The Machinery of Exosomes: Biogenesis, Release, and Uptake

Krylova, Sofia V.; Feng, Dingqing

doi:10.3390/ijms24021337

Cited by 171 publications

(92 citation statements)

References 118 publications

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“…One of them can be associated with an RNA-induced silencing complex, and the other is found in exosomes, microvesicles, and apoptotic bodies [ 42 ]. Exosomes contain different proteins, microRNAs, mRNAs, and DNA [ 42 , 43 ]. It was shown that exosomal NUCB1 is involved in the pathophysiology of cancer and participates in EMT and atypical migration in non-tumor cells [ 44 ].…”

Section: Discussionmentioning

confidence: 99%

The RNA-Binding and RNA-Melting Activities of the Multifunctional Protein Nucleobindin 1

Mikhaylina

Svoeglazova

Stolboushkina

et al. 2023

IJMS

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Nucleobindin 1 (NUCB1) is a ubiquitous multidomain protein that belongs to the EF-hand Ca2+-binding superfamily. NUCB1 interacts with Galphai3 protein, cyclooxygenase, amyloid precursor protein, and lipids. It is involved in stress response and human diseases. In addition, this protein is a transcription factor that binds to the DNA E-box motif. Using surface plasmon resonance and molecular beacon approaches, we first showed the RNA binding and RNA melting activities of NUCB1. We suggest that NUCB1 could induce local changes in structured RNAs via binding to the GGAUAU loop sequence. Our results demonstrate the importance of the multidomain structure of NUCB1 for its RNA-chaperone activity in vitro.

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Section: Discussionmentioning

confidence: 99%

The RNA-Binding and RNA-Melting Activities of the Multifunctional Protein Nucleobindin 1

Mikhaylina

Svoeglazova

Stolboushkina

et al. 2023

IJMS

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“…4.4 CD63 and VPS36 showed spatial-dependent colocalization and correlation after miRNA delivery Next, we verified the involvement of miR-125b-5p in the modulation of EV trafficking, focusing on the localization of EV-related proteins in pTr cells. VPS36 (miR-125b-5p target and marker of ESCRT-II complex present in endosomes/MVBs [47]) and CD63 (marker of EVs [48]) were selected based on our recent studies pointing to embryonic signals as triggers of EV trafficking in endometrial luminal epithelial cells [26]. Regions of interest (ROIs) were set based on the overlapping localization of both proteins present in the highly visible and well-shaped cell borders (white) in comparison to the whole cell (Fig.…”

Section: Mir-125b-5p Is Efficiently Delivered To Ptr Cells and Intern...mentioning

confidence: 99%

miR-125b-5p impacts extracellular vesicle biogenesis, trafficking, and EV subpopulation release in the porcine trophoblast by regulating ESCRT-dependent pathway

Guzewska

Witek

Karnas

et al. 2023

Preprint

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Intercellular communication is a critical process that ensures cooperation between distinct cell types at the embryo-maternal interface. Extracellular vesicles (EVs) are considered to be potent mediators of this communication by transferring biological information in their cargo (e.g. miRNAs) to the recipient cells. miRNAs are small non-coding RNAs that affect the function and fate of neighboring and distant cells by regulating gene expression. Focusing on the maternal side of the dialog, we recently revealed the impact of embryonic signals, including miRNAs, on EV-mediated cell-to-cell communication. In this study, we show the regulatory mechanism of the miR-125b-5p ESCRT-mediated EV biogenesis pathway and the further secretion of EVs by trophoblasts at the time when the crucial steps of implantation are taking place. To test the ability of miR-125b-5p to influence the expression of genes involved in the generation and release of EV subpopulations in porcine conceptuses, we used an ex vivo approach. Next, in silico and in vitro analyses were performed to confirm miRNA-mRNA interactions. Finally, EV trafficking and release were assessed using several imaging and particle analysis tools. Our results indicated that conceptus development and implantation are accompanied by changes in the abundance of EV biogenesis and trafficking machinery. ESCRT-dependent EV biogenesis and the further secretion of EVs were modulated by miR-125b-5p, specifically impacting the ESCRT-II complex (via VPS36) and EV trafficking in primary porcine trophoblast cells. The identified miRNA-ESCRT interplay led to the generation and secretion of specific subpopulations of EVs. miRNA present at the embryo-maternal interface governs EV-mediated communication between the mother and the developing conceptus, leading to the generation, trafficking, and release of characteristic subpopulations of EVs.

