Leishmania in 360: Guidelines for Exosomal Research

Leishmania parasites are a group of kinetoplastid pathogens that cause a variety of clinical forms while maintaining cell communication by secreting extracellular vesicles. Emerging technologies have been adapted for the studies of Leishmania-host-cell interactions to enable broad scale analysis of parasite extracellular vesicles. Leishmania extracellular vesicles (LEVS) are naturally released spheroidal nanoparticles of polydispersed suspensions surrounded by a lipid layer of membrane. Although LEVs have increasingly gained in importance, much is still unexplained, including bioavailability and function in the complex molecular mechanisms of pathogenesis. Considering the importance of LEVs in the parasite-host interaction and in the parasite-parasite relationships emerged during evolution, the current review aims at giving an overview of Leishmania summarizing knowledge and formulating guidelines for LEVs research. In the end, we report, direct methods for specific isolation of LEVs from promastigotes and amastigotes culture supernatant suitable for a range of different downstream applications increasing the compatibility and reproducibility to establish optimal and comparable isolation conditions and full LEVs characterization, and crucial immunomodulatory events triggered by this important group of parasites.

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Leishmania 360°: Guidelines for Exosomal Research

Microorganisms ◽

10.3390/microorganisms9102081 ◽

2021 ◽

Vol 9 (10) ◽

pp. 2081

Author(s):

Áurea Martins Gabriel ◽

Adan Galué-Parra ◽

Washington Luiz Assunção Pereira ◽

Ketil Winther Pedersen ◽

Edilene Oliveira da Silva

Keyword(s):

Extracellular Vesicles ◽

Molecular Mechanisms ◽

Lipid Membrane ◽

Cell Communication ◽

Direct Methods ◽

Complete Characterization ◽

Host Interaction ◽

History Of ◽

Host Cell Interactions ◽

And Function

Leishmania parasites are a group of kinetoplastid pathogens that cause a variety of clinical disorders while maintaining cell communication by secreting extracellular vesicles. Emerging technologies have been adapted for the study of Leishmania-host cell interactions, to enable the broad-scale analysis of the extracellular vesicles of this parasite. Leishmania extracellular vesicles (LEVs) are spheroidal nanoparticles of polydispersed suspensions surrounded by a layer of lipid membrane. Although LEVs have attracted increasing attention from researchers, many aspects of their biology remain unclear, including their bioavailability and function in the complex molecular mechanisms of pathogenesis. Given the importance of LEVs in the parasite-host interaction, and in the parasite-parasite relationships that have emerged during the evolutionary history of these organisms, the present review provides an overview of the available data on Leishmania, and formulates guidelines for LEV research. We conclude by reporting direct methods for the isolation of specific LEVs from the culture supernatant of the promastigotes and amastigotes that are suitable for a range of different downstream applications, which increases the compatibility and reproducibility of the approach for the establishment of optimal and comparable isolation conditions and the complete characterization of the LEV, as well as the critical immunomodulatory events triggered by this important group of parasites.

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Microglia extracellular vesicles: focus on molecular composition and biological function

Biochemical Society Transactions ◽

10.1042/bst20210202 ◽

2021 ◽

Vol 49 (4) ◽

pp. 1779-1790 ◽

Cited By ~ 1

Author(s):

Lorenzo Ceccarelli ◽

Chiara Giacomelli ◽

Laura Marchetti ◽

Claudia Martini

Keyword(s):

Extracellular Vesicles ◽

Molecular Mechanisms ◽

Biological Function ◽

Biological Effects ◽

Genetic Material ◽

Cell Communication ◽

Molecular Composition ◽

Cargo Proteins ◽

A Cell ◽

The Central Nervous System

Extracellular vesicles (EVs) are a heterogeneous family of cell-derived lipid bounded vesicles comprising exosomes and microvesicles. They are potentially produced by all types of cells and are used as a cell-to-cell communication method that allows protein, lipid, and genetic material exchange. Microglia cells produce a large number of EVs both in resting and activated conditions, in the latter case changing their production and related biological effects. Several actions of microglia in the central nervous system are ascribed to EVs, but the molecular mechanisms by which each effect occurs are still largely unknown. Conflicting functions have been ascribed to microglia-derived EVs starting from the neuronal support and ending with the propagation of inflammation and neurodegeneration, confirming the crucial role of these organelles in tuning brain homeostasis. Despite the increasing number of studies reported on microglia-EVs, there is also a lot of fragmentation in the knowledge on the mechanism at the basis of their production and modification of their cargo. In this review, a collection of literature data about the surface and cargo proteins and lipids as well as the miRNA content of EVs produced by microglial cells has been reported. A special highlight was given to the works in which the EV molecular composition is linked to a precise biological function.

