ABSTRACT
This work proposed a simple and ultrasensitive nanozyme-based immunoassay on a filtration device for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (NP). Gold core porous platinum shell nanoparticles (Au@Pt NPs) were synthesized with high catalytic activity to oxidize 3,3',5,5'-tetramethylbenzidine, leading to an oblivious color change. The filtration device was designed based on the size difference of magnetic beads, filter membrane pore, and Au@Pt NPs. A simple, rapid, and consistent washing procedure can be performed with the help of a plastic syringe. This detection method could realize the quantitative detection of SARS-CoV-2 NP within 80 min for point-of-care needs. The limit of detection for the SARS-CoV-2 antigen was 0.01 ng/mL in buffer. The coefficients of variation of the assay were 1.78% for 10 ng/mL SARS-CoV-2 antigen, 2.03% for 1 ng/mL SARS-CoV-2 antigen, and 2.34% for the negative sample, respectively. The specificity of the detection platform was verified by the detection of various respiratory viruses. This simple and effective detection system was expected to promote substantial progress in the development and application of virus immunodetection technology.
ABSTRACT
A photonic-assisted microwave frequency measurement (MFM) method based on optical heterodyne detection is proposed and experimentally demonstrated. In the proposed MFM system, a linearly chirped optical waveform (LCOW) from a three-electrode distributed Bragg reflector laser diode (DBR-LD) and a multi-wavelength signal from a Mach-Zehnder modulator (MZM), where the signal under test (SUT) is modulated on an optical carrier from a distributed feedback laser diode (DFB-LD), are heterodyne detected by the photodetector (PD). A bandpass filter then filters the detected signal, and the envelope is detected by an oscilloscope. Then, frequency-to-time mapping is realized, and the signal frequency is measured. Thanks to the fast tuning rate and large tuning range of the DBR-LD, the proposed MFM system has a high measurement speed and a broad instantaneous measurement bandwidth. In the experimental demonstration, a measurement error below 39.1â MHz is achieved at an instantaneous bandwidth of 20â GHz and a measurement speed of 1.12â GHz/µs. The MFM of a frequency-hopping signal is also experimentally demonstrated. The successful demonstration of the MFM system with a simple structure provides a new optical solution for realizing broadband and fast microwave frequency measurements.
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The global outbreak of monkeypox virus (MPXV) has highlighted the need for rapid molecular diagnostics techniques. In this study, a single-step recombinase polymerase amplification (RPA)-CRISPR/Cas12a system was developed for rapid and sensitive detection of MPXV. The limit of detection of this assay was 1 copy per µL of extracted nucleic acids. A heating lysis method was integrated to further simplify the sample processing workflow and shorten the assay time to 40 min from sample to result. The reaction mixture can be lyophilized to improve its accessibility in resource-limited settings. The analysis results of the proposed single-step RPA-CRISPR/Cas12a assay for clinical MPXV positive and negative samples were 100% consistent with standard PCR assay. These results demonstrate the feasibility and efficiency of this method for rapid and accurate MPXV detection in real-world settings, showcasing its potential utility in urgent and practical settings.
ABSTRACT
The emerging monkeypox virus (MPXV) has raised global health concern, thereby highlighting the need for rapid, sensitive, and easy-to-use diagnostics. Here, we develop a single-step CRISPR-based diagnostic platform, termed SCOPE (Streamlined CRISPR On Pod Evaluation platform), for field-deployable ultrasensitive detection of MPXV in resource-limited settings. The viral nucleic acids are rapidly released from the rash fluid swab, oral swab, saliva, and urine samples in 2 min via a streamlined viral lysis protocol, followed by a 10-min single-step recombinase polymerase amplification (RPA)-CRISPR/Cas13a reaction. A pod-shaped vest-pocket analysis device achieves the whole process for reaction execution, signal acquisition, and result interpretation. SCOPE can detect as low as 0.5 copies/µL (2.5 copies/reaction) of MPXV within 15 min from the sample input to the answer. We validate the developed assay on 102 clinical samples from male patients / volunteers, and the testing results are 100% concordant with the real-time PCR. SCOPE achieves a single-molecular level sensitivity in minutes with a simplified procedure performed on a miniaturized wireless device, which is expected to spur substantial progress to enable the practice application of CRISPR-based diagnostics techniques in a point-of-care setting.
