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1.
Cell Physiol Biochem ; 54(1): 88-109, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31990489

RESUMO

Extracellular vesicles (EVs) are important mediators of intercellular communication. Since EVs are also released during pathological conditions, there has been considerable interest in their potential as sensitive biomarkers of cellular stress and/or injury. In the context of kidney disease, urinary EVs are promising indicators of glomerular and tubular damage. In the present review we discuss the role of urinary EVs in kidney health and disease. Our focus is to explore urinary large EVs (lEVs, often referred to as microparticles or microvesicles) as direct and noninvasive early biomarkers of renal injury. In this regard, studies have been demonstrating altered levels of urinary lEVs, especially podocyte-derived lEVs, preceding the decrease of renal function assessed by classical markers. In addition, we discuss the role of small EVs (sEVs, often referred to as exosomes) and their contents in kidney pathophysiology. Even though results concerning the production of sEVs during diseased conditions are varied, there has been a consensus on the importance of urinary sEV content assessment in kidney disease. These mediators, including EV-released miRNAs and mRNAs, are responsible for EV-mediated signaling in the regulation of renal cellular homeostasis, pathogenesis and regeneration. Finally, steps necessary for the validation of EVs as reliable markers will be discussed.


Assuntos
Vesículas Extracelulares/patologia , Nefropatias/diagnóstico , Glomérulos Renais/patologia , Túbulos Renais/patologia , Animais , Biomarcadores/análise , Biomarcadores/urina , Humanos , Nefropatias/patologia , Nefropatias/urina
2.
Cancer Sci ; 111(1): 98-111, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31715081

RESUMO

The role of long noncoding RNAs (lncRNAs) in the epithelial-mesenchymal transition (EMT) in pancreatic ductal adenocarcinoma (PDAC) is unclear. Some lncRNAs can be transferred by extracellular vesicles (EVs) and have potential as biomarkers. Here, we identify an lncRNA that could serve as a biomarker for PDAC and show the functional roles of the lncRNA. Expression profiling of lncRNAs revealed that highly upregulated in liver cancer (HULC) was highly expressed, and induced, by transforming growth factor-ß in PDAC cells and their EVs. Knockdown of HULC decreased PDAC cell invasion and migration by inhibiting the EMT. Thus, HULC could be transferred by EVs, and promote EMT, invasion, and migration in recipient PDAC cells. To assess the roles of HULC, PDAC cell xenografts in nude mice were established. Knockdown of HULC in PDAC cells implanted in mice inhibited tumor growth. Moreover, microRNA-133b suppressed PDAC cell invasion and migration by inhibiting the EMT through targeting HULC. Furthermore, serum samples were obtained from 20 PDAC and 22 intraductal papillary mucinous neoplasm (IPMN) patients, as well as 21 healthy individuals. Analysis of serum EV HULC expression by digital PCR showed that HULC expression was significantly increased in PDAC patients compared to healthy individuals or IPMN patients. Additionally, HULC showed good predictive performance for discriminating PDAC, suggesting that the analysis of EV-encapsulated HULC would contribute to the diagnosis for human PDAC. Extracellular vesicle-transported HULC promotes cell invasion and migration by inducing the EMT, and microRNA-133b suppresses the EMT by targeting HULC. Extracellular vesicle-encapsulated HULC could be a potential circulating biomarker for human PDAC.


Assuntos
Biomarcadores Tumorais/sangue , Vesículas Extracelulares/patologia , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/patologia , Neoplasias Pancreáticas/sangue , Neoplasias Pancreáticas/patologia , Regulação para Cima/genética , Adenocarcinoma/sangue , Adenocarcinoma/genética , Adenocarcinoma/patologia , Idoso , Idoso de 80 Anos ou mais , Animais , Apoptose/genética , Carcinoma Ductal Pancreático/sangue , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Transição Epitelial-Mesenquimal/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , RNA Longo não Codificante/genética , Ativação Transcricional/genética
3.
J Cancer Res Clin Oncol ; 146(2): 467-475, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31734835

