Biology subjects

Gao, Y.

Publications and source records attributed to Gao, Y..

At least 253 records · Page 14Linked to original sources

bioRxiv2024

Chemically induced cell plasticity enables the generation of high-fidelity embryo model

Embryo models open exciting opportunities for improving our understanding of development and advancing medicine. However, current models depend on intricate procedures and the integration of separately generated cell types. Here, we aimed to establish a chemical-only strategy to induce cellular plasticity and generate embryo founder-like cells. Within 60 hours, chemical-induced pluripotent stem cells activated early lineage specifiers, Cdx2 and Gata6, while maintaining Oct4/ Pou5f1 expression. Following a second 36-hour chemical treatment, founder-like cells developed into all blastocyst fates and self-assembled into embryo-like structures. These progressed into high-fidelity late-streak gastrulation stages within 4.5 days, displaying primitive streak formation via epithelial-to-mesenchymal transition, along with germ-layer tissues, amnion, and ectoplacental cone. Fate trajectories and molecular signatures from founder-like cells to chemically-induced embryo model (ci-MO) were defined, revealing remarkable similarity up to Embryonic day (E) 7.5 embryo. Summarily, ci-MO offers a straightforward, rapid, small-molecule-only, and high-fidelity approach to study embryonic development and cell fate specification. HighlightsO_LIGeneration of a direct, efficient, and sophisticated chemically induced embryo model (ci-MO) C_LIO_LITwo-pulse chemical induction reprograms ESCs into founder-like cells capable of developing all embryo fates C_LIO_LIGastrulating ci-MOs display high morphological and molecular accuracy C_LIO_LIci-MOs reconstruct embryo developmental trajectories C_LI

cell biology↗

bioRxiv2024

Changes in urinary proteome in healthy individuals taking multi-vitamin/mineral supplements

Multivitamin/mineral supplements are widely used in many populations. Urine proteome has not been used to investigate its overall effects on healthy individuals. In this study, the urine proteome changed after taking compound nutrient supplements for 2 weeks and 4 weeks, and the differential proteins and their enriched pathways may be associated with nutrient supplementation. We noted that after taking the supplements for 2 weeks, the amount of erythropoietin receptor in urine was average 449 times higher than before, with four out of nine people experiencing changes from 0 to 14635, 11851, 11181, and 5930. The results of this study revealed that the urine proteome can provide new clues about the relatively short-term health effects of the supplements. It may help optimize guidelines and recommendations for the use of supplements.

biochemistry↗

bioRxiv2024

Urine Proteomics in Methamphetamine Addicted Patients

Drug addiction is a serious chronic relapsing brain disease, and methamphetamine dependence has a complex course and is difficult to treat, causing a serious public health burden. In this study, we compared and analyzed the urine proteome of methamphetamine-using patients, methamphetamine-withdrawing patients, healthy individuals. The urine proteome of methamphetamine-using patients and methamphetamine-withdrawing patients was significantly different from that of healthy individuals, and some differential proteins and their enriched biological functions showed that they were associated with addiction or neurotoxicity of methamphetamine. The fact, that the urine proteome of patients who withdrew methamphetamine for more than 3 months were significantly different from that of healthy patients, may reveal the reasons for the high rate of methamphetamine relapse and potential intervention targets. This study proved that urine proteome can reflect the effects of addiction on the body comprehensively, and has the potential to provide clues for addiction.

biochemistry↗

bioRxiv2024

The transcriptomic and spatial organization of telencephalic GABAergic neuronal types