Subject(s)
Exanthema , Monkeypox virus , Humans , Male , Biological Assay , Cell Death , Nucleotidyltransferases , Nucleic Acid Amplification Techniques , Sensitivity and Specificity , CRISPR-Cas Systems , RecombinasesABSTRACT
The ABO blood group plays an important role in blood transfusion, linkage analysis, individual identification, etc. Serologic methods of blood typing are gold standards for the time being, which require stable typing antisera and fresh blood samples and are labor intensive. At present, reliable determination of ABO blood group genotypes based on single-nucleotide polymorphisms (SNPs) among A, B, and O alleles remains necessary. Thus, in this work, CRISPR/Cas13a-mediated genotyping for the ABO blood group by detecting SNPs between different alleles was proposed. The ABO*O.01.01(c.261delG) allele (G for the A/B allele and del for the O allele) and ABO*B.01(c.796C > A) allele (C for the A/O allele and A for the B allele) were selected to determine the six genotypes (AA, AO, BB, BO, OO, and AB) of the ABO blood group. Multiplex PCR was adapted to simultaneously amplify the two loci. CRISPR/Cas13a was then used to specifically differentiate ABO*O.01.01(c.261delG) and ABO*B.01(c.796C > A) of A, B, and O alleles. Highly accurate determination of different genotypes was achieved with a limit of detection of 50 pg per reaction within 60 min. The reliability of this method was further validated based on its applicability in detecting buccal swab samples with six genotypes. The results were compared with those of serological and sequencing methods, with 100% accuracy. Thus, the CRISPR/Cas13a-mediated assay shows great application potential in the reliable identification of ABO blood group genotypes in a wide range of samples, eliminating the need to collect fresh blood samples in the traditional method.
Subject(s)
ABO Blood-Group System , Polymorphism, Single Nucleotide , ABO Blood-Group System/genetics , Reproducibility of Results , Clustered Regularly Interspaced Short Palindromic Repeats , Genotype , Multiplex Polymerase Chain ReactionABSTRACT
Ascorbic acid (AsA) is an important nutrient for human health and disease cures, and it is also a crucial indicator for the quality of fruit and vegetables. As a reductant, AsA plays a pivotal role in maintaining the intracellular redox balance throughout all the stages of plant growth and development, fruit ripening, and abiotic stress responses. In recent years, the de novo synthesis and regulation at the transcriptional level and post-transcriptional level of AsA in plants have been studied relatively thoroughly. However, a comprehensive and systematic summary about AsA-involved biochemical pathways, as well as AsA's physiological functions in plants, is still lacking. In this review, we summarize and discuss the multiple physiological and biochemical functions of AsA in plants, including its involvement as a cofactor, substrate, antioxidant, and pro-oxidant. This review will help to facilitate a better understanding of the multiple functions of AsA in plant cells, as well as provide information on how to utilize AsA more efficiently by using modern molecular biology methods.
Subject(s)
Antioxidants , Ascorbic Acid , Humans , Ascorbic Acid/metabolism , Antioxidants/metabolism , Stress, Physiological , Fruit/metabolism , Oxidation-Reduction , Gene Expression Regulation, PlantABSTRACT
Staphylococcus aureus (S. aureus) is a prevalent foodborne pathogen that could cause severe food poisoning. Thus, rapid, efficient, and ultrasensitive detection of S. aureus in food samples is urgently needed. Here, we report an efficient magnetic enrichment cascade single-step recombinase polymerase amplification (RPA)-CRISPR/Cas12a assay for the ultrasensitive detection of S. aureus. Magnetic beads (MBs) functionalized with S. aureus-specific antibodies were initially used for S. aureus enrichment from the complex matrix, with 98% capture efficiency in 5 min and 100-fold sensitivity improvement compared with unenriched S. aureus. Next, a single-step RPA-CRISPR/Cas12a-based diagnostic system with optimized extraction-free bacteria lysis was constructed. This assay could detect as low as 1 copy/µL (five copies/reaction) of extracted DNA template and 10 CFU/mL of S. aureus within 40 min. Furthermore, the assay could effectively detect S. aureus in real food samples such as lake water, orange juice, pork, and lettuce, with concordant results to qPCR assays. The proposed cascade signal-amplification assay eliminates the need for lengthy bacterial culture and complex sample preparation steps. Hence, the proposed assay shows great application potential for rapid, efficient, and ultrasensitive detection of pathogens in real food samples.
