Familial dysautonomia （FD） is a rare children neurodegenerative disease caused due to a point mutation in the IKBKAP gene that results in decreased IKK complex-associated protein （IKAP） protein production. The dis...Familial dysautonomia （FD） is a rare children neurodegenerative disease caused due to a point mutation in the IKBKAP gene that results in decreased IKK complex-associated protein （IKAP） protein production. The disease affects mostly the dorsal root ganglion （DRG） and the sympathetic ganglion. Recently, we found that the molecular mechanisms underlying neurodegeneration in FD patients are defects in axonal transport of nerve growth factors and microtubule stability in the DRG. Neurons are highly polarized cells with very long axons. In order to survive and maintain proper function, neurons depend on transport of proteins and other cellular components from the neuronal body along the axons. We further demonstrated that IKAP is necessary for axon maintenance and showed that phosphatidylserine acts as an HDAC6 inhib- itor to rescue neuronal function in FD cells. In this review, we will highlight our latest research findings.展开更多
Phosphatidylserine(PS)is the part of cell structure in the body and has many beneficial functions especially in brain-related aging diseases.Although daily foods can provide PS to human body,the amount is very limited...Phosphatidylserine(PS)is the part of cell structure in the body and has many beneficial functions especially in brain-related aging diseases.Although daily foods can provide PS to human body,the amount is very limited due to its poverty in most foods.To overcome the issue,numerous studies based on PS have been reported to develop PS-related supplements.In this review,PS was comprehensively and critically reviewed from the view of resources,functions,processing techniques,patents,and prospects.For resources,animal,plant,and microorganism origins were all covered with their differences in composition profiles.For functions,benefits regarding memory,cognitive enhancement,exercise performance,reducing Alzheimer’s disease,and attention-deficit hyperactivity disorder symptoms were covered as well as the functional differences among animal-,plant-,and microorganism-based PS-related supplements.For processing techniques,traditional extracting methods from animal,plant,and microorganism tissues were comparatively discussed with enzymatic synthesis based on different reaction systems.Finally,patents of PS-related supplements were evaluated as well as their applications.This review could provide scientific and valuable support for PS industry.展开更多
Sensitive monitoring of the target products during the biosynthesis process is crucial,and facile analytical approaches are urgently needed.Herein,phosphatidylserine(PS)was chosen as the model target,a colorimetric ap...Sensitive monitoring of the target products during the biosynthesis process is crucial,and facile analytical approaches are urgently needed.Herein,phosphatidylserine(PS)was chosen as the model target,a colorimetric aptasensor was developed for the rapid quantitation in biosynthesis samples.A chimeric aptamer was constructed with two homogeneous original PS aptamers.Specific recognition between the chimeric aptamer and PS results in the desorption of aptamer from the surface of the AuNPs nanozyme,and the peroxidase-like enzymatic activity of the AuNPs nanozyme was weakened in a relationship with the different concentrations.The developed aptasensor performed well when applied for analyzing PS in biosynthesis samples.The aptasensor offers good sensitivity and selectivity,under optimal conditions,achieving monitoring and quantitation of PS in the range of 2.5-80.0μmol/L,with a limit of detection at 536.2 nmol/L.Moreover,the aptasensor provides good accuracy,with comparison rates of 98.17%-106.40%,when compared with the HPLC-ELSD.This study provides a good reference for monitoring other biosynthesized products and promoting the development of aptamers and aptasensors in real-world applications.展开更多
We have recently demonstrated that liposomes composed of phosphatidylserine (PS-liposomes) suppressed nitric oxide and inflammatory cytokine productions following LPS stimulation in macrophages. In this study, we exam...We have recently demonstrated that liposomes composed of phosphatidylserine (PS-liposomes) suppressed nitric oxide and inflammatory cytokine productions following LPS stimulation in macrophages. In this study, we examined the effect of PS-liposomes on expressions of TLR-4 and MyD88, which are essential for the signal transduction in LPS stimulation. Expression of MyD88 was suppressed when macrophages were treated with PS-liposomes, but not with liposomes of phosphatidylcholine. No change in TLR-4 expression was observed. MyD88 suppression was restored to the control levels when cells were pre-treated with anti-TGF-β antibody, suggesting that TGF-β plays an important role in down-regulation of MyD88 following PS-liposome treatment.展开更多
Chemotherapeutic drugs,such as cisplatin and phenanthriplatin(PhenPt),as STING agonists to induce DNA damage and activate the cyclic GMP-AMP synthase-stimulator of interferon genes(cGAS-STING)signaling pathway provide...Chemotherapeutic drugs,such as cisplatin and phenanthriplatin(PhenPt),as STING agonists to induce DNA damage and activate the cyclic GMP-AMP synthase-stimulator of interferon genes(cGAS-STING)signaling pathway provides a potential strategy for clinical chemo-immunotherapy.However,treatment with Pt-based drugs leads to irreversible ectopia of phosphatidylserine(PS),a major component of the intracellular membrane,to the surface of the cancer cells by enzymes(Xkr8).Exposed PS can bind to immune cell receptors and inhibit the presentation of tumor antigens,leading to immunosuppression and attenuation of chemotherapy.Herein,we report a novel approach to enhance chemo-immunotherapy by constructing siRNA targeted Xkr8(siXkr8)-mediated tetrahedral framework nucleic acid nanogel structure concurrently loaded with PhenPt(siXkr8-FNG/PhenPt)for co-delivery of siRNA and Pt-based drugs.The results showed that siXkr8-FNG/PhenPt can not only be used as an efficient delivery carrier to deliver siXkr8,block the expression of Xkr8,reduce the exposure of PS on the cancer cells surface,but also act as an immune stimulant to activate cGAS-STING pathway,effectively improve the immunosuppressive microenvironment,produce antitumor immune response,and inhibit tumor growth and metastasis.Overall,this new delivery system is important for improving the effect of Pt-based drug chemotherapy,inducing immune enhancement and nucleic acid drug delivery.展开更多
Radiation-induced fibrosis(RIF)is a major complication following radiotherapy treatment.Macrophages are the key regulators of inflammatory responses and have emerged as critical targets for the prevention of fibrosis....Radiation-induced fibrosis(RIF)is a major complication following radiotherapy treatment.Macrophages are the key regulators of inflammatory responses and have emerged as critical targets for the prevention of fibrosis.This study evaluated the efficacy of PEGylated phosphatidylserine-containing liposomes(PEG-PSLs)in mitigating RIF by modulating macrophage activity.PEG-PSLs influenced the polarization of bone marrow-derived macrophages(BMDMs),reducing the expression levels of inflammatory cytokines and fibrosis-associated markers.These ef-fects were observed within a specific PEGylation range,with the greatest reduction achieved at 1-1.5mol%PEG.Analysis of the IL-4-induced JAK-STAT signaling pathway indicated that the PEG-PSL-mediated suppression occurred at or beyond the point of JAK-1 phosphorylation.In in vitro fibrosis assays,PEG-PSLs reduced fibroblast-to-myofibroblast and epithelial-to-mesenchymal transitions when co-cultured with BMDMs.In vivo studies using a rat model of esophageal irradiation demonstrated that PEG-PSLs effectively attenuated fibrotic progression,preserved tissue architecture,and enhanced muscle regeneration.These findings indicate the po-tential of partially PEGylated phosphatidylserine liposomes as therapeutic agents for mitigating post-irradiation esophageal fibrosis.展开更多
