Biology subjects

Wei, S.

Publications and source records attributed to Wei, S..

At least 73 records · Page 4Linked to original sources

bioRxiv2023

Spiral NeuroString: High-Density Soft Bioelectronic Fibers for Multimodal Sensing and Stimulation

Bioelectronic fibers hold promise for both research and clinical applications due to their compactness, ease of implantation, and ability to incorporate various functionalities such as sensing and stimulation. However, existing devices suffer from bulkiness, rigidity, limited functionality, and low density of active components. These limitations stem from the difficulty to incorporate many components on one-dimensional (1D) fiber devices due to the incompatibility of conventional microfabrication methods (e.g., photolithography) with curved, thin and long fiber structures. Herein, we introduce a fabrication approach, "spiral transformation'', to convert two-dimensional (2D) films containing microfabricated devices into 1D soft fibers. This approach allows for the creation of high density multimodal soft bioelectronic fibers, termed Spiral NeuroString (S-NeuroString), while enabling precise control over the longitudinal, angular, and radial positioning and distribution of the functional components. We show the utility of S-NeuroString for motility mapping, serotonin sensing, and tissue stimulation within the dynamic and soft gastrointestinal (GI) system, as well as for single-unit recordings in the brain. The described bioelectronic fibers hold great promises for next-generation multifunctional implantable electronics.

bioengineering↗

bioRxiv2023

Analysis of ductal carcinoma in situ by self-reported race reveals molecular differences related to outcome.

Ductal carcinoma in situ (DCIS) is a non-obligate precursor to invasive breast cancer (IBC). Studies have indicated differences in DCIS outcome based on race or ethnicity, but molecular differences have not been investigated. We examined the molecular profile of DCIS by self-reported race (SRR) and outcome groups in Black (n=99) and White (n=191) women with DCIS in a large DCIS case-control cohort with longitudinal follow up. Gene expression and pathway analyses indicated that different genes and pathways are involved in ipsilateral breast outcome (DCIS or IBC) after DCIS treatment in White versus Black women. We identified differences in ER and HER2 expression, tumor microenvironment composition, and copy number variations by SRR and outcome groups. Our results suggest that different molecular mechanisms drive subsequent ipsilateral breast events in Black versus White women.

cancer biology↗

bioRxiv2023

Rapid ionic current phenotyping (RICP) identifies mechanistic underpinnings of iPSC-CM AP heterogeneity

As a renewable, easily accessible, human-derived in vitro model, human induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) are a promising tool for studying arrhythmia-related factors, including cardiotoxicity and congenital proarrhythmia risks. An oft-mentioned limitation of iPSC-CMs is the abundant cell-to-cell variability in recordings of their electrical activity. Here, we develop a new method, rapid ionic current phenotyping (RICP), that utilizes a short (10 s) voltage clamp protocol to quantify cell-to-cell heterogeneity in key ionic currents. We correlate these ionic current dynamics to action potential recordings from the same cells and produce mechanistic insights into cellular heterogeneity. We present evidence that the L-type calcium current is the main determinant of upstroke velocity, rapid delayed rectifier K+ current is the main determinant of the maximal diastolic potential, and an outward current in the excitable range of slow delayed rectifier K+ is the main determinant of action potential duration. We measure an unidentified outward current in several cells at 6 mV that is not recapitulated by iPSC-CM mathematical models but contributes to determining action potential duration. In this way, our study both quantifies cell-to-cell variability in membrane potential and ionic currents, and demonstrates how the ionic current variability gives rise to action potential heterogeneity. Based on these results, we argue that iPSC-CM heterogeneity should not be viewed simply as a problem to be solved but as a model system to understand the mechanistic underpinnings of cellular variability. New & NoteworthyWe present rapid ionic current phenotyping (RICP), a current quantification approach based on an optimized voltage clamp protocol. The method captures a rich snapshot of ionic currents that provides quantitative information about multiple currents (e.g., ICaL, IKr) in the same cell. The protocol helped to identify key ionic determinants of cellular action potential heterogeneity in iPSC-CMs. This included unexpected results, such as the critical role of IKr in establishing the maximum diastolic potential.

bioengineering↗

bioRxiv2023

Shape-changing electrode array for minimally invasive large-scale intracranial brain activity mapping