Subject(s)
Recombinases , Staphylococcus aureus , CRISPR-Cas Systems , Biological Assay , Magnetic Phenomena , Nucleic Acid Amplification TechniquesABSTRACT
The RNA secondary structure is like a blueprint that holds the key to unlocking the mysteries of RNA function and 3D structure. It serves as a crucial foundation for investigating the complex world of RNA, making it an indispensable component of research in this exciting field. However, pseudoknots cannot be accurately predicted by conventional prediction methods based on free energy minimization, which results in a performance bottleneck. To this end, we propose a deep learning-based method called TransUFold to train directly on RNA data annotated with structure information. It employs an encoder-decoder network architecture, named Vision Transformer, to extract long-range interactions in RNA sequences and utilizes convolutions with lateral connections to supplement short-range interactions. Then, a post-processing program is designed to constrain the model's output to produce realistic and effective RNA secondary structures, including pseudoknots. After training TransUFold on benchmark datasets, we outperform other methods in test data on the same family. Additionally, we achieve better results on longer sequences up to 1600 nt, demonstrating the outstanding performance of Vision Transformer in extracting long-range interactions in RNA sequences. Finally, our analysis indicates that TransUFold produces effective pseudoknot structures in long sequences. As more high-quality RNA structures become available, deep learning-based prediction methods like Vision Transformer can exhibit better performance.
Subject(s)
Algorithms , RNA , Nucleic Acid Conformation , RNA/genetics , RNA/chemistry , Sequence Analysis, RNA/methods , Base SequenceABSTRACT
Kiwifruits are often exposed to various temperature fluctuations (TFs) during postharvest transportation and storage. To evaluate the effect of TFs on the qualities of kiwifruits during storage, kiwifruits were stored at 2 °C, 2 °C or 5 °C (TF2 °C-5 °C, alternating every 12 h), 2 °C or 7 °C (TF2 °C-7 °C, alternating every 12 h) for 3 d before long time storage at 2 °C. Observations revealed that kiwifruits stored at a constant 2 °C showed the lowest loss of weight and vitamin C because of minimized ethylene production and respiratory rate compared with that of TF2 °C-5 °C and TF2 °C-7 °C. Moreover, the results of RT-qPCR verified that the expression levels of genes encoding polygalacturonase, ß-galacturonidase, and pectin methylesterase were significantly increased by the treatment of TF. Hence, TF accelerated the degradation of cell walls, softening, translucency, and relative conductivity of the flesh of kiwifruits. In addition, the impact of TF2 °C-7 °C on kiwifruits was more significant relative to TF2 °C-5 °C. The present study provides a theoretical basis for kiwifruit during cold storage.
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AIMS: Intestinal ischemia reperfusion (II/R) is a common clinical emergency. Ferroptosis is reported to play a role in II/R injury. Our previous studies revealed that corilagin significantly attenuates intestinal ischemia/reperfusion injuries. However, the underlying molecular mechanism is unclear and requires further study. MATERIALS AND METHODS: DAO, GSSG/T-GSH, MDA, and Fe2+ were measured by assay kits, 4-HNE was assessed by IHC, and 15-LOX was measured by ELISA. Mitochondrial damage was observed by TEM and cellular oxidation levels were detected by C11-BODIPY 581/591 and DHE probes. LC3, p62, Beclin1, ACSL4, GPX4, NCOA4, and ferritin expression were examined by WB in vivo and in vitro. IF, co-IF, q-PCR, and constructed NCOA4-knock-down IEC-6 cells were used to evaluate the role of NCOA4 in the effect of corilagin against II/R injury. Temporal and nucleoplasmic variations with or without corilagin were observed by WB. Co-IP and molecular docking were used to investigate the NCOA4-ferritin interaction. KEY FINDINGS: Corilagin attenuated II/R-induced ferroptosis both in vitro and in vivo. Further study revealed that the anti-ferroptosis bioactivity of corilagin might be due to the modulation of iron homeostasis via inhibition of ferritinophagy in an NCOA4-dependent manner. SIGNIFICANCE: Corilagin might be a potential therapeutic agent for II/R-induced tissue injury.