The telencephalon of the mammalian brain comprises multiple regions and circuit pathways that play adaptive and integrative roles in a variety of brain functions. There is a wide array of GABAergic neurons in the telencephalon; they play a multitude of circuit functions, and dysfunction of these neurons has been implicated in diverse brain disorders. In this study, we conducted a systematic and in-depth analysis of the transcriptomic and spatial organization of GABAergic neuronal types in all regions of the mouse telencephalon and their developmental origins. This was accomplished by utilizing 611,423 single-cell transcriptomes from the comprehensive and high-resolution transcriptomic and spatial cell type atlas for the adult whole mouse brain we have generated, supplemented with an additional single-cell RNA-sequencing dataset containing 99,438 high-quality single-cell transcriptomes collected from the pre- and postnatal developing mouse brain. We present a hierarchically organized adult telencephalic GABAergic neuronal cell type taxonomy of 7 classes, 52 subclasses, 284 supertypes, and 1,051 clusters, as well as a corresponding developmental taxonomy of 450 clusters across different ages. Detailed charting efforts reveal extraordinary complexity where relationships among cell types reflect both spatial locations and developmental origins. Transcriptomically and developmentally related cell types can often be found in distant and diverse brain regions indicating that long-distance migration and dispersion is a common characteristic of nearly all classes of telencephalic GABAergic neurons. Additionally, we find various spatial dimensions of both discrete and continuous variations among related cell types that are correlated with gene expression gradients. Lastly, we find that cortical, striatal and some pallidal GABAergic neurons undergo extensive postnatal diversification, whereas septal and most pallidal GABAergic neuronal types emerge simultaneously during the embryonic stage with limited postnatal diversification. Overall, the telencephalic GABAergic cell type taxonomy can serve as a foundational reference for molecular, structural and functional studies of cell types and circuits by the entire community.

neuroscience↗

bioRxiv2024

A unifying model of species abundance distribution

The species abundance distribution (SAD) is one of the most fundamental and best-studied macroecological patterns at the core of any biodiversity theory. Remarkably, almost every community investigated to date shows a hollow curve, indicative of the presence of many rare species and a few abundant species. While the precise nature of SAD is believed to reflect fundamental ecological processes underlying community assembly, ecologists have yet to identify a single model that comprehensively explains all SADs. Recent studies using large datasets suggested that logseries best describes animal and plant communities1,2 while lognormal is the best model for microbes3, thereby challenging the notion of a unifying SAD model across the tree of life. Using a large dataset of [~]30,000 globally distributed communities spanning animals, plants and microbes from diverse environments, here we show that powerbend distribution, predicted by a maximum information entropy-based theory of ecology, emerges as a unifying model that accurately captures SADs of all life forms, habitats and abundance scales, supporting the existence of universal ecological principles. Our findings reject the notion of pure neutrality and support the idea that community assembly is driven by both random fluctuations and deterministic mechanisms, such as interspecific trait variation and resource competition. We also show that the previously estimated one trillion microbial species existing on Earth might be orders of magnitude off.

ecology↗

bioRxiv2024

Single-Cell Phenotyping of Extracellular Electron Transfer via Microdroplet Encapsulation

Electroactive organisms contribute to metal cycling, pollutant removal, and other redox-driven environmental processes. Studying this phenomenon in high-throughput is challenging since extracellular reduction cannot easily be traced back to its cell of origin within a mixed population. Here, we describe the development of a microdroplet emulsion system to enrich EET-capable organisms. We validated our system using the model electroactive organism S. oneidensis and describe the tooling of a benchtop microfluidic system for oxygen-limited processes. We demonstrated enrichment of EET-capable phenotypes from a mixed wild-type and EET-knockout population. As a proof-of-concept application, bacteria were collected from iron sedimentation from Town Lake (Austin, TX) and subjected to microdroplet enrichment. We observed an increase in EET-capable organisms in the sorted population that was distinct when compared to a population enriched in a bulk culture more closely akin to traditional techniques for discovering EET-capable bacteria. Finally, two bacterial species, C. sakazakii and V. fessus not previously shown to be electroactive, were further cultured and characterized for their ability to reduce channel conductance in an organic electrochemical transistor (OECT) and to reduce soluble Fe(III). We characterized two bacterial species not previously shown to exhibit electrogenic behavior. Our results demonstrate the utility of a microdroplet emulsions for identifying putative EET-capable bacteria and how this technology can be leveraged in tandem with existing methods.

bioengineering↗

bioRxiv2024

Increased pathogenicity and transmission of SARS-CoV-2 Omicron XBB.1.9 sublineage, including HK.3 and EG.5.1

