Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs fr...Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs from plants remains challenging.A variety of methods have been utilized to isolate plant EVs from apoplastic washing fluid(AWF).Here,we compare published plant EV isolation methods,and provide our recommended method for the isolation and purification of plant EVs.This method includes a detailed protocol for clean AWF collection from Arabidopsis thaliana leaves,followed by EV isolation via differential centrifugation.To further separate and purify specific subclasses of EVs from heterogeneous vesicle populations,density gradient ultracentrifugation and immunoaffinity capture are then utilized.We found that immunoaffinity capture is the most precise method for specific EV subclass isolation when suitable specific EV biomarkers and their corresponding antibodies are available.Overall,this study provides a guide for the selection and optimization of EV isolation methods for desired downstream applications.展开更多
Background Previous researches have indicated that glioma invasion may occur within a tumor-host microecology, and that fibronectin may be involved in glioma invasion as an important component of the extracellular mat...Background Previous researches have indicated that glioma invasion may occur within a tumor-host microecology, and that fibronectin may be involved in glioma invasion as an important component of the extracellular matrix. However, how the interaction between tumor cells and vascular endothelial cells affects glioma invasion is poorly understood. The aim of this study was to investigate the effects of the interaction between tumor cells and vascular endothelial cells on glioma invasion, and the relationship of this interaction to fibronectin. Methods The localization of fibronectin in different brain astrocytoma tissues was determined by immunohistochemistry Then, vascular endothelial cells and glioma cells were co-cultured in a Transwell co-culturing system. Fibronectin expression was detected by reverse transcriptase-polymerase chain reaction, immunocytochemistry, and enzyme-linked immunosorbent assay. Additionally, the influence of the interaction between tumor cells and vascular endothelial cells on glioma cell invasion was determined by an in vitro rapid invasion test. Results In brain astrocytoma tissues, fibronectin was present on the endothelial cells, in the extracellular matrix. Fibronectin expression was greater in higher grade tumors than in lower grade tumors. The interaction of glioma cells and vascular endothelial cells in vitro induced fibronectin release from vascular endothelial cells, which in turn stimulated glioma cell migration. This effect was inhibited by fibronectin blocking antibody. Conclusion Glioma cells may induce vascular epithelial cells to express fibronectin, and in turn fibronectin could promote glioma cell invasion.展开更多
Although microbes primarily are single-cell organisms,they are not isolated individuals.Microbes use various means to communicate with one another.Based on the communication,microbes establish a social interaction wit...Although microbes primarily are single-cell organisms,they are not isolated individuals.Microbes use various means to communicate with one another.Based on the communication,microbes establish a social interaction with their neighbors in a specific ecological niche,and cooperative behaviors are normally performed to provide benefits on the population and species levels.In the microbiome era,in order to better understand the behaviors of microbes,deep understanding of the social communication between microbes hence becomes a key to interpret microbe behaviors.Here we summarize the molecular mechanisms that underlie the cell-to-cell communication in prokaryotic and eukaryotic microorganisms,the recent discoveries and novel technologies in understanding the interspecies and interkingdom communication,and discuss new concepts of the sociomicrobiology.展开更多
Development of drug resistance represents the major cause of cancer therapy failure,determines disease progression and results in poor prognosis for cancer patients.Different mechanisms are responsible for drug resist...Development of drug resistance represents the major cause of cancer therapy failure,determines disease progression and results in poor prognosis for cancer patients.Different mechanisms are responsible for drug resistance.Intrinsic genetic modifications of cancer cells induce the alteration of expression of gene controlling specific pathways that regulate drug resistance:drug transport and metabolism;alteration of drug targets;DNA damage repair;and deregulation of apoptosis,autophagy,and pro-survival signaling.On the other hand,a complex signaling network among the entire cell component characterizes tumor microenvironment and regulates the pathways involved in the development of drug resistance.Gut microbiota represents a new player in the regulation of a patient’s response to cancer therapies,including chemotherapy and immunotherapy.In particular,commensal bacteria can regulate the efficacy of immune checkpoint inhibitor therapy by modulating the activation of immune responses to cancer.Commensal bacteria can also regulate the efficacy of chemotherapeutic drugs,such as oxaliplatin,gemcitabine,and cyclophosphamide.Recently,it has been shown that such bacteria can produce extracellular vesicles(EVs)that can mediate intercellular communication with human host cells.Indeed,bacterial EVs carry RNA molecules with gene expression regulatory ability that can be delivered to recipient cells of the host and potentially regulate the expression of genes involved in controlling the resistance to cancer therapy.On the other hand,host cells can also deliver human EVs to commensal bacteria and similarly,regulate gene expression.EVmediated intercellular communication between commensal bacteria and host cells may thus represent a novel research area into potential mechanisms regulating the efficacy of cancer therapy.展开更多
