Biology subjects

Dong, M.

Publications and source records attributed to Dong, M..

At least 37 records · Page 2Linked to original sources

bioRxiv2025

Hormetic elevation of taurine restrains inflammaging by deactivating the NLRP3 inflammasome.

Taurine, the most abundant sulfonic amino acid in humans is largely obtained from diets rich in animal proteins. However, taurine is dietary non-essential because it can be synthesized from cysteine by activation of transsulfuration pathway (TSP) when food consumption is low or if the diet is predominantly plant based. The decline of taurine was proposed as the driver of aging through an undefined mechanism. Here, we found that mild food restriction in humans for one year that resulted in 14% reduction of calorie intake elevated the hypotaurine and taurine concentration in adipose tissue. Therefore, we investigated whether elevated taurine mimics caloric-restrictions beneficial effects on inflammation, a key mechanism of aging. Interestingly, aging increased the circulating and tissue concentrations of taurine suggesting that elevated taurine may serve as a hormetic stress response metabolite that regulates mechanism of age-related inflammation. The elevated taurine protected mice against mortality from sepsis and inhibited inflammasome-driven inflammation and gasdermin-D (GSDMD) mediated pyroptosis. Mechanistically, danger signals including hypotonicity that activate NLRP3-inflammasome, caused upstream taurine efflux from macrophages, which triggered potassium (K+) release and downstream canonical NLRP3 inflammasome assembly, caspase-1 activation, GSDMD cleavage and IL-1{beta} and IL-18 secretion that was reversed by taurine restoration. Notably, taurine does not efflux from GSDMD pore and inhibited IL-1{beta} from macrophages independently of known transporters SLC6A6 and SLC36A1. Increased taurine in old mice promotes healthspan by inducing anti-inflammatory pathways previously linked to youthfulness. These findings demonstrate that taurine is an upstream metabolic sensor of cellular perturbations that control NLRP3 inflammasome and lowers age-related inflammation.

immunology↗

bioRxiv2025

CreER activation transiently impairs angiogenesis by slowing endothelial proliferation

Tamoxifen-inducible gene targeting in mice with estrogen receptor-dependent Cre (CreER) recombinase has enormously advanced vascular biology research. However, CreER activation under the control of vascular endothelial promoters is now recognized to cause off-target effects that impair angiogenesis in the widely used perinatal mouse retina model. Although ubiquitously expressed CreER is also used to study retinal angiogenesis, it remains unknown whether it causes similar or more severe off-target effects compared to endothelial-selective CreER activation. Moreover, the cellular processes disrupted by CreER-induced endothelial toxicity remain to be identified. Here, we demonstrate that ubiquitous CreER activation in postnatal mice decreases body growth throughout the period of retinal angiogenesis and impairs retinal angiogenesis in a tamoxifen dose-dependent manner. We further show that CreER activation from both endothelial and ubiquitously expressed CreER transgenes suppresses endothelial cell proliferation downstream of p21/CDKNA1 upregulation. By contrast, we find that p21/CDKNA1 is not upregulated in quiescent adult retinal endothelium, and that CreER-induced postnatal angiogenesis defects recover two weeks after tamoxifen discontinuation. Altogether, our findings indicate that ubiquitous promoters should be avoided for CreER expression when studying endothelial genes, and that short-term retinal angiogenesis studies require endothelial CreER toxicity controls that may be less critical for adult vascular studies.

developmental biology↗

bioRxiv2025

CD163+/Dab2+ Macrophages Alleviate Cardiac Hypertrophy via Nrg2/ErbB4-Mediated Mitochondrial Reprogramming

BackgroundPathological cardiac hypertrophy is a hallmark of numerous cardiovascular diseases, yet effective targeted therapies remain elusive in current clinical practice. Cardiac macrophages contribute to disease progression, but the underlying mechanisms have not been fully elucidated. MethodsSingle-nucleus RNA-sequencing, bulk RNA-sequencing, proteomics, metabolomics, and a mouse model of pressure overload were employed to investigate cardiac remodeling. We identified a macrophage subset co-expressing CD163 and Dab2 and examined its role using immunofluorescence, flow cytometry, and functional assays. We further assessed the Nrg2/ErbB4 signaling axis through genetic and pharmacological modulation. ResultsCD163/Dab2 macrophages were reduced in hypertrophic hearts and positively correlated with Nrg2 expression. These macrophages alleviated cardiomyocyte hypertrophy in vitro, an effect abolished by Nrg2 knockdown. In vivo, recombinant Nrg2 treatment mitigated cardiac hypertrophy, preserved mitochondrial structure, and restored bioenergetics via the ErbB4 receptor. Transcriptomic analyses confirmed enhanced expression of genes involved in mitochondrial oxidative phosphorylation. Furthermore, CD163+/Dab2+ macrophages improved mitochondrial dysfunction by the Nrg2/ErbB4 pathway in vitro. ConclusionsWe identified a CD163/Dab2 macrophage subset that protects against pathological cardiac hypertrophy by promoting mitochondrial function through Nrg2/ErbB4 signaling. This axis may offer a promising therapeutic target for interventions in pathological cardiac hypertrophy.