RESUMO

PURPOSE: The expression of active tissue factor (TF) on the surface of microvesicles (MVs) is essential for the activation of the coagulation system and transduction of the signaling pathways in cancer cells. In its use as a biomarker for cancer-associated venous thromboembolism (VTE), TF has shown high expression variability. As a contribution to this discussion, we present a study investigating plasma samples from patients with various progressive tumors at high risk for VTE. METHODS: Based on our previous study uncovering microvesicles (MVs), the larger ectosome-like extracellular vesicles (EV), as the major source of TF activity in EV preparations, we now determined TF activity on enriched MVs isolated from plasma of cancer patients and compared it with that on MVs from healthy individuals. RESULTS: We found considerably higher amounts of MVs as well as higher levels of MV-bound TF activities in the plasma of cancer patients. We also show that preparations from plasma of cancer patients have the potency to induce ERK phosphorylation in a human tumor cell line through proteinase-activated receptor two (PAR2) activation. CONCLUSION: We suggest that MVs instead of whole EV preparations, and TF activity rather than its antigenic quantification should be used in clinical studies for identifying patients with progressive tumors at high risk for VTE.


Assuntos
Tromboplastina/metabolismo , Idoso , Estudos de Casos e Controles , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/patologia , Feminino , Humanos , Sistema de Sinalização das MAP Quinases , Masculino , Neoplasias Pancreáticas/sangue , Neoplasias Pancreáticas/patologia , Fosforilação , Tromboplastina/biossíntese , Tromboembolia Venosa/sangue , Tromboembolia Venosa/patologia
4.
Biomed Res Int ; 2019: 9258075, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31309120

RESUMO

Cancer metastasis has been demonstrated as it is the culmination of a cascade of priming steps. Increasing evidence has shown that tumor-derived molecular components (TDMCs) are known as extra cellular vesicle and nonvesicle factors and serve as versatile intercellular communication vehicles which can mediate signaling in the tumor microenvironment while creating the premetastatic niche. Noncoding RNAs (ncRNAs) as one of the TDMCs have been proved in participating in the formation of the premetastatic niche. Understanding the premetastatic niche formation mechanisms through TDMCs, especially ncRNAs may open a new avenue for cancer metastasis therapeutic strategies. In this review, recent findings regarding ncRNAs function were summarized, and then the interaction with the premetastatic niche formation was studied, which highlight the potential of using ncRNAs for cancer diagnosis and therapeutic effect.


Assuntos
Comunicação Celular , Vesículas Extracelulares/metabolismo , Neoplasias/metabolismo , RNA Neoplásico/metabolismo , RNA não Traduzido/metabolismo , Transdução de Sinais , Microambiente Tumoral , Animais , Vesículas Extracelulares/patologia , Humanos , Metástase Neoplásica , Neoplasias/patologia
5.
Clin Chim Acta ; 497: 130-136, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31361990

RESUMO

Diabetes and diabetic vascular complications are now the leading cause of death in the world. The effects of traditional medical treatment are usually limited and accompanied by many side effects, such as hypoglycemia, obesity, liver and kidney damage, and gastrointestinal adverse reactions. Thus, it is urgent to explore some new strategies for the treatment of patients with diabetes. Recently, extracellular vesicles have received increased attention because of their emerging roles of cell-to-cell communication under physiological and pathological conditions. In addition, because of their abundant existence in almost all body fluids, as well as their plentiful cargos of bioactive proteins and miRNAs they carry, extracellular vesicles have a strong potential for therapeutic and diagnostic applications in many metabolic diseases, such as obesity and insulin resistance. Here, with the aim of providing the basis for the development of new treatments for diabetes, we review current understanding of extracellular vesicles and the critical roles it has played in the onset and progression of diabetes and diabetic complications.


Assuntos
Diabetes Mellitus/metabolismo , Vesículas Extracelulares/metabolismo , Diabetes Mellitus/patologia , Vesículas Extracelulares/patologia , Humanos
6.
Rev Cardiovasc Med ; 20(2): 59-71, 2019 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-31344998

RESUMO

Remote ischemic conditioning of the heart (including pre-, per-, and post-conditioning) is a phenomenon where short episodes of non-lethal ischemia in the distant vessels within the heart or distant organs from the heart protects the myocardium against sustained ischemia/reperfusion injury. Several pathways have been proposed to be involved in the mechanisms of Remote ischemic conditioning. While triggers of Remote ischemic conditioning act in preconditioned areas, its mediators transduce protective signals via humoral or neuronal pathways to the heart. Remote ischemic conditioning is mediated via receptor and nonreceptor signaling through secondary mediators, which transfer the signal within the cardiomyocyte and activate cardioprotective pathways that lead to higher resistance of the heart to ischemia/reperfusion. Apparently, identification of endogenous signal molecules involved in the mechanisms of Remote ischemic conditioning have therapeutic implications in the management of patients suffering from myocardial ischemia through the development of diverse beneficial effects. Recently, different non-coding RNAs such as microRNAs or long non-coding RNAs have been identified as emerging factors that trigger protective mechanisms in the heart. These non-coding RNAs are transferred to the heart via extracellular vesicles that exert remote cardioprotection. This review is intended to summarize the existing knowledge about the potential role of extracellular vesicles as humoral transmitters of Remote ischemic conditioning and emphasize the involvement of non-coding RNAs in the mechanism of cardioprotection by Remote ischemic conditioning.


