Anti‐SARS‐CoV‐2 effect of extracellular vesicles released from mesenchymal stem cells

Abstract

coronavirus disease 2019 caused by SARS-CoV-2 accounted for 267 million people with up to 5.3 million deaths worldwide (https://covid19.who.int). Since late 2019, much progress has been made in response to the COVID-19 pandemic, including the rapid developments of effective vaccines and the treatment guidelines consisting of antiviral drugs, immunomodulators, and critical care support (https://covid19.who.int). However, SARS-CoV-2 evolves over time as its genome has a high mutation rate that leads to reasonable concerns of breakthrough infection due to immune escape and resistant strain emergence under antiviral pressure (Lipsitch et al., 2021;Szemiel et al., 2021). A newly emerging Omicron (B.1.1.529) variant rings alarms around the globe that, perhaps, the COVID-19 war has just begun. Relentless efforts should be made to advance our knowledge and treatment regimens against COVID-19. These included studies of mesenchymal stem cell (MSC) therapy that aimed to mitigate cytokine storm and promote tissue repair in severely ill patients with COVID-19 pneumonia and acute respiratory distress syndrome (ARDS) (Hashemian et al., 2021;Meng et al., 2020;Zhu et al., 2021). Nevertheless, as extensively discussed in a recent review by Dr. Phillip W. Askenase of Yale University School of Medicine, the immunomodulatory and regenerative effects of MSC therapy are mediated through MSC-derived extracellular vesicles (MSC-EVs) (Askenase, 2020), while the use of MSC-EVs has less safety concerns of thromboembolism, arrhythmia and malignant transformation. In this direction, MSC-EV investigations for COVID-19 treatment would be more appealing and undeniable if MSC-EVs also exhibit anti-SARS-CoV-2 effects. A previous study revealed that MSC-EVs pertained antiviral activity against influenza virus in a preclinical model (Khatri et al., 2018). It is known that MSCs are highly resistant to viral infections (Wu et al., 2018), including SARS-CoV-2 (Avanzini et al., 2021). We, therefore, hypothesized that the EVs released from MSCs could inhibit SARS-CoV-2 infection.Accordingly, we applied in vitro anti-SARS-CoV-2 assays, which were previously developed by our group (Kanjanasirirat et al., 2020;Kongsomros et al., 2021;Sa-Ngiamsuntorn et al., 2021), to determine the dose dependent anti-SARS-CoV-2 effect of MSC-EVs. In this study, MSC-EV was referred to as the exosome (or small EV) subpopulation of extracellular vesicles. Full details of the methods used in this study were provided in Supplementary material.MSC-EVs were isolated from 100 ml culture media (supplemented with exosome-depleted foetal bovine serum) of human umbilical cord-derived MSCs (ATCC®PCS-500-010) by the combination of step-wise centrifugation and 0.2 μm filtration (to remove microvesicles and apoptotic bodies), the 100-kDa cutoff ultrafiltration (to concentrate the culture medium), and the Izon qEV size exclusion chromatography (to separate extracellular vesicles from soluble proteins) (Figure 1a). The final volume of the MSC-EV isolate was 500 μl. Ten aliquots (50 μl each)...

Extracellular Vesicle-Based Method for Detecting MYCN Amplification Status of Pediatric Neuroblastoma

“…EVs are lipid bilayer-encapsulated nanoparticles released from all cells and present in all kinds of biofluids. EVs are known to play crucial roles in cell-to-cell communication and physiological regulation in both healthy and disease states, some of which have therapeutic potential [ 16 , 17 , 18 , 19 ]. EVs contain molecular signatures of original cells including lipids, proteins and nucleic acids, and thus serve as a promising source for biomarker discovery [ 15 , 16 ].…”

“…EVs contain molecular signatures of original cells including lipids, proteins and nucleic acids, and thus serve as a promising source for biomarker discovery [ 15 , 16 ]. EVs can be classified into three subtypes based on their biogenesis pathways and bio-physico-chemical properties [ 18 , 19 , 20 ]. Exosomes (Exo) are the small EVs (40–150 nm in diameter), which originate from the endosomal pathway and multivesicular bodies, and are enriched with common exosome markers, e.g., Alix, Tsg101, Hsp70 and tetraspanins [ 18 , 19 , 20 ].…”

“…EVs can be classified into three subtypes based on their biogenesis pathways and bio-physico-chemical properties [ 18 , 19 , 20 ]. Exosomes (Exo) are the small EVs (40–150 nm in diameter), which originate from the endosomal pathway and multivesicular bodies, and are enriched with common exosome markers, e.g., Alix, Tsg101, Hsp70 and tetraspanins [ 18 , 19 , 20 ]. Microvesicles (MVs) are the large EVs (150–1000 nm in diameter) produced from plasma membrane budding and thus share the membrane and cytosolic proteins of parent cells [ 18 , 19 , 20 ].…”

“…Exosomes (Exo) are the small EVs (40–150 nm in diameter), which originate from the endosomal pathway and multivesicular bodies, and are enriched with common exosome markers, e.g., Alix, Tsg101, Hsp70 and tetraspanins [ 18 , 19 , 20 ]. Microvesicles (MVs) are the large EVs (150–1000 nm in diameter) produced from plasma membrane budding and thus share the membrane and cytosolic proteins of parent cells [ 18 , 19 , 20 ]. Apoptotic-derived EVs or apoptotic bodies (>1000 nm in diameter) are generated from plasma membrane blebbing during cell apoptosis [ 18 , 19 , 20 ].…”

“…Microvesicles (MVs) are the large EVs (150–1000 nm in diameter) produced from plasma membrane budding and thus share the membrane and cytosolic proteins of parent cells [ 18 , 19 , 20 ]. Apoptotic-derived EVs or apoptotic bodies (>1000 nm in diameter) are generated from plasma membrane blebbing during cell apoptosis [ 18 , 19 , 20 ]. Exo and MVs are the main focus of the EV-based method development for several cancers, i.e., head and neck [ 16 ], brain [ 21 ], lung [ 22 ], colon [ 23 ], bladder [ 24 ], endometrial [ 25 ], ovarian [ 26 ] and prostate [ 27 ].…”