Plasma-Derived Extracellular Vesicles as Potential Biomarkers in Heart Transplant Patient with Chronic Chagas Disease

Cortes-Serra, Núria; Mendes, Maria Tays; Mazagatos, Clara; Seguí-Barber, Joan; Ellis, Cameron; Ballart, Cristina; García, Ana Martín; Gállego, Montserrat; Gascón, Joaquim; Almeida, Igor C.; Pinazo, María Jesús; Fernández-Becerra, Carmen

doi:10.3201/eid2608.191042

Cited by 14 publications

(23 citation statements)

References 15 publications

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“…In parasites, proteins from the murine malarial parasite Plasmodium yoelii have been detected in proteomics analysis of EVs from infected reticulocytes ( Martin-Jaular et al, 2011 ). Similar findings have been recently observed in EVs isolated from plasma of a patient with chronic Chagas disease ( Cortes-Serra et al, 2020 ). The use of EVs as biomarkers has been better explored in cancer biology.…”

Section: Extracellular Vesicle Components As Disease Biomarkers and Cell Biology Markerssupporting

confidence: 90%

Omics Approaches for Understanding Biogenesis, Composition and Functions of Fungal Extracellular Vesicles

et al. 2021

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Extracellular vesicles (EVs) are lipid bilayer structures released by organisms from all kingdoms of life. The diverse biogenesis pathways of EVs result in a wide variety of physical properties and functions across different organisms. Fungal EVs were first described in 2007 and different omics approaches have been fundamental to understand their composition, biogenesis, and function. In this review, we discuss the role of omics in elucidating fungal EVs biology. Transcriptomics, proteomics, metabolomics, and lipidomics have each enabled the molecular characterization of fungal EVs, providing evidence that these structures serve a wide array of functions, ranging from key carriers of cell wall biosynthetic machinery to virulence factors. Omics in combination with genetic approaches have been instrumental in determining both biogenesis and cargo loading into EVs. We also discuss how omics technologies are being employed to elucidate the role of EVs in antifungal resistance, disease biomarkers, and their potential use as vaccines. Finally, we review recent advances in analytical technology and multi-omic integration tools, which will help to address key knowledge gaps in EVs biology and translate basic research information into urgently needed clinical applications such as diagnostics, and immuno- and chemotherapies to fungal infections.

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Section: Extracellular Vesicle Components As Disease Biomarkers and Cell Biology Markerssupporting

confidence: 90%

Omics Approaches for Understanding Biogenesis, Composition and Functions of Fungal Extracellular Vesicles

et al. 2021

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“…The presence of parasite is hardly detected in chronic patients. Only two studies have been performed to assess circulating EVs from patients with CCD: One concerning a single patient with reactivation after a heart transplant (Cortes‐Serra et al, 2020) and another that assessed immune responses and disease stage stratification but did not focus on the isolation method per se (Madeira et al, 2021). Therefore, the aim of this study is to describe the effect of different anticoagulants and the combination of two purification methods for isolation and characterization of circulating EVs from blood of CCD patients according to current ISEV guidelines (Madeira et al, 2021; Théry et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Isolation and molecular characterization of circulating extracellular vesicles from blood of chronic Chagas disease patients