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“…Exosomes are the smallest extracellular vesicles (EVs) which also include two other larger subtypes (microvesicles and apoptotic bodies) . EVs contain a distinctive signature of macromolecules derived from parental cells, such as lipids, nucleic acids, proteins, and metabolites, and many of them are present both in the lumen and on the membrane of an EV. − Initially, EVs were regarded as remnants of cellular decay or indicative of waste elimination processes associated with cell death. ,, However, subsequent research has indicated that these vesicles are actively secreted by donor cells into the extracellular milieu, undertaking diverse biological roles, such as intercellular communication and exchange of proteins and genetic materials between an originating cell and its neighboring cells. ,− These EVs are taken up into recipient cells through one of three processes: (1) endocytosis, (2) interaction between exosomal surface proteins and plasma membrane receptors, or (3) fusion with the recipient cell’s membrane . Interestingly, recent research suggests that MSC-secreted exosomes can substitute stem cell-based treatments in numerous wound-healing animal models. , Because of their distinctive properties, for instance, low immunogenicity, intrinsic stability, and excellent cell and tissue penetration capacity, exosomes have been explored as delivery vehicles to transport therapeutic agents including small molecules, nucleic acids, and proteins to cells .…”

Section: Introductionmentioning

confidence: 99%

Significant Enhancement of Fibroblast Migration, Invasion, and Proliferation by Exosomes Loaded with Human Fibroblast Growth Factor 1

Hade,

Suire,

Suo

2024

ACS Appl. Mater. Interfaces

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Exosomes possess several inherent properties that make them ideal for biomedical applications, including robust stability, biocompatibility, minimal immunogenicity, and the ability to cross biological barriers. These natural nanoparticles have recently been developed as drug delivery vesicles. To do so, therapeutic molecules must be efficiently loaded into exosomes first. Very recently, we developed a cell-penetrating peptide (CPP)-based platform for loading of nucleic acids and small molecules into exosomes by taking advantage of the membrane-penetration power of CPPs. Here, we extended this simple but effective platform by loading a protein cargo into exosomes isolated from either mesenchymal stem cells from three different sources or two different cancer cell lines. The protein cargo is a fusion protein YARA−FGF1−GFP through the covalent conjugation of a model CPP called YARA to human fibroblast growth factor 1 (FGF1) and green fluorescence protein (GFP). Loading of YARA− FGF1−GFP into exosomes was time-dependent and reached a maximum of about 1600 YARA−FGF1−GFP molecules in each exosome after 16 h. The ladened exosomes were effectively internalized by mammalian cells, and subsequently, the loaded protein cargo YARA−FGF1−GFP was delivered intracellularly. In comparison to YARA, YARA−FGF1−GFP, the unloaded exosomes, and the exosomes loaded with YARA, the exosomes loaded with YARA−FGF1−GFP substantially promoted the migration, proliferation, and invasion capabilities of mouse and human fibroblasts, which are important factors for wound repair. The work extended our CPP-based exosomal cargo loading platform and established a foundation for developing novel wound-healing therapies using exosomes loaded with FGF1 and other growth factors.

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The Machinery of Exosomes: Biogenesis, Release, and Uptake

Cited by 171 publications

References 118 publications

The RNA-Binding and RNA-Melting Activities of the Multifunctional Protein Nucleobindin 1

The RNA-Binding and RNA-Melting Activities of the Multifunctional Protein Nucleobindin 1

miR-125b-5p impacts extracellular vesicle biogenesis, trafficking, and EV subpopulation release in the porcine trophoblast by regulating ESCRT-dependent pathway

Significant Enhancement of Fibroblast Migration, Invasion, and Proliferation by Exosomes Loaded with Human Fibroblast Growth Factor 1

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