With the SARS-CoV-2 Omicron XBB.1.9 sublineage circulating worldwide, two XBB.1.9 variants, EG.5.1 and HK.3 spread rapidly and became dominant from middle 2023. However, the spike features, pathogenicity, and transmissibility of HK.3 are largely unknown. Here, we performed multiscale investigations to reveal the virological features of XBB.1.9 subvariants, especially the newly emerging HK.3. HK.3 revealed high replication efficiency in vitro. The HK.3 spike exhibited enhanced processing, although its infectivity, fusogenicity, and hACE2 binding affinity were comparable to those of the EG.5 and XBB.1 spikes. All XBB.1.9.1, EG.5.1 and HK.3 strains demonstrated efficient transmission in hamsters, although XBB.1.9.1 exhibited stronger fitness in the upper airways. HK.3 and EG.5.1 exhibited greater pathogenicity than XBB.1.9.1 and BA.2 in H11-K18-hACE2 hamsters. Our studies provide insight into the newly emerging pathogens HK.3 and EG.5.1. ImportanceIn animal models, the ongoing attenuated pathogenicity and poor transmission of Omicron subvariants seems to reach a consensus. However, our results revealed that Omicron XBB.1.9 subvariants, including one of the key variants of interest, EG.5 with its another key subvariant HK.3, universally exhibited both increased pathogenicity and highly transmission. This study reemphasized the importance of surveillance in characteristics of epidemic Omicron subvariants.

microbiology↗

bioRxiv2024

How does the ion concentration affect the functions of kinesin BimC

BimC family proteins are bipolar motor proteins belonging to the kinesin superfamily which promote mitosis by crosslinking and sliding apart antiparallel microtubules. Understanding the binding mechanism between the kinesin and the microtubule is crucial for researchers to make advances in the treatment of cancer and other malignancies. Experimental research has shown that the ion concentration affects the function of BimC significantly. But the insights of the ion-dependent function of BimC remain unclear. By combining molecular dynamics (MD) simulations with a series of computational approaches, we studied the electrostatic interactions at the binding interfaces of BimC and the microtubule under different KCl concentrations. We found the electrostatic interaction between BimC and microtubule is stronger at 0 mM KCl compared to 150 mM KCl, which is consistent with experimental conclusions. Furthermore, important salt bridges and residues at the binding interfaces of the complex were identified, which illustrates the details of the BimC-microtubule interactions. Molecular dynamics analyses of salt bridges identified that the important residues on the binding interface of BimC are positively charged, while those residues on the binding interface of the tubulin heterodimer are negatively charged. The finding in this work reveals some important mechanisms of kinesin-microtubule binding, which helps the future drug design for cancer therapy.

biophysics↗

bioRxiv2024

Use of machine learning for quantification of retinal pigment epithelium tight junctions improves assay sensitivity

The retinal pigment epithelium (RPE) is critical for maintaining outer retinal barrier homeostasis. In age-related macular degeneration (AMD), the RPE can undergo a dedifferentiation process that includes tight junction (TJ) loss and displacement of zonula occludens-1 (ZO-1), which may impair structural and functional integrity of the RPE barrier and contribute to disease pathogenesis. Our objective was to develop an automated and sensitive quantification method for TJ aberrations in an RPE immunofluorescence imaging assay, following treatment with TNF or TGF{beta}2. However, quantifying ZO-1 morphological changes in the RPE using standard image analysis methods did not provide a satisfactory assay window. To address this challenge, we developed an imaging assay to quantify ZO-1 changes using a machine learning approach, enabling enhanced phenotypic characterization of the ZO-1 changes in RPE cells and improved assay sensitivity. We were also able to capture and quantify the reversal of these changes using etanercept, an TNF inhibitor, with this imaging assay. Our findings indicated that this machine learning ZO-1 quantification assay could serve as a potential phenotypic readout for RPE dedifferentiation and enabling large-scale mechanistic studies.

bioinformatics↗

bioRxiv2024

Effect of autonomous consumption of different sweet substances on the urinary proteome of mice