Assuntos
Vesículas Extracelulares/metabolismo , Pós-Condicionamento Isquêmico/métodos , Precondicionamento Isquêmico/métodos , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/metabolismo , RNA não Traduzido/metabolismo , Animais , Vesículas Extracelulares/genética , Vesículas Extracelulares/patologia , Regulação da Expressão Gênica , Humanos , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/patologia , RNA não Traduzido/genética , Transdução de Sinais
7.
Zhonghua Gan Zang Bing Za Zhi ; 27(6): 407-410, 2019 Jun 20.
Artigo em Chinês | MEDLINE | ID: mdl-31357753

RESUMO

Liver fibrosis is a pathophysiological process characterized by abnormal accumulation of connective tissues in the liver caused by chronic liver injuries, in which the activation and migration of hepatic stellate cells (HSCs) play a central role. Extracellular vesicles (EVs) are a class of nanoscale, bilayer lipid enveloped vesicles secreted by almost all cells. EVs are of great interest in liver pathology because they have been found to mediate the communication between cells and regulate cellular microenvironment via horizontal transfer of their cargoes. EVs carry bioactive cargoes including proteins, lipids and RNA molecules, and are involved in the activation of HSCs during liver fibrogenesis.


Assuntos
Vesículas Extracelulares , Cirrose Hepática , Pesquisa , Vesículas Extracelulares/patologia , Células Estreladas do Fígado , Humanos , Cirrose Hepática/patologia , Pesquisa/tendências
8.
Immunohorizons ; 3(6): 186-193, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-31356164

RESUMO

Accumulating evidence shows that extracellular vesicles (EVs) secreted by immune cells play an important role in intercellular communication. In the current report, we show that EVs released from wild-type bone marrow-derived dendritic cells (BMDCs) transfer TLRs to TLR4-knockout (TLR4KO) BMDCs and increase cellular responsiveness to LPS in recipient cells. The transferred EVs have exosomal characteristics and induce the activation of NF-κB signaling pathways in recipient cells. We further show that BMDC-derived EVs can promote LPS-induced inflammation in TLR4KO mice in vivo. These results indicate that functional TLR4 can be transferred from wild-type to TLR4KO BMDCs through exosome-like EVs.


Assuntos
Células Dendríticas/imunologia , Exossomos/metabolismo , Vesículas Extracelulares/metabolismo , Inflamação/imunologia , Receptor 4 Toll-Like/metabolismo , Animais , Diferenciação Celular , Células Cultivadas , Exossomos/patologia , Vesículas Extracelulares/patologia , Lipopolissacarídeos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/genética
9.
J Exp Clin Cancer Res ; 38(1): 317, 2019 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-31319863

RESUMO

BACKGROUND: Molecular markers for prostate cancer (PCa) are required to improve the early definition of patient outcomes. Atypically large extracellular vesicles (EVs), referred as "Large Oncosomes" (LO), have been identified in highly migratory and invasive PCa cells. We recently developed and characterized the DU145R80 subline, selected from parental DU145 cells as resistant to inhibitors of mevalonate pathway. DU145R80 showed different proteomic profile compared to parental DU145 cells, along with altered cytoskeleton dynamics and a more aggressive phenotype. METHODS: Immunofluorescence staining and western blotting were used to identify blebbing and EVs protein cargo. EVs, purified by gradient ultra-centrifugations, were analyzed by tunable resistive pulse sensing and multi-parametric flow cytometry approach coupled with high-resolution imaging technologies. LO functional effects were tested in vitro by adhesion and invasion assays and in vivo xenograft model in nude mice. Xenograft and patient tumor tissues were analyzed by immunohistochemistry. RESULTS: We found spontaneous blebbing and increased shedding of LO from DU145R80 compared to DU145 cells. LO from DU145R80, compared to those from DU145, carried increased amounts of key-molecules involved in PCa progression including integrin alpha V (αV-integrin). By incubating DU145 cells with DU145R80-derived LO we demonstrated that αV-integrin on LO surface was functionally involved in the increased adhesion and invasion of recipient cells, via AKT. Indeed either the pre-incubation of LO with an αV-integrin blocking antibody, or a specific AKT inhibition in recipient cells are able to revert the LO-induced functional effects. Moreover, DU145R80-derived LO also increased DU145 tumor engraftment in a mice model. Finally, we identified αV-integrin positive LO-like structures in tumor xenografts as well as in PCa patient tissues. Increased αV-integrin tumor expression correlated with high Gleason score and lymph node status. CONCLUSIONS: Overall, this study is the first to demonstrate the critical role of αV-integrin positive LO in PCa aggressive features, adding new insights in biological function of these large EVs and suggesting their potential use as PCa prognostic markers.