The endoplasmic reticulum quality control(ER-QC)is a conserved mechanism in surveillance of secreted signaling factors during cell-to-cell communication in eukaryotes.Recent data show that the ER-QC plays important ro...The endoplasmic reticulum quality control(ER-QC)is a conserved mechanism in surveillance of secreted signaling factors during cell-to-cell communication in eukaryotes.Recent data show that the ER-QC plays important roles in diverse cell-to-cell signaling processes during immune response,vegetative and reproductive development in plants.Pollen tube guidance is a precisely guided cell-cell communication process between the male and female gametophytes during plant reproduction.Recently,the female signal has been identified as small secreted peptides,but how the pollen tube responds to this signal is still unclear.In this review,we intend to summarize the role of ER-QC in plants and discuss the recent advances regarding our understanding of the mechanism of pollen tube response to the female signals.展开更多
Exosomes are 40–100 nm nano-sized vesicles that are released from many cell types into the extracellular space. Such vesicles are widely distributed in various body fluids. Recently,m RNAs and micro RNAs（mi RNAs） h...Exosomes are 40–100 nm nano-sized vesicles that are released from many cell types into the extracellular space. Such vesicles are widely distributed in various body fluids. Recently,m RNAs and micro RNAs（mi RNAs） have been identified in exosomes, which can be taken up by neighboring or distant cells and subsequently modulate recipient cells. This suggests an active sorting mechanism of exosomal mi RNAs, since the mi RNA profiles of exosomes may differ from those of the parent cells. Exosomal mi RNAs play an important role in disease progression, and can stimulate angiogenesis and facilitate metastasis in cancers. In this review, we will introduce the origin and the trafficking of exosomes between cells, display current research on the sorting mechanism of exosomal mi RNAs, and briefly describe how exosomes and their mi RNAs function in recipient cells.Finally, we will discuss the potential applications of these mi RNA-containing vesicles in clinical settings.展开更多
Meiosis is a highly complex process significantly influenced by transcriptional regulation.However,studies on the mechanisms that govern transcriptomic changes during meiosis,especially in prophase I,are limited.Here,...Meiosis is a highly complex process significantly influenced by transcriptional regulation.However,studies on the mechanisms that govern transcriptomic changes during meiosis,especially in prophase I,are limited.Here,we performed single-cell ATAC-seq of human testis tissues and observed reprogramming during the transition from zygotene to pachytene spermatocytes.This event,conserved in mice,involved the deactivation of genes associated with meiosis after reprogramming and the activation of those related to spermatogenesis before their functional onset.Furthermore,we identified 282 transcriptional regulators(TRs)that underwent activation or deactivation subsequent to this process.Evidence suggested that physical contact signals from Sertoli cells may regulate these TRs in spermatocytes,while secreted ENHO signals may alter metabolic patterns in these cells.Our results further indicated that defective transcriptional reprogramming may be associated with non-obstructive azoospermia(NOA).This study revealed the importance of both physical contact and secreted signals between Sertoli cells and germ cells in meiotic progression.展开更多
Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical...Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical concern,exacerbating the current organ shortage and detrimentally affecting the prognosis of recipients.Unfortunately,the perplexing hepatic heterogeneity has impeded characterization of the cellular traits and molecular events that contribute to EAD.Herein,we constructed a pioneering single-cell transcriptomic landscape of human transplanted livers derived from non-EAD and EAD patients,with 12 liver samples collected from 7 donors during the cold perfusion and portal reperfusion stages.Comparison of the 75231 cells of non-EAD and EAD patients revealed an EAD-associated immune niche comprising mucosal-associated invariant T cells,granzyme B^(+)(GZMB^(+))granzyme K^(+)(GZMK^(+))natural killer cells,and S100 calcium binding protein A12^(+)(S100A12^(+))neutrophils.Moreover,we verified this immune niche and its association with EAD occurrence in two independent cohorts.Our findings elucidate the cellular characteristics of transplanted livers and the EAD-associated pathogenic immune niche at the single-cell level,thus,offering valuable insights into EAD onset.展开更多
基金supported by grants from the National Natural Science Foundation of China(32070288)supported by grants from the National Institute of Health(R35 GM136379)+3 种基金the National Science Foundation(IOS2017314)the United States Department of Agriculture National Institute of Food and Agriculture(2021-67013-34258 and 2019-70016-29067)the Australian Research Council Industrial Transformation Research Hub(IH190100022)the CIFAR Fungal Kingdom fellowship to H.J。
文摘Plant extracellular vesicles(EVs)play critical roles in the cross-kingdom trafficking of molecules from hosts to interacting microbes,most notably in plant defense responses.However,the isolation of pure,intact EVs from plants remains challenging.A variety of methods have been utilized to isolate plant EVs from apoplastic washing fluid(AWF).Here,we compare published plant EV isolation methods,and provide our recommended method for the isolation and purification of plant EVs.This method includes a detailed protocol for clean AWF collection from Arabidopsis thaliana leaves,followed by EV isolation via differential centrifugation.To further separate and purify specific subclasses of EVs from heterogeneous vesicle populations,density gradient ultracentrifugation and immunoaffinity capture are then utilized.We found that immunoaffinity capture is the most precise method for specific EV subclass isolation when suitable specific EV biomarkers and their corresponding antibodies are available.Overall,this study provides a guide for the selection and optimization of EV isolation methods for desired downstream applications.
