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https://www.readbyqxmd.com/read/28936266/a-novel-method-for-the-isolation-of-extracellular-vesicles-and-rna-from-urine
#1
Anna Markowska, R Scott Pendergrast, J Stephen Pendergrast, P Shannon Pendergrast
The discovery of urinary extracellular biomarkers has been impeded by the lack of efficient methods for the isolation of extracellular vesicles (EVs: exosomes and microvesicles) and extracellular nucleic acid (RNA and DNA) from urine. Ultracentrifugation (UC), considered the gold standard for vesicle isolation from many biofluids, is efficacious but laborious, and, like most commercially available methods, is unable to isolate enough material from small volumes for protein or RNA-based biomarker discovery. We have developed a novel precipitation method for the isolation of EVs and nucleic acids from urine...
January 2017: J Circ Biomark
https://www.readbyqxmd.com/read/28934507/a-comprehensive-cell-specific-microrna-catalogue-of-human-peripheral-blood
#2
Simonas Juzenas, Geetha Venkatesh, Matthias Hübenthal, Marc P Hoeppner, Zhipei Gracie Du, Maren Paulsen, Philip Rosenstiel, Philipp Senger, Martin Hofmann-Apitius, Andreas Keller, Limas Kupcinskas, Andre Franke, Georg Hemmrich-Stanisak
With this study, we provide a comprehensive reference dataset of detailed miRNA expression profiles from seven types of human peripheral blood cells (NK cells, B lymphocytes, cytotoxic T lymphocytes, T helper cells, monocytes, neutrophils and erythrocytes), serum, exosomes and whole blood. The peripheral blood cells from buffy coats were typed and sorted using FACS/MACS. The overall dataset was generated from 450 small RNA libraries using high-throughput sequencing. By employing a comprehensive bioinformatics and statistical analysis, we show that 3' trimming modifications as well as composition of 3' added non-templated nucleotides are distributed in a lineage-specific manner-the closer the hematopoietic progenitors are, the higher their similarities in sequence variation of the 3' end...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28923936/isolation-of-exosomes-from-whole-blood-by-integrating-acoustics-and-microfluidics
#3
Mengxi Wu, Yingshi Ouyang, Zeyu Wang, Rui Zhang, Po-Hsun Huang, Chuyi Chen, Hui Li, Peng Li, David Quinn, Ming Dao, Subra Suresh, Yoel Sadovsky, Tony Jun Huang
Exosomes are nanoscale extracellular vesicles that play an important role in many biological processes, including intercellular communications, antigen presentation, and the transport of proteins, RNA, and other molecules. Recently there has been significant interest in exosome-related fundamental research, seeking new exosome-based biomarkers for health monitoring and disease diagnoses. Here, we report a separation method based on acoustofluidics (i.e., the integration of acoustics and microfluidics) to isolate exosomes directly from whole blood in a label-free and contact-free manner...
September 18, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28919558/cellular-uptake-of-extracellular-vesicles-is-mediated-by-clathrin-independent-endocytosis-and-macropinocytosis
#4
Helena Costa Verdera, Jerney J Gitz-Francois, Raymond M Schiffelers, Pieter Vader
Recent evidence has established that extracellular vesicles (EVs), including exosomes and microvesicles, form an endogenous transport system through which biomolecules, including proteins and RNA, are exchanged between cells. This endows EVs with immense potential for drug delivery and regenerative medicine applications. Understanding the biology underlying EV-based intercellular transfer of cargo is of great importance for the development of EV-based therapeutics. Here, we sought to characterize the cellular mechanisms involved in EV uptake...
September 14, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28918518/micro-rna-profiling-of-exosomes-from-marrow-derived-mesenchymal-stromal-cells-in-patients-with-acute-myeloid-leukemia-implications-in-leukemogenesis
#5
Juliana Barrera-Ramirez, Jessie R Lavoie, Harinad B Maganti, William L Stanford, Caryn Ito, Mitchell Sabloff, Marjorie Brand, Michael Rosu-Myles, Yevgeniya Le, David S Allan
Gene regulatory networks in AML may be influenced by microRNAs (miRs) contained in exosomes derived from bone marrow mesenchymal stromal cells (MSCs). We sequenced miRs from exosomes isolated from marrow-derived MSCs from patients with AML (n = 3) and from healthy controls (n = 3; not age-matched). Known targets of mIRs that were significantly different in AML-derived MSC exosomes compared to controls were identified. Of the five candidate miRs identified by differential packaging in exosomes, only miR-26a-5p and miR-101-3p were significantly increased in AML-derived samples while miR-23b-5p, miR-339-3p and miR-425-5p were significantly decreased...
