation and maintaining proper membrane tension to facilitate VEGFR2 endocytosis and downstream signaling. Using both developmental and pathological models, we demonstrate that the interaction of Kindlin-2 and Moesin is enhanced in high angiogenic conditions, and endothelial Kindlin-2 deletion reduces angiogenesis. Furthermore, mutation of Moesin at N62 phenocopies the effects of Kindlin-2 loss. Together, these findings uncover a previously unrecognized mechanism linking membrane tension regulation to angiogenesis and provide new insights into targeting the Kindlin-2-Moesin axis for therapeutic intervention in neovascular diseases.

developmental biology↗

Prime-InDel version 1: A web tool providing InDel markers for targeted regions narrowing

While high-density SNP genotyping enables genome-wide assays in rice (Oryza sativa L.) and other crops, PCR-based markers--particularly those derived from insertion-deletion (InDel) variations--remain crucial for fine-mapping. Currently, readily available primer information for high-density InDel rice markers is still limited. We present the first version of PrimeInDel (https://rfgb.rmbreeding.cn/search/variation/primeIndel), an online tool integrating three sets of InDel markers: an 8K-set and a 316K-set from the 3K Rice Genomes (3K-RG) project, and a 22K-set compiled from published literatures. In user cases, primers from PrimeInDel proved highly effective, narrowing a cold-tolerance QTL (qSR2) from 2.7 Mb to 200 kb and a heading-date QTL (qDeh1.1) from 239.5 kb to 83.6 kb. Our work provides the community with a robust, PCR-based InDel primer, accessible via an online data set, which will facilitate germplasm genotyping and gene identification in rice breeding.

plant biology↗