Assuntos
Vesículas Extracelulares/patologia , Integrina alfaV/metabolismo , Neoplasias da Próstata/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Adesão Celular , Linhagem Celular Tumoral , Transição Epitelial-Mesenquimal , Vesículas Extracelulares/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Camundongos , Camundongos Nus , Gradação de Tumores , Invasividade Neoplásica , Transplante de Neoplasias , Neoplasias da Próstata/metabolismo , Proteômica/métodos , Regulação para Cima
10.
Int J Cancer ; 145(9): 2521-2534, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31216364

RESUMO

The major cause of melanoma mortality is metastasis to distant organs, including lungs and brain. Reciprocal interactions of metastasizing tumor cells with stromal cells in secondary sites play a critical role in all stages of tumorigenesis and metastasis. Changes in the metastatic microenvironment were shown to precede clinically relevant metastases, and may occur prior to the arrival of disseminated tumor cells to the distant organ, thus creating a hospitable "premetastatic niche." Exosomes secreted by tumor cells were demonstrated to play an important role in the preparation of a hospitable metastatic niche. However, the functional role of melanoma-derived exosomes on metastatic niche formation, and the downstream pathways activated in stromal cells at the metastatic niche are largely unresolved. Here we show that extracellular vesicles (EVs) secreted by metastatic melanoma cells that spontaneously metastasize to lungs and to brain, activate proinflammatory signaling in lung fibroblasts and in astrocytes. Interestingly, unlike paracrine signaling by melanoma cells, EVs secreted by metastatic melanoma cells instigated a proinflammatory gene signature in lung fibroblasts but did not activate wound-healing functions, suggesting that tumor cell-secreted EVs activate distinct CAF characteristics and tumor-promoting functions. Moreover, melanoma-secreted EVs also activated proinflammatory signaling in astrocytes, indicating that EV-mediated reprogramming of stromal cells is a general mechanism of modulating the metastatic niche in multiple distant organs. Thus, our study demonstrates that melanoma-derived EVs reprogram tumor-promoting functions in stromal cells in a distinct manner, implicating a central role for tumor-derived EV signaling in promoting the formation of an inflammatory metastatic niche.


Assuntos
Vesículas Extracelulares/patologia , Inflamação/patologia , Melanoma/patologia , Transdução de Sinais/fisiologia , Microambiente Tumoral/fisiologia , Animais , Astrócitos/patologia , Exossomos/patologia , Fibroblastos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , Comunicação Parácrina/fisiologia , Células Estromais/patologia
11.
PLoS One ; 14(6): e0218270, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31211806

RESUMO

Extracellular vesicles (EVs) are increasingly recognized as important mediators of intercellular communication that carry protein, lipids, and nucleic acids via the circulation to target cells whereupon they mediate physiological changes. In pregnancy, EVs are released in high quantities from the placenta and have been postulated to target multiple cell types, including those of the vascular and immune systems. However, most studies of pregnancy-associated EVs have used clinical samples and in vitro models; to date, few studies have taken advantage of murine models in which pregnancy can be precisely timed and manipulated. In this study, we used a murine model to determine whether the quantity of EVs is altered during healthy pregnancy and during inflammation-associated preterm birth. To facilitate data analysis, we developed a novel software package, tidyNano, an R package that provides functions to import, clean, and quickly summarize raw data generated by the nanoparticle tracking device, NanoSight (Malvern Panalytical). We also developed shinySIGHT, a Shiny web application that allows for interactive exploration and visualization of EV data. In mice, EV concentration in blood increased linearly across pregnancy, with significant rises at GD14.5 and 17.5 relative to EV concentrations in nonpregnant females. Additionally, lipopolysaccharide treatment resulted in a significant reduction in circulating EV concentrations relative to vehicle-treated controls at GD16.5 within 4 hours. Use of tidyNano facilitated rapid analysis of EV data; importantly, this package provides a straightforward framework by which diverse types of large datasets can be simply and efficiently analyzed, is freely available under the MIT license, and is hosted on GitHub (https://nguyens7.github.io/tidyNano/). Our data highlight the utility of the mouse as a model of EV biology in pregnancy, and suggest that placental dysfunction is associated with reduced circulating EVs.