基金This work was supported by a grant from the National Natural Science Foundation of China （No.30973083）. We are not aware of any conflict of financial interest for any author.Acknowledgement： We thank Prof. HU Zhi-jian for assistance in data processing and statistical analysis.
文摘Background Previous researches have indicated that glioma invasion may occur within a tumor-host microecology, and that fibronectin may be involved in glioma invasion as an important component of the extracellular matrix. However, how the interaction between tumor cells and vascular endothelial cells affects glioma invasion is poorly understood. The aim of this study was to investigate the effects of the interaction between tumor cells and vascular endothelial cells on glioma invasion, and the relationship of this interaction to fibronectin. Methods The localization of fibronectin in different brain astrocytoma tissues was determined by immunohistochemistry Then, vascular endothelial cells and glioma cells were co-cultured in a Transwell co-culturing system. Fibronectin expression was detected by reverse transcriptase-polymerase chain reaction, immunocytochemistry, and enzyme-linked immunosorbent assay. Additionally, the influence of the interaction between tumor cells and vascular endothelial cells on glioma cell invasion was determined by an in vitro rapid invasion test. Results In brain astrocytoma tissues, fibronectin was present on the endothelial cells, in the extracellular matrix. Fibronectin expression was greater in higher grade tumors than in lower grade tumors. The interaction of glioma cells and vascular endothelial cells in vitro induced fibronectin release from vascular endothelial cells, which in turn stimulated glioma cell migration. This effect was inhibited by fibronectin blocking antibody. Conclusion Glioma cells may induce vascular epithelial cells to express fibronectin, and in turn fibronectin could promote glioma cell invasion.
基金supported by the National Natural Science Foundation of China(31571288)CAS Interdisciplinary Innovation Team and the Newton Advanced Fellowship(NA140085)from the Royal Society
文摘Although microbes primarily are single-cell organisms,they are not isolated individuals.Microbes use various means to communicate with one another.Based on the communication,microbes establish a social interaction with their neighbors in a specific ecological niche,and cooperative behaviors are normally performed to provide benefits on the population and species levels.In the microbiome era,in order to better understand the behaviors of microbes,deep understanding of the social communication between microbes hence becomes a key to interpret microbe behaviors.Here we summarize the molecular mechanisms that underlie the cell-to-cell communication in prokaryotic and eukaryotic microorganisms,the recent discoveries and novel technologies in understanding the interspecies and interkingdom communication,and discuss new concepts of the sociomicrobiology.
文摘Development of drug resistance represents the major cause of cancer therapy failure,determines disease progression and results in poor prognosis for cancer patients.Different mechanisms are responsible for drug resistance.Intrinsic genetic modifications of cancer cells induce the alteration of expression of gene controlling specific pathways that regulate drug resistance:drug transport and metabolism;alteration of drug targets;DNA damage repair;and deregulation of apoptosis,autophagy,and pro-survival signaling.On the other hand,a complex signaling network among the entire cell component characterizes tumor microenvironment and regulates the pathways involved in the development of drug resistance.Gut microbiota represents a new player in the regulation of a patient’s response to cancer therapies,including chemotherapy and immunotherapy.In particular,commensal bacteria can regulate the efficacy of immune checkpoint inhibitor therapy by modulating the activation of immune responses to cancer.Commensal bacteria can also regulate the efficacy of chemotherapeutic drugs,such as oxaliplatin,gemcitabine,and cyclophosphamide.Recently,it has been shown that such bacteria can produce extracellular vesicles(EVs)that can mediate intercellular communication with human host cells.Indeed,bacterial EVs carry RNA molecules with gene expression regulatory ability that can be delivered to recipient cells of the host and potentially regulate the expression of genes involved in controlling the resistance to cancer therapy.On the other hand,host cells can also deliver human EVs to commensal bacteria and similarly,regulate gene expression.EVmediated intercellular communication between commensal bacteria and host cells may thus represent a novel research area into potential mechanisms regulating the efficacy of cancer therapy.