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Extracellular Vesicles: An Emerging Mechanism Governing the Secretion and Biological Roles of Tenascin-C

Frontiers in Immunology ◽

10.3389/fimmu.2021.671485 ◽

2021 ◽

Vol 12 ◽

Author(s):

Lucas Albacete-Albacete ◽

Miguel Sánchez-Álvarez ◽

Miguel Angel del Pozo

Keyword(s):

Extracellular Vesicles ◽

Molecular Mechanisms ◽

Cell Communication ◽

Cell Types ◽

Target Cells ◽

Tenascin C ◽

Signalling Molecules ◽

Inflammatory Processes ◽

Bona Fide ◽

Cellular Components

ECM composition and architecture are tightly regulated for tissue homeostasis. Different disorders have been associated to alterations in the levels of proteins such as collagens, fibronectin (FN) or tenascin-C (TnC). TnC emerges as a key regulator of multiple inflammatory processes, both during physiological tissue repair as well as pathological conditions ranging from tumor progression to cardiovascular disease. Importantly, our current understanding as to how TnC and other non-collagen ECM components are secreted has remained elusive. Extracellular vesicles (EVs) are small membrane-bound particles released to the extracellular space by most cell types, playing a key role in cell-cell communication. A broad range of cellular components can be transported by EVs (e.g. nucleic acids, lipids, signalling molecules and proteins). These cargoes can be transferred to target cells, potentially modulating their function. Recently, several extracellular matrix (ECM) proteins have been characterized as bona fide EV cargoes, exosomal secretion being particularly critical for TnC. EV-dependent ECM secretion might underpin diseases where ECM integrity is altered, establishing novel concepts in the field such as ECM nucleation over long distances, and highlighting novel opportunities for diagnostics and therapeutic intervention. Here, we review recent findings and standing questions on the molecular mechanisms governing EV–dependent ECM secretion and its potential relevance for disease, with a focus on TnC.

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Cell-to-Cell Communication in Learning and Memory: From Neuro- and Glio-Transmission to Information Exchange Mediated by Extracellular Vesicles

International Journal of Molecular Sciences ◽

10.3390/ijms21010266 ◽

2019 ◽

Vol 21 (1) ◽

pp. 266 ◽

Cited By ~ 9

Author(s):

Gabriella Schiera ◽

Carlo Maria Di Liegro ◽

Italia Di Liegro

Keyword(s):

Learning And Memory ◽

Extracellular Vesicles ◽

Glial Cells ◽

Information Exchange ◽

System Development ◽

Cell Communication ◽

Nervous System Development ◽

The Body ◽

Pathological Conditions ◽

And Function

Most aspects of nervous system development and function rely on the continuous crosstalk between neurons and the variegated universe of non-neuronal cells surrounding them. The most extraordinary property of this cellular community is its ability to undergo adaptive modifications in response to environmental cues originating from inside or outside the body. Such ability, known as neuronal plasticity, allows long-lasting modifications of the strength, composition and efficacy of the connections between neurons, which constitutes the biochemical base for learning and memory. Nerve cells communicate with each other through both wiring (synaptic) and volume transmission of signals. It is by now clear that glial cells, and in particular astrocytes, also play critical roles in both modes by releasing different kinds of molecules (e.g., D-serine secreted by astrocytes). On the other hand, neurons produce factors that can regulate the activity of glial cells, including their ability to release regulatory molecules. In the last fifteen years it has been demonstrated that both neurons and glial cells release extracellular vesicles (EVs) of different kinds, both in physiologic and pathological conditions. Here we discuss the possible involvement of EVs in the events underlying learning and memory, in both physiologic and pathological conditions.

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Tiny Actors in the Big Cellular World: Extracellular Vesicles Playing Critical Roles in Cancer

International Journal of Molecular Sciences ◽

10.3390/ijms21207688 ◽

2020 ◽

Vol 21 (20) ◽

pp. 7688 ◽

Cited By ~ 2

Author(s):

Ancuta Jurj ◽

Cecilia Pop-Bica ◽

Ondrej Slaby ◽

Cristina D. Ştefan ◽

William C. Cho ◽

...

