Intestine epithelial cell-Derived Extracellular Vesicles alleviate Inflammation Induced by Clostridioides Difficile TcdB through the Activity of 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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Intestine epithelial cell-derived extracellular vesicles alleviate inflammation induced by Clostridioides difficile TcdB through the activity of TGF-β1

10.21203/rs.3.rs-240505/v1 ◽

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.

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Alimentary and Pharmaceutical Approach to Natural Antimicrobials against Clostridioides difficile Gastrointestinal Infection

Foods ◽

10.3390/foods10051124 ◽

2021 ◽

Vol 10 (5) ◽

pp. 1124

Author(s):

Miguel Tortajada-Girbés ◽

Alejandro Rivas ◽

Manuel Hernández ◽

Ana González ◽

Maria A. Ferrús ◽

...

Keyword(s):

Toxin Production ◽

Gastrointestinal Infection ◽

Natural Antimicrobials ◽

Gastrointestinal Microbiome ◽

Clostridioides Difficile ◽

Clostridioides Difficile Infection ◽

Successful Control ◽

Marine Bioactives

Incidence of Clostridioides difficile infection (CDI) has been increasing in recent decades due to different factors, namely (i) extended use of broad-spectrum antibiotics, (ii) transmission within asymptomatic and susceptible patients, and (iii) unbalanced gastrointestinal microbiome and collateral diseases that favor C. difficile gastrointestinal domination and toxin production. Although antibiotic therapies have resulted in successful control of CDI in the last 20 years, the development of novel strategies is urged in order to combat the capability of C. difficile to generate and acquire resistance to conventional treatments and its consequent proliferation. In this regard, vegetable and marine bioactives have emerged as alternative and effective molecules to fight against this concerning pathogen. The present review examines the effectiveness of natural antimicrobials from vegetable and algae origin that have been used experimentally in in vitro and in vivo settings to prevent and combat CDI. The aim of the present work is to contribute to accurately describe the prospective use of emerging antimicrobials as future nutraceuticals and preventive therapies, namely (i) as dietary supplement to prevent CDI and reduce CDI recurrence by means of microbiota modulation and (ii) administering them complementarily to other treatments requiring antibiotics to prevent C. difficile gut invasion and infection progression.

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Mechanism of Enhanced MerTK-Dependent Macrophage Efferocytosis by Extracellular Vesicles

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.119.313115 ◽

2019 ◽

Vol 39 (10) ◽

pp. 2082-2096 ◽

Cited By ~ 6

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.

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Extracellular vesicles produced by bone marrow mesenchymal stem cells attenuate renal fibrosis, in part by inhibiting the RhoA/ROCK pathway, in a UUO rat model

10.21203/rs.3.rs-17275/v1 ◽

2020 ◽

Author(s):

Zhengzhou Shi ◽

Qi Wang ◽

Youbo Zhang ◽

Dapeng Jiang

Keyword(s):