基金by grants from the Ministry of Science and Technology of China(No.2007CB947600)National Natural Science Foundation of China(Grant Nos.30830063 and 30921003)to W.C.Y.
文摘The endoplasmic reticulum quality control(ER-QC)is a conserved mechanism in surveillance of secreted signaling factors during cell-to-cell communication in eukaryotes.Recent data show that the ER-QC plays important roles in diverse cell-to-cell signaling processes during immune response,vegetative and reproductive development in plants.Pollen tube guidance is a precisely guided cell-cell communication process between the male and female gametophytes during plant reproduction.Recently,the female signal has been identified as small secreted peptides,but how the pollen tube responds to this signal is still unclear.In this review,we intend to summarize the role of ER-QC in plants and discuss the recent advances regarding our understanding of the mechanism of pollen tube response to the female signals.
基金supported by the Projects of International Cooperation and Exchanges from the National Natural Science Foundation of China(Grant No.31161120358)the National Basic Research Program from the Ministry of Science and Technology of China(973 program+3 种基金Grant Nos20111CB510106 and 2015CB910603)the Open Project of State Key Laboratory of Biomembrane and Membrane Biotechnologythe Scientific Research Foundation for Returned Scholars from the Ministry of Education of ChinaML was supported by National Natural Science Foundation of China(Grant No.31400741)
文摘Exosomes are 40–100 nm nano-sized vesicles that are released from many cell types into the extracellular space. Such vesicles are widely distributed in various body fluids. Recently,m RNAs and micro RNAs（mi RNAs） have been identified in exosomes, which can be taken up by neighboring or distant cells and subsequently modulate recipient cells. This suggests an active sorting mechanism of exosomal mi RNAs, since the mi RNA profiles of exosomes may differ from those of the parent cells. Exosomal mi RNAs play an important role in disease progression, and can stimulate angiogenesis and facilitate metastasis in cancers. In this review, we will introduce the origin and the trafficking of exosomes between cells, display current research on the sorting mechanism of exosomal mi RNAs, and briefly describe how exosomes and their mi RNAs function in recipient cells.Finally, we will discuss the potential applications of these mi RNA-containing vesicles in clinical settings.
基金supported by the National Natural Science Foundation of China(82271645)National Key Research and Development Program of China(2021YFC2700200 to F.S.)。
文摘Meiosis is a highly complex process significantly influenced by transcriptional regulation.However,studies on the mechanisms that govern transcriptomic changes during meiosis,especially in prophase I,are limited.Here,we performed single-cell ATAC-seq of human testis tissues and observed reprogramming during the transition from zygotene to pachytene spermatocytes.This event,conserved in mice,involved the deactivation of genes associated with meiosis after reprogramming and the activation of those related to spermatogenesis before their functional onset.Furthermore,we identified 282 transcriptional regulators(TRs)that underwent activation or deactivation subsequent to this process.Evidence suggested that physical contact signals from Sertoli cells may regulate these TRs in spermatocytes,while secreted ENHO signals may alter metabolic patterns in these cells.Our results further indicated that defective transcriptional reprogramming may be associated with non-obstructive azoospermia(NOA).This study revealed the importance of both physical contact and secreted signals between Sertoli cells and germ cells in meiotic progression.
基金supported by the National Natural Science Foundation of China(82200725)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202002)+4 种基金the Fundamental Research Funds for the Central Universities(226-2023-00114,226-2022-00226,and 226-2023-00059)the Key Program of National Natural Science Foundation of China(81930016)the Key Research and Development Program of China(2021YFA1100500)the Major Research Plan of the National Natural Science Foundation of China(92159202)the Ningbo Top Medical and Health Research Program(2022030309).
文摘Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical concern,exacerbating the current organ shortage and detrimentally affecting the prognosis of recipients.Unfortunately,the perplexing hepatic heterogeneity has impeded characterization of the cellular traits and molecular events that contribute to EAD.Herein,we constructed a pioneering single-cell transcriptomic landscape of human transplanted livers derived from non-EAD and EAD patients,with 12 liver samples collected from 7 donors during the cold perfusion and portal reperfusion stages.Comparison of the 75231 cells of non-EAD and EAD patients revealed an EAD-associated immune niche comprising mucosal-associated invariant T cells,granzyme B^(+)(GZMB^(+))granzyme K^(+)(GZMK^(+))natural killer cells,and S100 calcium binding protein A12^(+)(S100A12^(+))neutrophils.Moreover,we verified this immune niche and its association with EAD occurrence in two independent cohorts.Our findings elucidate the cellular characteristics of transplanted livers and the EAD-associated pathogenic immune niche at the single-cell level,thus,offering valuable insights into EAD onset.