Large-scale brain activity mapping is important for understanding the neural basis of behaviour. Electrocorticograms (ECoGs) have high spatiotemporal resolution, bandwidth, and signal quality. However, the invasiveness and surgical risks of electrode array implantation limit its application scope. We developed an ultrathin, flexible shape-changing electrode array (SCEA) for large-scale ECoG mapping with minimal invasiveness. SCEAs were inserted into cortical surfaces in compressed states through small openings in the skull or dura and fully expanded to cover large cortical areas. MRI and histological studies on rats proved the minimal invasiveness of the implantation process and the high chronic biocompatibility of the SCEAs. High-quality micro-ECoG activities mapped with SCEAs from rodent brains during seizures and canine brains during the emergence period revealed the spatiotemporal organization of different brain states with resolution and bandwidth that cannot be achieved using existing noninvasive techniques. The biocompatibility and ability to map large-scale physiological and pathological cortical activities with high spatiotemporal resolution, bandwidth, and signal quality in a minimally invasive manner offer SCEAs as a superior tool for applications ranging from fundamental brain research to brain-machine interfaces.

neuroscience↗

bioRxiv2023

HPC-based genome variant calling workflow (HPC-GVCW)

A high-performance computing genome variant calling workflow was designed to run GATK on HPC platforms. This workflow efficiently called an average of 27.3 M, 32.6 M, 168.9 M, and 16.2 M SNPs for rice, sorghum, maize, and soybean, respectively, on the most recently released high-quality reference sequences. Analysis of a rice pan-genome reference panel revealed 2.1 M novel SNPs that have yet to be publicly released.

genomics↗

bioRxiv2023

Prognostic Gene Signature for lung adenocarcinoma with a Higher Risk for Treatment Failure and Accelerated cell cycle and cell proliferation Pathway Activity

Lung cancer is the most common of malignant cancers which represents poor prognosis and high invasive. However, the reliable prognosis genes signature and the differential between the genetic and epigenetic are remain unclear. Here, we confirmed 78 genes of signature A to construct a model to predicted the prognosis and the activity of tumorigenesis-related pathways. The score based on the 78 genes expression is high correlation with the cell cycle and cell proliferation activity. Based on the median of score, the lung adenocarcinoma patients were divided into two cohorts which presents different prognosis, tumorigenesis-related pathways activity. The high score group was enriched in the E2F target, mtorc1 signaling and MYC targets. The low score group can be identified as better prognosis subtype. Also, we used the GDSC data to predicted the PIM inhibitor (AZD1208) as the effective compounds for high score group patients, which is supported by a higher PIM1 and PIM2 expression in the score group. An integrative analysis of multi-omics data identified ECT2 as key nodes in a regulatory network related to the prognostic phenotype. In conclusion, our data identified a molecular classifier for the different score groups patients, and the high score group patients might benefit from treatment with PIM inhibitor.

cancer biology↗

bioRxiv2023

Transcriptional profiling of mucus production and modification in rhesus macaque endocervical cells under hormonal regulation

ObjectiveEndocervical mucus production is a key regulator of fertility throughout the menstrual cycle. With cycle-dependent variability in mucus quality and quantity, cervical mucus can either facilitate or block sperm ascension into the upper female reproductive tract. This study seeks to identify genes involved in the hormonal regulation of mucus production, modification, and regulation through profiling the transcriptome of endocervical cells from the non-human primate, the Rhesus Macaque (Macaca mulatta). DesignExperimental. SettingTranslational science laboratory. InterventionWe treated differentiated primary endocervical cultures with estradiol (E2) and progesterone (P4) to mimic peri-ovulatory and luteal-phase hormonal changes. Using RNA-sequencing, we identified differential expression of gene pathways and mucus producing and modifying genes in cells treated with E2 compared to hormone-free conditions and E2 compared to E2-primed cells treated with P4. Main Outcome MeasuresWe pursued differential gene expression analysis on RNA-sequenced cells. Sequence validation was done using qPCR. ResultsOur study identified 158 genes that show significant differential expression in E2-only conditions compared to hormone-free control, and 250 genes that show significant differential expression in P4-treated conditions compared to E2-only conditions. From this list, we found hormone-induced changes in transcriptional profiles for genes across several classes of mucus production, including ion channels and enzymes involved in post-translational mucin modification that have not previously been described as hormonally regulated. ConclusionOur study is the first to use an in vitro culture system to create an epithelial-cell specific transcriptome of the endocervix. As a result, our study identifies new genes and pathways that are altered by sex-steroids in cervical mucus production.