cell biology↗

bioRxiv2025

Flexible sensitivity to inputs during skilled tongue movements

A hallmark of complex goal-directed movement sequences is the ability to rapidly switch motor programs by integrating incoming sensations with action context1,2, yet the underlying neural implementation remains elusive. Here, we demonstrate a network mechanism for flexibly adjusting the sensitivity of ongoing motor execution to external inputs in different sensorimotor contexts. We trained mice to make sequences of licks directed at a moving target. In a random subset of trials, they made "backtracking" licks by abruptly switching motor programs based on tactile feedback. We divided sessions into alternating cued blocks of trials; sensory-driven backtracking was required during one block-type, but not in the other. Targeted optogenetic stimulation of tongue/jaw somatosensory cortical inputs to the tongue premotor (anterolateral motor, or ALM) cortex that were precisely timed to arrive near the motor switching point reliably induced licking movements resembling those in backtracking trials. This effect was more readily induced during blocks with sensory-driven backtracking and was accompanied by larger optically-evoked deviations in neural activity. Population activity patterns could be separated along a latent axis that discriminated between block-types. On single trials, the location of population activity along this axis correlated with the impact of optogenetic stimulation and influenced the speed of sensory-driven motor switching. Our findings provide causal evidence for how external inputs are integrated with internal context signals to achieve flexible motor control.

neuroscience↗

bioRxiv2025

Point-Of-Need One-Pot Multiplexed RT-LAMP Test For Detecting Three Common Respiratory Viruses In Saliva

Respiratory viral infections pose a significant global public health challenge, partly due to the difficulty in rapidly and accurately distinguishing between viruses with similar symptoms at the point of care, hindering timely and appropriate treatment and limiting effective infection control and prevention efforts. Here, we developed a multiplexed, non- invasive saliva-based, reverse transcription loop-mediated isothermal amplification (RT- LAMP) test that enables the simultaneous detection of three of the most common respiratory infections, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Influenza (Flu), and respiratory syncytial virus (RSV), in a single reaction via specific probes and monitored in real-time by a machine-learning-enabled compact analyzer. Our results demonstrate that the multiplexed assay can effectively detect three target RNAs with high accuracy. Further, testing with spiked saliva samples showed strong agreement with reverse transcription polymerase chain reaction (RT-PCR) assay, with area under the curve (AUC) values of 0.82, 0.93, and 0.96 for RSV, Influenza, and SARS-CoV-2, respectively. By enabling the rapid detection of respiratory infections from easily collected saliva samples at the point of care, the device presented here offers a practical and efficient tool for improving outcomes and helping prevent the spread of contagious diseases. SignificanceThis research presents an innovative approach to respiratory infection diagnostics by combining a one-pot isothermal molecular test with machine learning-based analysis to simultaneously detect SARS-CoV-2, Influenza, and RSV in saliva samples. The battery- powered portable analyzer features novel machine-learning-assisted fluorescence detection for multiplexed reporter quantification, eliminating the need for traditional filter- based optical components and enabling adaptation to new targets without hardware changes. The test demonstrates high accuracy in detecting single and co-infections in spiked saliva samples, providing a rapid, cost-effective point-of-need solution. This tool can expand testing access, improve patient outcomes, and support more effective disease control, particularly in resource-limited or decentralized healthcare settings.

bioengineering↗

bioRxiv2025

RNA-triggered fluorescence controlled by RNA switches for real-time RNA expression tracking in living plants