Background:Understanding how the tumor microenvironment is shaped by various factors is important for the development of new therapeutic strategies.Tumor cells often undergo spontaneous apoptotic cell death in tumor m...Background:Understanding how the tumor microenvironment is shaped by various factors is important for the development of new therapeutic strategies.Tumor cells often undergo spontaneous apoptotic cell death in tumor microen-vironment,these apoptotic cells are histologically co-localized with immunosup-pressive macrophages.However,the mechanism by which tumor cell apoptosis modulates macrophage polarization is not fully understood.In this study,we aimed to explore the tumor promoting effects of apoptotic tumor cells and the signal pathways involved.Methods:Apoptotic cells and macrophages in tumors were detected by immunohistochemical staining.Morphological analysis was performed with Giemsa staining.Lipids generated from apoptotic cells were detected by liq-uid chromatography-mass spectrometry.Phosphatidylserine-containing lipo-somes were prepared to mimic apoptotic cells.The expression of protein was determined by real-time PCR,immunohistochemistry enzyme-linked immunosorbent assay and Western blotting.Mouse malignant ascites and subcu-taneous tumor models were designed for in vivo analysis.Transgenic mice with specific genes knocked out and inhibitors specific to certain proteins were used for the mechanistic studies.Results:The location and the number of apoptotic cells were correlated with that of macrophages in several types of carcinomas.Phosphatidylserine,a lipid molecule generated in apoptotic cells,induced polarization and accumulation of M2-like macrophages in vivo and in vitro.Moreover,sustained administration of phosphoserine promoted tumor growth in the malignant ascites and subcuta-neous tumor models.Further analyses suggested that phosphoserine induced a M2-like phenotype in macrophages,which was related to the activation of phosphoserine receptors including T-cell immunoglobin mucin 4(TIM4)and the FAK-SRC-STAT3 signaling pathway as well as elevated the expression of the histone demethylase Jumonji domain-containing protein 3(JMJD3).Adminis-tration of specific inhibitors of these pathways could reduce tumor progression.Conclusions:This study suggest that apoptotic cell-generated phosphoserine might be a notable signal for immunosuppressive macrophages in tumors,and the related pathways might be potential therapeutic targets for cancer therapy.展开更多
The role of surfactant protein A(SP-A)in the recognition and clearance of apoptotic cells is well established,but to date,it is still not clear which surface molecules of apoptotic cells are involved in the process.He...The role of surfactant protein A(SP-A)in the recognition and clearance of apoptotic cells is well established,but to date,it is still not clear which surface molecules of apoptotic cells are involved in the process.Here we present evidence that phosphatidylserine(PS)is a relevant binding molecule for human SP-A.The binding is Ca^(2+)-dependent and is not inhibited by mannose,suggesting that the sugar-binding site of the carbohydrate recognition domain(CRD)of SP-A is not involved.Flow cytometry studies on apoptotic Jurkat cells revealed apparent inhibition of annexin V binding by increasing concentrations of SP-A in late apoptotic but not early apoptotic cells,and this was consistent for Jurkat cells and neutrophils.Supporting these data,confocal microscopy results show a co-localisation of annexin V and SP-A in late apoptotic but not early apoptotic cells.However,we cannot conclude that this inhibition is exclusively due to the binding of SP-A to PS on the cell surface,as annexin V is not wholly specific for PS and SP-A also interacts with other phospholipids that might become exposed on the apoptotic cell surface.展开更多
One of the hallmarks of live cells is the asymmetric distribution of lipids across their plasma membrane.Changes in this asymmetry due to lipid"scrambling"result in phosphatidylserine exposure at the cell su...One of the hallmarks of live cells is the asymmetric distribution of lipids across their plasma membrane.Changes in this asymmetry due to lipid"scrambling"result in phosphatidylserine exposure at the cell surface that is detected by annexin V staining.This alteration is observed during cell death processes such as apoptosis,and during physiological responses such as platelet degranulation and membrane repair.Previous studies have shown that activation of NK cells is accompanied by exposure of phosphatidylserine at the cell surface.While this response was thought to be indicative of ongoing NK cell death,it may also reflect the regulation of NK cell activation in the absence of cell death.Herein,we found that NK cell activation was accompanied by rapid phosphatidylserine exposure to an extent proportional to the degree of NK cell activation.Through enforced expression of a lipid scramblase,we provided evidence that activation-induced lipid scrambling in NK cells is reversible and does not lead to cell death.In contrast,lipid scrambling attenuates NKcell activation.This response was accompanied by reduced cell surface expression of activating receptors such as 2B4,and by loss of binding of Src family protein tyrosine kinases Fyn and Lck to the inner leaflet of the plasma membrane.Hence,lipid scrambling during NK cell activation is,at least in part,a physiological response that reduces the NK cell activation level.This effect is due to the ability of lipid scrambling to alter the distribution of membrane-associated receptors and kinases required for NK cell activation.展开更多
Exosomes are membrane bound extracellular vesicles that play an important role in many biological processes.While they have great application value,exosome isolation is still considered a major scientific challenge.In...Exosomes are membrane bound extracellular vesicles that play an important role in many biological processes.While they have great application value,exosome isolation is still considered a major scientific challenge.In the present study,a novel separation strategy for exosomes is proposed based on the specific interaction between immobilized peptide ligands and phosphatidylserine moieties which are highly abundant on the surface of exosomes.With the new affinity method,intact model exosomes can be recovered with a high yield in a short processing time.The purity of exosome samples enriched from serum by the affinity method is far higher than that isolated by ultrafiltration,and similar to that obtained by density gradient centrifugation and ultracentrifugation.Moreover,the variety of contaminants co-isolated by the affinity method is relatively low due to its specific separation principle.Proteomics analysis of exosomes isolated by the affinity method from the serum of healthy,hepatocellular carcinoma patients,and intrahepatic cholangiocarcinoma patients was performed to prove the applicability of this method.In conclusion,our novel strategy shows characteristics of easy preparation,high specificity,and cost-effectiveness,and provides a promising approach for exosome isolation which should have wide applications.展开更多