ObjectiveTo explore the possible effects on the organism by analysing changes in the urinary proteome of mice after autonomous consumption of different sweet substances. MethodsUrine samples were collected from C57BL/6l mice before and after active consumption of sweet substances. The sweet substances included sucrose, stevia glycosides, acesulfame, and sucralose, which are more widely used worldwide and can elicit a preference response in mice, and the concentrations of the non-nutritive sweeteners were chosen to be those that have been shown by existing studies to have the strongest preference response in mice. The analyses were performed by the non-labelled quantitative proteomics technique of high performance liquid chromatography tandem mass spectrometry (LC-MS/MS), and groups were screened for differential proteins in the urinary proteome for the analysis of protein functions and biological processes; comparisons of urinary proteomes before and after consumption of sweeteners by a single mouse were carried out; and side-by-side comparisons of different sweeteners were made. Results and ConclusionsUrine proteome could reflect the changes in the organism of mice after voluntary consumption of sweeteners, and the effects of different sweeteners on the urine proteome were not the same. Among the four sweeteners, sucralose and sucrose induced the most similar changes in the organism, and steviol glycosides induced the furthest changes in the organism; the changes induced by sucrose, acesulfame, and sucralose were similar, and steviol glycosides induced changes different from those of the other sweeteners. Stevia glycosides induced changes that were different from the other sweeteners. The urinary proteomic proteins that differed among the four sweeteners consumed autonomously by mice included proteins that have been reported to be associated with brain reward circuits, whereas only the urinary proteomic proteins that differed among the four sweeteners consumed autonomously by mice were associated with metabolic processes in large quantities after the voluntary consumption of sucrose, acesulfame, and sucralose, and the urinary proteomic proteins that differed among the four sweeteners consumed autonomously by mice were associated with the assembly of nucleosomes, gene expression, and other processes after the voluntary consumption of stevia glycosides.

biochemistry↗

bioRxiv2024

Interleukin-1 prevents SARS-CoV-2-induced membrane fusion to restrict viral transmission via induction of actin bundles

Innate immune responses triggered by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection play pivotal roles in the pathogenesis of COVID-19, while host factors including pro-inflammatory cytokines are critical for viral containment. By utilizing quantitative and qualitative models, we discovered that soluble factors secreted by human monocytes potently inhibit SARS-CoV-2-induced cell-cell fusion in viral-infected cells. Through cytokine screening, we identified that interleukin-1{beta} (IL-1{beta}), a key mediator of inflammation, inhibits syncytia formation mediated by various SARS-CoV-2 strains. Mechanistically, IL-1{beta} activates RhoA/ROCK signaling through a non-canonical IL-1 receptor-dependent pathway, which drives the enrichment of actin bundles at the cell-cell junctions, thus prevents syncytia formation. Notably, in vivo infection experiments in mice confirms that IL-1{beta} significantly restricted SARS-CoV-2 spreading in the lung epithelia. Together, by revealing the function and underlying mechanism of IL-1{beta} on SARS-CoV-2-induced cell-cell fusion, our study highlights an unprecedented antiviral function for cytokines during viral infection.

immunology↗

bioRxiv2024

Unbiased metastatic niche-labeling identifies estrogen receptor-positive macrophages as a barrier of T cell infiltration during bone colonization