molecular biology↗

bioRxiv2023

The ecological characteristics of the safe sites for early-stage establishment of Chamaecyparis obtusa var. formosana seedlings in Taiwan

Chamaecyparis obtusa var. formosana is an ecologically and economically important species in Taiwan, with a high affinity for fog immersion. Our study aims to identify possible stress factors that induced seedling mortality and investigate how different ecological factors influence early-stage safe site requirements of the seedlings. We focused on the effect of large-scale climatic variables, small-scale microhabitat conditions, and biotic interactions on seedling survival and establishment by applying seasonal seedling survival monitoring and establishment survey on both regional and local scale. We identified two alternative ways of seedling death, by environmental-induced mortality and by herbivory. Opposite effects of the same environmental factors on different causes of mortality showed that seedlings might need to balance the risks posed by both causes to optimize their growing conditions. On a regional scale, we observed limited effect of regional climatic variables (namely fog frequency) on seedlings establishment and survival but noted a similar seasonal survival pattern among regions. We hypothesize that short-duration droughts during the transition from Plum rain to typhoon season is one of the key mechanisms of environmental-induced mortality. On a local scale, we found that decayed coarse wood debris (CWD) facilitates seedling establishment by providing a "safe site", likely due to increased colonization of small-stature bryophytes and decreased litterfall accumulation. The effect of bryophytes on seedling establishment varies depending on their thickness, with thicker ones having stronger negative effects. Aside from the bryophytes, the accumulation of litter significantly hindered seedling establishment. We argue that to safeguard the regeneration of Chamaecyparis obtusa var. formosana population, preserving CWD in the forest floor as a safe site for the seedlings after tree-replacing disturbance in natural forests is essential, particularly under ongoing climate change where more frequent and prolonged drought events are predicted. HighlightsO_LIOn a local scale, decayed coarse wood debris (CWD) provides "safe site" for the establishment of Chamaecypairs obtusa var. formosana seedlings. C_LIO_LIRegional climatic variables had limited effects on seedlings, but all regions had similar seasonal patterns of seedling mortatlity. C_LIO_LIFacilitation effect of small-stature bryophytes and litterfall avoidance might be the underlying mechanisms behind CWD safe sites. C_LIO_LIPreserving CWD for seedlings is important in the context of predicted prolong drought events under ongoing climate change. C_LI

ecology↗

bioRxiv2023

GPR158 in pyramidal neurons mediates social novelty behavior via modulating synaptic transmission in mice

Social novelty impairment is a hallmark feature of autism spectrum disorder associated with synaptic dysfunction. While G-protein coupled receptor 158 (GPR158) has been shown to be essential for synaptic neurotransmission, its role in modulating social novelty remains unknown. Here, we investigated the impact of GPR158 on social behavior in mice and observed that both constitutive and cell/tissue-specific knockout of Gpr158 in pyramidal neurons or the medial prefrontal cortex (mPFC) result in impaired novelty preference, but not sociability. Notably, we found a significant decline in excitatory synaptic transmission and glutamate vesicles in the mPFC synapses of global Gpr158 knockouts. Mechanistically, we identified that constitutive loss of Gpr158 led to suppressed Vglut1 distribution, possibly resulting from altered expression of vesicular V-ATPases and SNAREs by Gpr158 ablation in pyramidal neurons. Our findings suggest that GPR158 in pyramidal neurons specifically modulates social novelty and may be a potential therapeutic target for treating social disorders. Graphic Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/536148v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@1013ca9org.highwire.dtl.DTLVardef@359faaorg.highwire.dtl.DTLVardef@856371org.highwire.dtl.DTLVardef@143c866_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIKnockout of Gpr158 causes social novelty deficit in mice. C_LIO_LIKnockout of Gpr158 in pyramidal neurons leads to disrupted synaptic transmission and an E-I imbalance in the mPFC. C_LIO_LIDisturbed E-I homeostasis in the mPFC is likely due to reduced density of glutamate vesicles caused by Gpr158 knockout. C_LI

neuroscience↗

bioRxiv2023

A nanobody-based strategy for rapid and scalable purification of native human protein complexes