Hearing relies on the structural and functional integrity of cochlear hair cells,particularly their apical F-actin-filled stereocilia.Phospholipid scramblases are important for maintaining membrane asymmetry,but their...Hearing relies on the structural and functional integrity of cochlear hair cells,particularly their apical F-actin-filled stereocilia.Phospholipid scramblases are important for maintaining membrane asymmetry,but their roles in the stereocilia and auditory functions are not fully understood.Here,we identify Plscr5 as a downstream target of the transcription factor POU4F3 essential for hair cell function,whose mutation causes human DFNA15 deafness.Plscr5 knockout mice exhibit progressive hearing loss due to stereocilia degeneration and hair cell loss.Functional analyses reveal that PLSCR5 contributes to phosphatidylserine externalization in hair cell apical membranes,particularly in inner hair cells,and is important for outer hair cell and stereocilia maintenance.Our findings highlight PLSCR5 as an important downstream effector of POU4F3 and regulator of phosphatidylserine externalization and membrane dynamics required for auditory functions.展开更多
INTRODUCTIONApoptosis,described by Kerr in 1972,plays a keyrole in all types of regulated cellular processes inmulticellular,organisms.It is defined as amorphologic change,including fragmentation of theDNA,cell shrink...INTRODUCTIONApoptosis,described by Kerr in 1972,plays a keyrole in all types of regulated cellular processes inmulticellular,organisms.It is defined as amorphologic change,including fragmentation of theDNA,cell shrinkage,dilation of the endoplasmaticreticulum,cell fragmentation and formation展开更多
We question whether,in men with an abnormal rate of sperm DNA fragmentation,the magnetic-activated cell sorting(MACS)could select spermatozoa with lower rates of DNA fragmentation as well as spermatozoa with unbalance...We question whether,in men with an abnormal rate of sperm DNA fragmentation,the magnetic-activated cell sorting(MACS)could select spermatozoa with lower rates of DNA fragmentation as well as spermatozoa with unbalanced chromosome content.Cryopreserved spermatozoa from six males were separated into nonapoptotic and apoptotic populations.We determined the percentages of spermatozoa with(i)externalization of phosphatidylserine(EPS)by annexin V-Fluorescein isothiocyanate(FITC)labeling,(ii)DNA fragmentation by TdT-mediated-dUTP nick-end labeling(TUNEL),and(iii)numerical abnormalities for chromosomes X,Y,13,18,and 21 by fluorescence in situ hybridization(FISH),on the whole ejaculate and selected spermatozoa in the same patient.Compared to the nonapoptotic fraction,the apoptotic fraction statistically showed a higher number of spermatozoa with EPS,with DNA fragmentation,and with numerical chromosomal abnormalities.Compared to the whole ejaculate,we found a significant decrease in the percentage of spermatozoa with EPS and decrease tendencies of the DNA fragmentation rate and the sum of disomy levels in the nonapoptotic fraction.Conversely,we observed statistically significant higher rates of these three parameters in the apoptotic fraction.MACS may help to select spermatozoa with lower rates of DNA fragmentation and unbalanced chromosome content in men with abnormal rates of sperm DNA fragmentation.展开更多
Objective: The human cluster of differentiation(CD)300A, a type-I transmembrane protein with immunoreceptor tyrosine-based inhibitory motifs, was investigated as a potential immune checkpoint for human natural killer(...Objective: The human cluster of differentiation(CD)300A, a type-I transmembrane protein with immunoreceptor tyrosine-based inhibitory motifs, was investigated as a potential immune checkpoint for human natural killer(NK) cells targeting hematologic malignancies(HMs).Methods: We implemented a stimulation system involving the CD300A ligand, phosphatidylserine(PS), exposed to the outer surface of malignant cells. Additionally, we utilized CD300A overexpression, a CD300A blocking system, and a xenotransplantation model to evaluate the impact of CD300A on NK cell efficacy against HMs in in vitro and in vivo settings. Furthermore, we explored the association between CD300A and HM progression in patients.Results: Our findings indicated that PS hampers the function of NK cells. Increased CD300A expression inhibited HM lysis by NK cells. CD300A overexpression shortened the survival of HM-xenografted mice by impairing transplanted NK cells. Blocking PS–CD300A signals with antibodies significantly amplified the expression of lysis function-related proteins and effector cytokines in NK cells, thereby augmenting the ability to lyse HMs. Clinically, heightened CD300A expression correlated with shorter survival and an “exhausted” phenotype of intratumoral NK cells in patients with HMs or solid tumors.Conclusions: These results propose CD300A as a potential target for invigorating NK cell-based treatments against HMs.展开更多
As people live longer,the burden of aging-related brain diseases,especially dementia,is increasing.Brain aging increases the risk of cognitive impairment,which manifests as a progressive loss of neuron function caused...As people live longer,the burden of aging-related brain diseases,especially dementia,is increasing.Brain aging increases the risk of cognitive impairment,which manifests as a progressive loss of neuron function caused by the impairment of synaptic plasticity via disrupting lipid homeostasis.Therefore,supplemental dietary lipids have the potential to prevent brain aging.This review summarizes the important roles of dietary lipids in brain function from both structure and mechanism perspectives.Epidemiological and animal studies have provided evidence of the functions of polyunsaturated fatty acids(PUFAs)in brain health.The results of interventions indicate that phospholipids—including phosphatidylcholine,phosphatidylserine,and plasmalogen—are efficient in alleviating cognitive impairment during aging,with plasmalogen exhibiting higher efficacy than phosphatidylserine.Plasmalogen is a recognized nutrient used in clinical trials due to its special vinyl ether bonds and abundance in the postsynaptic membrane of neurons.Future research should determine the dose-dependent effects of plasmalogen in alleviating brain-aging diseases and should develop extraction and storage procedures for its clinical application.展开更多
