Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles

Yuan Liu; Sherry Wu; Yeonjong Koo; An Yang; Yanwan Dai; Htet Khant; Samantha R. Osman; Mamur Chowdhury; Haichao Wei; Yang Li; Karem Court; Elaine Hwang; Yunfei Wen; Santosh K. Dasari; Michael Nguyen; E. Chia Cheng Tang; E. Wassim Chehab; Natalia de Val; Janet Braam; Anil K. Sood

doi:10.1016/j.nano.2020.102271

Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles

Yuan Liu, Sherry Wu, Yeonjong Koo, An Yang, Yanwan Dai, Htet Khant, Samantha R. Osman, Mamur Chowdhury, Haichao Wei, Yang Li, Karem Court, Elaine Hwang, Yunfei Wen, Santosh K. Dasari, Michael Nguyen, E. Chia Cheng Tang, E. Wassim Chehab, Natalia de Val, Janet Braam, Anil K. Sood

Gynecological Oncology & Reproductive Medicine

Research output: Contribution to journal › Article › peer-review

49 Scopus citations

Abstract

Mammalian small extracellular vesicles (sEVs) can deliver diverse molecules to target cells. However, they are difficult to obtain in large quantities and can activate host immune responses. Plant-derived vesicles may help to overcome these challenges. We optimized isolation methods for two types of plant vesicles, nanovesicles from disrupted leaf and sEVs from the extracellular apoplastic space of Arabidopsis thaliana. Both preparations yielded intact vesicles of uniform size, and a mean membrane charge of approximately −25 mV. We also demonstrated applicability of these preparative methods using Brassicaceae vegetables. Proteomic analysis of a subset of vesicles with a density of 1.1-1.19 g mL⁻¹ sheds light on the likely cellular origin and complexity of the vesicles. Both leaf nanovesicles and sEVs were taken up by cancer cells, with sEVs showing an approximately three-fold higher efficiency compared to leaf nanovesicles. These results support the potential of plant-derived vesicles as vehicles for therapeutic delivery.

Original language	English (US)
Article number	102271
Journal	Nanomedicine: Nanotechnology, Biology, and Medicine
Volume	29
DOIs	https://doi.org/10.1016/j.nano.2020.102271
State	Published - Oct 2020

Keywords

Cancer cell uptake
Isolation method
Leaf nanovesicles
Plant sEV and nanovesicle proteome
Plant sEVs
Therapeutic delivery

ASJC Scopus subject areas

Bioengineering
Medicine (miscellaneous)
Molecular Medicine
Biomedical Engineering
General Materials Science
Pharmaceutical Science

MD Anderson CCSG core facilities

Proteomics Facility
Flow Cytometry and Cellular Imaging Facility
High Resolution Electron Microscopy Facility

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10.1016/j.nano.2020.102271

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Liu, Y., Wu, S., Koo, Y., Yang, A., Dai, Y., Khant, H., Osman, S. R., Chowdhury, M., Wei, H., Li, Y., Court, K., Hwang, E., Wen, Y., Dasari, S. K., Nguyen, M., Tang, E. C. C., Chehab, E. W., de Val, N., Braam, J., & Sood, A. K. (2020). Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles. Nanomedicine: Nanotechnology, Biology, and Medicine, 29, Article 102271. https://doi.org/10.1016/j.nano.2020.102271

Liu, Y, Wu, S, Koo, Y, Yang, A, Dai, Y, Khant, H, Osman, SR, Chowdhury, M, Wei, H, Li, Y, Court, K, Hwang, E, Wen, Y, Dasari, SK, Nguyen, M, Tang, ECC, Chehab, EW, de Val, N, Braam, J & Sood, AK 2020, 'Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles', Nanomedicine: Nanotechnology, Biology, and Medicine, vol. 29, 102271. https://doi.org/10.1016/j.nano.2020.102271

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title = "Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles",

abstract = "Mammalian small extracellular vesicles (sEVs) can deliver diverse molecules to target cells. However, they are difficult to obtain in large quantities and can activate host immune responses. Plant-derived vesicles may help to overcome these challenges. We optimized isolation methods for two types of plant vesicles, nanovesicles from disrupted leaf and sEVs from the extracellular apoplastic space of Arabidopsis thaliana. Both preparations yielded intact vesicles of uniform size, and a mean membrane charge of approximately −25 mV. We also demonstrated applicability of these preparative methods using Brassicaceae vegetables. Proteomic analysis of a subset of vesicles with a density of 1.1-1.19 g mL−1 sheds light on the likely cellular origin and complexity of the vesicles. Both leaf nanovesicles and sEVs were taken up by cancer cells, with sEVs showing an approximately three-fold higher efficiency compared to leaf nanovesicles. These results support the potential of plant-derived vesicles as vehicles for therapeutic delivery.",

keywords = "Cancer cell uptake, Isolation method, Leaf nanovesicles, Plant sEV and nanovesicle proteome, Plant sEVs, Therapeutic delivery",

author = "Yuan Liu and Sherry Wu and Yeonjong Koo and An Yang and Yanwan Dai and Htet Khant and Osman, {Samantha R.} and Mamur Chowdhury and Haichao Wei and Yang Li and Karem Court and Elaine Hwang and Yunfei Wen and Dasari, {Santosh K.} and Michael Nguyen and Tang, {E. Chia Cheng} and Chehab, {E. Wassim} and {de Val}, Natalia and Janet Braam and Sood, {Anil K.}",

