scholarly journals tiRNA signaling via stress-regulated vesicle transfer in the hematopoietic niche

2021 ◽  
Author(s):  
Youmna S. Kfoury ◽  
Fei Ji ◽  
Michael Mazzola ◽  
David B. Sykes ◽  
Allison K. Scherer ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stress Responses ◽  
Protein Translation ◽  
Extracellular Vesicle ◽  
Osteoblastic Cells ◽  
Myeloid Differentiation ◽  
Signaling Axis ◽  
Post Radiation ◽  
Hematopoietic Niche

AbstractExtracellular vesicles transfer complex biologic material between cells, whose role in in-vivo organismal physiology is poorly defined. Here, we demonstrate that osteoblastic cells in the bone marrow elaborate extracellular vesicles that are taken up by hematopoietic progenitor cells in vivo. Genotoxic or infectious stress rapidly increased stromal-derived extracellular vesicle transfer to granulocyte-monocyte progenitors. Stimulating osteoblastic cells with parathyroid hormone or activating its receptor enhanced extracellular vesicle transfer, myeloid recovery post radiation and improved animal survival from Candida sepsis. The extracellular vesicles contained tiRNAs known to modulate protein translation. 5’-ti-Pro-CGG-1 was preferentially abundant in osteoblast-derived extracellular vesicles and when transferred to granulocyte macrophage progenitors, increased protein translation, cell proliferation and myeloid differentiation. Therefore, EV-mediated tiRNA transfer provides a stress modulated signaling axis distinct from conventional cytokine-driven stress responses.One sentence summaryStress regulated tiRNA transfer alters hematopoiesis

scholarly journals Microvilli-derived Extracellular Vesicles Govern Morphogenesis in Drosophila wing epithelium

2020 ◽  
Author(s):  
Ilse Hurbain ◽  
Anne-Sophie Macé ◽  
Maryse Romao ◽  
Lucie Sengmanivong ◽  
Laurent Ruel ◽  
...  
Keyword(s):  
Long Range ◽  
Extracellular Vesicles ◽  
Cell Biology ◽  
Extracellular Vesicle ◽  
Wing Disc ◽  
Cellular Mechanisms ◽  
Long Distance ◽  
Drosophila Wing ◽  
And Function

ABSTRACTThe regulation and coordination of developmental processes involves the secretion of morphogens and membrane carriers, including extracellular vesicles, which facilitate their transport over long distance. The long-range activity of the Hedgehog morphogen is conveyed by extracellular vesicles. However, the site and the molecular basis of their biogenesis remains unknown. By combining fluorescence and electron microscopy combined with genetics and cell biology approaches, we investigated the origin and the cellular mechanisms underlying extracellular vesicle biogenesis, and their contribution to Drosophila wing disc development, exploiting Hedgehog as a long-range morphogen. We show that microvilli of Drosophila wing disc epithelium are the site of generation of small extracellular vesicles that transport Hedgehog across the tissue. This process requires the Prominin-like protein, whose activity, together with interacting cytoskeleton components and lipids, is critical for maintaining microvilli integrity and function in secretion. Our results provide the first evidence that microvilli-derived extracellular vesicles contribute to Hedgehog long-range signaling activity highlighting their physiological significance in tissue development in vivo.

scholarly journals Therapeutic plasma exchange clears circulating soluble PD-L1 and PD-L1-positive extracellular vesicles

2020 ◽  
Vol 8 (2) ◽  
pp. e001113
Author(s):  
Jacob J Orme ◽  
Elizabeth Ann L Enninga ◽  
Fabrice Lucien-Matteoni ◽  
Heather Dale ◽  
Edwin Burgstaler ◽  
...  
Keyword(s):  
Plasma Exchange ◽  
Extracellular Vesicles ◽  
Poor Prognosis ◽  
Checkpoint Inhibitors ◽  
Clinical Intervention ◽  
Extracellular Vesicle ◽  
Therapeutic Plasma Exchange ◽  
Programmed Death Ligand 1 ◽  
Programmed Death