The interaction of oviductal epithelial cells （OECs） with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatoz...The interaction of oviductal epithelial cells （OECs） with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatozoa previously selected by density gradient in OEC and determinate some sperm characteristics that could explain the results obtained. In this study, we assessed in vitro fertilization （IVF）, tyrosine phosphorylation, phosphatidylserine translocation, nuclear DNA fragmentation, and chromatin decondensation. Three experimental sperm groups, previously selected by Percoll gradient, were established according to the origin of the sperm used for IVF： （i） W30 group： spermatozoa were incubated with oocytes in the absence of OEC; （ii） NB group： after sperm incubation in OEC, the unbound spermatozoa were incubated with oocytes, in the absence of OEC; and （iii） B group： after sperm incubation with OEC, the bound spermatozoa were incubated with oocytes in the OEC plates. The results showed that sperm from the NB group led to a lower IVF yield, accompanied by low penetration rates （NB： 19.6%, B： 94.9%, and W30： 62.9%; P 〈 0.001） and problems of nuclear decondensation. Moreover, higher levels of tyrosine phosphorylation were observed in the NB group compared with the W30 and B groups （NB： 58.7%, B： 2.5%, and W30： 4.5%; P 〈 0.01）. A similar trend was observed in phosphatidylserine translocation （NB： 93.7%, B. 5.7%, and W30： 44.2%; P 〈 0.01）. These results demonstrate that the OEC exerts a rigorous degree of sperm selection, even within an already highly selected population of spermatozoa, and can capture the best functional spermatozoa for fertilization.展开更多
Injuries to the central or peripheral nervous system frequently cause long-term disabilities because damaged neurons are unable to efficiently self-repair.This inherent deficiency necessitates the need for new treatme...Injuries to the central or peripheral nervous system frequently cause long-term disabilities because damaged neurons are unable to efficiently self-repair.This inherent deficiency necessitates the need for new treatment options aimed at restoring lost function to patients.Compared to humans,a number of species possess far greater regenerative capabilities,and can therefore provide important insights into how our own nervous systems can be repaired.In particular,several invertebrate species have been shown to rapidly initiate regeneration post-injury,allowing separated axon segments to re-join.This process,known as axonal fusion,represents a highly efficient repair mechanism as a regrowing axon needs to only bridge the site of damage and fuse with its separated counterpart in order to re-establish its original structure.Our recent findings in the nematode Caenorhabditis elegans have expanded the promise of axonal fusion by demonstrating that it can restore complete function to damaged neurons.Moreover,we revealed the importance of injury-induced changes in the composition of the axonal membrane for mediating axonal fusion,and discovered that the level of axonal fusion can be enhanced by promoting a neuron＇s intrinsic growth potential.A complete understanding of the molecular mechanisms controlling axonal fusion may permit similar approaches to be applied in a clinical setting.展开更多
Amyotrophic lateral sclerosis is the most common adult-onset neurodegenerative disease affecting motor neurons. Its defining feature is progressive loss of motor neuron function in the cortex, brainstem, and spinal co...Amyotrophic lateral sclerosis is the most common adult-onset neurodegenerative disease affecting motor neurons. Its defining feature is progressive loss of motor neuron function in the cortex, brainstem, and spinal cord, leading to paralysis and death. Despite major advances in identifying genes that can cause disease when mutated and model the disease in animals and cellular models, it still remains unclear why motor symptoms suddenly appear after a long pre-symptomatic phase of apparently normal function. One hypothesis is that age-related deregulation of specific proteins within key cell types, especially motor neurons themselves, initiates disease symptom appearance and may also drive progressive degeneration. Genome-wide in vivo cell-type-specific screening tools are enabling identification of candidates for such proteins. In this minireview, we first briefly discuss the methodology used in a recent study that applied a motor neuron-specific RNASeq screening approach to a standard model of TAR DNA-binding protein-43(TDP-43)-driven amyotrophic lateral sclerosis. A key finding of this study is that synaptogyrin-4 and pleckstrin homology domain-containing family B member 1 are also deregulated at the protein level within motor neurons of two unrelated mouse models of mutant TDP-43 driven amyotrophic lateral sclerosis. Guided by what is known about molecular and cellular functions of these proteins and their orthologs, we outline here specific hypotheses for how changes in their levels might potentially alter cellular physiology of motor neurons and detrimentally affect motor neuron function. Where possible, we also discuss how this information could potentially be used in a translational context to develop new therapeutic strategies for this currently incurable, devastating disease.展开更多
Cells and exosomes derived from them are extensively used as biological carrier systems.Cells demonstrate superior targeting specificity and prolonged circulation facilitated by their rich array of surface proteins,wh...Cells and exosomes derived from them are extensively used as biological carrier systems.Cells demonstrate superior targeting specificity and prolonged circulation facilitated by their rich array of surface proteins,while exosomes,due to their small size,cross barriers and penetrate tumors efficiently.However,challenges remain,cells’large size restricts tissue penetration,and exosomes have limited targeting accuracy and short circulation times.To address these challenges,we developed a novel concept termed exosomal spheres.This approach involved incorporating platelet-derived exosomes shielded with phosphatidylserine(PS)and linked via pH-sensitive bonds for drug delivery applications.The study demonstrated that,compared with exosomes,the exosomal spheres improved blood circulation through the upregulation of CD47 expression and shielding of phosphatidylserine,thereby minimizing immune clearance.Moreover,the increased expression of P-selectin promoted adhesion to circulating tumor cells,thereby enhancing targeting efficiency.Upon reaching the tumor site,the hydrazone bonds of exosome spheres were protonated in the acidic tumor microenvironment,leading to disintegration into uniform-sized exosomes capable of deeper tumor penetration compared to platelets.These findings suggested that exosome spheres addressed the challenges and offered significant potential for efficient and precise drug delivery.展开更多
基金provided by grants from the Dysautonomia Foundation.Israel Science Foundation(ISF)[142/13,1439/14]by Teva Pharmaceutical Industries Ltd as part of the Israeli National Network of Excellence in Neuroscience(NNE)[1234944]+2 种基金supported by grants from the Israel Science Foundation(ISF)[561/11]the European Research Council(ERC)[309377]supported by grants from Teva Pharmaceutical Industries Ltd.under the Israeli National Network of Excellence in Neuroscience
文摘Familial dysautonomia （FD） is a rare children neurodegenerative disease caused due to a point mutation in the IKBKAP gene that results in decreased IKK complex-associated protein （IKAP） protein production. The disease affects mostly the dorsal root ganglion （DRG） and the sympathetic ganglion. Recently, we found that the molecular mechanisms underlying neurodegeneration in FD patients are defects in axonal transport of nerve growth factors and microtubule stability in the DRG. Neurons are highly polarized cells with very long axons. In order to survive and maintain proper function, neurons depend on transport of proteins and other cellular components from the neuronal body along the axons. We further demonstrated that IKAP is necessary for axon maintenance and showed that phosphatidylserine acts as an HDAC6 inhib- itor to rescue neuronal function in FD cells. In this review, we will highlight our latest research findings.