Microenvironment niches determine cellular fates of metastatic cancer cells. However, robust and unbiased approaches to identify niche components and their molecular profiles are lacking. We established Sortase A-Based Microenvironment Niche Tagging (SAMENT), which selectively labels cells encountered by cancer cells during metastatic colonization. SAMENT was applied to multiple cancer models colonizing the same organ and the same cancer to different organs. Common metastatic niche features include macrophage enrichment and T cell depletion. Macrophage niches are phenotypically diverse between different organs. In bone, macrophages express the estrogen receptor alpha (ER) and exhibit active ER signaling in male and female hosts. Conditional knockout of Esr1 in macrophages significantly retarded bone colonization by allowing T cell infiltration. ER expression was also discovered in human bone metastases of both genders. Collectively, we identified a unique population of ER+ macrophages in the metastatic niche and functionally tied ER signaling in macrophages to T cell exclusion during metastatic colonization. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=165 SRC="FIGDIR/small/593016v1_ufig1.gif" ALT="Figure 1"> View larger version (60K): org.highwire.dtl.DTLVardef@1cd65b6org.highwire.dtl.DTLVardef@1d099e2org.highwire.dtl.DTLVardef@26cd91org.highwire.dtl.DTLVardef@181f5f3_HPS_FORMAT_FIGEXP M_FIG C_FIG HIGHLIGHTSO_LISAMENT is a robust metastatic niche-labeling approach amenable to single-cell omics. C_LIO_LIMetastatic niches are typically enriched with macrophages and depleted of T cells. C_LIO_LIDirect interaction with cancer cells induces ER expression in niche macrophages. C_LIO_LIKnockout of Esr1 in macrophages allows T cell infiltration and retards bone colonization. C_LI

cancer biology↗

bioRxiv2024

Single cell profiling of bone metastasis ecosystems from multiple cancer types reveals convergent and divergent mechanisms of bone colonization.

Bone represents congenial soil for metastatic seeds and is frequently affected by metastasis of multiple cancer types. The histological and molecular characteristics of bone metastases (BMs) are diverse but poorly understood. Herein, we performed single-cell RNA-seq on 34 BMs from 6 cancer types and identified 3 ecosystem archetypes characterized by enrichment of macrophages/osteoclasts (M{varphi}-OC), regulatory/exhausted T cells (Treg-Tex), and monocytes (Mono), respectively. Breast cancer BMs are mostly the M{varphi}-OC archetype driven by the osteolytic vicious cycle, whereas kidney cancers BMs mainly belong to the Treg-Tex archetype that lacks osteoclasts. Lung cancers BMs evenly distributed across all archetypes. Further analyses revealed parallel mechanisms of immunosuppression and bone remodeling. Elevated estrogen signaling distinguishes macrophages in the M{varphi}-OC subtype, which was investigated in a companion study. Together, we elucidated that divergent mechanisms toward bone colonization and that BMs of different origins can adopt the same mechanism through convergent evolution or adaptation. HIGHLIGHTSO_LIAnalyses of bone metastases from 6 cancer types revealed three immune archetypes. C_LIO_LIArchetypes diverge on immune trajectories, and features of tumor and stromal cells. C_LIO_LIDominant cell type in each archetype undergoes convergent evolution. C_LIO_LIRegulatory networks converge on osteoclasts and Tregs to drive archetype formation. C_LI Graphic Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=166 SRC="FIGDIR/small/593027v1_ufig1.gif" ALT="Figure 1"> View larger version (61K): org.highwire.dtl.DTLVardef@82cef6org.highwire.dtl.DTLVardef@1e1b021org.highwire.dtl.DTLVardef@1f2881eorg.highwire.dtl.DTLVardef@1c6874a_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2024

Observing one-divalent-metal-ion dependent and histidine-promoted His-Me family I-PpoI nuclease catalysis in crystallo

Metal-ion-dependent nucleases play crucial roles in cellular defense and biotechnological applications. Time-resolved crystallography has resolved catalytic details of metal-ion-dependent DNA hydrolysis and synthesis, uncovering the essential roles of multiple metal ions during catalysis. The histidine-metal (His-Me) superfamily nucleases are renowned for binding one divalent metal ion and requiring a conserved histidine to promote catalysis. Many His-Me family nucleases, including homing endonucleases and Cas9 nuclease, have been adapted for biotechnological and biomedical applications. However, it remains unclear how the single metal ion in His-Me nucleases, together with the histidine, promotes water deprotonation, nucleophilic attack, and phosphodiester bond breakage. By observing DNA hydrolysis in crystallo with His-Me I-PpoI nuclease as a model system, we proved that only one divalent metal ion is required during its catalysis. Moreover, we uncovered several possible deprotonation pathways for the nucleophilic water. Interestingly, binding of the single metal ion and water deprotonation are concerted during catalysis. Our results reveal catalytic details of His-Me nucleases, which is distinct from multi-metal-ion-dependent DNA polymerases and nucleases.