Real-time visualization of RNA dynamics across spatial scales in living plants is desirable yet remains challenging, hindered by the absence of imaging tools that simultaneously offer high sensitivity and deployability in whole living plants. Here, we engineered a modular, RNA-triggered fluorescence (RTF) reporter system de novo for spatiotemporal imaging of RNA from cellular to whole-plant scales in vivo. This system integrates three functional modules: a target-specific allosteric RNA switch, a degradable adapter for background suppression, and a fluorescent reporter. All components are stably expressed from a single vector and assemble in vivo, enabling uniform deployment throughout entire plants. The modified RTF (RTFst) achieves single-molecule sensitivity (signal-to-noise ratio >60) for dynamic tracking of developmentally regulated, tissue-specific, circadian, and stress-responsive mRNAs. Crucially, it enables the first direct, real-time observation of graft-transmissible mRNA trafficking across tissue scales and cross-kingdom transfer of the aphid-secreted long non-coding RNA Ya1 to plant cells. RTF establishes a versatile platform for studying gene regulation, signaling, and transcript mobility in plants, with broad applicability in synthetic biology and crop improvement.

molecular biology↗

bioRxiv2025

Identification of a divergent botulinum neurotoxin like gene cluster in Furfurilactobacillus

Botulinum neurotoxins (BoNTs) are among the most potent biological toxins and are traditionally associated with Clostridium species. However, recent discoveries have identified BoNT-like proteins in diverse bacterial genera, revealing an expanding family of neurotoxins with unique evolutionary and functional traits. In this study, we report the identification of a novel BoNT-like gene cluster in Furfurilactobacillus sp. OKN36, encoding a toxin we tentatively designate as "Furfuritoxin". Sequence and structural analyses indicate that Furfuritoxin shares key domains with clostridial neurotoxins, including a light chain zinc metalloprotease domain and heavy chain translocase and binding domains, while exhibiting significant sequence divergence. Phylogenetic analysis places Furfuritoxin within a divergent lineage alongside previously reported BoNT/Wo, suggesting a shared ancestral relationship. Gene neighborhood analysis reveals features shared with other BoNT gene clusters including ORFX-related genes and conjugation-associated elements, indicating horizontal gene transfer may have facilitated its distribution. This study adds to the growing family of BoNT-like toxins, providing insights into their evolution and diversity.

bioinformatics↗

bioRxiv2024

Perceptual choice and motor signals in mouse somatosensory cortex

Somatosensory cortex activity relates both to sensation and movement, reflecting their intimate relationship, but the extent and nature of sensory-motor interactions in the somatosensory cortex remain unclear. Here, we investigated perception-related sensory and motor signals in the whisker areas of mouse primary (wS1) and secondary (wS2) somatosensory cortices. We recorded neuronal activity while mice performed a whisker detection task using two alternative lickports, one each to indicate the presence or absence of a whisker deflection on a given trial. One group of mice reported the presence of the whisker stimulus by licking at the port on the same ("congruent") side of the animal as the stimulated whisker, whereas a second group of mice did so by licking at the opposite ("incongruent") side. Activity of single neurons in wS1 and wS2 correlated with perceptual choice. This choice-related activity was enhanced when responding to the congruent side. wS2 neurons projecting along two output pathways--to wS1 or to whisker secondary motor cortex, wM2--also showed choice-related activity, but differed in their dependence on congruence and in the effects of optogenetic manipulation. Thus, somatosensory cortex contains pathway- and action-specific choice-related activity.

neuroscience↗

bioRxiv2024

Organic opto-nanobiointerface enables multiscale biomodulation

Virtually all organic material on Earth has been produced converting solar energy through photosynthesis in chloroplasts, a sack-like, double membrane organelle in plants and algae, where transmembrane electron transfer occurs from lumen to stroma. Although animals hardly harness the power of photosynthesis, their bioelectrical signals extensively regulate complex electrophysiological behaviors, rendering it a superior target for biomedical innovation. Here, a crude structural mimicry of chloroplast has led us to discover that hollow sphere graphitic carbon nitride nanoparticles (hg-C3N4 NPs) endowed non-genetic, subcellular and intercellular photo-modulation of various excitable and non-excitable cells, accumulatively achieving modulation at tissue/organ function level. The homogeneous hg-C3N4 NPs showed responsiveness to light via both photoelectrochemical and photothermal mechanisms. The hg-C3N4 NPs can be spontaneously internalized with excellent cytocompatibility. Using a focusing laser, the hg-C3N4 NPs enable intracellular optical stimulation with subcellular resolution, inducing calcium transient release in multiple cells and propagation in primary cardiomyocytes and cardiac fibroblasts. At multicellular scale, optical pacing and synchronization of cardiomyocyte beating is readily achieved by LED. Further, we demonstrate that hg-C3N4 nanoparticles can be safely delivered into the mouse eye and elicit measurable cortical and behavioral light responses in a subset of animals in a model of advanced retinal degeneration. Finally, application of hg-C3N4 NPs to porcine retinal tissue ex vivo confirmed their modulation capability to directly activate RGCs activity under LED photostimulation. Taken together, these nanostructured biomimetic photocatalytic NPs offer high resolution, leadless optical probing, non-invasive delivery and great biocompatibility, serving as a versatile tool for addressing a range of complex biomedical challenges through subcellular, intercellular and tissue-level photo-modulation across a broad spectrum of scales.