基金financially supported by the Innovative Funds Plan of Henan University of Technology(2020ZKCJ10)Cultivation Programme for Young Backbone Teachers in Henan University of Technology.
文摘Phosphatidylserine(PS)is the part of cell structure in the body and has many beneficial functions especially in brain-related aging diseases.Although daily foods can provide PS to human body,the amount is very limited due to its poverty in most foods.To overcome the issue,numerous studies based on PS have been reported to develop PS-related supplements.In this review,PS was comprehensively and critically reviewed from the view of resources,functions,processing techniques,patents,and prospects.For resources,animal,plant,and microorganism origins were all covered with their differences in composition profiles.For functions,benefits regarding memory,cognitive enhancement,exercise performance,reducing Alzheimer’s disease,and attention-deficit hyperactivity disorder symptoms were covered as well as the functional differences among animal-,plant-,and microorganism-based PS-related supplements.For processing techniques,traditional extracting methods from animal,plant,and microorganism tissues were comparatively discussed with enzymatic synthesis based on different reaction systems.Finally,patents of PS-related supplements were evaluated as well as their applications.This review could provide scientific and valuable support for PS industry.
基金supported by the National Natural Science Foundation of China(31922072)the Natural Science Foundation of Shandong Province(ZR2020JQ15)the Taishan Scholar Project of Shandong Province(tsqn201812020)。
文摘Sensitive monitoring of the target products during the biosynthesis process is crucial,and facile analytical approaches are urgently needed.Herein,phosphatidylserine(PS)was chosen as the model target,a colorimetric aptasensor was developed for the rapid quantitation in biosynthesis samples.A chimeric aptamer was constructed with two homogeneous original PS aptamers.Specific recognition between the chimeric aptamer and PS results in the desorption of aptamer from the surface of the AuNPs nanozyme,and the peroxidase-like enzymatic activity of the AuNPs nanozyme was weakened in a relationship with the different concentrations.The developed aptasensor performed well when applied for analyzing PS in biosynthesis samples.The aptasensor offers good sensitivity and selectivity,under optimal conditions,achieving monitoring and quantitation of PS in the range of 2.5-80.0μmol/L,with a limit of detection at 536.2 nmol/L.Moreover,the aptasensor provides good accuracy,with comparison rates of 98.17%-106.40%,when compared with the HPLC-ELSD.This study provides a good reference for monitoring other biosynthesized products and promoting the development of aptamers and aptasensors in real-world applications.
文摘We have recently demonstrated that liposomes composed of phosphatidylserine (PS-liposomes) suppressed nitric oxide and inflammatory cytokine productions following LPS stimulation in macrophages. In this study, we examined the effect of PS-liposomes on expressions of TLR-4 and MyD88, which are essential for the signal transduction in LPS stimulation. Expression of MyD88 was suppressed when macrophages were treated with PS-liposomes, but not with liposomes of phosphatidylcholine. No change in TLR-4 expression was observed. MyD88 suppression was restored to the control levels when cells were pre-treated with anti-TGF-β antibody, suggesting that TGF-β plays an important role in down-regulation of MyD88 following PS-liposome treatment.
基金supported by the Natural Science Foundation of Shandong province(ZR2023QH204 and ZR2024ME024,China).
文摘Chemotherapeutic drugs,such as cisplatin and phenanthriplatin(PhenPt),as STING agonists to induce DNA damage and activate the cyclic GMP-AMP synthase-stimulator of interferon genes(cGAS-STING)signaling pathway provides a potential strategy for clinical chemo-immunotherapy.However,treatment with Pt-based drugs leads to irreversible ectopia of phosphatidylserine(PS),a major component of the intracellular membrane,to the surface of the cancer cells by enzymes(Xkr8).Exposed PS can bind to immune cell receptors and inhibit the presentation of tumor antigens,leading to immunosuppression and attenuation of chemotherapy.Herein,we report a novel approach to enhance chemo-immunotherapy by constructing siRNA targeted Xkr8(siXkr8)-mediated tetrahedral framework nucleic acid nanogel structure concurrently loaded with PhenPt(siXkr8-FNG/PhenPt)for co-delivery of siRNA and Pt-based drugs.The results showed that siXkr8-FNG/PhenPt can not only be used as an efficient delivery carrier to deliver siXkr8,block the expression of Xkr8,reduce the exposure of PS on the cancer cells surface,but also act as an immune stimulant to activate cGAS-STING pathway,effectively improve the immunosuppressive microenvironment,produce antitumor immune response,and inhibit tumor growth and metastasis.Overall,this new delivery system is important for improving the effect of Pt-based drug chemotherapy,inducing immune enhancement and nucleic acid drug delivery.
基金supported by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Develop-ment Institute(KHIDI),funded by the Ministry of Health&Welfare,Republic of Korea(HI22C1323)supported by the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(2021R1A6A1A03039462).
