Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation

Zhaogang Yang; Junfeng Shi; Jing Xie; Yifan Wang; Jingyao Sun; Tongzheng Liu; Yarong Zhao; Xiuting Zhao; Xinmei Wang; Yifan Ma; Veysi Malkoc; Chiling Chiang; Weiye Deng; Yuanxin Chen; Yuan Fu; Kwang J. Kwak; Yamin Fan; Chen Kang; Changcheng Yin; June Rhee; Paul Bertani; Jose Otero; Wu Lu; Kyuson Yun; Andrew S. Lee; Wen Jiang; Lesheng Teng; Betty Y.S. Kim; L. James Lee

doi:10.1038/s41551-019-0485-1

Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation

Zhaogang Yang, Junfeng Shi, Jing Xie, Yifan Wang, Jingyao Sun, Tongzheng Liu, Yarong Zhao, Xiuting Zhao, Xinmei Wang, Yifan Ma, Veysi Malkoc, Chiling Chiang, Weiye Deng, Yuanxin Chen, Yuan Fu, Kwang J. Kwak, Yamin Fan, Chen Kang, Changcheng Yin, June RheePaul Bertani, Jose Otero, Wu Lu, Kyuson Yun, Andrew S. Lee, Wen Jiang, Lesheng Teng, Betty Y.S. Kim, L. James Lee^*

^*此作品的通信作者

生物藥學研究所

研究成果 › 同行評審

588 引文斯高帕斯（Scopus）

摘要

Exosomes are attractive as nucleic-acid carriers because of their favourable pharmacokinetic and immunological properties and their ability to penetrate physiological barriers that are impermeable to synthetic drug-delivery vehicles. However, inserting exogenous nucleic acids, especially large messenger RNAs, into cell-secreted exosomes leads to low yields. Here we report a cellular-nanoporation method for the production of large quantities of exosomes containing therapeutic mRNAs and targeting peptides. We transfected various source cells with plasmid DNAs and stimulated the cells with a focal and transient electrical stimulus that promotes the release of exosomes carrying transcribed mRNAs and targeting peptides. Compared with bulk electroporation and other exosome-production strategies, cellular nanoporation produced up to 50-fold more exosomes and a more than 10³-fold increase in exosomal mRNA transcripts, even from cells with low basal levels of exosome secretion. In orthotopic phosphatase and tensin homologue (PTEN)-deficient glioma mouse models, mRNA-containing exosomes restored tumour-suppressor function, enhanced inhibition of tumour growth and increased survival. Cellular nanoporation may enable the use of exosomes as a universal nucleic-acid carrier for applications requiring transcriptional manipulation.

原文	English
頁（從 - 到）	69-83
頁數	15
期刊	Nature Biomedical Engineering
卷	4
發行號	1
DOIs	https://doi.org/10.1038/s41551-019-0485-1
出版狀態	Published - 1 1月 2020

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Yang, Z., Shi, J., Xie, J., Wang, Y., Sun, J., Liu, T., Zhao, Y., Zhao, X., Wang, X., Ma, Y., Malkoc, V., Chiang, C., Deng, W., Chen, Y., Fu, Y., Kwak, K. J., Fan, Y., Kang, C., Yin, C., ... Lee, L. J. (2020). Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation. Nature Biomedical Engineering, 4(1), 69-83. https://doi.org/10.1038/s41551-019-0485-1

@article{1c59008cd1eb43d08d92597e248c8824,

title = "Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation",

abstract = "Exosomes are attractive as nucleic-acid carriers because of their favourable pharmacokinetic and immunological properties and their ability to penetrate physiological barriers that are impermeable to synthetic drug-delivery vehicles. However, inserting exogenous nucleic acids, especially large messenger RNAs, into cell-secreted exosomes leads to low yields. Here we report a cellular-nanoporation method for the production of large quantities of exosomes containing therapeutic mRNAs and targeting peptides. We transfected various source cells with plasmid DNAs and stimulated the cells with a focal and transient electrical stimulus that promotes the release of exosomes carrying transcribed mRNAs and targeting peptides. Compared with bulk electroporation and other exosome-production strategies, cellular nanoporation produced up to 50-fold more exosomes and a more than 103-fold increase in exosomal mRNA transcripts, even from cells with low basal levels of exosome secretion. In orthotopic phosphatase and tensin homologue (PTEN)-deficient glioma mouse models, mRNA-containing exosomes restored tumour-suppressor function, enhanced inhibition of tumour growth and increased survival. Cellular nanoporation may enable the use of exosomes as a universal nucleic-acid carrier for applications requiring transcriptional manipulation.",

author = "Zhaogang Yang and Junfeng Shi and Jing Xie and Yifan Wang and Jingyao Sun and Tongzheng Liu and Yarong Zhao and Xiuting Zhao and Xinmei Wang and Yifan Ma and Veysi Malkoc and Chiling Chiang and Weiye Deng and Yuanxin Chen and Yuan Fu and Kwak, \{Kwang J.\} and Yamin Fan and Chen Kang and Changcheng Yin and June Rhee and Paul Bertani and Jose Otero and Wu Lu and Kyuson Yun and Lee, \{Andrew S.\} and Wen Jiang and Lesheng Teng and Kim, \{Betty Y.S.\} and Lee, \{L. James\}",