Native isolation of proteins in high yield and purity is a major bottleneck for analysis of their three- dimensional structure, function, and interactome. Here, we present a streamlined workflow for the rapid production of proteins or protein complexes using lentiviral transduction of human suspension cells, combined with highly-specific nanobody-mediated purification and proteolytic elution. (1) First, generation of a plasmid coding for a protein of interest fused to an N- or C- terminal GFP or ALFA peptide tag is rapidly achieved using the lentiviral plasmid toolkit we have designed. (2) Human suspension cell lines stably expressing the tagged fusion protein can be generated in <5 days using lentiviral transduction. (3) Leveraging the picomolar affinity of the GFP and ALFA nanobodies for their respective tags, proteins expressed even at low levels can be specifically captured from the resulting cell lysate in a variety of conditions, including detergents and mild denaturants. (4) Finally, rapid and specific elution of tagged or untagged proteins under native conditions is achieved within minutes at 4{degrees}C using an engineered SUMO protease. We demonstrate the wide applicability of the method by purifying multiple challenging soluble and membrane protein complexes to high purity from human cells. Our strategy is also directly compatible with many widely used GFP expression plasmids, cell lines and transgenic model organisms; is faster than alternative approaches, requiring [~]8 days from cloning to purification; and results in substantially improved yields and purity.

biochemistry↗

bioRxiv2023

A novel chimeric RNA originating from BmCPV S4 and Bombyx mori HDAC11 transcripts regulates virus proliferation

Polymerases encoded by segmented negative-strand RNA viruses cleave 5-m7G-capped host transcripts to prime viral mRNA synthesis ( cap-snatching) to generate chimeric RNA, and trans-splicing occurs between viral and cellular transcripts. Bombyx mori cytoplasmic polyhedrosis virus (BmCPV), an RNA virus belonging to Reoviridae, is a major pathogen of silkworm (B. mori). The genome of BmCPV consists of 10 segmented double-stranded RNAs (S1-S10) from which viral RNAs encoding a protein are transcribed. In this study, chimeric silkworm-BmCPV RNAs, in which the sequence derived from the silkworm transcript could fuse with both the 5 end and the 3 end of viral RNA, were identified in the midgut of BmCPV-infected silkworms by RNA_seq and further confirmed by PCR and Sanger sequencing. A novel chimeric RNA, HDAC11-S4 RNA 4, derived from silkworm histone deacetylase 11 (HDAC11) and the BmCPV S4 transcript encoding viral structural protein 4 (VP4), was selected for validation by in situ hybridization and Northern blotting. Interestingly, our results indicated that HDAC11-S4 RNA 4 was generated in a BmCPV RNA-dependent RNA polymerase (RdRp)-independent manner and could be translated into a truncated BmCPV VP4 with a silkworm HDAC11-derived N-terminal extension. Moreover, it was confirmed that HDAC11-S4 RNA 4 inhibited BmCPV proliferation, decreased the level of H3K9me3 and increased the level of H3K9ac. These results indicated that during infection with BmCPV, a novel mechanism, different from that described in previous reports, allows the genesis of chimeric silkworm-BmCPV RNAs with biological functions. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=130 SRC="FIGDIR/small/527451v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@1b3b949org.highwire.dtl.DTLVardef@4972c0org.highwire.dtl.DTLVardef@cc15aorg.highwire.dtl.DTLVardef@16a0a19_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2022

DeepCCI: a deep learning framework for identifying cell-cell interactions from single-cell RNA sequencing data

With the rapid development of high throughput single-cell RNA sequencing (scRNA-seq) technologies, it is of high importance to identify Cell-cell interactions (CCIs) from the ever-increasing scRNA-seq data. However, limited by the algorithmic constraints, current computational methods based on statistical strategies ignore some key latent information contained in scRNA-seq data with high sparsity and heterogeneity. To address the issue, here, we developed a deep learning framework named DeepCCI to identify meaningful CCIs from scRNA-seq data. Applications of DeepCCI to a wide range of publicly available datasets from diverse technologies and platforms demonstrate its ability to predict significant CCIs accurately and effectively.

bioinformatics↗

bioRxiv2022

Leak current, even with gigaohm seals, can cause misinterpretation of stem-cell derived cardiomyocyte action potential recordings

Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) have become an essential tool to study arrhythmia mechanisms. Much of the foundational work on these cells, and the computational models built from the resultant data, has overlooked the contribution of seal-leak current on the immature and heterogeneous phenotype that has come to define these cells. Here, we use in silico and in vitro studies to demonstrate how seal-leak current depolarises action potentials (APs), substantially affecting their morphology, even with seal resistances (Rseal) above 1 G{Omega}. We show that compensation of this leak current is difficult due to challenges with recording accurate measures of Rseal during an experiment. Using simulation, we show that Rseal measures: 1) change during an experiment, invalidating the use of pre-rupture values, and 2) are polluted by the presence of transmembrane currents at every voltage. Finally, we posit the background sodium current in baseline iPSC-CM models imitates the effects of seal-leak current and is increased to a level that masks the effects of seal-leak current on iPSC-CMs. Based on these findings, we make three recommendations to improve iPSC-CM AP data acquisition, interpretation, and model-building. Taking these recommendations into account will improve our understanding of iPSC-CM physiology and the descriptive ability of models built from such data. Key pointsO_LIHuman induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are an essential tool in the study of cardiac arrhythmia mechanisms. C_LIO_LITheir immature and heterogeneous action potential phenotype complicates the interpretation of experimental data, and has slowed their acceptance in industry and academia. C_LIO_LIWe suggest that a leak current caused by an imperfect pipette-membrane seal during single-cell patch-clamp experiments is partly responsible for inducing this phenotype. C_LIO_LIUsing in vitro experiments and computational modelling, we show that this seal-leak current affects iPSC-CM AP morphology, even under ideal experimental conditions. C_LIO_LIBased on these findings, we make recommendations that should be considered when interpreting, analysing and fitting iPSC-CM data. C_LI

physiology↗

bioRxiv2022

Targeted Protein Degradation via Nanoparticles

Strategies that hijack selective proteins of interest (POIs) to the intracellular protein recycling machinery for targeted protein degradation (TPD) have recently emerged as powerful tools for undruggable targets in biomedical research and the pharmaceutical industry. However, targeting any new POI with current TPD tools requires laborious case-by-case design for different diseases and cell types, especially for those extracellular targets. Here, we observed that nanoparticles (NPs) can mediate the receptor-free internalization of hijacked protein and further developed a generic paradigm for both intra- and extracellular POI degradation, by making full use of clinically approved components. The phenomenon is general, as we found nanostructures such as lipid nanoparticle (LNP), liposomes, exosomes, polymeric nanoparticles, inorganic nanoparticles and their hybrid nanoparticles modified with POI-recognizing moiety (antibody, peptides, small molecule drugs) can mediate TPD for a wide range of extracellular/membrane and intracellular targets. The super flexible and feasible-to-synthesize TPD-NPs paradigm may revolutionize the current TPD tools development landscape and it provides fundamental knowledge to receptor mediated drug therapies. HighlightsO_LINanoparticle mediated targeted protein degradation (TPD-NP) can be constructed by "Mix-and-Match" and do not require de novo synthesis or specific internalization design. C_LIO_LITPD-NPs can be equipped with specific cell type targeting capacity, loading and controlled release of therapeutic cargos, as well as biological barrier penetration capacity. C_LIO_LIAssembling components can be clinical-approved or biodegradable for translational medicine. C_LIO_LITPD-NP highly boosted current application platforms of nano-delivery and TPD. C_LI

bioengineering↗

bioRxiv2022

Trade-off between conservation of biological variation and batch effect removal in deep generative modeling for single-cell transcriptomics

Single-cell RNA sequencing (scRNA-seq) technology has contributed significantly to diverse research areas in biology, from cancer to development. Since scRNA-seq data is high-dimensional, a common strategy is to learn low-dimensional latent representations better to understand overall structure in the data. In this work, we build upon scVI, a powerful deep generative model which can learn biologically meaningful latent representations, but which has limited explicit control of batch effects. Rather than prioritizing batch effect removal over conservation of biological variation, or vice versa, our goal is to provide a birds eye view of the trade-offs between these two conflicting objectives. Specifically, using the well established concept of Pareto front from economics and engineering, we seek to learn the entire trade-off curve between conservation of biological variation and removal of batch effects. A multi-objective optimisation technique known as Pareto multi-task learning (Pareto MTL) is used to obtain the Pareto front between conservation of biological variation and batch effect removal. Our results indicate Pareto MTL can obtain a better Pareto front than the naive scalarization approach typically encountered in the literature. In addition, we propose to measure batch effect by applying a neural-network based estimator called Mutual Information Neural Estimation (MINE) and show benefits over the more standard Maximum Mean Discrepancy (MMD) measure. The Pareto front between conservation of biological variation and batch effect removal is a valuable tool for researchers in computational biology. Our results demonstrate the efficacy of applying Pareto MTL to estimate the Pareto front in conjunction with applying MINE to measure the batch effect.