文摘Radiation-induced fibrosis(RIF)is a major complication following radiotherapy treatment.Macrophages are the key regulators of inflammatory responses and have emerged as critical targets for the prevention of fibrosis.This study evaluated the efficacy of PEGylated phosphatidylserine-containing liposomes(PEG-PSLs)in mitigating RIF by modulating macrophage activity.PEG-PSLs influenced the polarization of bone marrow-derived macrophages(BMDMs),reducing the expression levels of inflammatory cytokines and fibrosis-associated markers.These ef-fects were observed within a specific PEGylation range,with the greatest reduction achieved at 1-1.5mol%PEG.Analysis of the IL-4-induced JAK-STAT signaling pathway indicated that the PEG-PSL-mediated suppression occurred at or beyond the point of JAK-1 phosphorylation.In in vitro fibrosis assays,PEG-PSLs reduced fibroblast-to-myofibroblast and epithelial-to-mesenchymal transitions when co-cultured with BMDMs.In vivo studies using a rat model of esophageal irradiation demonstrated that PEG-PSLs effectively attenuated fibrotic progression,preserved tissue architecture,and enhanced muscle regeneration.These findings indicate the po-tential of partially PEGylated phosphatidylserine liposomes as therapeutic agents for mitigating post-irradiation esophageal fibrosis.
基金NationalNatural Science Foundation of China:National Science Foundation for ExcellentYoung Scholars,Grant/Award Number:32122052NationalNatural Sci-ence Foundation of China:NationalNat-ural Science FoundationRegional Inno-vation and Development,Grant/Award Number:U19A2003NationalNatural Science Foundation ofChina:National Science Foundation forYoung Scholars,Grant/Award Number:81902662。
文摘Background:Understanding how the tumor microenvironment is shaped by various factors is important for the development of new therapeutic strategies.Tumor cells often undergo spontaneous apoptotic cell death in tumor microen-vironment,these apoptotic cells are histologically co-localized with immunosup-pressive macrophages.However,the mechanism by which tumor cell apoptosis modulates macrophage polarization is not fully understood.In this study,we aimed to explore the tumor promoting effects of apoptotic tumor cells and the signal pathways involved.Methods:Apoptotic cells and macrophages in tumors were detected by immunohistochemical staining.Morphological analysis was performed with Giemsa staining.Lipids generated from apoptotic cells were detected by liq-uid chromatography-mass spectrometry.Phosphatidylserine-containing lipo-somes were prepared to mimic apoptotic cells.The expression of protein was determined by real-time PCR,immunohistochemistry enzyme-linked immunosorbent assay and Western blotting.Mouse malignant ascites and subcu-taneous tumor models were designed for in vivo analysis.Transgenic mice with specific genes knocked out and inhibitors specific to certain proteins were used for the mechanistic studies.Results:The location and the number of apoptotic cells were correlated with that of macrophages in several types of carcinomas.Phosphatidylserine,a lipid molecule generated in apoptotic cells,induced polarization and accumulation of M2-like macrophages in vivo and in vitro.Moreover,sustained administration of phosphoserine promoted tumor growth in the malignant ascites and subcuta-neous tumor models.Further analyses suggested that phosphoserine induced a M2-like phenotype in macrophages,which was related to the activation of phosphoserine receptors including T-cell immunoglobin mucin 4(TIM4)and the FAK-SRC-STAT3 signaling pathway as well as elevated the expression of the histone demethylase Jumonji domain-containing protein 3(JMJD3).Adminis-tration of specific inhibitors of these pathways could reduce tumor progression.Conclusions:This study suggest that apoptotic cell-generated phosphoserine might be a notable signal for immunosuppressive macrophages in tumors,and the related pathways might be potential therapeutic targets for cancer therapy.
基金This work was financially supported by the Medical Research Council,UK.
文摘The role of surfactant protein A(SP-A)in the recognition and clearance of apoptotic cells is well established,but to date,it is still not clear which surface molecules of apoptotic cells are involved in the process.Here we present evidence that phosphatidylserine(PS)is a relevant binding molecule for human SP-A.The binding is Ca^(2+)-dependent and is not inhibited by mannose,suggesting that the sugar-binding site of the carbohydrate recognition domain(CRD)of SP-A is not involved.Flow cytometry studies on apoptotic Jurkat cells revealed apparent inhibition of annexin V binding by increasing concentrations of SP-A in late apoptotic but not early apoptotic cells,and this was consistent for Jurkat cells and neutrophils.Supporting these data,confocal microscopy results show a co-localisation of annexin V and SP-A in late apoptotic but not early apoptotic cells.However,we cannot conclude that this inhibition is exclusively due to the binding of SP-A to PS on the cell surface,as annexin V is not wholly specific for PS and SP-A also interacts with other phospholipids that might become exposed on the apoptotic cell surface.
基金supported by grants from the Canadian Institutes of Health Research(CIHR)MT-14429,MOP-82906,and FDN-143338 to A.V.NSFC-31870863 from the National Natural Science Foundation of China to N.W.supported by a Postdoctoral Fellowship from Fonds de recherche du Quebec Santr(FRQS)。
文摘One of the hallmarks of live cells is the asymmetric distribution of lipids across their plasma membrane.Changes in this asymmetry due to lipid"scrambling"result in phosphatidylserine exposure at the cell surface that is detected by annexin V staining.This alteration is observed during cell death processes such as apoptosis,and during physiological responses such as platelet degranulation and membrane repair.Previous studies have shown that activation of NK cells is accompanied by exposure of phosphatidylserine at the cell surface.While this response was thought to be indicative of ongoing NK cell death,it may also reflect the regulation of NK cell activation in the absence of cell death.Herein,we found that NK cell activation was accompanied by rapid phosphatidylserine exposure to an extent proportional to the degree of NK cell activation.Through enforced expression of a lipid scramblase,we provided evidence that activation-induced lipid scrambling in NK cells is reversible and does not lead to cell death.In contrast,lipid scrambling attenuates NKcell activation.This response was accompanied by reduced cell surface expression of activating receptors such as 2B4,and by loss of binding of Src family protein tyrosine kinases Fyn and Lck to the inner leaflet of the plasma membrane.Hence,lipid scrambling during NK cell activation is,at least in part,a physiological response that reduces the NK cell activation level.This effect is due to the ability of lipid scrambling to alter the distribution of membrane-associated receptors and kinases required for NK cell activation.
基金supported by the National Natural Science Foundation of China(81874307,21874088)Shanghai Science and Technology Commission Scientific Research Project(18142200700,19142203100,20142200400)+1 种基金the Open Project Program of Engineering Research Center of Cell&Therapeutic Antibody,Ministry of Education,Shanghai Jiao Tong University(19X110020009-005)Startup Fund for Youngman Research at SJTU(19X100040029).