September 16, 2017: Stem Cell Reviews
https://www.readbyqxmd.com/read/28910400/293texosome-mediated-mir-146a-transfer-suppresses-type-i-interferon-response-and-facilitates-ev71-infection
#6
Yuxuan Fu, Li Zhang, Fang Zhang, Ting Tang, Qi Zhou, Chunhong Feng, Yu Jin, Zhiwei Wu
Exosomes can transfer genetic materials between cells. Their roles in viral infections are beginning to be appreciated. Researches have shown that exosomes released from virus-infected cells contain a variety of viral and host cellular factors that are able to modulate recipient's cellular response and result in productive infection of the recipient host. Here, we showed that EV71 infection resulted in upregulated exosome secretion and differential packaging of the viral genomic RNA and miR-146a into exosomes...
September 14, 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28906477/microrna-signaling-in-embryo-development
#7
REVIEW
Nicole Gross, Jenna Kropp, Hasan Khatib
Expression of microRNAs (miRNAs) is essential for embryonic development and serves important roles in gametogenesis. miRNAs are secreted into the extracellular environment by the embryo during the preimplantation stage of development. Several cell types secrete miRNAs into biological fluids in the extracellular environment. These fluid-derived miRNAs have been shown to circulate the body. Stable transport is dependent on proper packaging of the miRNAs into extracellular vesicles (EVs), including exosomes. These vesicles, which also contain RNA, DNA and proteins, are on the forefront of research on cell-to-cell communication...
September 14, 2017: Biology
https://www.readbyqxmd.com/read/28898757/extracellular-matrix-derived-extracellular-vesicles-promote-cardiomyocyte-growth-and-electrical-activity-in-engineered-cardiac-atria
#8
Minae An, Kihwan Kwon, Junbeom Park, Dong-Ryeol Ryu, Jung-A Shin, Jihee Lee Kang, Ji Ha Choi, Eun-Mi Park, Kyung Eun Lee, Minna Woo, Minsuk Kim
Extracellular matrix (ECM) plays a critical role in the provision of the necessary microenvironment for the proper regeneration of the cardiac tissue. However, specific mechanisms that lead to ECM-mediated cardiac regeneration are not well understood. To elucidate the potential mechanisms, we investigated ultra-structures of the cardiac ECM using electron microscopy. Intriguingly, we observed large quantities of micro-vesicles from decellularized right atria. RNA and protein analyses revealed that these contained exosomal proteins and microRNAs (miRNAs), which we referred to herein as ECM-derived extracellular vesicles (ECM-EVs)...
September 4, 2017: Biomaterials
https://www.readbyqxmd.com/read/28895353/-circulating-long-noncoding-rnas-as-biomarkers-in-tumor-diagnosis
#9
Nan Jiang, Haihua Tian, Jinchang Pan, Zhaohui Gong
Long noncoding RNAs (lncRNAs) are involved in vital life processes of gene expression, epigenetic regulation and X-chromosome inactivation. lncRNAs are also closely associated with tumor initiation and progression. Moreover, lncRNAs may enter human circulation system in the form of microvesicle or exosome, or in combination with RNA binding protein. Interestingly, the circulating lncRNAs are widely existed in body fluids, such as blood and urine. We review the origin of circulating lncRNAs, and the detection methods as potential biomarkers...
June 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28889388/quantification-of-micrornas-in-urine-derived-specimens
#10
Susanne Fuessel, Andrea Lohse-Fischer, Dana Vu Van, Karsten Salomo, Kati Erdmann, Manfred P Wirth
MicroRNAs are small noncoding RNAs which regulate the expression of genes involved in a multitude of cellular processes. Dysregulation of microRNAs and-in consequence-of the affected pathways is frequently observed in numerous pathologies including cancers. Therefore, tumor-related alterations in microRNA expression and function can reflect molecular processes of tumor onset and progression qualifying microRNAs as potential diagnostic and prognostic biomarkers.In particular, microRNAs with differential expression in bladder cancer (BCa) might represent promising tools for noninvasive tumor detection in urine...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28883156/structural-insights-into-the-interaction-of-the-nuclear-exosome-helicase-mtr4-with-the-pre-ribosomal-protein-nop53
#11
Sebastian Falk, Jan-Niklas Tants, Jerôme Basquin, Matthias Thoms, Ed Hurt, Michael Sattler, Elena Conti
The nuclear exosome and the associated RNA helicase Mtr4 participate in the processing of several ribonucleoprotein particles (RNP), including the maturation of the large ribosomal subunit (60S). S. cerevisiae Mtr4 interacts directly with Nop53, a ribosomal biogenesis factor present in late pre-60S particles containing precursors of the 5.8S rRNA. The Mtr4-Nop53 interaction plays a pivotal role in the maturation of the 5.8S rRNA, providing a physical link between the nuclear exosome and the pre-60S RNP. An analogous interaction between Mtr4 and another ribosome biogenesis factor, Utp18, directs the exosome to an earlier pre-ribosomal particle...