Assuntos
Nanopartículas/química , Nanotecnologia , Placenta/metabolismo , Nascimento Prematuro/epidemiologia , Animais , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/genética , Visualização de Dados , Modelos Animais de Doenças , Exossomos/metabolismo , Exossomos/patologia , Vesículas Extracelulares/efeitos dos fármacos , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/patologia , Feminino , Humanos , Sistema Imunitário/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Camundongos , Placenta/patologia , Gravidez , Nascimento Prematuro/patologia , Trofoblastos/metabolismo , Trofoblastos/patologia
12.
Int J Mol Sci ; 20(9)2019 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-31052525

RESUMO

Liver diseases are perpetuated by the orchestration of hepatocytes and other hepatic non-parenchymal cells. These cells communicate and regulate with each other by secreting mediators such as peptides, hormones, and cytokines. Extracellular vesicles (EVs), small particles secreted from cells, contain proteins, DNAs, and RNAs as cargos. EVs have attracted recent research interests since they can communicate information from donor cells to recipient cells thereby regulating physiological events via delivering of specific cargo mediators. Previous studies have demonstrated that liver cells secrete elevated numbers of EVs during diseased conditions, and those EVs are internalized into other liver cells inducing disease-related reactions such as inflammation, angiogenesis, and fibrogenesis. Reactions in recipient cells are caused by proteins and RNAs carried in disease-derived EVs. This review summarizes cell-to-cell communication especially via EVs in the pathogenesis of liver diseases and their potential as a novel therapeutic target.


Assuntos
Comunicação Celular , Vesículas Extracelulares/patologia , Hepatócitos/patologia , Hepatopatias/patologia , Animais , Vesículas Extracelulares/metabolismo , Hepatócitos/metabolismo , Humanos , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Cirrose Hepática/terapia , Hepatopatias/metabolismo , Hepatopatias/terapia , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Neovascularização Patológica/terapia
13.
Int J Mol Sci ; 20(10)2019 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-31130715

RESUMO

The primary cause of mortality among patients with cancer is the progression of the tumor, better known as cancer invasion and metastasis. Cancer progression involves a series of biologically important steps in which the cross-talk between cancer cells and the cells in the surrounding environment is positioned as an important issue. Notably, angiogenesis is a key tumorigenic phenomenon for cancer progression. Cancer-related extracellular vesicles (EVs) commonly contribute to the modulation of a microenvironment favorable to cancer cells through their function of cell-to-cell communication. Vascular-related cells such as endothelial cells (ECs) and platelets activated by cancer cells and cancer-derived EVs develop procoagulant and proinflammatory statuses, which help excite the tumor environment, and play major roles in tumor progression, including in tumor extravasation, tumor cell microthrombi formation, platelet aggregation, and metastasis. In particular, cancer-derived EVs influence ECs, which then play multiple roles such as contributing to tumor angiogenesis, loss of endothelial vascular barrier by binding to ECs, and the subsequent endothelial-to-mesenchymal transition, i.e., extracellular matrix remodeling. Thus, cell-to-cell communication between cancer cells and ECs via EVs may be an important target for controlling cancer progression. This review describes the current knowledge regarding the involvement of EVs, especially exosomes derived from cancer cells, in EC-related cancer progression.