note = "Funding Information: Funding: This work was supported by the National Institutes of Health (grant numbers R35 CA209904 , P50 CA217685 , P50 CA098258 , and CA016672 ); the Blanton-Davis Ovarian Cancer Research Program ; the American Cancer Society Research Professor Award ; and the Frank McGraw Memorial Chair in Cancer Research . The Electron Microscopy work performed by the Center for Molecular Microscopy was funded by FNLCR Contract HHSN261200800001E . Funding Information: We used the High Resolution Electron Microscope Facility (supported by CCSG grant NIH P30 CA016672 ); Proteomic and Metabolomics Facility and Flow Cytometry Facility (supported by grant P30 CA16672 ) at MD Anderson. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",

year = "2020",

month = oct,

doi = "10.1016/j.nano.2020.102271",

language = "English (US)",

volume = "29",

journal = "Nanomedicine: Nanotechnology, Biology, and Medicine",

issn = "1549-9634",

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T1 - Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles

AU - Liu, Yuan

AU - Wu, Sherry

AU - Koo, Yeonjong

AU - Yang, An

AU - Dai, Yanwan

AU - Khant, Htet

AU - Osman, Samantha R.

AU - Chowdhury, Mamur

AU - Wei, Haichao

AU - Li, Yang

AU - Court, Karem

AU - Hwang, Elaine

AU - Wen, Yunfei

AU - Dasari, Santosh K.

AU - Nguyen, Michael

AU - Tang, E. Chia Cheng

AU - Chehab, E. Wassim

AU - de Val, Natalia

AU - Braam, Janet

AU - Sood, Anil K.

N1 - Funding Information: Funding: This work was supported by the National Institutes of Health (grant numbers R35 CA209904 , P50 CA217685 , P50 CA098258 , and CA016672 ); the Blanton-Davis Ovarian Cancer Research Program ; the American Cancer Society Research Professor Award ; and the Frank McGraw Memorial Chair in Cancer Research . The Electron Microscopy work performed by the Center for Molecular Microscopy was funded by FNLCR Contract HHSN261200800001E . Funding Information: We used the High Resolution Electron Microscope Facility (supported by CCSG grant NIH P30 CA016672 ); Proteomic and Metabolomics Facility and Flow Cytometry Facility (supported by grant P30 CA16672 ) at MD Anderson. Publisher Copyright: © 2020 Elsevier Inc.

PY - 2020/10

Y1 - 2020/10

N2 - Mammalian small extracellular vesicles (sEVs) can deliver diverse molecules to target cells. However, they are difficult to obtain in large quantities and can activate host immune responses. Plant-derived vesicles may help to overcome these challenges. We optimized isolation methods for two types of plant vesicles, nanovesicles from disrupted leaf and sEVs from the extracellular apoplastic space of Arabidopsis thaliana. Both preparations yielded intact vesicles of uniform size, and a mean membrane charge of approximately −25 mV. We also demonstrated applicability of these preparative methods using Brassicaceae vegetables. Proteomic analysis of a subset of vesicles with a density of 1.1-1.19 g mL−1 sheds light on the likely cellular origin and complexity of the vesicles. Both leaf nanovesicles and sEVs were taken up by cancer cells, with sEVs showing an approximately three-fold higher efficiency compared to leaf nanovesicles. These results support the potential of plant-derived vesicles as vehicles for therapeutic delivery.

AB - Mammalian small extracellular vesicles (sEVs) can deliver diverse molecules to target cells. However, they are difficult to obtain in large quantities and can activate host immune responses. Plant-derived vesicles may help to overcome these challenges. We optimized isolation methods for two types of plant vesicles, nanovesicles from disrupted leaf and sEVs from the extracellular apoplastic space of Arabidopsis thaliana. Both preparations yielded intact vesicles of uniform size, and a mean membrane charge of approximately −25 mV. We also demonstrated applicability of these preparative methods using Brassicaceae vegetables. Proteomic analysis of a subset of vesicles with a density of 1.1-1.19 g mL−1 sheds light on the likely cellular origin and complexity of the vesicles. Both leaf nanovesicles and sEVs were taken up by cancer cells, with sEVs showing an approximately three-fold higher efficiency compared to leaf nanovesicles. These results support the potential of plant-derived vesicles as vehicles for therapeutic delivery.

KW - Cancer cell uptake

KW - Isolation method

KW - Leaf nanovesicles

KW - Plant sEV and nanovesicle proteome

KW - Plant sEVs

KW - Therapeutic delivery

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DO - 10.1016/j.nano.2020.102271

M3 - Article

C2 - 32702466

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SN - 1549-9634

VL - 29

JO - Nanomedicine: Nanotechnology, Biology, and Medicine

JF - Nanomedicine: Nanotechnology, Biology, and Medicine

M1 - 102271

ER -

Characterization of and isolation methods for plant leaf nanovesicles and small extracellular vesicles

Abstract

Keywords

ASJC Scopus subject areas

MD Anderson CCSG core facilities

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