September 7, 2017: RNA
https://www.readbyqxmd.com/read/28880477/exosomes-as-new-players-in-metabolic-organ-cross-talk
#12
REVIEW
Claudiane Guay, Romano Regazzi
Blood glucose homeostasis requires a constant communication between insulin-secreting and insulin-sensitive cells. A wide variety of circulating factors, including hormones, cytokines and chemokines work together to orchestrate the systemic response of metabolic organs to changes in the nutritional state. Failure in the coordination between these organs can lead to a rise in blood glucose levels and to the appearance of metabolic disorders such as diabetes mellitus. Exosomes are small extracellular vesicles (EVs) that are produced via the endosomal pathway and are released from the cells upon fusion of multivesicular bodies with the plasma membrane...
September 2017: Diabetes, Obesity & Metabolism
https://www.readbyqxmd.com/read/28878075/the-characterization-of-the-quasi-enveloped-hepatitis-e-virus-particles-released-by-the-cellular-exosomal-pathway
#13
Shigeo Nagashima, Masaharu Takahashi, Tominari Kobayashi, Tanggis, Tsutomu Nishizawa, Takashi Nishiyama, Putu Prathiwi Primadharsini, Hiroaki Okamoto
Our previous studies demonstrated that membrane-associated hepatitis E virus (HEV) particles-now considered "quasi-enveloped particles"-are present in the multivesicular body with intraluminal vesicles (exosomes) in infected cells, and that the release of HEV virions is related to the exosomal pathway. In this study, we characterized exosomes purified from culture supernatant of HEV-infected PLC/PRF/5 cells. Purified CD63-, CD9-, or CD81-positive exosomes derived from the culture supernatant of HEV-infected cells that had been cultivated in serum-free medium were found to contain HEV RNA and the viral capsid (ORF2) and ORF3 proteins, as determined by a RT-PCR and Western blotting, respectively...
September 6, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28877463/mpp6-incorporation-in-the-nuclear-exosome-contributes-to-rna-channeling-through-the-mtr4-helicase
#14
Sebastian Falk, Fabien Bonneau, Judith Ebert, Alexander Kögel, Elena Conti
The RNA-degrading exosome mediates the processing and decay of many cellular transcripts. In the yeast nucleus, the ubiquitous 10-subunit exosome core complex (Exo-9-Rrp44) functions with four conserved cofactors (Rrp6, Rrp47, Mtr4, and Mpp6). Biochemical and structural studies to date have shed insights into the mechanisms of the exosome core and its nuclear cofactors, with the exception of Mpp6. We report the 3.2-Å resolution crystal structure of a S. cerevisiae Exo-9-Mpp6 complex, revealing how linear motifs in the Mpp6 middle domain bind Rrp40 via evolutionary conserved residues...
September 5, 2017: Cell Reports
https://www.readbyqxmd.com/read/28862291/trans-activation-response-element-rna-is-detectable-in-the-plasma-of-a-subset-of-aviremic-hiv-1-infected-patients
#15
Anžej Hladnik, Jana Ferdin, Katja Goričar, G Steven Deeks, M Boris Peterlin, Ana Plemenitaš, Dolžan Vita, Lenassi Metka
Determining the HIV-1 reservoir size in infected individuals is of great importance for improvement of their treatment. Plasma trans-activation response element (TAR) RNA has been suggested as one of the possible biomarkers. TAR RNA is produced during non-processive transcription in HIV-1 productively infected and latent T cells. Here, plasma samples and paired exosome samples of 55 subjects from the observational SCOPE cohort were analysed for the presence of TAR RNA. First, a PCR-based assay was optimized, which provided 100% specificity and 100% sensitivity in differentiating HIV-1 infected non-controllers from uninfected individuals...