Subject(s)
Ferroptosis , Mesenteric Ischemia , Reperfusion Injury , Animals , Mice , Molecular Docking Simulation , Reperfusion Injury/drug therapy , Ferritins , IschemiaABSTRACT
Employing deep learning models to design high-performance metasurfaces has garnered significant attention due to its potential benefits in terms of accuracy and efficiency. A deep learning-based metasurface design framework typically comprises a forward prediction path for predicting optical responses and a backward retrieval path for generating geometrical configurations. In the forward design path, a specific geometrical configuration corresponds to a unique optical response. However, in the inverse design path, a single performance metric can correspond to multiple potential designs. This one-to-many mapping poses a significant challenge for deep learning models and can potentially impede their performance. Although representing the inverse path as a probabilistic distribution is a widely adopted method for tackling this problem, accurately capturing the posterior distribution to encompass all potential solutions remains an ongoing challenge. Furthermore, in most pioneering works, the forward and backward paths are captured using separate models. However, the knowledge acquired from the forward path does not contribute to the training of the backward model. This separation of models adds complexity to the system and can hinder the overall efficiency and effectiveness of the design framework. Here, we utilized an invertible neural network (INN) to simultaneously model both the forward and inverse process. Unlike other frameworks, INN focuses on the forward process and implicitly captures a probabilistic model for the inverse process. Given a specific optical response, the INN enables the recovery of the complete posterior over the parameter space. This capability allows for the generation of novel designs that are not present in the training data. Through the integration of the INN with the angular spectrum method, we have developed an efficient and automated end-to-end metasurface design and evaluation framework. This novel approach eliminates the need for human intervention and significantly speeds up the design process. Utilizing this advanced framework, we have effectively designed high-efficiency metalenses and dual-polarization metasurface holograms. This approach extends beyond dielectric metasurface design, serving as a general method for modeling optical inverse design problems in diverse optical fields.
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Nowadays, Spark Streaming, a computing framework based on Spark, is widely used to process streaming data such as social media data, IoT sensor data or web logs. Due to the extensive utilization of streaming media data analysis, performance optimization for Spark Streaming has gradually developed into a popular research topic. Several methods for enhancing Spark Streaming's performance include task scheduling, resource allocation and data skew optimization, which primarily focus on how to manually tune the parameter configuration. However, it is indeed very challenging and inefficient to adjust more than 200 parameters by means of continuous debugging. In this paper, we propose an improved dueling double deep Q-network (DQN) technique for parameter tuning, which can significantly improve the performance of Spark Streaming. This approach fuses reinforcement learning and Gaussian process regression to cut down on the number of iterations and speed convergence dramatically. The experimental results demonstrate that the performance of the dueling double DQN method with Gaussian process regression can be enhanced by up to 30.24%.
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BACKGROUND: In recent years, there has been growing interest in exploring the relationship between activities of daily living (ADL) and cardiovascular diseases. This retrospective cross-sectional study aimed to investigate the association of ADL measured by Barthel index (BI) with periprocedural myocardial infarction (PMI) and injury following percutaneous coronary intervention (PCI). METHODS: Enrolled patients were stratified into impaired and unimpaired ADL groups according to their BI scores. Logistic regressions were conducted to explore the association of ADL on admission with periprocedural myocardial injury and infarction. Restricted cubic spline (RCS) curve and subgroup analysis were also performed. RESULTS: Totally, 16.4% of patients suffered from PMI; the mean age was 65.8 ± 10.4 years old. RCS analysis showed that the morbidity of periprocedural myocardial infarction and injury showed a downward tendency with increasing BI scores. Multivariable logistic regression analysis demonstrated that impaired ADL was an independent risk factor for periprocedural myocardial infarction (OR = 1.190, 95% CI [1.041, 1.360], P = 0.011) and injury (OR = 1.131, 95% CI [1.017, 1.257], P = 0.023). Subgroup analysis showed that the association between ADL and PMI was founded in several subgroups, while the association between ADL and periprocedural myocardial injury was founded only in BMI ≥ 24 kg/m2 subgroup. CONCLUSION: Impaired ADL at hospital admission was an independent risk factor for periprocedural myocardial infarction and injury among patients following PCI.
