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

Nanomedicine. 2020 Oct:29:102271. doi: 10.1016/j.nano.2020.102271. Epub 2020 Jul 21.

Authors

Yuan Liu¹, Sherry Wu², Yeonjong Koo³, An Yang⁴, Yanwan Dai⁵, Htet Khant⁶, Samantha R Osman⁷, Mamur Chowdhury⁸, Haichao Wei⁹, Yang Li¹⁰, Karem Court¹¹, Elaine Hwang¹², Yunfei Wen¹³, Santosh K Dasari¹⁴, Michael Nguyen¹⁵, E Chia-Cheng Tang¹⁶, E Wassim Chehab¹⁷, Natalia de Val¹⁸, Janet Braam¹⁹, Anil K Sood²⁰

Affiliations

¹ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX; BioSciences, Rice University, Houston, TX. Electronic address: yliu32@mdanderson.org.
² Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: sherry.wu@uq.edu.au.
³ BioSciences, Rice University, Houston, TX. Electronic address: yeonjong@gmail.com.
⁴ BioSciences, Rice University, Houston, TX; State Key Laboratory of Vegetation and Environmental Change, Institute of Botany, Chinese Academy of Sciences, Beijing, People's Republic of China. Electronic address: anyang@ibcas.ac.cn.
⁵ BioSciences, Rice University, Houston, TX. Electronic address: Yanwan.Dai@rice.edu.
⁶ Center for Molecular Microscopy, Center for Cancer Research, National Cancer Institute, Frederick, MD; Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Inc., Frederick, MD. Electronic address: htet.khant@nih.gov.
⁷ BioSciences, Rice University, Houston, TX. Electronic address: sro3@rice.edu.
⁸ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: MAChowdhury@mdanderson.org.
⁹ The Vivian L. Smith Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX. Electronic address: Haichao.Wei@uth.tmc.edu.
¹⁰ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: YLi52@mdanderson.org.
¹¹ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: karemcourt@gmail.com.
¹² BioSciences, Rice University, Houston, TX. Electronic address: esh3@rice.edu.
¹³ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: YWen2@mdanderson.org.
¹⁴ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: SKDasari@mdanderson.org.
¹⁵ BioSciences, Rice University, Houston, TX. Electronic address: mhn2@rice.edu.
¹⁶ BioSciences, Rice University, Houston, TX. Electronic address: ct44@rice.edu.
¹⁷ BioSciences, Rice University, Houston, TX. Electronic address: ewchehab@rice.edu.
¹⁸ Center for Molecular Microscopy, Center for Cancer Research, National Cancer Institute, Frederick, MD; Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Inc., Frederick, MD. Electronic address: natalia.devalalda@nih.gov.
¹⁹ BioSciences, Rice University, Houston, TX. Electronic address: Braam@rice.edu.
²⁰ Department of Gynecologic Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. Electronic address: asood@mdanderson.org.

PMID: 32702466
DOI: 10.1016/j.nano.2020.102271

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / chemistry*
Arabidopsis / genetics
Drug Delivery Systems*
Extracellular Vesicles / chemistry*
Extracellular Vesicles / genetics
Humans
Plant Leaves / chemistry*
Plant Leaves / genetics
Proteomics / methods

Abstract

Publication types

MeSH terms

Grants and funding