Oxidative Stress ◽

Extracellular Vesicles ◽

Renal Fibrosis ◽

Sham Group ◽

Rock Inhibitor ◽

Marrow Mesenchymal Stem Cells ◽

Hk2 Cells ◽

Tgf Β1

Abstract Background Renal interstitial fibrosis is a critical symptom of chronic kidney disease (CKD) that is associated with high incidence. Extracellular vesicles produced by bone marrow mesenchymal stem cells (BMSC-EVs) can play important roles in the repair of injured tissues. However, no reports have investigated the role and mechanism of BMSC-EVs in renal fibrosis. Thus, we hypothesized that BMSC-EVs containing milk fat globule-EGF factor-8 (MFG-E8) could attenuate renal fibrosis by inhibiting the RhoA/ROCK pathway.Methods We investigated whether BMSC-EVs have antifibrotic effects in a rat model of renal fibrosis-rats subjected to unilateral ureteral obstruction (UUO) - as well as in cultured HK2 cells. In vivo, Sprague-Dawley (SD) rats were randomly divided into 6 groups: Sham group, Sham + EVs group, UUO group, UUO + EVs group, UUO + EVs Ctrl group, and UUO + EVs shMFGE8 group. In vitro, extracellular vesicles from BMSCs were collected and co-cultured with HK2 cells during transforming growth factor-β1 (TGF-β1) treatment. Besides, HK2 cells co-cultured with TGF-β1 were also treated with the ROCK inhibitor, Y-27632. Results Compared with the Sham group, UUO rats displayed fibrotic abnormalities, accompanied by increased expression of α-SMA and Fibronectin, and decreased expression of E-cadherin. Both molecular and pathological changes suggested an increased inflammation in damaged kidneys. Oxidative stress, as evidenced by decreased levels of SOD1 and Catalase, was also observed in UUO kidneys. In addition, activation of cleaved caspase-3 and PARP1 and increased apoptosis in the proximal tubules confirmed tubular cell apoptosis in the UUO group. All of these phenotypes exhibited by UUO rats were suppressed by treatment with BMSC-EVs. However, the protective effect of BMSC-EVs was completely abolished by inhibition of MFG-E8. Consistent with the in vivo results, treatment with BMSC-EVs reduced inflammation, oxidative stress, apoptosis, and fibrosis in HK-2 cells stimulated with TGF-β1 in vitro . Interestingly, treatment with Y-27632, a ROCK inhibitor, protected HK-2 cells against inflammation and fibrosis, although oxidative stress and apoptosis were unchanged. Conclusions In summary, our results show that BMSC-EVs containing MFG-E8 attenuate renal fibrosis, partly through RhoA/ROCK pathway inhibition.

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TGF-β1 in extracellular vesicles from HIV-infected plasma and macrophages linked to cardiopulmonary dysfunction

10.1101/2020.09.09.20191338 ◽

2020 ◽

Author(s):

Balaji Krishnamachary ◽

Stuti Agarwal ◽

Aatish Mahajan ◽

Ashok Kumar ◽

Aradhana Mohan ◽

...

Keyword(s):

Extracellular Vesicles ◽

Troponin I ◽

Vascular Dysfunction ◽

Systolic Pressure ◽

Cell Communication ◽

People Living With Hiv ◽

Potential Biomarker ◽

Tgf Β1

Rationale: Extracellular vesicles (EVs) have emerged as important mediators in cell-cell communication and disease pathogenesis; however, their relevance in pulmonary hypertension (PH) secondary to HIV infection is yet to be explored. Objective: To examine the role of circulating small EVs and monocyte-derived macrophage (MDM) EVs in the development of HIV-associated PH Methods: EVs isolated from plasma of HIV-infected drug users and non-users with/without PH and from supernatants of HIV-infected MDMs treated with/without second hit of cocaine were studied for their effect on vascular dysfunction both in vitro and in vivo. Measurements and Main Results: We report significantly higher numbers of plasma derived EVs (PEVs) carrying higher levels of TGF-β1 in people living with HIV (PLWH) that had PH compared to non-PH PLWH. Importantly, levels of these TGF-β1 loaded PEVs correlated with pulmonary arterial systolic pressures, CD4 counts, but not with diffusion capacity for carbon monoxide or viral load. Correspondingly, enhanced TGF-β1-dependent pulmonary endothelial injury and smooth muscle hyperplasia was observed. Cocaine treatment of HIV-1 infected-MDMs resulted in increased number of TGF-β1 high-EVs. Intravenous injection of these EVs in rats led to increased right ventricle systolic pressure accompanied with myocardial injury and increased levels of serum endothelin-1, TNF-α, and cardiac Troponin-I. Conversely, pretreatment of rats with TGFβ-Receptor 1 inhibitor prevented these EV-mediated changes. Conclusion: Findings define the ability of macrophage-derived small EVs to cause pulmonary vascular modeling and PH via modulation of TGF-β signaling and suggest clinical implications of circulating TGF-β high-EVs as a potential biomarker of HIV-PH.

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A Systematic Review on Extracellular Vesicles-Enriched Fat Grafting: A Shifting Paradigm

Aesthetic Surgery Journal ◽

10.1093/asj/sjaa362 ◽

2020 ◽

Author(s):

Mohammad Ghiasloo ◽

Laura De Wilde ◽

Kashika Singh ◽

Patrick Tonnard ◽

Alexis Verpaele ◽

...