bioengineering↗

bioRxiv2024

Electromagnetic Stimulated Microfillamented Hydrogels for Guided Neuronal Morphogenesis

Biohybrid materials that integrate structural and bioelectronic cues across scales offer new routes to regulate neural development and repair. We present a hybrid, hydrogel-electromagnetic stimulation platform with a 3D hierarchically anisotropic structure, combining FLight bioprinted hydrogel microfilaments with a melt-electrowritten wireless electromagnetic stimulation system, to guide neural morphogenesis across central and peripheral systems at cellular and tissue scale. In the central nervous system, the soft microfilaments selectively promoted healthy, white-matter-like neurite extension rather than glioblastoma-derived outgrowth in human cortical brain tissues, while the electrically stimulation further enhanced the hippocampal neurosphere networking along the microfilaments. As a peripheral nerve conduit, the system directs dorsal root ganglion alignment in vitro and, in rat sciatic defects, drives a coordinated pro-regenerative response that improves axonal guidance, remyelination, and functional recovery. This multiscale biohybrid strategy demonstrates how synergistic integration of soft biomaterials and systematic, wireless bioelectric cues can non-invasively regulate neural morphogenesis, establishing new design principles for next-generation bioelectronic interfaces and regenerative medicine.

bioengineering↗

bioRxiv2024

Pertpy: an end-to-end framework for perturbation analysis

Advances in single-cell technology have enabled the measurement of cell-resolved molecular states across a variety of cell lines and tissues under a plethora of genetic, chemical, environmental, or disease perturbations. Current methods focus on differential comparison or are specific to a particular task in a multi-condition setting with purely statistical perspectives. The quickly growing number, size, and complexity of such studies requires a scalable analysis framework that takes existing biological context into account. Here, we present pertpy, a Python-based modular framework for the analysis of large-scale perturbation single-cell experiments. Pertpy provides access to harmonized perturbation datasets and metadata databases along with numerous fast and user-friendly implementations of both established and novel methods such as automatic metadata annotation or perturbation distances to efficiently analyze perturbation data. As part of the scverse ecosystem, pertpy interoperates with existing libraries for the analysis of single-cell data and is designed to be easily extended.

bioinformatics↗

bioRxiv2024

Freestanding rGO electrodes with tunable porous structures for improved neural recording

Invasive neural electrodes prepared from materials with miniaturized geometrical size could improve the longevity of implants by reducing the chronic inflammatory response. Graphene-based microfibers with tunable porous structures have a large electrochemical surface area (ESA)/geometrical surface area (GSA) ratio that has been reported to possess low impedance and high charge injection capacity (CIC), yet the control of the porous structure remains to be fully investigated. In this study, we introduce wet-spun graphene-based electrodes with pores tuned by sucrose concentrations in the coagulation bath. The electrochemical properties of thermally reduced rGO were optimized by adjusting the ratio of rGO to sucrose, resulting in significantly lower impedance, higher CIC, and higher charge storage capacity (CSC) than platinum microwires. Tensile and insertion tests confirmed that optimized electrodes had sufficient strength to ensure a 100% insertion success rate with low angle shift, thus allowing precise implantation without the need for additional mechanical enhancement. Acute in-vivo recordings from the auditory cortex found low impedance benefits from the recorded amplitude of spikes, leading to an increase in the signal-to-noise ratio (SNR). Ex-vivo recordings from hippocampal brain slices demonstrate that it is possible to record and/or stimulate with graphene-based electrodes with good fidelity compared with conventional electrodes.

neuroscience↗

bioRxiv2024

Interpreting mammalian evolutionary constraint at synonymous sites in light of the unwanted transcript hypothesis