Subject(s)
Heart Injuries , Myocardial Infarction , Percutaneous Coronary Intervention , Humans , Middle Aged , Aged , Activities of Daily Living , Percutaneous Coronary Intervention/adverse effects , Cross-Sectional Studies , Retrospective Studies , Myocardial Infarction/diagnosis , Myocardial Infarction/epidemiology , Myocardial Infarction/etiologyABSTRACT
Background: Coronary artery disease (CAD) is a multifactorial immune disease, but research into the specific immune mechanism is still needed. The present study aimed to identify novel immune-related markers of CAD. Methods: Three CAD-related datasets (GSE12288, GSE98583, GSE113079) were downloaded from the Gene Expression Integrated Database. Gene ontology annotation, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis and weighted gene co-expression network analysis were performed on the common significantly differentially expressed genes (DEGs) of these three data sets, and the most relevant module genes for CAD obtained. The immune cell infiltration of module genes was evaluated with the CIBERSORT algorithm, and characteristic genes accompanied by their diagnostic effectiveness were screened by the machine-learning algorithm least absolute shrinkage and selection operator (LASSO) regression analysis. The expression levels of characteristic genes were evaluated in the peripheral blood mononuclear cells of CAD patients and healthy controls for verification. Results: A total of 204 upregulated and 339 downregulated DEGs were identified, which were mainly enriched in the following pathways: "Apoptosis", "Th17 cell differentiation", "Th1 and Th2 cell differentiation", "Glycerolipid metabolism", and "Fat digestion and absorption". Five characteristic genes, LMAN1L, DOK4, CHFR, CEL and CCDC28A, were identified by LASSO analysis, and the results of the immune cell infiltration analysis indicated that the proportion of immune infiltrating cells (activated CD8 T cells and CD56 DIM natural killer cells) in the CAD group was lower than that in the control group. The expressions of CHFR, CEL and CCDC28A in the peripheral blood of the healthy controls and CAD patients were significantly different. Conclusion: We identified CHFR, CEL and CCDC28A as potential biomarkers related to immune infiltration in CAD based on public data sets and clinical samples. This finding will contribute to providing a potential target for early noninvasive diagnosis and immunotherapy of CAD.
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Recent years have witnessed significant progress in quantum communication and quantum internet with the emerging quantum photonic chips, whose characteristics of scalability, stability, and low cost, flourish and open up new possibilities in miniaturized footprints. Here, we provide an overview of the advances in quantum photonic chips for quantum communication, beginning with a summary of the prevalent photonic integrated fabrication platforms and key components for integrated quantum communication systems. We then discuss a range of quantum communication applications, such as quantum key distribution and quantum teleportation. Finally, the review culminates with a perspective on challenges towards high-performance chip-based quantum communication, as well as a glimpse into future opportunities for integrated quantum networks.
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Coronary perforation (CP) is a rare complication of percutaneous coronary intervention (PCI) and can lead to pericardial tamponade. Prolonged balloon inflation is a reasonable treatment for CP, but there is no standard recommendation on the preferable choice between the balloon site for prolonged balloon inflation (ie, proximal and in situ of the perforation). We present a rare case of successful prolonged balloon inflation at the proximal site of the CP after the failure of balloon inflation at the site of perforation. The patient developed CP during balloon inflation post-stent, rapidly progressing to cardiac tamponade. In situ prolonged balloon inflation (3 times) failed to close the CP, but proximal inflation could manage the CP. The take-home message from this case is that balloon expansion at the proximal site could be better than in situ of perforation in patients with CP after PCI.
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The carbon dioxide reduction reaction (CO2RR) is a promising method to both reduce greenhouse gas carbon dioxide (CO2) concentrations and provide an alternative to fossil fuel by converting water and CO2 into high-energy-density chemicals. Nevertheless, the CO2RR suffers from high chemical reaction barriers and low selectivity. Here we demonstrate that 4 nm gap plasmonic nano-finger arrays provide a reliable and repeatable plasmon-resonant photocatalyst for multiple-electrons reactions: the CO2RR to generate higher-order hydrocarbons. Electromagnetics simulation shows that hot spots with 10,000 light intensity enhancement can be achieved using nano-gap fingers under a resonant wavelength of 638 nm. From cryogenic 1H-NMR spectra, formic acid and acetic acid productions are observed with a nano-fingers array sample. After 1 h laser irradiation, we only observe the generation of formic acid in the liquid solution. While increasing the laser irradiation period, we observe both formic and acetic acid in the liquid solution. We also observe that laser irradiation at different wavelengths significantly affected the generation of formic acid and acetic acid. The ratio, 2.29, of the product concentration generated at the resonant wavelength 638 nm and the non-resonant wavelength 405 nm is close to the ratio, 4.93, of the generated hot electrons inside the TiO2 layer at different wavelengths from the electromagnetics simulation. This shows that product generation is related to the strength of localized electric fields.