Keyword(s):

Systematic Review ◽

Graft Survival ◽

Extracellular Vesicles ◽

Retention Rate ◽

Fat Grafting ◽

Retention Rates ◽

Fat Graft ◽

Cochrane Central Register

Abstract Background Recent evidence confirms that mesenchymal stem cells (MSCs) facilitate angiogenesis mainly through paracrine function. Extracellular vesicles (EVs) are regarded as key components of the cell secretome, possessing functional properties of their source cells. Subsequently, MSC-EVs have emerged as a novel cell-free approach to improve fat graft retention rate. Objectives To provide a systematic review of all studies reporting the use of MSC-EVs to improve graft retention rate. Methods A systematic search was undertaken using the Embase, PubMed and the Cochrane Central Register of Controlled Trials databases. Outcome measures included donor/receptor organism of the fat graft, study model, intervention groups, evaluation intervals, EV research data, in vitro and in vivo results. Results Of the total 1717 articles, 62 full-texts were screened. Seven studies reporting on 294mice were included. Overall, EV treated groups showed higher graft retention rates compared to untreated groups. Notably, retention rate was similar following EV- and MSC-treatment. In addition to reduced inflammation, graft enrichment with EVs resulted in early revascularization and better graft integrity. Interestingly, hypoxic preconditioning of MSCs improved their beneficial paracrine effects and led to a more proangiogenic EV population, as observed by both in vitro and in vivo results. Conclusions MSC-EVs appear to offer an interesting cell-free alternative to improve fat graft survival. While their clinical relevance remains to be determined, it is clear that not the cells, but their secretome is essential for graft survival. Thus, a paradigm shift from cell-assisted lipotransfer towards ‘secretome-assisted lipotransfer’ is well on its way.

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Plant-Derived Extracellular Vesicles: Current Findings,Challenges, and Future Applications

Membranes ◽

10.3390/membranes11060411 ◽

2021 ◽

Vol 11 (6) ◽

pp. 411

Author(s):

Nader Kameli ◽

Anya Dragojlovic-Kerkache ◽

Paul Savelkoul ◽

Frank R. Stassen

Keyword(s):

Human Health ◽

Extracellular Vesicles ◽

Preliminary Results ◽

In Vivo Experiments ◽

Health And Disease ◽

Conducting Research ◽

Protocol Standardization ◽

Insight Into

In recent years, plant-derived extracellular vesicles (PDEVs) have gained the interest of many experts in fields such as microbiology and immunology, and research in this field has exponentially increased. These nano-sized particles have provided researchers with a number of interesting findings, making their application in human health and disease very promising. Both in vitro and in vivo experiments have shown that PDEVs can exhibit a multitude of effects, suggesting that these vesicles may have many potential future applications, including therapeutics and nano-delivery of compounds. While the preliminary results are promising, there are still some challenges to face, such as a lack of protocol standardization, as well as knowledge gaps that need to be filled. This review aims to discuss various aspects of PDEV knowledge, including their preliminary findings, challenges, and future uses, giving insight into the complexity of conducting research in this field.

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Regorafenib-Attenuated, Bleomycin-Induced Pulmonary Fibrosis by Inhibiting the TGF-β1 Signaling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms22041985 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1985

Author(s):

Xiaohe Li ◽

Ling Ma ◽

Kai Huang ◽

Yuli Wei ◽

Shida Long ◽

...

Keyword(s):

Pulmonary Fibrosis ◽

Signaling Pathway ◽

Transforming Growth Factor ◽

Kinase Inhibitor ◽

In Vivo Experiments ◽

Age Related ◽

And Migration ◽

Tgf Β1

Idiopathic pulmonary fibrosis (IPF) is a fatal and age-related pulmonary disease. Nintedanib is a receptor tyrosine kinase inhibitor, and one of the only two listed drugs against IPF. Regorafenib is a novel, orally active, multi-kinase inhibitor that has similar targets to nintedanib and is applied to treat colorectal cancer and gastrointestinal stromal tumors in patients. In this study, we first identified that regorafenib could alleviate bleomycin-induced pulmonary fibrosis in mice. The in vivo experiments indicated that regorafenib suppresses collagen accumulation and myofibroblast activation. Further in vitro mechanism studies showed that regorafenib inhibits the activation and migration of myofibroblasts and extracellular matrix production, mainly through suppressing the transforming growth factor (TGF)-β1/Smad and non-Smad signaling pathways. In vitro studies have also indicated that regorafenib could augment autophagy in myofibroblasts by suppressing TGF-β1/mTOR (mechanistic target of rapamycin) signaling, and could promote apoptosis in myofibroblasts. In conclusion, regorafenib attenuates bleomycin-induced pulmonary fibrosis by suppressing the TGF-β1 signaling pathway.