biochemistry↗

bioRxiv2024

Amblyopic deficits in monocular processing and binocular interactions revealed by submillimeter 7T fMRI and EEG frequency tagging

Disruption of retinal input early in life can lead to amblyopia, a condition characterized by reduced visual acuity despite corrected optics. Although extensive losses of neural activity have been found in the early visual cortex, it remains unclear whether they reflect deficits in feedforward or feedback processing, or abnormal binocular interactions. Combining submillimeter 7T fMRI and EEG frequency tagging, our study revealed the precise neural deficits in monocular processing and binocular interactions in human adults with unilateral amblyopia. Cortical depth-dependent fMRI revealed monocular response deficits in cortical layers of the primary visual cortex (V1) receiving thalamic input, which carried over to the downstream areas (V2-V4) in feedforward processing. Binocular stimulation produced a greater signal loss in the superficial layers of V1, consistent with suppression from the fellow eye by lateral inhibition. EEG data further demonstrate reduced suppression from the amblyopic eye, weakened binocular integration, and delayed monocular and binocular processing. Our results support attenuated and delayed monocular processing in V1 layers receiving thalamic input in human amblyopia, followed by imbalanced binocular suppression and weakened binocular integration in the superficial layers, which further reduced signal strength and processing speed. These precise neural deficits can help developing more targeted and effective treatments for the vision disorder. SignificanceCortical depth-dependent 7T fMRI and EEG frequency tagging revealed attenuated and delayed neural activity in monocular processing and binocular interactions in human amblyopia. Neural deficits in monocular processing arise from the input layers of V1, followed by imbalanced binocular suppression and weakened binocular integration in the superficial layers. The precise neural deficits at high spatiotemporal resolution can help developing more targeted and effective treatment for amblyopia. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=159 SRC="FIGDIR/small/591076v1_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@c174d8org.highwire.dtl.DTLVardef@de51eborg.highwire.dtl.DTLVardef@1d443c0org.highwire.dtl.DTLVardef@ac267a_HPS_FORMAT_FIGEXP M_FIG C_FIG This figure illustrates the precise neural deficits of amblyopia in monocular processing and binocular interactions based on the main findings of the current study. Attenuated and delayed neural activity arises from V1 cortical layers receiving thalamic input in monocular processing, followed by imbalanced binocular suppression and weakened binocular integration in the superficial layers, further reducing visual signal strength and processing speed.

neuroscience↗

bioRxiv2024

Exploring differences between depression and bipolar disorder through the urinary proteome

How to differentiate the diagnosis of depression and bipolar disorder has always been an important problem that needs to be solved urgently in clinical practice. In this study, from the perspective of urine proteomics, urine samples of similar age were collected from two hospitals to investigate the candidate biomarkers for differentiating the diagnosis of depression and bipolar disorder using both group analysis and one-to-many analysis(1 patient: many control samples). The experimental results of the paired group analysis showed that 108 differential proteins were identified in the depressed group compared to the bipolar group under strict screening conditions with screening criteria of FC [≥] 2 or [≤] 0.5 and a two-tailed unpaired t-test of P < 0.01, with an average of 3.7 randomly generated differential proteins, and a confidence level of 96.6 % for the correlation between these proteins and the disease difference. In the one-to-many analysis, 24 differential proteins were co-identified by the samples of 13 depressed patients, 16 of which showed a completely consistent trend of expression changes in all depressed patients studied, and 6 of which were associated with immunoglobulins; 41 differential proteins were co-identified by the samples of 12 depressed patients out of 13, and 19 of which showed a completely consistent trend of expression change in the These results reflect the strong consistency of differential proteins between the two groups of patients. 12 or more samples from depressed patients were enriched for differential proteins related to multiple biological processes and signaling pathways associated with the immune system, which is consistent with previous studies: immune mechanisms may be one of the pathogenetic mechanisms of major depression and that drugs with major immune targets can improve depressive symptoms. In the future, it may be possible to observe the immune status of patients with depression to provide direction and basis for the precise treatment of depression. The results of this paper show that urine proteomics can differentiate between depression and bipolar disorder, suggest possible mechanisms and potential targets for the treatment of depression and bipolar disorder, and provide a tool for future differential diagnosis and precision treatment of the diseases.

