“…3 A) [ 50 ]. The EVs containing PDGFR-RVG, hnRNPA2B1, and mirSilencer, demonstrated an effective silencing function against ATXN3, known as the causative factor of Machado-Joseph disease (MJD) [ 51 ]. Similarly, to promote the targeting and internalization using PDGFR-RVG in this study, lentivirus was employed to express RVG and BDNF in HEK293T cells.…”
Section: Engineering Methods Of Ev and Applications For Regenerative ...mentioning
confidence: 99%
“… Engineered materials Method Targeting cell Ref. 1 PDGFR-RVG, hnRNPA2B1expressed/mirSilencer lentivirus HEK293T [ 51 ] 2 RVG/BDNF lentivirus HEK293T [ 52 ] 3 SERPINA5 protein lentivirus HEK293 [ 57 ] 4 Serpin protein lentivirus HEK293 [ 58 ] 5 miR-146a lentivirus PMSC [ 56 ] 6 miR-214-3p lentivirus UCMSC [ 55 ] 7 miR-133-3p lentivirus UCMSC [ 54 ] 8 Tat protein Lipofectamine HEK293T [ 59 ] 9 Ldlr mRNA Lipofectamine HEK293T [ 60 ] 10 Protein drug (srIkB) Effectene Transfection Reagent HEK293T [ 61 ] 11 Protein drug (srIkB) Effectene Transfection Reagent HEK293T [ 62 ] 12 Protein drug (srIkB) Effectene Transfection Reagent HEK293T [ 63 ] Fig. 3 Gene editing approaches for EV engineering.…”
Section: Engineering Methods Of Ev and Applications For Regenerative ...mentioning
confidence: 99%
“…A The ribonucleoprotein A2B1 (hnRNPA2B1) and RVG decorated EVs production using gene transfection. Reproduced with permission from [ 51 ]. Copyright 2023 Cell Press.…”
Section: Engineering Methods Of Ev and Applications For Regenerative ...mentioning
Extracellular vesicles (EVs) are nanosized particles that are released from cells and reflect the characteristics of the mother cell. Recently, the EVs have been used in several types of studies across many different fields. In the field of EV research, multiple cell culture and EV isolation techniques have been highlighted in importance. Various strategies, including exclusive component culture media, three-dimensional (3D) cultures, and hypoxic conditions, have been proposed for the cell culture to control function of the EVs. Ultracentrifugation, ultrafiltration, precipitation, and tangential flow filtration (TFF) have been utilized for EV isolation. Although isolated EVs have their own functionalities, several researchers are trying to functionalize EVs by applying various engineering approaches. Gene editing, exogenous, endogenous, and hybridization methods are the four well-known types of EV functionalization strategies. EV engineered through these processes has been applied in the field of regenerative medicine, including kidney diseases, osteoarthritis, rheumatoid arthritis, nervous system-related diseases, and others. In this review, it was focused on engineering approaches for EV functionalization and their applications in regenerative medicine.Graphical Abstract
“…3 A) [ 50 ]. The EVs containing PDGFR-RVG, hnRNPA2B1, and mirSilencer, demonstrated an effective silencing function against ATXN3, known as the causative factor of Machado-Joseph disease (MJD) [ 51 ]. Similarly, to promote the targeting and internalization using PDGFR-RVG in this study, lentivirus was employed to express RVG and BDNF in HEK293T cells.…”
Section: Engineering Methods Of Ev and Applications For Regenerative ...mentioning
confidence: 99%
“… Engineered materials Method Targeting cell Ref. 1 PDGFR-RVG, hnRNPA2B1expressed/mirSilencer lentivirus HEK293T [ 51 ] 2 RVG/BDNF lentivirus HEK293T [ 52 ] 3 SERPINA5 protein lentivirus HEK293 [ 57 ] 4 Serpin protein lentivirus HEK293 [ 58 ] 5 miR-146a lentivirus PMSC [ 56 ] 6 miR-214-3p lentivirus UCMSC [ 55 ] 7 miR-133-3p lentivirus UCMSC [ 54 ] 8 Tat protein Lipofectamine HEK293T [ 59 ] 9 Ldlr mRNA Lipofectamine HEK293T [ 60 ] 10 Protein drug (srIkB) Effectene Transfection Reagent HEK293T [ 61 ] 11 Protein drug (srIkB) Effectene Transfection Reagent HEK293T [ 62 ] 12 Protein drug (srIkB) Effectene Transfection Reagent HEK293T [ 63 ] Fig. 3 Gene editing approaches for EV engineering.…”
Section: Engineering Methods Of Ev and Applications For Regenerative ...mentioning
confidence: 99%
“…A The ribonucleoprotein A2B1 (hnRNPA2B1) and RVG decorated EVs production using gene transfection. Reproduced with permission from [ 51 ]. Copyright 2023 Cell Press.…”
Section: Engineering Methods Of Ev and Applications For Regenerative ...mentioning
Extracellular vesicles (EVs) are nanosized particles that are released from cells and reflect the characteristics of the mother cell. Recently, the EVs have been used in several types of studies across many different fields. In the field of EV research, multiple cell culture and EV isolation techniques have been highlighted in importance. Various strategies, including exclusive component culture media, three-dimensional (3D) cultures, and hypoxic conditions, have been proposed for the cell culture to control function of the EVs. Ultracentrifugation, ultrafiltration, precipitation, and tangential flow filtration (TFF) have been utilized for EV isolation. Although isolated EVs have their own functionalities, several researchers are trying to functionalize EVs by applying various engineering approaches. Gene editing, exogenous, endogenous, and hybridization methods are the four well-known types of EV functionalization strategies. EV engineered through these processes has been applied in the field of regenerative medicine, including kidney diseases, osteoarthritis, rheumatoid arthritis, nervous system-related diseases, and others. In this review, it was focused on engineering approaches for EV functionalization and their applications in regenerative medicine.Graphical Abstract
“…Transfection e ciency was monitored by the co-expression of EGFP and mCherry. Lentiviral vectors from these plasmids were produced at ViraVector (University of Coimbra, Portugal), as previously described [34].…”
Chronic stress exerts profound negative effects on cognitive and emotional behaviours and is a major risk factor for the development of neuropsychiatric disorders. However, the molecular links between chronic stress and its deleterious effects on neuronal and synaptic function remain elusive. Here, using a combination of in vitro and in vivo approaches, we demonstrate that the upregulation of miR-186-5p triggered by chronic stress may be a key mediator of such changes, leading to synaptic dysfunction. Our results show that the expression levels of miR-186-5p are increased both in the prefrontal cortex (PFC) of mice exposed to chronic stress and in cortical neurons chronically exposed to dexamethasone. Additionally, viral overexpression of miR-186-5p in the PFC of naïve mice induces anxiety- and depressive-like behaviours. The upregulation of miR-186-5p through prolonged glucocorticoid receptor activation differentially affects glutamatergic and GABAergic synaptic transmission, causing an imbalance in excitation/inhibition and leading to altered neuronal network activity. At glutamatergic synapses, we observed both a reduction in synaptic AMPAR and synaptic transmission, along with a switch in their subunit composition, whereas GABAergic synaptic transmission was strengthened. These changes could be rescued by a miR-186-5p inhibitor. Overall, our results establish a novel molecular link between chronic glucocorticoid receptor activation, the upregulation of miR-186-5p and the synaptic changes induced by chronic stress, that may be amenable to therapeutic intervention.
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