Cigarette Smoke-Induced Pulmonary Inflammation Becomes Systemic by Circulating Extracellular Vesicles Containing Wnt5a and Inflammatory Cytokines

Feller, D.; Kun, József; Rácz, I; Rapp, Judit; Sárosi, Veronika; Kvell, Krisztián; Helyes, Zsuzsanna; Pongrácz, Judit E.

doi:10.3389/fimmu.2018.01724

Cited by 37 publications

(36 citation statements)

References 67 publications

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“…In vitro experiments were performed using the TEP1 primary-derived (BALB/c) thymic epithelial cell lines or the A549 human lung epithelial cell line (A549 served as control producer cell compared to TEP1). The Wnt4 over-expressing version of TEP1 and the Wnt5a over-expressing version of A549 were generated via lentiviral transfection linked to the green fluorescent protein (GFP) as published previously (Wnt5a served as control compared to Wnt4) (29, 30). Cells were maintained in DMEM (Dulbecco's Modified Eagle's medium, Lonza) supplemented with 10% FBS (EuroClone), Penicillin-Streptomycin, L-glutamine, Hepes buffer, non-essential amino acids (Lonza), and β-mercapto-ethanol (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Transgenic Exosomes for Thymus Regeneration

et al. 2019

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During senescence, Wnt4 expression is down-regulated (unlike their Frizzled receptors), while PPARgamma expression increases in the thymus. Together, these changes allow for thymic degeneration to occur, observed as adipose involution. However, when restored, Wnt4 can efficiently counteract PPARgamma and prevent thymic senescence from developing. The Wnt-pathway activator miR27b has also been reported to inhibit PPARgamma. Our goal was to evaluate the Wnt4 and miR27b levels of Wnt4-transgenic thymic epithelial cell (TEC)-derived exosomes, show their regenerative potential against age-related thymic degeneration, and visualize their binding and distribution both in vitro and in vivo . First, transgenic exosomes were harvested from Wnt4 over-expressing TECs and analyzed by transmission electron microscopy. This unveiled exosomes ranging from 50 to 100 nm in size. Exosomal Wnt4 protein content was assayed by ELISA, while miR27b levels were measured by TaqMan qPCR, both showing elevated levels in transgenic exosomes relative to controls. Of note, kit-purified TEI (total exosome isolate) outperformed UC (ultracentrifugation)-purified exosomes in these parameters. In addition, a significant portion of exosomal Wnt4 proved to be displayed on exosomal surfaces. For functional studies, steroid (Dexamethasone or DX)-induced TECs were used as cellular aging models in which DX-triggered cellular aging was efficiently prevented by transgenic exosomes. Finally, DiI lipid-stained exosomes were applied on the mouse thymus sections and also iv-injected into mice, for in vitro binding and in vivo tracking, respectively. We have observed distinct staining patterns using DiI lipid-stained transgenic exosomes on sections of young and aging murine thymus samples. Moreover, in vivo injected DiI lipid-stained transgenic exosomes showed detectable homing to the thymus. Of note, Wnt4-transgenic exosome homing outperformed control (Wnt5a-transgenic) exosome homing. In summary, our findings indicate that exosomal Wnt4 and miR27b can efficiently counteract thymic adipose involution. Although extrapolation of mouse results to the human setting needs caution, our results appoint transgenic TEC exosomes as promising tools of immune rejuvenation and contribute to the characterization of the immune-modulatory effects of extracellular vesicles in the context of regenerative medicine.

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Section: Methodsmentioning

confidence: 99%

Transgenic Exosomes for Thymus Regeneration

et al. 2019

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“…The role of lung epithelial cell-derived EVs in the pathogenesis of COPD has also been well reported [120]. Cigarette smoke exposure leads to exaggerated release of exosomes from damaged epithelial cells and promotes emphysema [120]. It has been shown that human bronchial epithelial cells (BEAS-2B) exposed to cigarette smoke demonstrate increased Rab27A-dependent release of EVs [121].…”

Section: Lung Cancermentioning

confidence: 99%

“…In COPD induced by cigarette smoke exposure, alveolar macrophages and alveolar epithelial cells are directly exposed to the toxic components of cigarette smoke [120]. The role of lung epithelial cell-derived EVs in the pathogenesis of COPD has also been well reported [120]. Cigarette smoke exposure leads to exaggerated release of exosomes from damaged epithelial cells and promotes emphysema [120].…”