September 2017: Acta Chimica Slovenica
https://www.readbyqxmd.com/read/28859135/identification-of-mir-16-as-an-endogenous-reference-gene-for-the-normalization-of-urinary-exosomal-mirna-expression-data-from-ckd-patients
#16
Tim Lange, Sylvia Stracke, Rainer Rettig, Uwe Lendeckel, Jana Kuhn, Rabea Schlüter, Volkhard Rippe, Karlhans Endlich, Nicole Endlich
Chronic kidney disease (CKD) is a severe disorder with an increasing incidence worldwide. An early detection may help to prevent its progression and to minimize the risk of cardiovascular diseases as one of the major comorbidities. Recently, extracellular miRNAs like urinary exosomal miRNAs became of great interest as non-invasive biomarkers which can be determined by RT-qPCR. But until now, there is no consensus regarding the normalization of miRNAs isolated from body fluids. The present study analyzed the miRNAs miR-16, miR-92a, miR-21, miR-124a and the small nuclear RNA RNU6B for their applicability as an endogenous reference gene in expression studies of exosomal miRNAs isolated from CKD patients...
2017: PloS One
https://www.readbyqxmd.com/read/28855728/meta-signature-of-human-endometrial-receptivity-a-meta-analysis-and-validation-study-of-transcriptomic-biomarkers
#17
Signe Altmäe, Mariann Koel, Urmo Võsa, Priit Adler, Marina Suhorutšenko, Triin Laisk-Podar, Viktorija Kukushkina, Merli Saare, Agne Velthut-Meikas, Kaarel Krjutškov, Lusine Aghajanova, Parameswaran G Lalitkumar, Kristina Gemzell-Danielsson, Linda Giudice, Carlos Simón, Andres Salumets
Previous transcriptome studies of the human endometrium have revealed hundreds of simultaneously up- and down-regulated genes that are involved in endometrial receptivity. However, the overlap between the studies is relatively small, and we are still searching for potential diagnostic biomarkers. Here we perform a meta-analysis of endometrial-receptivity associated genes on 164 endometrial samples (76 from 'pre-receptive' and 88 from mid-secretory, 'receptive' phase endometria) using a robust rank aggregation (RRA) method, followed by enrichment analysis, and regulatory microRNA prediction...
August 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28841829/hypoxic-exosomes-facilitate-bladder-tumor-growth-and-development-through-transferring-long-non-coding-rna-uca1
#18
Mei Xue, Wei Chen, An Xiang, Ruiqi Wang, He Chen, Jingjing Pan, Huan Pang, Hongli An, Xiang Wang, Huilian Hou, Xu Li
BACKGROUND: To overcome the hostile hypoxic microenvironment of solid tumors, tumor cells secrete a large number of non-coding RNA-containing exosomes that facilitate tumor development and metastasis. However, the precise mechanisms of tumor cell-derived exosomes during hypoxia are unknown. Here, we aim to clarify whether hypoxia affects tumor growth and progression by transferring long non-coding RNA-urothelial cancer-associated 1 (lncRNA-UCA1) enriched exosomes secreted from bladder cancer cells...
August 25, 2017: Molecular Cancer
https://www.readbyqxmd.com/read/28841135/ubiquitination-dependent-control-of-sexual-differentiation-in-fission-yeast
#19
Fabrizio Simonetti, Tito Candelli, Sebastien Leon, Domenico Libri, Mathieu Rougemaille
In fission yeast, meiosis-specific transcripts are selectively eliminated during vegetative growth by the combined action of the YTH-family RNA-binding protein Mmi1 and the nuclear exosome. Upon nutritional starvation, the master regulator of meiosis Mei2 inactivates Mmi1, thereby allowing expression of the meiotic program. Here, we show that the E3 ubiquitin ligase subunit Not4/Mot2 of the evolutionarily conserved Ccr4-Not complex, which associates with Mmi1, promotes suppression of meiotic transcripts expression in mitotic cells...
August 25, 2017: ELife
https://www.readbyqxmd.com/read/28835418/stem-cell-derived-exosomes-a-novel-vector-for-tissue-repair-and-diabetic-therapy
#20
William Newton, Joseph Kim, John Luo, LuGuang Luo
Exosomes are extracellular vesicles (EVs) secreted from a majority of cell types. Exosomes play a role in healthy and pathogenic intercellular interactions via the transfer of proteins, lipids, and RNA. The contents and effects of exosomes vary depending on the properties of the originating cell. Exosomes secreted from some cell types, including stem cells, carry biological factors implicated in the protection, regeneration, and angiogenesis of damaged tissues. Due to these properties, exosomes have attracted attention as a novel vector for regenerative therapies...
August 23, 2017: Journal of Molecular Endocrinology
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