Assuntos
Células Endoteliais/patologia , Vesículas Extracelulares/patologia , Neoplasias/patologia , Animais , Progressão da Doença , Exossomos/patologia , Humanos , Metástase Neoplásica/patologia , Neoplasias/irrigação sanguínea , Neovascularização Patológica/patologia
14.
Breast J ; 25(4): 691-695, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31079422

RESUMO

Currently, tumor biopsies are used for breast cancer molecular subtyping. Biopsies are associated with various pathological changes and are thought to contribute to the dissemination of tumor cells. Extracellular vesicles shed by tumor cells into circulation exhibit the molecular signature of the parent cells. Herein, we show that proteomic analysis of circulating EV can discriminate BC patients from healthy subjects and indicate stage of the disease. Also, we performed a correlation between the BC molecular subtype using plasma EV and immunohistochemistry of tumor biopsies. Circulating EV may represent a useful, non-invasive tool to study the molecular makeup of BC tumors.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias da Mama/patologia , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/patologia , Proteína ADAM12/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/sangue , Estudos de Casos e Controles , Linhagem Celular Tumoral , Receptor alfa de Estrogênio/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Proteômica , Receptor ErbB-2/metabolismo , Receptores de Progesterona/metabolismo , beta Catenina/metabolismo
15.
Cell Mol Life Sci ; 76(12): 2463-2476, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31028424

RESUMO

Extracellular vesicles (EVs) are membrane-surrounded structures that transmit biologically important molecules from the releasing to target cells, thus providing a novel intercellular communication mechanism. Since EVs carry their cargo in a protected form and their secretion is generally increased in tumorigenesis, EVs hold a great potential for early cancer diagnosis. By 3D culturing, we provide evidence that colorectal cancer (CRC) patient-derived organoids, representing a state-of-the-art established and essential approach for studying human CRC, is a suitable model for EV analysis. When testing the effects of major factors promoting CRC progression on EV release in the organoid model, we observed that Apc mutation, leading to uncontrolled Wnt activation and thus to tumorigenesis in the vast majority in CRC patients, critically induces EV release by activating the Wnt pathway. Furthermore, the extracellular matrix component collagen, known to accumulate in tumorigenesis, enhances EV secretion as well. Importantly, we show that fibroblast-derived EVs induce colony formation of CRC organoid cells under hypoxia. In contrast, there was no major effect of tumor cell-derived EVs on the activation of fibroblasts. Collectively, our results with CRC and Apc-mutant adenoma organoids identify Apc mutation and collagen deposition as critical factors for increasing EV release from tumors. Furthermore, we provide evidence that stromal fibroblast-derived EVs contribute to tumorigenesis under unfavorable conditions in CRC.


Assuntos
Proteína da Polipose Adenomatosa do Colo/genética , Neoplasias Colorretais/patologia , Vesículas Extracelulares/patologia , Intestinos/patologia , Organoides/patologia , Animais , Carcinogênese/genética , Carcinogênese/metabolismo , Carcinogênese/patologia , Linhagem Celular Tumoral , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Progressão da Doença , Vesículas Extracelulares/genética , Humanos , Camundongos Endogâmicos C57BL , Mutação , Organoides/metabolismo , Células Tumorais Cultivadas , Via de Sinalização Wnt
16.
Oncogene ; 38(21): 4182-4196, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30936459

RESUMO

Insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) is a multifunctional RNA-binding protein with an oncofetal pattern of expression shown to be implicated in the development of a variety of malignancies. In this study, we explored the role and mechanisms of IGF2BP1 in melanoma development and progression. In two different in vivo models, we showed that although genetic deletion or shRNA-mediated suppression of IGF2BP1 did not affect primary tumor formation, it drastically suppressed lung metastasis. Here we demonstrated that extracellular vesicles (EVs) secreted by melanoma cells mediate the effects of IGF2BP1 on metastasis: EVs from the IGF2BP1 knockdown melanoma cells failed to promote metastasis, whereas EVs isolated from IGF2BP1-overexpressed melanoma cells further accelerated EV-induced metastasis. Moreover, the EVs from IGF2BP1 knockdown melanoma cells inhibited fibronectin deposition and accumulation of CD45+ cells in the lungs compared with control EVs, thus blocking the pre-metastatic niche formation potential of EVs. IGF2BP1 knockdown did not affect size, number, or protein/RNA concentration of secreted EVs or their uptake by recipient cells in vitro or in vivo. However, RNA-sequencing and proteomics analysis of the EVs revealed differential expression in a number of mRNA, proteins, and miRNAs. This suggested that IGF2BP1 is intimately involved in the regulation of the cargo of EVs, thereby affecting the pro-metastatic function of melanoma-derived EVs. To the best of our knowledge, this is the first study that demonstrates the role of RNA-binding protein IGF2BP1 in EV-mediated promotion of melanoma metastasis and may provide novel avenues for the development of metastatic inhibitors.