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SCOPE: Intestinal ischemia-reperfusion (II/R) injury is a common pathological process with high morbidity and mortality. Effective prevention and treatment therapies for II/R are clinically necessary. 6-Shogaol (6-SG), the main active ingredient in dried ginger, behaviors multiple biological activities, including anti-inflammation, antioxidation, and anti-apoptosis. This study aims to elucidate the protective effects and mechanism of 6-SG against II/R-induced injury. METHODS AND RESULTS: Sprague-Dawley rats are pre-treated orally with 6-SG and subjected to II/R injury by clamping superior mesenteric artery for 1 h and reperfusion for 2 h. Caco-2 cells are challenged by hypoxia/reoxygenation to mimic II/R in vitro. 6-SG pre-treatment protects against II/R injury by reducing intestinal morphological damage and intestinal barrier injury via inhibiting cell apoptosis. Network pharmacology and molecular docking analyses reveal that 6-SG has a high affinity with brain-derived neurotrophic factor (BDNF) formed homodimer or heterodimer with NT4 instead of the monomer, and thus the dimer configuration is stabilized, activating BDNF/TrkB/PI3K/AKT signaling pathway and inhibiting II/R-induced cell apoptosis. The outcome is further validated both in vivo and in vitro. CONCLUSION: 6-Shogaol protects against II/R injury by inhibiting cell apoptosis through the BDNF/TrkB/PI3K/AKT pathway. This study offers a new understanding of the protection mechanism of 6-SG against II/R-induced injury.
Subject(s)
Intestinal Diseases , Reperfusion Injury , Zingiber officinale , Humans , Rats , Animals , Proto-Oncogene Proteins c-akt/metabolism , Brain-Derived Neurotrophic Factor/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Rats, Sprague-Dawley , Caco-2 Cells , Molecular Docking Simulation , Reperfusion Injury/drug therapy , Reperfusion Injury/prevention & control , Reperfusion , IschemiaABSTRACT
The continued emergence of SARS-CoV-2 variants of concern (VOCs) has raised great challenges for epidemic prevention and control. A rapid, sensitive, and on-site SARS-CoV-2 genotyping technique is urgently needed for individual diagnosis and routine surveillance. Here, a field-deployable ultrasensitive CRISPR-based diagnostics system, called Chemical additive-Enhanced Single-Step Accurate CRISPR/Cas13 Testing system (CESSAT), for simultaneous screening of SARS-CoV-2 and its five VOCs (Alpha, Beta, Gamma, Delta, and Omicron) within 40 min was reported. In this system, a single-step reverse transcription recombinase polymerase amplification-CRISPR/Cas13a assay was incorporated with optimized extraction-free viral lysis and reagent lyophilization, which could eliminate complicated sample processing steps and rigorous reagent storage conditions. Remarkably, 10% glycine as a chemical additive could improve the assay sensitivity by 10 times, making the limit of detection as low as 1 copy/µL (5 copies/reaction). A compact optic fiber-integrated smartphone-based device was developed for sample lysis, assay incubation, fluorescence imaging, and result interpretation. CESSAT could specifically differentiate the synthetic pseudovirus of SARS-CoV-2 and its five VOCs. The genotyping results for 40 clinical samples were in 100% concordance with standard method. We believe this simple but efficient enhancement strategy can be widely incorporated with existing Cas13a-based assays, thus leading a substantial progress in the development and application of rapid, ultrasensitive, and accurate nucleic acid analysis technology.
Subject(s)
Biosensing Techniques , COVID-19 , Humans , COVID-19/diagnosis , CRISPR-Cas Systems/genetics , Genotype , SARS-CoV-2/genetics , RNA, Viral/geneticsABSTRACT
Liver fibrosis is characterised by the activation of hepatic stellate cells (HSCs) and matrix deposition. Accumulating evidence has revealed that the oncogenic protein tyrosine phosphatase Src homology 2 domain-containing phosphatase 2 (SHP2) acts as a therapeutic target of fibrosis. Although several SHP2 inhibitors have reached early clinical trials, there are currently no FDA-approved drugs that target SHP2. In this study, we aimed to identify novel SHP2 inhibitors from an in-house natural product library to treat liver fibrosis. Out of the screened 800 compounds, a furanogermacrane sesquiterpene, linderalactone (LIN), significantly inhibited SHP2 dephosphorylation activity in vitro. Cross-validated enzymatic assays, bio-layer interferometry (BLI) assays, and site-directed mutagenesis were used to confirm that LIN directly binds to the catalytic PTP domain of SHP2. In vivo administration of LIN significantly ameliorated carbon tetrachloride (CCl4)-induced HSC activation and liver fibrosis by inhibiting the TGFß/Smad3 pathway. Thus, LIN or its derivatives could be considered potential therapeutic agents against SHP2-related diseases, such as liver fibrosis or NASH.