bioinformatics↗

bioRxiv2022

A low-cost and open-source solution to automate imaging and analysis of cyst nematode infection assays for Arabidopsis thaliana

BackgroundCyst nematodes are one of the major groups of plant-parasitic nematode, responsible for considerable crop losses worldwide. Improving genetic resources, and therefore resistant cultivars, is an ongoing focus of many pest management strategies. One of the major bottlenecks in identifying the plant genes that impact the infection, and thus the yield, is phenotyping. The current available screening method is slow, has unidimensional quantification of infection limiting the range of scorable parameters, and does not account for phenotypic variation of the host. The ever-evolving field of computer vision may be the solution for both the above-mentioned issues. To utilise these tools, a specialised imaging platform is required to take consistent images of nematode infection in quick succession. ResultsHere, we describe an open-source, easy to adopt, imaging hardware and trait analysis software method based on a pre-existing nematode infection screening method in axenic culture. A cost-effective, easy-to-build and -use, 3D-printed imaging device was developed to acquire images of the root system of Arabidopsis thaliana infected with the cyst nematode Heterodera schachtii, replacing costly microscopy equipment. Coupling the output of this device to simple analysis scripts allowed the measurement of some key traits such as nematode number and size from collected images, in a semi-automated manner. Additionally, we used this combined solution to quantify an additional trait, root area before infection, and showed both the confounding relationship of this trait on nematode infection and a method to account for it. ConclusionTaken together, this manuscript provides a low-cost and open-source method for nematode phenotyping that includes the biologically relevant nematode size as a scorable parameter, and a method to account for phenotypic variation of the host. Together these tools highlight great potential in aiding our understanding of nematode parasitism.

plant biology↗

bioRxiv2022

Identification and calibration of ultrabright localizations to eliminate quantification error in SMLM

Single molecule localization microscopy (SMLM) is irreplaceable among super-resolution microscopies in revealing biological ultra-structures, given its unmatched high resolution. However, its sub-optimal quantitative capability, which is critical for characterizing true biomolecular organization of ultra-structures in cells, has hindered its widest application in biomedical research. Here, in SMLM imaging of cellular structures such as lipid rafts and microtubules with saturation labelling, we identified ultra-bright localizations, each of which is contributed by simultaneous emission of multiple molecules within a diffraction-limit region and has been regarded before as a regular localization from single molecule. Consistently, ultra-bright localizations are also observed in simulated SMLM imaging of endoplasmic reticulum or microtubules from public resource. Furthermore, after calibrating each ultrabright localization into multiple single-molecule localizations using the photon-number-based models, the density of total localizations shows linear correlation with the true molecule density, presenting SMLM with new reconstruction method as a quantitative analysis approach. Therefore, identification and dissection of ultra-bright localizations in SMLM enable the close and quantitative estimate of the true biomolecular organization.

biophysics↗

bioRxiv2022

Domain fusion TLR2-4 enhances the autophagy-dependent clearance of Staphylococcus aureus in the genetic engineering goat generated by CRISPR/Cas9

Staphylococcus aureus infections pose a potential threat to livestock production and public health. A novel strategy is needed to control S. aureus infection due to its adaptive evolution to antibiotics. Autophagy plays a key role in degrading bacteria for innate immune cells. In order to promote S. aureus clearance via TLR induced autophagy pathway, the domain fusion TLR2-4 with the extracellular domain of TLR2, specific recognizing S. aureus, and transmembrane and intracellular domains of TLR4 is assembled, then the goats expressing TLR2-4 is generated. TLR2-4 substantially augments the removal of S. aureus within macrophages by elevating autophagy level. Phosphorylated JNK/ERK1/2 promote LC3- puncta in TLR2-4 macrophages during S. aureus-induced autophagy via MyD88-mediated the TAK1 signaling cascade. Meantime, the TRIF-dependent TBK1-TFEB-OPTN signaling is involved in TLR2-4-triggered autophagy after S. aureus challenge. Moreover, the transcript of ATG5 and ATG12 is significantly increased via cAMP-PKA-NF-{kappa}B signaling, which facilitates S. aureus-induced autophagy in TLR2-4 macrophages. Overall, the novel receptor TLR2-4 enhances the autophagy-dependent clearance of S. aureus in macrophages via TAK1/TBK1-JNK/ERK, TBK1-TFEB-OPTN and cAMP-PKA-NF-{kappa}B-ATGs signaling pathways, which provide an alternative approach to resistant against S. aureus infection.

immunology↗