BackgroundTrans-acting programmed death-ligand 1 (PD-L1) derives from malignant cells in three known forms. High levels of secreted splice variant PD-L1 (sPD-L1), ADAM10/ADAM17-shed sPD-L1, and PD-L1-positive extracellular vesicles (evPD-L1) each predict poor prognosis and limited response to PD-(L)1 checkpoint inhibitors in cancer. To our knowledge, no clinical intervention has reduced any of these circulating forms of extracellular PD-L1. Here, we explore therapeutic plasma exchange (TPE) as a treatment to reduce circulating extracellular PD-L1.ResultsIn patients with melanoma, sPD-L1 levels above 0.277 ng/mL predicted inferior overall survival. In patients undergoing TPE for non-malignant indications, each TPE session removed a mean 70.8% sPD-L1 and 73.1% evPD-L1 detectable in plasma. TPE also reduced total and ADAM10-positive extracellular vesicles.ConclusionHere, we report the first known clinical intervention to remove either sPD-L1 or evPD-L1 from plasma in vivo. TPE reduces plasma sPD-L1 and evPD-L1 in vivo and may have a role in treatment with immunotherapy. TPE may also prove useful in patients with other extracellular vesicle-related conditions.

scholarly journals Extracellular Vesicle-Mediated RNA Release in Histoplasma capsulatum

mSphere ◽  
2019 ◽  
Vol 4 (2) ◽  
Author(s):  
Lysangela R. Alves ◽  
Roberta Peres da Silva ◽  
David A. Sanchez ◽  
Daniel Zamith-Miranda ◽  
Marcio L. Rodrigues ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stress Responses ◽  
Histoplasma Capsulatum ◽  
Pathogenic Fungi ◽  
Extracellular Vesicle ◽  
Fungal Species ◽  
Cellular Communication ◽  
Content Type ◽  
Rna Molecules ◽  
Rna Content

ABSTRACT Eukaryotic cells, including fungi, release extracellular vesicles (EVs). These lipid bilayered compartments play essential roles in cellular communication and pathogenesis. EV composition is complex and includes proteins, glycans, pigments, and RNA. RNAs with putative roles in pathogenesis have been described in EVs produced by fungi. Here we describe the RNA content in EVs produced by the G186AR and G217B strains of Histoplasma capsulatum, an important human-pathogenic fungal pathogen. A total of 124 mRNAs were identified in both strains. In this set of RNA classes, 93 transcripts were enriched in EVs from the G217B strain, whereas 31 were enriched in EVs produced by the G186AR strain. This result suggests that there are important strain-specific properties in the mRNA composition of fungal EVs. We also identified short fragments (25 to 40 nucleotides in length) that were strain specific, with a greater number identified in EVs produced by the G217B strain. Remarkably, the most highly enriched processes were stress responses and translation. Half of these fragments aligned to the reverse strand of the transcript, suggesting the occurrence of microRNA (miRNA)-like molecules in fungal EVs. We also compared the transcriptome profiles of H. capsulatum with the RNA composition of EVs, and no correlation was observed. Taking the results together, our study provided information about the RNA molecules present in H. capsulatum EVs and about the differences in composition between the strains. In addition, we found no correlation between the most highly expressed transcripts in the cell and their presence in the EVs, reinforcing the idea that the RNAs were directed to the EVs by a regulated mechanism. IMPORTANCE Extracellular vesicles (EVs) play important roles in cellular communication and pathogenesis. The RNA molecules in EVs have been implicated in a variety of processes. EV-associated RNA classes have recently been described in pathogenic fungi; however, only a few reports of studies describing the RNAs in fungal EVs are available. Improved knowledge of EV-associated RNA will contribute to the understanding of their role during infection. In this study, we described the RNA content in EVs produced by two isolates of Histoplasma capsulatum. Our results add this important pathogen to the current short list of fungal species with the ability to use EVs for the extracellular release of RNA.

scholarly journals Extracellular vesicle-mediated RNA release inHistoplasma capsulatum

2019 ◽  
Author(s):  
Lysangela R. Alves ◽  
Roberta Peres da Silva ◽  
David A. Sanchez ◽  
Daniel Zamith-Miranda ◽  
Marcio L. Rodrigues ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stress Responses ◽  
Histoplasma Capsulatum ◽  
Pathogenic Fungi ◽  
Extracellular Vesicle ◽  
Fungal Species ◽  
Cellular Communication ◽  
Rna Molecules ◽  
Rna Content ◽  
Reverse Strand

