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    Home > Biochemistry News > Biotechnology News > 【Research Dynamics】Professor Guo Anyuan's team at the School of Life Sciences of Huazhong University of Science and Technology identified RNA features in a single extracellular vesicle for the first time

    【Research Dynamics】Professor Guo Anyuan's team at the School of Life Sciences of Huazhong University of Science and Technology identified RNA features in a single extracellular vesicle for the first time

    • Last Update: 2022-09-21
    • Source: Internet
    • Author: User
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    Extracellular vesicles (EVs) are cells secreted by cells with lipid bilayer structures, carrying biologically active molecules such as DNA, RNA, proteins and lipids, and are important carriers of intercellular communication, which can transport various active biomolecules from EVs to related recipient cells
    .
    This capability allows EVs to be used as novel drug delivery vehicles to overcome problems with
    other drug delivery systems.
    In recent years, a number of studies have confirmed that RNA is a key functional molecule for EVs involved in regulating a variety of physiological and pathological processes, and has great clinical application potential
    in biomarkers.
    Existing EVs-related RNA studies have analyzed
    a large number of EVs as a whole.
    At present, there is a lack of high-throughput analysis methods to analyze the RNA carried by a single EV at the single EV level, and the type, quantity, and heterogeneity of RNA in a single EV are not clear
    .

    On September 6, 2022, small Methods magazine published online the findings of a study
    titled Transcriptomic Features in a Single Extracellular Vesicle via Single-Cell RNA Sequencing.
    The study is the first to utilize improved experimental and analytical methods to study transcriptome characteristics
    of EVs at the single EV level based on the 10x Genomics platform.
    Professor Guo Anyuan of the School of Life Science and Technology of Huazhong University of Science and Technology and Lei Qian, assistant researcher of Zhongnan Hospital of Wuhan University, are co-corresponding authors, and Dr.
    Luo Tao is the first author
    of the article.

    Figure 1 Article information

    This study proposes a high-throughput sequencing method for EVs based on the 10x Genomics platform, which explores
    the characteristics of the EVs transcriptome at the single EV level.
    The authors labeled intact EVs from K562 cells and mesenchymal stem cell sources using The Calcein-AM dye and detected the concentration
    of EVs samples by flow cytometry.
    The isolated EVs are sequenced
    using the 10x Genomics platform for single EVs.
    In the data analysis section, the authors tried and improved and found that the CB2 algorithm using adaptive thresholds can effectively distinguish real EVs
    from the background.
    Remove low-quality data using reliable quality control parameters and remove potential doublets
    using The DolbletFinder software.
    Finally, transcriptome data for 2088, 3935 and 1603 EVs were obtained in three different EV samples
    , respectively.

    Analysis of the EV transcriptome found that the average number of mRNA genes contained in a single EV was 52, with different EVs ranging
    from 6 to 148.
    A higher proportion of EVs contain ribosome genes, mitochondrial genes, and EEF1A1 genes
    .
    K562 cell-derived EVs contain high hemoglobin genes while mesenchymal stem cell-derived EVs are rich in cytoskeletal-related genes
    .
    In addition, through dimensionality reduction and clustering analysis of the data, the authors confirmed that even EVs of all-species cell origin are highly heterogeneous and can be divided into multiple distinct subpopulations
    of EVs with specific genes.

    This study reveals the transcriptome characteristics and heterogeneity of EVs at the single EV level for the first time through a high-throughput method, refreshing people's understanding of RNA contents in individual EVs and providing an important basis
    for functional research and modified applications of EVs.


    Figure 2 Single EV transcriptome analysis process based on single-cell sequencing technology

    The research has been funded by the National Key R&D Program, the National Natural Science Foundation of China, the Natural Science Foundation of Hubei Province and the Outstanding Doctor program of Zhongnan Hospital, and has been strongly supported
    by Wuhan Biological Sample Bank Co.
    , Ltd.

    Prof.
    Anyuan Guo's team has long focused on the study of tumor cells, immune cells and extracellular vesicles in the tumor microenvironment, especially for extracellular vesicles, and has developed a series of methods (EVAtool, Briefings in Bioinformatics 2022) and databases (EVmiRNA and EVAtlas, Nucleic Acids Research 2019, 2022), The analysis found and validated the important functions of extracellular vesicles and their key molecules (Science Translational Medicine 2021, Cancer Letter 2022, Theranostics 2019, 2017, OncoImmunology 2018, Cancer Research 2016
    , etc.
    ).

    References: Luo T, Chen SY, Qiu ZX, Miao YR, Ding Y, Pan XY, Li Y, Lei Q, Guo AY.
    Transcriptomic Features in a Single Extracellular Vesicle via Single-Cell RNA Sequencing.
    Small Methods.
    2022 Sep 6:e2200881.

    Full-text links: https://onlinelibrary.
    wiley.
    com/doi/10.
    1002/smtd.
    202200881

     

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