note = "Publisher Copyright: {\textcopyright} 2019, The Author(s), under exclusive licence to Springer Nature Limited.",

year = "2020",

month = jan,

day = "1",

doi = "10.1038/s41551-019-0485-1",

language = "English",

volume = "4",

pages = "69--83",

journal = "Nature Biomedical Engineering",

issn = "2157-846X",

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Yang, Z, Shi, J, Xie, J, Wang, Y, Sun, J, Liu, T, Zhao, Y, Zhao, X, Wang, X, Ma, Y, Malkoc, V, Chiang, C, Deng, W, Chen, Y, Fu, Y, Kwak, KJ, Fan, Y, Kang, C, Yin, C, Rhee, J, Bertani, P, Otero, J, Lu, W, Yun, K, Lee, AS, Jiang, W, Teng, L, Kim, BYS & Lee, LJ 2020, 'Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation', Nature Biomedical Engineering, 卷 4, 編號 1, 頁 69-83. https://doi.org/10.1038/s41551-019-0485-1

TY - JOUR

T1 - Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation

AU - Yang, Zhaogang

AU - Shi, Junfeng

AU - Xie, Jing

AU - Wang, Yifan

AU - Sun, Jingyao

AU - Liu, Tongzheng

AU - Zhao, Yarong

AU - Zhao, Xiuting

AU - Wang, Xinmei

AU - Ma, Yifan

AU - Malkoc, Veysi

AU - Chiang, Chiling

AU - Deng, Weiye

AU - Chen, Yuanxin

AU - Fu, Yuan

AU - Kwak, Kwang J.

AU - Fan, Yamin

AU - Kang, Chen

AU - Yin, Changcheng

AU - Rhee, June

AU - Bertani, Paul

AU - Otero, Jose

AU - Lu, Wu

AU - Yun, Kyuson

AU - Lee, Andrew S.

AU - Jiang, Wen

AU - Teng, Lesheng

AU - Kim, Betty Y.S.

AU - Lee, L. James

PY - 2020/1/1

Y1 - 2020/1/1

N2 - Exosomes are attractive as nucleic-acid carriers because of their favourable pharmacokinetic and immunological properties and their ability to penetrate physiological barriers that are impermeable to synthetic drug-delivery vehicles. However, inserting exogenous nucleic acids, especially large messenger RNAs, into cell-secreted exosomes leads to low yields. Here we report a cellular-nanoporation method for the production of large quantities of exosomes containing therapeutic mRNAs and targeting peptides. We transfected various source cells with plasmid DNAs and stimulated the cells with a focal and transient electrical stimulus that promotes the release of exosomes carrying transcribed mRNAs and targeting peptides. Compared with bulk electroporation and other exosome-production strategies, cellular nanoporation produced up to 50-fold more exosomes and a more than 103-fold increase in exosomal mRNA transcripts, even from cells with low basal levels of exosome secretion. In orthotopic phosphatase and tensin homologue (PTEN)-deficient glioma mouse models, mRNA-containing exosomes restored tumour-suppressor function, enhanced inhibition of tumour growth and increased survival. Cellular nanoporation may enable the use of exosomes as a universal nucleic-acid carrier for applications requiring transcriptional manipulation.

AB - Exosomes are attractive as nucleic-acid carriers because of their favourable pharmacokinetic and immunological properties and their ability to penetrate physiological barriers that are impermeable to synthetic drug-delivery vehicles. However, inserting exogenous nucleic acids, especially large messenger RNAs, into cell-secreted exosomes leads to low yields. Here we report a cellular-nanoporation method for the production of large quantities of exosomes containing therapeutic mRNAs and targeting peptides. We transfected various source cells with plasmid DNAs and stimulated the cells with a focal and transient electrical stimulus that promotes the release of exosomes carrying transcribed mRNAs and targeting peptides. Compared with bulk electroporation and other exosome-production strategies, cellular nanoporation produced up to 50-fold more exosomes and a more than 103-fold increase in exosomal mRNA transcripts, even from cells with low basal levels of exosome secretion. In orthotopic phosphatase and tensin homologue (PTEN)-deficient glioma mouse models, mRNA-containing exosomes restored tumour-suppressor function, enhanced inhibition of tumour growth and increased survival. Cellular nanoporation may enable the use of exosomes as a universal nucleic-acid carrier for applications requiring transcriptional manipulation.

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U2 - 10.1038/s41551-019-0485-1

DO - 10.1038/s41551-019-0485-1

M3 - Article

C2 - 31844155

AN - SCOPUS:85076495753

SN - 2157-846X

VL - 4

SP - 69

EP - 83

JO - Nature Biomedical Engineering

JF - Nature Biomedical Engineering

IS - 1

ER -

Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation

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