文摘Exosomes are membrane bound extracellular vesicles that play an important role in many biological processes.While they have great application value,exosome isolation is still considered a major scientific challenge.In the present study,a novel separation strategy for exosomes is proposed based on the specific interaction between immobilized peptide ligands and phosphatidylserine moieties which are highly abundant on the surface of exosomes.With the new affinity method,intact model exosomes can be recovered with a high yield in a short processing time.The purity of exosome samples enriched from serum by the affinity method is far higher than that isolated by ultrafiltration,and similar to that obtained by density gradient centrifugation and ultracentrifugation.Moreover,the variety of contaminants co-isolated by the affinity method is relatively low due to its specific separation principle.Proteomics analysis of exosomes isolated by the affinity method from the serum of healthy,hepatocellular carcinoma patients,and intrahepatic cholangiocarcinoma patients was performed to prove the applicability of this method.In conclusion,our novel strategy shows characteristics of easy preparation,high specificity,and cost-effectiveness,and provides a promising approach for exosome isolation which should have wide applications.
基金supported by the National Natural Science Foundation of China(82171136 and 92368110 to G.W.,82201291 to G.-J.Z.,82192861 to Z.X.,81970884 and 82192862 to X.G.)the Natural Science Foundation of Jiangsu Province(BK20220188 to Q.L.,BK20220189 to G.-J.Z.)the Fundamental Research Funds for the Central Universities(021414380533 to G.W.).
文摘Hearing relies on the structural and functional integrity of cochlear hair cells,particularly their apical F-actin-filled stereocilia.Phospholipid scramblases are important for maintaining membrane asymmetry,but their roles in the stereocilia and auditory functions are not fully understood.Here,we identify Plscr5 as a downstream target of the transcription factor POU4F3 essential for hair cell function,whose mutation causes human DFNA15 deafness.Plscr5 knockout mice exhibit progressive hearing loss due to stereocilia degeneration and hair cell loss.Functional analyses reveal that PLSCR5 contributes to phosphatidylserine externalization in hair cell apical membranes,particularly in inner hair cells,and is important for outer hair cell and stereocilia maintenance.Our findings highlight PLSCR5 as an important downstream effector of POU4F3 and regulator of phosphatidylserine externalization and membrane dynamics required for auditory functions.
基金a grant from the Science and Technology Committee of Jiangsu Province,No.BJ98110
文摘INTRODUCTIONApoptosis,described by Kerr in 1972,plays a keyrole in all types of regulated cellular processes inmulticellular,organisms.It is defined as amorphologic change,including fragmentation of theDNA,cell shrinkage,dilation of the endoplasmaticreticulum,cell fragmentation and formation
文摘We question whether,in men with an abnormal rate of sperm DNA fragmentation,the magnetic-activated cell sorting(MACS)could select spermatozoa with lower rates of DNA fragmentation as well as spermatozoa with unbalanced chromosome content.Cryopreserved spermatozoa from six males were separated into nonapoptotic and apoptotic populations.We determined the percentages of spermatozoa with(i)externalization of phosphatidylserine(EPS)by annexin V-Fluorescein isothiocyanate(FITC)labeling,(ii)DNA fragmentation by TdT-mediated-dUTP nick-end labeling(TUNEL),and(iii)numerical abnormalities for chromosomes X,Y,13,18,and 21 by fluorescence in situ hybridization(FISH),on the whole ejaculate and selected spermatozoa in the same patient.Compared to the nonapoptotic fraction,the apoptotic fraction statistically showed a higher number of spermatozoa with EPS,with DNA fragmentation,and with numerical chromosomal abnormalities.Compared to the whole ejaculate,we found a significant decrease in the percentage of spermatozoa with EPS and decrease tendencies of the DNA fragmentation rate and the sum of disomy levels in the nonapoptotic fraction.Conversely,we observed statistically significant higher rates of these three parameters in the apoptotic fraction.MACS may help to select spermatozoa with lower rates of DNA fragmentation and unbalanced chromosome content in men with abnormal rates of sperm DNA fragmentation.
基金supported by the National Key R&D Program of China (2019YFA0508502/3 and 2021YFC2300604)the Natural Science Foundation of China (Reference numbers 82388201, 82241216, and 32270963)+1 种基金the Research Funds of Center for Advanced Interdisciplinary Science and Biomedicine of IHM (QYZD20220008)the Anhui Key Research and Development Plan (Reference number 2023z04020011)。
文摘Objective: The human cluster of differentiation(CD)300A, a type-I transmembrane protein with immunoreceptor tyrosine-based inhibitory motifs, was investigated as a potential immune checkpoint for human natural killer(NK) cells targeting hematologic malignancies(HMs).Methods: We implemented a stimulation system involving the CD300A ligand, phosphatidylserine(PS), exposed to the outer surface of malignant cells. Additionally, we utilized CD300A overexpression, a CD300A blocking system, and a xenotransplantation model to evaluate the impact of CD300A on NK cell efficacy against HMs in in vitro and in vivo settings. Furthermore, we explored the association between CD300A and HM progression in patients.Results: Our findings indicated that PS hampers the function of NK cells. Increased CD300A expression inhibited HM lysis by NK cells. CD300A overexpression shortened the survival of HM-xenografted mice by impairing transplanted NK cells. Blocking PS–CD300A signals with antibodies significantly amplified the expression of lysis function-related proteins and effector cytokines in NK cells, thereby augmenting the ability to lyse HMs. Clinically, heightened CD300A expression correlated with shorter survival and an “exhausted” phenotype of intratumoral NK cells in patients with HMs or solid tumors.Conclusions: These results propose CD300A as a potential target for invigorating NK cell-based treatments against HMs.
基金supported by the National Key Research and Development Program of China(2022YFD2101003)the 111 Project from the Ministry of Education of the People’s Republic of China(B18053).