Keyword(s):

Extracellular Vesicles ◽

Molecular Mechanisms ◽

Recipient Cell ◽

Cell Communication ◽

Therapeutic Agents ◽

Bioactive Molecules ◽

Cellular Functions ◽

Membrane Bound

Communications among cells can be achieved either via direct interactions or via secretion of soluble factors. The emergence of extracellular vesicles (EVs) as entities that play key roles in cell-to-cell communication offer opportunities in exploring their features for use in therapeutics; i.e., management and treatment of various pathologies, such as those used for cancer. The potential use of EVs as therapeutic agents is attributed not only for their cell membrane-bound components, but also for their cargos, mostly bioactive molecules, wherein the former regulate interactions with a recipient cell while the latter trigger cellular functions/molecular mechanisms of a recipient cell. In this article, we highlight the involvement of EVs in hallmarks of a cancer cell, particularly focusing on those molecular processes that are influenced by EV cargos. Moreover, we explored the roles of RNA species and proteins carried by EVs in eliciting drug resistance phenotypes. Interestingly, engineered EVs have been investigated and proposed as therapeutic agents in various in vivo and in vitro studies, as well as in several clinical trials.

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Neuron-Derived Extracellular Vesicles Modulate Microglia Activation and Function

Biology ◽

10.3390/biology10100948 ◽

2021 ◽

Vol 10 (10) ◽

pp. 948

Author(s):

Hui Peng ◽

Brock T. Harvey ◽

Christopher I. Richards ◽

Kimberly Nixon

Keyword(s):

Extracellular Vesicles ◽

Cortical Neurons ◽

Molecular Mechanisms ◽

Proinflammatory Response ◽

Tnf Α ◽

Rat Cortical Neurons ◽

The Central Nervous System ◽

And Function ◽

And Control ◽

Brain Homeostasis

Microglia act as the immune cells of the central nervous system (CNS). They play an important role in maintaining brain homeostasis but also in mediating neuroimmune responses to insult. The interactions between neurons and microglia represent a key process for neuroimmune regulation and subsequent effects on CNS integrity. However, the molecular mechanisms of neuron-glia communication in regulating microglia function are not fully understood. One recently described means of this intercellular communication is via nano-sized extracellular vesicles (EVs) that transfer a large diversity of molecules between neurons and microglia, such as proteins, lipids, and nucleic acids. To determine the effects of neuron-derived EVs (NDEVs) on microglia, NDEVs were isolated from the culture supernatant of rat cortical neurons. When NDEVs were added to primary cultured rat microglia, we found significantly improved microglia viability via inhibition of apoptosis. Additionally, application of NDEVs to cultured microglia also inhibited the expression of activation surface markers on microglia. Furthermore, NDEVs reduced the LPS-induced proinflammatory response in microglia according to reduced gene expression of proinflammatory cytokines (TNF-α, IL-6, MCP-1) and iNOS, but increased expression of the anti-inflammatory cytokine, IL-10. These findings support that neurons critically regulate microglia activity and control inflammation via EV-mediated neuron–glia communication. (Supported by R21AA025563 and R01AA025591).

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The Aquatic Invertebrate Hydra vulgaris Releases Molecular Messages Through Extracellular Vesicles

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.788117 ◽

2021 ◽

Vol 9 ◽

Author(s):

Maria Moros ◽

Eugenio Fergola ◽

Valentina Marchesano ◽

Margherita Mutarelli ◽

Giuseppina Tommasini ◽

...

Keyword(s):

Extracellular Vesicles ◽

Cell Communication ◽

Population Level ◽

Target Cells ◽

Chemical Information ◽

Hydra Vulgaris ◽

Physiological Processes ◽

Developmental Signalling ◽

And Function ◽

Cell Cell

Recent body of evidence demonstrates that extracellular vesicles (EVs) represent the first language of cell-cell communication emerged during evolution. In aquatic environments, transferring signals between cells by EVs offers protection against degradation, allowing delivering of chemical information in high local concentrations to the target cells. The packaging of multiple signals, including those of hydrophobic nature, ensures target cells to receive the same EV-conveyed messages, and the coordination of a variety of physiological processes across cells of a single organisms, or at the population level, i.e., mediating the population’s response to changing environmental conditions. Here, we purified EVs from the medium of the freshwater invertebrate Hydra vulgaris, and the molecular profiling by proteomic and transcriptomic analyses revealed multiple markers of the exosome EV subtype, from structural proteins to stress induced messages promoting cell survival. Moreover, positive and negative regulators of the Wnt/β-catenin signaling pathway, the major developmental pathway acting in body axial patterning, were identified. Functional analysis on amputated polyps revealed EV ability to modulate both head and foot regeneration, suggesting bioactivity of the EV cargo and opening new perspectives on the mechanisms of developmental signalling. Our results open the path to unravel EV biogenesis and function in all cnidarian species, tracing back the origin of the cell-cell, cross-species or cross-kingdom communication in aquatic ecosystems.