Assuntos
Vesículas Extracelulares/genética , Melanoma/genética , Melanoma/patologia , Proteínas de Ligação a RNA/genética , Animais , Linhagem Celular Tumoral , Movimento Celular/genética , Vesículas Extracelulares/patologia , Feminino , Fibronectinas/genética , Regulação Neoplásica da Expressão Gênica/genética , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C3H , MicroRNAs/genética , RNA Mensageiro/genética
17.
Hypertension ; 73(5): 1112-1119, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30929513

RESUMO

NEP (neprilysin) is a widely expressed membrane-bound metalloprotease, which binds and cleaves a variety of peptides including vasodilators, natriuretics, and diuretics. Higher levels of NEP result in hypertension-a cardinal feature of the placental disease preeclampsia. Syncytiotrophoblast-derived extracellular vesicles (EVs), comprising microvesicles and exosomes, are released into the peripheral circulation in pregnancy and are postulated as a key mechanism coupling placental dysfunction and maternal phenotype in preeclampsia. We aimed to determine whether higher levels of active NEP are found in syncytiotrophoblast-derived EVs in preeclampsia compared with normal pregnancy. Using immunostaining and Western blotting, we first demonstrated that NEP levels are greater not only in preeclampsia placental tissue but also in syncytiotrophoblast-derived microvesicles and exosomes isolated from preeclampsia placentas ( P<0.05, n=5). We confirmed placental origin using antibody-coated magnetic beads to isolate NEP-bound vesicles, finding that they stain for placental alkaline phosphatase. NEP on syncytiotrophoblast-derived EVs is active and inhibited by thiorphan ( P<0.01, n=3; specific inhibitor). Syncytiotrophoblast-derived microvesicles, isolated from peripheral plasma, demonstrated higher NEP expression in preeclampsia using flow cytometry ( P<0.05, n=8). We isolated plasma exosomes using size-exclusion chromatography and showed greater NEP activity in preeclampsia ( P<0.05, n=8). These findings show that the placenta releases active NEP into the maternal circulation on syncytiotrophoblast-derived EVs, at significantly greater levels in preeclampsia. NEP has pathological roles in hypertension, heart failure, and amyloid deposition, all of which are features of preeclampsia. Circulating syncytiotrophoblast-derived EV-bound NEP thus may contribute to the pathogenesis of this disease.


Assuntos
Vesículas Extracelulares/metabolismo , Neprilisina/biossíntese , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Trofoblastos/metabolismo , Adulto , Pressão Sanguínea/fisiologia , Western Blotting , Vesículas Extracelulares/patologia , Feminino , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Placenta/patologia , Pré-Eclâmpsia/patologia , Pré-Eclâmpsia/fisiopatologia , Gravidez , Trofoblastos/patologia
18.
Autoimmun Rev ; 18(6): 593-606, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30959208

RESUMO

Systemic vasculitis is diverse group of autoimmune disorders which are characterized by inflammation of blood vessel walls with deep aching and burning pain. Their underlying etiology and pathophysiology still remain poorly understood. Extracellular vesicles (EVs), including exosomes, microvesicles (MVs), and apoptotic bodies, are membrane vesicular structures that are released either during cell activation, or when cells undergo programmed cell death, including apoptosis, necroptosis, and pyroptosis. Although EVs were thought as cell dusts, but now they have been found to be potently active since they harbor bioactive molecules, such as proteins, lipids, nucleic acids, or multi-molecular complexes. EVs can serve as novel mediators for cell-to-cell communications by delivery bioactive molecules from their parental cells to the recipient cells. Earlier studies mainly focused on MVs budding from membrane surface. Recent studies demonstrated that EVs may also carry molecules from cytoplasm or even from nucleus of their parental cells, and these EVs may carry autoantigens and are important in vasculitis. EVs may play important roles in vasculitis through their potential pathogenic involvements in inflammation, autoimmune responses, procoagulation, endothelial dysfunction/damage, angiogenesis, and intimal hyperplasia. EVs have also been used as specific biomarkers for diagnostic use or disease severity monitoring. In this review, we have focused on the aspects of EV biology most relevant to the pathogenesis of vasculitis, discussed their perspective insights, and summarized the exist literature on EV relevant studies in vasculitis, therefore provides an integration of current knowledge regarding the novel role of EVs in systemic vasculitis.