The unwanted transcript hypothesis presents a potential explanation for cryptic evolutionary constraint at synonymous sites in species with low effective population sizes, such as humans and other mammals. Selection for higher GC content and against mutations that alter splicing in native transcripts is predicted to shape synonymous site content and protect against unwanted transcripts. Here, we interpret mammalian synonymous site constraint in this context. Utilising the largest alignment of 240 placental mammal genomes and single-base resolution constraint scores, we show that 20.8% of four-fold degenerate sites are under significant constraint across mammals. There is a strong bias for guanine (G) and cytosine (C) at constrained sites, marked constraint near splice sites, and variation in human populations shows a bias against mutations that reduce synonymous site GC content. We find evidence for higher constraint on four-fold degenerate sites in species with small historic effective population sizes and high young transposable element genome content. Genes enriched for synonymous site constraint, including those forming CpG sites, are tightly regulated and integral to organismal viability through their involvement in embryo development and transcriptional regulation.

evolutionary biology↗

bioRxiv2024

Characterization of the SARS-CoV-2 BA.5 Variants in H11-K18-hACE2 Hamsters

This study aims to comprehensively characterize the SARS-CoV-2 BA.5 variants using K18 hACE2 transgenic mice and golden hamsters as model organisms. Previous research on SARS-CoV-2 has utilized both mouse and hamster models, leading to conflicting results concerning the viruss lethality. In our study, the finding suggests that H11-K18 hACE2 golden hamsters closely mimic the disease progression observed in human COVID-19 cases caused by BA.5 variants, demonstrating consistent severity and symptoms comparable to severe infections. Additionally, hamsters exhibit heightened respiratory viral replication, accurately reflecting the clinical viral kinetics observed in humans. The study emphasizes the critical importance of selecting an appropriate animal model for SARS-CoV-2 research, while also providing robust support for the hypothesis that BA.5 variants contribute to fatal outcomes in COVID-19 cases. These findings highlight the pivotal role of the golden hamster model in advancing our understanding of the pathogenic mechanisms underlying SARS-CoV-2 variants, as well as in the development of targeted therapeutic strategies. Significance StatementOur research work explores groundbreaking insights that could reshape our understanding of COVID-19 and pave the way for targeted therapies. We use golden hamster models to express the possibility of different animal models could contribute to human diseases. We hope this finding could clarify some conflicts existed, and help further development of medication for COVID.

immunology↗

bioRxiv2024

High Shear Stress Reduces ERG Causing Endothelial-Mesenchymal Transition and Pulmonary Arterial Hypertension

Pathological high shear stress (HSS, 100 dyn/cm2) is generated in distal pulmonary arteries (PA) (100-500 m) in congenital heart defects and in progressive PA hypertension (PAH) with inward remodeling and luminal narrowing. Human PA endothelial cells (PAEC) were subjected to HSS versus physiologic laminar shear stress (LSS, 15 dyn/cm2). Endothelial-mesenchymal transition (EndMT), a feature of PAH not previously attributed to HSS, was observed. H3K27ac peaks containing motifs for an ETS-family transcription factor (ERG) were reduced, as was ERG-Kruppel-like factors (KLF)2/4 interaction and ERG expression. Reducing ERG by siRNA in PAEC during LSS caused EndMT; transfection of ERG in PAEC under HSS prevented EndMT. An aorto-caval shunt was preformed in mice to induce HSS and progressive PAH. Elevated PA pressure, EndMT and vascular remodeling were reduced by an adeno-associated vector that selectively replenished ERG in PAEC. Agents maintaining ERG in PAEC should overcome the adverse effect of HSS on progressive PAH.

cell biology↗

bioRxiv2024

Arsinothricin Biosynthesis Involving a Noncanonical Radical SAM Enzyme for C-As Bond Formation

Arsinothricin is a potent antibiotic secreted by soil bacteria. The biosynthesis of Arsinothricin was proposed to involve two steps. The first step is C-As bond formation between trivalent As and the 3-amino-3-carboxypropyl (ACP) group of S-adenosyl-L-methionine (SAM), which is catalyzed by the protein ArsL. However, the reaction has not been verified in vitro, and ArsL has not been characterized in detail. Interestingly, ArsL contains a CxxxCxxC motif and thus belongs to the radical SAM enzyme superfamily, the members of which cleave SAM and generate a 5'-deoxyadenosyl radical. Here, we found that ArsL cleaves the C{gamma},Met-S bond of SAM and generates an ACP radical that resembles Dph2, a noncanonical radical SAM enzyme involved in diphthamid biosynthesis. As Dph2 does not contain the CxxxCxxC motif, ArsL is a unique noncanonical radical SAM enzyme that contains this motif but generates an ACP radical. Together with the methyltransferase ArsM, we successfully reconstituted arsinothricin biosynthesis in vitro. ArsL has a conserved RCCLKC motif in the C-terminal sequence and belongs to the RCCLKC-tail radical SAM protein subfamily. By truncation, we showed that this motif binds to the substrate arsenite and is highly important for its activity. Our results suggested that ArsL is a noncanonical radical SAM enzyme with a canonical radical SAM enzyme motif, implying that more noncanonical radical SAM chemistry may exist within the radical SAM enzyme superfamily. TOC O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=40 SRC="FIGDIR/small/577332v1_ufig1.gif" ALT="Figure 1"> View larger version (8K): org.highwire.dtl.DTLVardef@14547e2org.highwire.dtl.DTLVardef@172753eorg.highwire.dtl.DTLVardef@16f4a99org.highwire.dtl.DTLVardef@18ab4c8_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2024