AbstractEukaryotic cells, including fungi, release extracellular vesicles (EVs). These lipid bilayered compartments play essential roles in cellular communication and pathogenesis. EV composition is complex and includes proteins, glycans, pigments, and RNA. RNA classes with putative roles in pathogenesis have been described in EVs produced by fungi. Here we describe the RNA content in EVs produced by the G186AR and G217B strains ofHistoplasma capsulatum, an important human fungal pathogen. A total of 124 mRNA were identified in both strains. In this set of RNA classes, 93 transcripts were enriched in EVs from the G217B strain, while 31 enriched in EVs produced by the G186AR strain. This result suggests that there are important strain-specific properties in the mRNA composition of fungal EVs. We also identified short fragments (25-40 long) that were strain-specific, with a greater number of them identified in EVs produced by the G217B strain. Remarkably, the most enriched processes were stress responses and translation. Half of these fragments aligned to the reverse strand of the transcript, suggesting the occurrence of miRNA-like molecules in fungal EVs. We also compared the transcriptome profiles ofH. capsulatumwith the RNA composition of EVs and no correlation was observed. Altogether, our study provided information about the RNA molecules present inH. capsulatumEVs, and the differences in composition between the G186AR and G217B strains. In addition, we showed that the correlation between the most expressed transcripts in the cell and their presence in the EVs, reinforcing the idea that the RNAs were directed to the EVs by a regulated mechanism.ImportanceExtracellular vesicles (EVs) play important roles in cellular communication and pathogenesis. The RNA molecules in EVs have been implicated in a variety of processes. In pathogenic fungi, EV-associated RNA classes have recently been described; however, only a few studies describing the RNA in fungal EVs are available. An improved knowledge on EV-associated RNA will contribute to the understanding of their role during infection. In this study, we described the RNA content in EVs produced by two isolates ofHistoplasma capsulatum. Our results add this important pathogen to the current short list of fungal species with the ability to use EVs for the extracellular release of RNA.

scholarly journals Ciliary Rab28 and the BBSome negatively regulate extracellular vesicle shedding

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Jyothi S Akella ◽  
Stephen P Carter ◽  
Ken Nguyen ◽  
Sofia Tsiropoulou ◽  
Ailis L Moran ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
G Protein ◽  
Extracellular Vesicles ◽  
Autosomal Recessive ◽  
Extracellular Vesicle ◽  
Cell Tissue ◽  
Specific Manner ◽  
Communication Devices ◽  
Vesicle Shedding

Cilia both receive and send information, the latter in the form of extracellular vesicles (EVs). EVs are nano-communication devices that influence cell, tissue, and organism behavior. Mechanisms driving ciliary EV biogenesis are almost entirely unknown. Here, we show that the ciliary G-protein Rab28, associated with human autosomal recessive cone-rod dystrophy, negatively regulates EV levels in the sensory organs of Caenorhabditis elegans in a cilia specific manner. Sequential targeting of lipidated Rab28 to periciliary and ciliary membranes is highly dependent on the BBSome and the prenyl-binding protein phosphodiesterase 6 subunit delta (PDE6D), respectively, and BBSome loss causes excessive and ectopic EV production. We also find that EV defective mutants display abnormalities in sensory compartment morphogenesis. Together, these findings reveal that Rab28 and the BBSome are key in vivo regulators of EV production at the periciliary membrane and suggest that EVs may mediate signaling between cilia and glia to shape sensory organ compartments. Our data also suggest that defects in the biogenesis of cilia-related EVs may contribute to human ciliopathies.

Extracellular vesicle secretion is tissue-dependent ex vivo and skeletal muscle myofiber extracellular vesicles reach the circulation in vivo.

2021 ◽  
Author(s):  
Andrea L. Estrada ◽  
Zackary J. Valenti ◽  
Gabriella Hehn ◽  
Adam J. Amorese ◽  
Nicholas S. Williams ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Vesicles ◽  
Direct Evidence ◽  
Ex Vivo ◽  
Extracellular Vesicle ◽  
Fluorescent Reporter ◽  
Human Protein Atlas ◽  
Health And Disease ◽  
Vesicle Secretion