Assuntos
Doenças Autoimunes/patologia , Vesículas Extracelulares/patologia , Vasculite Sistêmica/patologia , Humanos
19.
BMC Cancer ; 19(1): 373, 2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-31014274

RESUMO

BACKGROUND: There is a paucity of plasma-based biomarkers that prospectively segregate the outcome of patients with head and neck squamous-cell carcinoma (HNSCC) treated with chemoradiation therapy (CRT). Plasma extracellular vesicles (EVs) might be an alternative source for discovery of new specific markers present in patients with HNSCC, which could help to re-direct patients to appropriate curative therapies without delay. METHODS: In order to identify new markers in plasma compartments, Cholerae toxin B chain (CTB) and Annexin V (AV) were used to isolate EVs from pooled plasma samples from patients with locally advanced HNSCC who responded (CR, n = 6) or presented incomplete response (NR, n = 6) to CRT. The crude plasma and EVs cargo were screened by antibody array. RESULTS: Of the 370 polypeptides detected, 119 proteins were specific to NR patients while 38 were exclusive of the CR subjects. The Gene Set Enrichment Analysis (GSEA) and Search Tool for the Retrieval of Interacting Genes (STRING) database analysis indicated that the content of circulating plasma EVs might have a relevant function for the tumor intercellular communication in the HNSCC patients. CONCLUSION: This study provides a list of potential markers present in plasma compartments that might contribute to the development of tools for prediction and assessment of CRT response and potentially guide therapeutic decisions in this context.


Assuntos
Biomarcadores Tumorais/sangue , Carcinoma de Células Escamosas/patologia , Quimiorradioterapia/métodos , Vesículas Extracelulares/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Adulto , Idoso , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/terapia , Ensaios Clínicos Fase II como Assunto , Vesículas Extracelulares/patologia , Feminino , Seguimentos , Neoplasias de Cabeça e Pescoço/sangue , Neoplasias de Cabeça e Pescoço/terapia , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Análise Serial de Proteínas , Mapas de Interação de Proteínas
20.
Prostate ; 79(9): 1032-1042, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31018022

RESUMO

BACKGROUND: Proteomic profiling of extracellular vesicles (EVs) from prostate cancer (PCa) and normal prostate cell lines, led to the identification of new candidate PCa markers. These proteins included the nuclear exportin proteins XPO1 (also known as CRM1), the EV-associated PDCD6IP (also known as ALIX), and the previously published fatty acid synthase FASN. In this study, we investigated differences in expression of XPO1 and PDCD6IP on well-characterized prostate cancer cohorts using mass spectrometry and tissue microarray (TMA) immunohistochemistry to determine their diagnostic and prognostic value. METHODS: Protein fractions from 67 tissue samples (n = 33 normal adjacent prostate [NAP] and n = 34 PCa) were analyzed by mass spectrometry (nano-LC-MS-MS). Label-free quantification of EVs was performed to identify differentially expressed proteins between PCa and NAP. Prognostic evaluation of the candidate markers was performed with a TMA, containing 481 radical prostatectomy samples. Samples were stained for the candidate markers and correlated with patient information and clinicopathological outcome. RESULTS: XPO1 was higher expressed in PCa compared to NAP in the MS data analysis (P > 0.0001). PDCD6IP was not significantly higher expressed (P = 0.0501). High cytoplasmic XPO1 staining in the TMA immunohistochemistry, correlated in a multivariable model with high Gleason scores (P = 0.002) and PCa-related death (P = 0.009). CONCLUSION: High expression of cytoplasmic XPO1 shows correlation with prostate cancer and has added clinical value in tissue samples. Furthermore, as an extracellular vesicles-associated protein, it might be a novel relevant liquid biomarker.


Assuntos
Biomarcadores Tumorais/biossíntese , Proteínas de Ligação ao Cálcio/biossíntese , Proteínas de Ciclo Celular/biossíntese , Complexos Endossomais de Distribuição Requeridos para Transporte/biossíntese , Vesículas Extracelulares/metabolismo , Carioferinas/biossíntese , Neoplasias da Próstata/metabolismo , Receptores Citoplasmáticos e Nucleares/biossíntese , Idoso , Vesículas Extracelulares/patologia , Ácido Graxo Sintase Tipo I/biossíntese , Humanos , Imuno-Histoquímica , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Gradação de Tumores , Neoplasias da Próstata/patologia , Análise Serial de Tecidos
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