Multi-state kinetics of the syringe-like injection mechanism of Tc toxins

Tc toxins are virulence factors of many insects and human pathogenic bacteria. They attach as soluble prepores to receptors on host cells and following acidification in the late endosome, perforate the cell membrane like a syringe to translocate toxic enzymes into the host cell through their pore-forming channel. Although this complex transformation has been structurally well studied, the functional aspects of this large-scale rearrangement, such as the reaction pathway with possible intermediate states and the resulting temporal evolution have remained elusive. Here, we used an integrated biophysical approach to monitor the prepore-to-pore transition and found that it takes [~]28 h when induced by high pH in the absence of other factors. In the presence of liposomes, an increasingly high pH or receptors, such as heparin or Vsg, the probability to transform prepores to pores increases by a factor of up to 4. This effect can also be mimicked by biotinylation or site-directed mutagenesis of the shell, demonstrating that shell destabilization is a crucial step in prepore-to-pore transition. We show that shell opening is a heterogeneous process with transition times ranging from 60 ms to 1.6 s and resolve three sequential intermediate states: an initial transient intermediate during shell destabilization, a first stable intermediate where the receptor-binding domains on the shell rearrange and a second stable intermediate with an open shell. In contrast, the ejection of the pore-forming channel from the open shell is highly cooperative with a transition time of < 60 ms. This detailed knowledge of the Tc toxin mechanism of action, even in the absence of receptors, is important for the future application of Tc toxins as biomedical devices or biopesticides.

biochemistry↗

bioRxiv2023

Targeting endothelial FOXO1 protects diabetic β-cells and improves wound healing

The forkhead box O1 (FOXO1) transcription factor plays critical roles in regulating not only metabolic activity but also angiogenesis in the vascular endothelium1-4. Our previous studies show that epsin endocytic adaptors can regulate both angiogenesis and lymphangiogenesis5-7. Endothelial cells (ECs) lining the inside of blood vessels are continuously exposed to circulating insulin and insulin-like growth factors (IGFs). Emerging evidences suggest that ECs can affect {beta}-cell function8-11. Excessive IGF2, especially elevated local IGF2 levels in islets, may represent a risk factor for developing diabetes12-15; however, the underlying molecular mechanisms by which aberrant angiogenesis and endothelium-derived factors regulate pancreatic {beta}-cell function in diabetes remain unclear. Here, we report that the pancreas of diabetic patients as well as the pancreas, skin, and plasma of streptozotocin/high fat diet (STZ/HFD)-induced diabetic mice and db/db mice contains excess IGF2, which can lead to {beta}-cell dysfunction and apoptosis. Single-cell transcriptomics combined with mass spectrometry analysis reveal that endothelial-specific knockout of FOXO1 increases circulating soluble and cell-membrane or intracellular expression levels of IGF type 2 receptor (IGF2R) and CCCTC-binding factor (CTCF), while decreasing IGF2 levels in diabetes. Both IGFR215-17 and CTCF18-21 can reduce IGF2 levels and may ameliorate {beta}-cell decline associated with excess IGF2 in diabetes. Furthermore, depletion of FOXO1, epsins, or knockdown of ULK1 inhibits autophagy formation in ECs, preventing degradation of vascular endothelial growth factor receptor 2 (VEGFR2) to promote angiogenesis and improve wound healing in diabetes. Our findings reveal that endothelial FOXO1 regulates epsin-dependent angiogenesis and affects {beta}-cell function and fate through CTCF and IGF2-IGF2R, providing a potential strategy for ameliorating diabetes and accelerating cutaneous wound healing.

cell biology↗