Extracellular vesicles (EVs) are biomarkers and modifiers of human disease. EVs secreted by insulin-responsive tissues like skeletal muscle (SkM) and white adipose (WAT) contribute to metabolic health and disease but the relative abundance of EVs from these tissues has not been directly examined. Human Protein Atlas data and directly measuring EV secretion in mouse SkM and WAT using an ex vivo tissue explant model confirmed that SkM tissue secretes more EVs than WAT. Differences in EV secretion between SkM and WAT were not due to SkM contraction but may be explained by differences in tissue metabolic capacity. We next examined how many EVs secreted from SkM tissue ex vivo and in vivo are myofiber-derived. To do this, a SkM myofiber-specific dual fluorescent reporter mouse was created. Spectral flow cytometry revealed that SkM myofibers are a major source of SkM tissue-derived EVs ex vivo and EV immunocapture indicate that ~5% of circulating tetraspanin-positive EVs are derived from SkM myofibers in vivo. Our findings demonstrate that 1) SkM secretes more EVs than WAT, 2) many SkM tissue EVs are derived from SkM myofibers and 3) SkM myofiber-derived EVs reach the circulation in vivo. These findings advance our understanding of EV secretion between metabolically active tissues and provide direct evidence that SkM myofibers secrete EVs that can reach the circulation in vivo.

scholarly journals Intestine epithelial cell-derived extracellular vesicles alleviate inflammation induced by Clostridioides difficile TcdB through the activity of TGF-β1

2021 ◽  
Author(s):  
Shuangshuang Wan ◽  
Guangzhong Song ◽  
Hui Hu ◽  
Yaqing Xu ◽  
Peng Zeng ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Extracellular Vesicles ◽  
Gradient Centrifugation ◽  
Sucrose Density Gradient ◽  
Extracellular Vesicle ◽  
Sucrose Density Gradient Centrifugation ◽  
Clostridioides Difficile ◽  
Intestinal Immune System

Abstract Objective: Clostridioides difficile infection (CDI) has been primarily associated with the toxin B (TcdB), which can activate the intestinal immune system and lead to pathological damage. Even though the biological functions of intestine epithelial cell-derived extracellular vesicles (I-Evs) have been well documented, the role of I-Evs in the process of CDI is still unknown. Methods: I-Evs were isolated from mouse intestine tissues by ultracentrifugation protocol, identified by electron microscopy, nanoparticle tracking, sucrose density gradient centrifugation, and western blotting. Intestinal pathological damage was measured after intraperitoneal injection of TcdB into mice. Results: We isolated I-Evs ranging from 100–200 nm in mean diameter, with a density of 1.09-1.17 g/mL. These I-Evs expressed the extracellular vesicle-associated specific surface markers, CD63 and TSG101. In vitro, 50 µg I-Evs decreased the expression of IL-6, TNF-a, IL-1β, and IL-22 induced by 0.8 ng/mL C. difficile TcdB, and increased expression of TGF-b1. In vivo, I-Evs also promoted regulatory T cell induction, which improved the survival rate of mice up to 80% relative to C. difficile TcdB mice, dependent on the TGF-b1 signalling pathway. Conclusion: As an emerging immunotherapy, I-Evs can reduce the intraperitoneal infection induced by C. difficile TcdB and improve survival in mice.

scholarly journals Mechanism of Enhanced MerTK-Dependent Macrophage Efferocytosis by Extracellular Vesicles

2019 ◽  
Vol 39 (10) ◽  
pp. 2082-2096 ◽  
Author(s):  
Geoffrey de Couto ◽  
Ervin Jaghatspanyan ◽  
Matthew DeBerge ◽  
Weixin Liu ◽  
Kristin Luther ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Extracellular Vesicles ◽  
Apoptotic Cell ◽  
Extracellular Vesicle ◽  
Complement Factor ◽  
Irreversible Damage ◽  
Phagocytic Capacity ◽  
Cell Clearance

Objective: Extracellular vesicles secreted by cardiosphere-derived cells (CDC ev ) polarize macrophages toward a distinctive phenotype with enhanced phagocytic capacity (M CDCev ). These changes underlie cardioprotection by CDC ev and by the parent CDCs, notably attenuating the no-reflow phenomenon following myocardial infarction, but the mechanisms are unclear. Here, we tested the hypothesis that M CDCev are especially effective at scavenging debris from dying cells (ie, efferocytosis) to attenuate irreversible damage post-myocardial infarction. Approach and Results: In vitro efferocytosis assays with bone marrow-derived macrophages, and in vivo transgenic rodent models of myocardial infarction, demonstrate enhanced apoptotic cell clearance with M CDCev . CDC ev exposure induces sustained MerTK expression in M CDCev through extracellular vesicle transfer of microRNA-26a (via suppression of Adam17 ); the cardioprotective response is lost in animals deficient in MerTK. Single-cell RNA-sequencing revealed phagocytic pathway activation in M CDCev , with increased expression of complement factor C1qa , a phagocytosis facilitator. Conclusions: Together, these data demonstrate that extracellular vesicle modulation of MerTK and C1qa expression leads to enhanced macrophage efferocytosis and cardioprotection.