Madeira

Meneghetti

Barros

et al. 2022

Cell Biology International

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Extracellular vesicles (EVs) are lipid bilayer envelopes that encase several types of molecules. Their contents mostly reflect their cell origin and possible targets at other locations in the organism and can be modified in pathological conditions to interfere with intercellular communication, thus promoting disease establishment and development. These characteristics, in addition to their presence in virtually all body fluids, make such vesicles ideal for biomarker discovery in human diseases.Here, we describe the effect of different anticoagulants and the combination of two purification methods for isolation and characterization of circulating EVs from blood of chronic Chagas disease (CCD) patients. We illustrated this procedure by studying a population of patients with Chagas disease at the indeterminate chronic stage, in which the Trypanosoma cruzi is very scarce in circulation. EVs were harvested from blood collected without or with different anticoagulants. Protein and nanoparticle tracking analysis was used to measure EVs size and concentration. The EVs were purified by ultracentrifugation, followed by size-exclusion chromatography and characterized by chemiluminescent enzyme-linked immunosorbent assay and dot blot using antibodies that recognized parasite-derived EVs, such as hyperimmune sera, polyclonal and monoclonal antibodies against trans-sialidase and mucins. In parallel, antibodies against classical human EV markers CD9, CD63, CD81, and CD82, were also analyzed. The results showed that anticoagulants did not interfere with the analyzed parameters and circulating EVs from CCD patients contain T. cruzi

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“…The large and small vesicles (LVs and SVs) secreted by Giardia were isolated by differential density- gradient centrifugation, as described above, and subjected to proteomic analysis by one-dimensional liquid chromatography-tandem mass spectrometry (1D-LC-MS/MS) ( Szempruch et al., 2016 ; Ribeiro et al., 2018 ; Cronemberger-Andrade et al., 2020 ; Cortes-Serra et al., 2020 ). Briefly, 1 μL of digested peptides was injected into a custom packed AQUA 5μm,125Å, C18 (Phenomenex, cat no.…”

Section: Methodsmentioning

confidence: 99%

Giardial lipid rafts share virulence factors with secreted vesicles and participate in parasitic infection in mice

Grajeda

Chatterjee

Villalobos

et al. 2022

Front. Cell. Infect. Microbiol.

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Giardia lamblia, a protozoan parasite, is a major cause of waterborne infection, worldwide. While the trophozoite form of this parasite induces pathological symptoms in the gut, the cyst form transmits the infection. Since Giardia is a noninvasive parasite, the actual mechanism by which it causes disease remains elusive. We have previously reported that Giardia assembles cholesterol and GM1 glycosphingolipid-enriched lipid rafts (LRs) that participate in encystation and cyst production. To further delineate the role of LRs in pathogenesis, we isolated LRs from Giardia and subjected them to proteomic analysis. Various cellular proteins including potential virulence factors—e.g., giardins, variant surface proteins, arginine deaminases, elongation factors, ornithine carbomyltransferases, and high cysteine-rich membrane proteins—were found to be present in LRs. Since Giardia secretes virulence factors encapsulated in extracellular vesicles (EVs) that induce proinflammatory responses in hosts, EVs released by the parasite were isolated and subjected to nanoparticle tracking and proteomic analysis. Two types of EV—i.e., small vesicles (SVs; <100 nm, exosome-like particles) and large vesicles (LVs; 100–400 nm, microvesicle-like particles)—were identified and found to contain a diverse group of proteins including above potential virulence factors. Although pretreatment of the parasite with two giardial lipid raft (gLR) disruptors, nystatin (27 μM) and oseltamivir (20 μM), altered the expression profiles of virulence factors in LVs and SVs, the effects were more robust in the case of SVs. To examine the potential role of rafts and vesicles in pathogenicity, Giardia-infected mice were treated with oseltamivir (1.5 and 3.0 mg/kg), and the shedding of cysts were monitored. We observed that this drug significantly reduced the parasite load in mice. Taken together, our results suggest that virulence factors partitioning in gLRs, released into the extracellular milieu via SVs and LVs, participate in spread of giardiasis and could be targeted for future drug development.

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Plasma-Derived Extracellular Vesicles as Potential Biomarkers in Heart Transplant Patient with Chronic Chagas Disease

Cited by 14 publications

References 15 publications

Omics Approaches for Understanding Biogenesis, Composition and Functions of Fungal Extracellular Vesicles

Omics Approaches for Understanding Biogenesis, Composition and Functions of Fungal Extracellular Vesicles

Isolation and molecular characterization of circulating extracellular vesicles from blood of chronic Chagas disease patients

Giardial lipid rafts share virulence factors with secreted vesicles and participate in parasitic infection in mice

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