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Molecular Mechanisms and Therapeutics for SBMA/Kennedy’s Disease

Neurotherapeutics ◽

10.1007/s13311-019-00790-9 ◽

2019 ◽

Vol 16 (4) ◽

pp. 928-947 ◽

Cited By ~ 2

Author(s):

Frederick J. Arnold ◽

Diane E. Merry

Keyword(s):

Motor Neurons ◽

Molecular Mechanisms ◽

Muscular Atrophy ◽

Structure And Function ◽

Loss Of Function ◽

Mechanisms Of Toxicity ◽

Current Review ◽

And Function ◽

Positive Results ◽

Polyq Expansion

Abstract Spinal and bulbar muscular atrophy (SBMA) is a neuromuscular disease caused by a polyglutamine (polyQ) expansion in the androgen receptor (AR). Despite the fact that the monogenic cause of SBMA has been known for nearly 3 decades, there is no effective treatment for this disease, underscoring the complexity of the pathogenic mechanisms that lead to a loss of motor neurons and muscle in SBMA patients. In the current review, we provide an overview of the system-wide clinical features of SBMA, summarize the structure and function of the AR, discuss both gain-of-function and loss-of-function mechanisms of toxicity caused by polyQ-expanded AR, and describe the cell and animal models utilized in the study of SBMA. Additionally, we summarize previously conducted clinical trials which, despite being based on positive results from preclinical studies, proved to be largely ineffective in the treatment of SBMA; nonetheless, these studies provide important insights as researchers develop the next generation of therapies.

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Extracellular vesicles: Exosomes, microvesicles, and friends

The Journal of Cell Biology ◽

10.1083/jcb.201211138 ◽

2013 ◽

Vol 200 (4) ◽

pp. 373-383 ◽

Cited By ~ 3421

Author(s):

Graça Raposo ◽

Willem Stoorvogel

Keyword(s):

Plasma Membrane ◽

Extracellular Vesicles ◽

Molecular Mechanisms ◽

Membrane Vesicles ◽

Vesicle Release ◽

Physiological Relevance ◽

And Function ◽

Extracellular Environment

Cells release into the extracellular environment diverse types of membrane vesicles of endosomal and plasma membrane origin called exosomes and microvesicles, respectively. These extracellular vesicles (EVs) represent an important mode of intercellular communication by serving as vehicles for transfer between cells of membrane and cytosolic proteins, lipids, and RNA. Deficiencies in our knowledge of the molecular mechanisms for EV formation and lack of methods to interfere with the packaging of cargo or with vesicle release, however, still hamper identification of their physiological relevance in vivo. In this review, we focus on the characterization of EVs and on currently proposed mechanisms for their formation, targeting, and function.

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Extracellular Vesicles: Potential Mediators of Psychosocial Stress Contribution to Osteoporosis?

International Journal of Molecular Sciences ◽

10.3390/ijms22115846 ◽

2021 ◽

Vol 22 (11) ◽

pp. 5846

Author(s):

Yangyang He ◽

Karin Wuertz-Kozak ◽

Linn K. Kuehl ◽

Pia-Maria Wippert

Keyword(s):

Extracellular Vesicles ◽

Psychosocial Stress ◽

Cell Communication ◽

Target Cells ◽

Low Bone Mass ◽

Endocrine Factors ◽

Sympathetic Nervous ◽

Mediate Cell ◽

And Function ◽

Stress Contribution

Osteoporosis is characterized by low bone mass and damage to the bone tissue’s microarchitecture, leading to increased fracture risk. Several studies have provided evidence for associations between psychosocial stress and osteoporosis through various pathways, including the hypothalamic-pituitary-adrenocortical axis, the sympathetic nervous system, and other endocrine factors. As psychosocial stress provokes oxidative cellular stress with consequences for mitochondrial function and cell signaling (e.g., gene expression, inflammation), it is of interest whether extracellular vesicles (EVs) may be a relevant biomarker in this context or act by transporting substances. EVs are intercellular communicators, transfer substances encapsulated in them, modify the phenotype and function of target cells, mediate cell-cell communication, and, therefore, have critical applications in disease progression and clinical diagnosis and therapy. This review summarizes the characteristics of EVs, their role in stress and osteoporosis, and their benefit as biological markers. We demonstrate that EVs are potential mediators of psychosocial stress and osteoporosis and may be beneficial in innovative research settings.

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