文摘As people live longer,the burden of aging-related brain diseases,especially dementia,is increasing.Brain aging increases the risk of cognitive impairment,which manifests as a progressive loss of neuron function caused by the impairment of synaptic plasticity via disrupting lipid homeostasis.Therefore,supplemental dietary lipids have the potential to prevent brain aging.This review summarizes the important roles of dietary lipids in brain function from both structure and mechanism perspectives.Epidemiological and animal studies have provided evidence of the functions of polyunsaturated fatty acids(PUFAs)in brain health.The results of interventions indicate that phospholipids—including phosphatidylcholine,phosphatidylserine,and plasmalogen—are efficient in alleviating cognitive impairment during aging,with plasmalogen exhibiting higher efficacy than phosphatidylserine.Plasmalogen is a recognized nutrient used in clinical trials due to its special vinyl ether bonds and abundance in the postsynaptic membrane of neurons.Future research should determine the dose-dependent effects of plasmalogen in alleviating brain-aging diseases and should develop extraction and storage procedures for its clinical application.
文摘The interaction of oviductal epithelial cells （OECs） with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatozoa previously selected by density gradient in OEC and determinate some sperm characteristics that could explain the results obtained. In this study, we assessed in vitro fertilization （IVF）, tyrosine phosphorylation, phosphatidylserine translocation, nuclear DNA fragmentation, and chromatin decondensation. Three experimental sperm groups, previously selected by Percoll gradient, were established according to the origin of the sperm used for IVF： （i） W30 group： spermatozoa were incubated with oocytes in the absence of OEC; （ii） NB group： after sperm incubation in OEC, the unbound spermatozoa were incubated with oocytes, in the absence of OEC; and （iii） B group： after sperm incubation with OEC, the bound spermatozoa were incubated with oocytes in the OEC plates. The results showed that sperm from the NB group led to a lower IVF yield, accompanied by low penetration rates （NB： 19.6%, B： 94.9%, and W30： 62.9%; P 〈 0.001） and problems of nuclear decondensation. Moreover, higher levels of tyrosine phosphorylation were observed in the NB group compared with the W30 and B groups （NB： 58.7%, B： 2.5%, and W30： 4.5%; P 〈 0.01）. A similar trend was observed in phosphatidylserine translocation （NB： 93.7%, B. 5.7%, and W30： 44.2%; P 〈 0.01）. These results demonstrate that the OEC exerts a rigorous degree of sperm selection, even within an already highly selected population of spermatozoa, and can capture the best functional spermatozoa for fertilization.
基金supported by National Health and Medical Research Council(NHMRC) Project Grant 1101974 to BN
文摘Injuries to the central or peripheral nervous system frequently cause long-term disabilities because damaged neurons are unable to efficiently self-repair.This inherent deficiency necessitates the need for new treatment options aimed at restoring lost function to patients.Compared to humans,a number of species possess far greater regenerative capabilities,and can therefore provide important insights into how our own nervous systems can be repaired.In particular,several invertebrate species have been shown to rapidly initiate regeneration post-injury,allowing separated axon segments to re-join.This process,known as axonal fusion,represents a highly efficient repair mechanism as a regrowing axon needs to only bridge the site of damage and fuse with its separated counterpart in order to re-establish its original structure.Our recent findings in the nematode Caenorhabditis elegans have expanded the promise of axonal fusion by demonstrating that it can restore complete function to damaged neurons.Moreover,we revealed the importance of injury-induced changes in the composition of the axonal membrane for mediating axonal fusion,and discovered that the level of axonal fusion can be enhanced by promoting a neuron＇s intrinsic growth potential.A complete understanding of the molecular mechanisms controlling axonal fusion may permit similar approaches to be applied in a clinical setting.
基金supported in part by funding from the Else Kr?ner Fresenius Stiftung(Co-PI)the Werner Otto Stiftung(PI)(to KED)。
文摘Amyotrophic lateral sclerosis is the most common adult-onset neurodegenerative disease affecting motor neurons. Its defining feature is progressive loss of motor neuron function in the cortex, brainstem, and spinal cord, leading to paralysis and death. Despite major advances in identifying genes that can cause disease when mutated and model the disease in animals and cellular models, it still remains unclear why motor symptoms suddenly appear after a long pre-symptomatic phase of apparently normal function. One hypothesis is that age-related deregulation of specific proteins within key cell types, especially motor neurons themselves, initiates disease symptom appearance and may also drive progressive degeneration. Genome-wide in vivo cell-type-specific screening tools are enabling identification of candidates for such proteins. In this minireview, we first briefly discuss the methodology used in a recent study that applied a motor neuron-specific RNASeq screening approach to a standard model of TAR DNA-binding protein-43(TDP-43)-driven amyotrophic lateral sclerosis. A key finding of this study is that synaptogyrin-4 and pleckstrin homology domain-containing family B member 1 are also deregulated at the protein level within motor neurons of two unrelated mouse models of mutant TDP-43 driven amyotrophic lateral sclerosis. Guided by what is known about molecular and cellular functions of these proteins and their orthologs, we outline here specific hypotheses for how changes in their levels might potentially alter cellular physiology of motor neurons and detrimentally affect motor neuron function. Where possible, we also discuss how this information could potentially be used in a translational context to develop new therapeutic strategies for this currently incurable, devastating disease.
基金support from National Key R&D Program of China(No.2022YFE0111600)National Natural Science Foundation of China(No.82273874)Liaoning Revitalization Talents Program(No.XLYC22202019,China).
文摘Cells and exosomes derived from them are extensively used as biological carrier systems.Cells demonstrate superior targeting specificity and prolonged circulation facilitated by their rich array of surface proteins,while exosomes,due to their small size,cross barriers and penetrate tumors efficiently.However,challenges remain,cells’large size restricts tissue penetration,and exosomes have limited targeting accuracy and short circulation times.To address these challenges,we developed a novel concept termed exosomal spheres.This approach involved incorporating platelet-derived exosomes shielded with phosphatidylserine(PS)and linked via pH-sensitive bonds for drug delivery applications.The study demonstrated that,compared with exosomes,the exosomal spheres improved blood circulation through the upregulation of CD47 expression and shielding of phosphatidylserine,thereby minimizing immune clearance.Moreover,the increased expression of P-selectin promoted adhesion to circulating tumor cells,thereby enhancing targeting efficiency.Upon reaching the tumor site,the hydrazone bonds of exosome spheres were protonated in the acidic tumor microenvironment,leading to disintegration into uniform-sized exosomes capable of deeper tumor penetration compared to platelets.These findings suggested that exosome spheres addressed the challenges and offered significant potential for efficient and precise drug delivery.