scholarly journals Biodistribution of Mesenchymal Stem Cell-Derived Extracellular Vesicles in a Radiation Injury Bone Marrow Murine Model

2019 ◽  
Vol 20 (21) ◽  
pp. 5468 ◽  
Author(s):  
Sicheng Wen ◽  
Mark Dooner ◽  
Elaine Papa ◽  
Michael Del Tatto ◽  
Mandy Pereira ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Extracellular Vesicles ◽  
Fibroblast Cell ◽  
Irradiation Damage ◽  
Hematopoietic Tissue ◽  
Human Lung Fibroblast ◽  
Post Radiation

We have previously shown that injury induced by irradiation to murine marrow can be partially or completely reversed by exposure to human or murine mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs). Investigation of the biodistribution of EVs in vivo is essential for understanding EV biology. In this study, we evaluated the DiD lipid dye labeled MSC-EV biodistribution in mice under different conditions, including different MSC-EV doses and injection schedules, time post MSC-EV injection, and doses of radiation. DiD-labeled MSC-EVs appeared highest in the liver and spleen; lower in bone marrow of the tibia, femur, and spine; and were undetectable in the heart, kidney and lung, while a predominant EV accumulation was detected in the lung of mice infused with human lung fibroblast cell derived EVs. There was significantly increased MSC-EV accumulation in the spleen and bone marrow (tibia and femur) post radiation appearing with an increase of MSC-EV uptake by CD11b+ and F4/80+ cells, but not by B220 cells, compared to those organs from non-irradiated mice. We further demonstrated that increasing levels of irradiation caused a selective increase in vesicle homing to marrow. This accumulation of MSC-EVs at the site of injured bone marrow could be detected as early as 1 h after MSC- EV injection and was not significantly different between 2 and 24 h post MSC-EV injection. Our study indicates that irradiation damage to hematopoietic tissue in the spleen and marrow targets MSC-EVs to these tissues.

scholarly journals Intestine epithelial cell-Derived Extracellular Vesicles alleviate Inflammation Induced by Clostridioides Difficile TcdB through the Activity of TGF- β1

2021 ◽  
Vol 4 (1) ◽  
pp. 01-09
Author(s):  
Dazhi Jin ◽  
Shuangshuang Wan ◽  
Guangzhong Song ◽  
Hui Hu ◽  
Yaqing Xu ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Extracellular Vesicles ◽  
Extracellular Vesicle ◽  
Clostridioides Difficile ◽  
Intestinal Immune System ◽  
Cell Induction ◽  
Clostridioides Difficile Infection ◽  
Tgf Β1

Background: Clostridioides difficile infection (CDI) has been primarily associated with the toxin B (TcdB), which can activate the intestinal immune system and lead to pathological damage. Even though the biological functions of intestine epithelial cell- derived extracellular vesicles (I-Evs) have been well documented, the role of I-Evs in the process of CDI is still unknown. Results: We isolated I-Evs ranging from 100–200 nm in mean diameter, with a density of 1.09-1.17 g/mL. These I-Evs expressed the extracellular vesicle-associated specific surface markers, CD63 and TSG101. In vitro, 50 µg I-Evs decreased the expression of IL-6, TNF- β, IL-1β, and IL-22 in MC38 induced by 0.8 ng/mL C. difficile TcdB, and increased expression of TGF- β1. In vivo, I-Evs also promoted regulatory T cell induction, which improved inflammation of mice up to 80% relative to C. difficile TcdB infected mice, depending on the TGF- β1 signal pathway. Conclusion: Our study firstly demonstrated that I-Evs originated from intestine epithelial cells is potentially a novel treatment endogenous candidate to effectively reduce the local infection induced by C. difficile TcdB.

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