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Lipoproteins Are Responsible for the Pro-Inflammatory Property of Staphylococcus aureus Extracellular Vesicles

International Journal of Molecular Sciences ◽

10.3390/ijms22137099 ◽

2021 ◽

Vol 22 (13) ◽

pp. 7099

Author(s):

Pradeep Kumar Kopparapu ◽

Meghshree Deshmukh ◽

Zhicheng Hu ◽

Majd Mohammad ◽

Marco Maugeri ◽

...

Keyword(s):

Extracellular Vesicles ◽

Inflammatory Responses ◽

Surface Proteins ◽

Host Cells ◽

Immune Stimulation ◽

Domains Of Life ◽

Major Surface ◽

Staphylococcal Aureus

Staphylococcal aureus (S. aureus), a Gram-positive bacteria, is known to cause various infections. Extracellular vesicles (EVs) are a heterogeneous array of membranous structures secreted by cells from all three domains of life, i.e., eukaryotes, bacteria, and archaea. Bacterial EVs are implied to be involved in both bacteria–bacteria and bacteria–host interactions during infections. It is still unclear how S. aureus EVs interact with host cells and induce inflammatory responses. In this study, EVs were isolated from S. aureus and mutant strains deficient in either prelipoprotein lipidation (Δlgt) or major surface proteins (ΔsrtAB). Their immunostimulatory capacities were assessed both in vitro and in vivo. We found that S. aureus EVs induced pro-inflammatory responses both in vitro and in vivo. However, this activity was dependent on lipidated lipoproteins (Lpp), since EVs isolated from the Δlgt showed no stimulation. On the other hand, EVs isolated from the ΔsrtAB mutant showed full immune stimulation, indicating the cell wall anchoring of surface proteins did not play a role in immune stimulation. The immune stimulation of S. aureus EVs was mediated mainly by monocytes/macrophages and was TLR2 dependent. In this study, we demonstrated that not only free Lpp but also EV-imbedded Lpp had high pro-inflammatory activity.

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Concurrent YAP/TAZ and SMAD signaling mediate vocal fold fibrosis

Scientific Reports ◽

10.1038/s41598-021-92871-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ryosuke Nakamura ◽

Nao Hiwatashi ◽

Renjie Bing ◽

Carina P. Doyle ◽

Ryan C. Branski

Keyword(s):

Vocal Fold ◽

Vocal Folds ◽

Nuclear Accumulation ◽

Smad Pathway ◽

Surgical Interventions ◽

Smad Signaling ◽

Iatrogenic Damage ◽

Tgf Β1

AbstractVocal fold (VF) fibrosis is a major cause of intractable voice-related disability and reduced quality of life. Excision of fibrotic regions is suboptimal and associated with scar recurrence and/or further iatrogenic damage. Non-surgical interventions are limited, putatively related to limited insight regarding biochemical events underlying fibrosis, and downstream, the lack of therapeutic targets. YAP/TAZ integrates diverse cell signaling events and interacts with signaling pathways related to fibrosis, including the TGF-β/SMAD pathway. We investigated the expression of YAP/TAZ following vocal fold injury in vivo as well as the effects of TGF-β1 on YAP/TAZ activity in human vocal fold fibroblasts, fibroblast-myofibroblast transition, and TGF-β/SMAD signaling. Iatrogenic injury increased nuclear localization of YAP and TAZ in fibrotic rat vocal folds. In vitro, TGF-β1 activated YAP and TAZ in human VF fibroblasts, and inhibition of YAP/TAZ reversed TGF-β1-stimulated fibroplastic gene upregulation. Additionally, TGF-β1 induced localization of YAP and TAZ in close proximity to SMAD2/3, and nuclear accumulation of SMAD2/3 was inhibited by a YAP/TAZ inhibitor. Collectively, YAP and TAZ were synergistically activated with the TGF-β/SMAD pathway, and likely essential for the fibroplastic phenotypic shift in VF fibroblasts. Based on these data, YAP/TAZ may evolve as an attractive therapeutic target for VF fibrosis.

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