Section: Lung Cancermentioning

confidence: 99%

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Extracellular vesicles: novel communicators in lung diseases

et al. 2020

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The lung is the organ with the highest vascular density in the human body. It is therefore perceivable that the endothelium of the lung contributes significantly to the circulation of extracellular vesicles (EVs), which include exosomes, microvesicles, and apoptotic bodies. In addition to the endothelium, EVs may arise from alveolar macrophages, fibroblasts and epithelial cells. Because EVs harbor cargo molecules, such as miRNA, mRNA, and proteins, these intercellular communicators provide important insight into the health and disease condition of donor cells and may serve as useful biomarkers of lung disease processes. This comprehensive review focuses on what is currently known about the role of EVs as markers and mediators of lung pathologies including COPD, pulmonary hypertension, asthma, lung cancer and ALI/ARDS. We also explore the role EVs can potentially serve as therapeutics for these lung diseases when released from healthy progenitor cells, such as mesenchymal stem cells. Extracellular vesicles Cell-to-cell communication is essential for nearly all physiologic and metabolic processes. This intercellular conveyance is achieved through receptor ligands, signaling molecules, hormones, and extracellular vesicles (EVs). Historically, secretion of the cell in the form of EVs was considered as unimportant waste material, cellular "garbage bags," or dust particles [1-5]. However, in recent years, this so-called "waste" is now known to be of profound importance in various biological systems, creating a boon in their exploration across the scientific community. Lipid bilayer membrane-enclosed vesicles are secreted by both prokaryotic and eukaryotic cells [4-9]. Although, the term "extracellular vesicle" is sometimes used in reference to exosomes, it is actually a very broad term that encompasses all different types of vesicles secreted outside the cells [10]. Regardless, the function of all these vesicles appears to be all the same: communication between the cells within an organism or between species [11, 12]. In addition, it is not necessary that all vesicles secreted from cells are functional or have any role in some kind of biological process. Sometimes, they just act as "dustcart" to remove the waste from cells [13]. Of note, there have been discrepancies in the classification of these vesicles in the literature. Some studies divide EVs into two major categories: I) exosomes, defined as vesicles released by exocytosis of the multivesicular bodies; and II) ectosomes, defined as the vesicles which are assembled and released by the plasma membrane [14]. However, most recent studies categorize EVs as either exosome, microvesicles, microparticles, or apoptotic bodies based on vesicle size and how they are formed [15-21] (Fig. 1). Exosomes Exosomes are small EVs with sizes ranging between 30 and 150 nm in diameter that originate from the internal vesicles of multivesicular bodies (MVB) of nearly all cell types. Exosomes originating from different cell types have different composition; however, there are certain

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“… 8 Moreover, further investigation demonstrated that CSE exposure increased Wnt5a expression in conjunction with inflammatory cytokines production in the model of COPD. 9 …”

Section: Introductionmentioning

confidence: 99%

<p>PM2.5 Induces the Expression of Inflammatory Cytokines via the Wnt5a/Ror2 Pathway in Human Bronchial Epithelial Cells</p>

Zou

Wang

et al. 2020

COPD

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Background and Purpose Recently, fine particulate matter (PM2.5) was identified as the main exposure risk for COPD, and inflammation is central to the development of COPD. In this study, we investigated whether PM2.5 can induce the secretion of interleukin-6 (IL-6), IL-8 and IL-1β in human bronchial epithelial cells (HBECs) in vitro via the wingless-related integration site 5A (Wnt5a)/receptor tyrosine kinase-like orphan receptor 2 (Ror2) signaling. Methods The expression of Wnt5a and Ror2 was assessed by immunohistochemistry in motor vehicle exhaust (MVE)-induced Sprague-Dawley rats. HBECs were transfected with small interfering RNA (siRNA) targeting Wnt5a or Ror2 and subsequently stimulated with PM2.5.The secretion of IL-6, IL-8 and IL-1β was assessed by ELISAs, and the expression of Wnt5a/Ror2 signaling were assessed by RT-PCR and Western blotting. Results Both Wnt5a and Ror2 protein were increased in the lung of MVE-induced rats. HBECs exposed to PM2.5 for 24 h significantly upregulated Wnt5a and Ror2 expression and subsequently promoted the nuclear translocation of NF-κB, which increased the production of IL-1β, IL-6 and IL-8. Wnt5a siRNA prevented these outcomes. Wnt5a antagonist (BOX5) also prevented inflammatory effects. Furthermore, Ror2 siRNA blocked the NF-κB activity and inhibited the release of IL-6, IL-8 and IL-1β from PM2.5-exposed HBECs. Conclusion PM2.5 induces the secretion of IL-6, IL-8 and IL-1β in HBECs via the Wnt5a/Ror2 signaling, demonstrating a novel mechanism for PM2.5-associated airway inflammation.

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Cigarette Smoke-Induced Pulmonary Inflammation Becomes Systemic by Circulating Extracellular Vesicles Containing Wnt5a and Inflammatory Cytokines

Cited by 37 publications

References 67 publications

Transgenic Exosomes for Thymus Regeneration

Transgenic Exosomes for Thymus Regeneration

Extracellular vesicles: novel communicators in lung diseases

<p>PM2.5 Induces the Expression of Inflammatory Cytokines via the Wnt5a/Ror2 Pathway in Human Bronchial Epithelial Cells</p>

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