biochemistry↗

bioRxiv2024

The splicing factor kinase SRPK1 is a therapeutic target for Peripheral Vascular Disease

IntroductionIn peripheral arterial disease (PAD) anti-angiogenic VEGF-A165b isoform overexpression in monocytes contributes to impaired collateralisation. Serine-arginine protein-kinase-1 (SRPK1) regulates VEGF splicing. To determine whether SRPK1 controlled monocytic VEGF, impairing collateralisation, we investigated SRPK1 inhibition and monocyte-specific knockout in mouse models of and in human monocytes from PAD. MethodsVEGF-A165b activity was measured by co-culture of PAD patients monocytes with endothelial cells with SRPK1 inhibition. Mice with impaired revascularisation due to soluble-frizzled-related-protein-5 knockout (Sfrp5-/-), monocyte-specific Wnt5a gain-of-function (LysM-Wnt5aGOF), or obese mice on a high-fat high-sucrose (HF/HS) diet were subjected to femoral artery ligation and treated with SRPK1 inhibitor. We generated an SRPK1 conditional knockout and crossed it with monocyte-specific (LysM-Cre) driver line to specifically knockout SRPK1 in monocyte lineages. Blood flow was measured by Laser Speckle Imaging before, and for 28 days after surgery. ResultsMonocytes from PAD patients significantly inhibited endothelial cell migration, which was reversed by an anti-VEGF-A165b antibody. Surprisingly, migration was stimulated by SRPK1 inhibition, switching splicing from VEGF-A165b to VEGF-A165a. In Sfrp5-/-, LysM-Wnt5aGOF and HF/HS mouse models of PAD, blood flow was improved by SRPK1 inhibition. Impaired revascularisation in LysM-Wnt5aGOF mice was rescued in LysM-Wnt5aGOF:SRPK1MoKO mice, which had a phenotypic shift towards M2 macrophages. Impaired blood flow recovery was also rescued in obese-SRPK1MoKO mice. ConclusionVEGF splicing in monocytes is regulated differently from VEGF splicing in epithelial or cancer cells suggesting that control of splicing is dependent on cell type and/or environment. SRPK1 inhibition enhances collateralisation in mice, and in human in vitro models of monocyte-dependent impaired angiogenesis. New and NoteworthyA novel potential treatment for peripheral arterial disease (PAD) is described. Inhibition of SRPK1, or knockout in monocytes, induces angiogenesis by preventing splicing to anti-angiogenic VEGF (VEGF-A165b) in patients and animal models. In PAD, monocyte splicing control is different from other cell types and SRPK1 inhibition by drug like compounds can alter macrophage phenotype and reverse PAD in mice using a new cell specific SRPK1-LoxP mouse.

physiology↗

bioRxiv2024

Effect of different odors on the rat urine proteome

Do rats have corresponding changes in their urinary proteome when smelling different odors? In this study, urine samples were collected from six rats after smelling sesame oil and essential balm for three days. And samples were collected before and on the third and fourth days. Comparing the urinary protein groups of Day0 and Day4 of the sesame oil group, 143 differential proteins were identified, and the average number of randomly generated differential proteins was 7.3, which means that about 95% of the differential proteins could not be randomly generated. in the sesame oil group, differential proteins such as low-density lipoprotein receptor-related protein 2 and fetuin B, a biomarker of COPD, which are associated with olfaction, were identified. While uteroglobulin, trichothecene factor 3, and visfatin 2 were identified in the essential balm group, which had significant changes and were related to the production of olfactory sensation. It is noteworthy that we identified odor-binding protein 2A in the essential balm group, which was present in the e-cigarette model. This study demonstrates that odor can affect rat urinary proteome, with different odors affecting it differently. This provides a new approach to explore the biological process of olfaction.

biochemistry↗