Body fluid derived exosomes as a novel template for clinical diagnostics

By Sascha Keller, J. Ridinger, Anne-Kathleen Rupp, Johannes W. G. Janssen, P. Altevogt. Journal of Translational Medicine, 2011

697 citations recorded by the source corpus.

BackgroundExosomes are small membrane vesicles with a size of 40-100 nm that are released by different cell types from a late endosomal cellular compartment. They can be found in various body fluids including plasma, malignant ascites, urine, amniotic fluid and saliva. Exosomes contain proteins, miRNAs and mRNAs (exosome shuttle RNA, esRNA) that could serve as novel platform for diagnosis.MethodWe isolated exosomes from amniotic fluid, saliva and urine by differential centrifugation on sucrose gradients. Marker proteins were identified by Western blot and FACS analysis after adsorption of exosomes to latex beads. We extracted esRNA from exosomes, carried out RT-PCR, and analyzed amplified products by restriction length polymorphism.ResultsExosomes were positive for the marker proteins CD24, CD9, Annexin-1 and Hsp70 and displayed the correct buoyant density and orientation of antigens. In sucrose gradients the exosomal fractions contained esRNA that could be isolated with sufficient quantity for further analysis. EsRNAs were protected in exosomes from enzymatic degradation. Amniotic fluid esRNA served as template for the typing of the CD24 single nucleotide polymorphism (rs52812045). It also allowed sex determination of the fetus based on the detection of the male specific ZFY gene product.ConclusionsOur data demonstrate that exosomes from body fluids carry esRNAs which can be analyzed and offers access to the transcriptome of the host organism. The exosomal lipid bilayer protects the genetic information from degradation. As the isolation of exosomes is a minimally invasive procedure, this technique opens new possibilities for diagnostics.

Subfields: Cancer Research, Molecular Biology, Obstetrics and Gynecology

View Paper: https://www.semanticscholar.org/paper/21ccecbd071a28e5758fb3103e65b23ff2261c0c

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Citations in the corpus

Focus on Extracellular Vesicles: Introducing the Next Small Big Thing (2016, background): “EVs have been isolated from many biological fluids, including blood, milk, saliva, malignant ascites, amniotic fluid and urine [7–9].”
Exosomes in Tumor Microenvironment Influence Cancer Progression and Metastasis (2013, background): “Apart from serum, exosomes can be detected in further body fluids such as urine or saliva and provide non-invasive diagnostic markers for cancer [60, 61].”
The biology and function of exosomes in cancer. (2016, background): “Exosomes have been detected in many biological fluids, including urine, semen, saliva, amniotic fluid, cerebrospinal fluid, bile, ascites, tears, breast milk, and blood (2, 11, 37, 38).”
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Exosome Theranostics: Biology and Translational Medicine (2018, background): “Additionally, mRNA that encodes the zinc finger protein ZFY and is contained in exosomes of amniotic fluid can be used to determine the sex of the fetus [20].”
Characterization of human plasma-derived exosomal RNAs by deep sequencing (2013, background): “Compared to cellular RNAs, exosomal RNAs are more stable [49], and are reportedly resistant to physical degradation such as prolonged storage and freeze/ thaw cycles [50].”
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Exosomal cell-to-cell transmission of alpha synuclein oligomers (2012, background): “To investigate whether membrane integrity is important for uptake, exosomes were subjected to sonication which is known to disrupt lipid bilayer integrity [69-71].”
Engineering exosomes for targeted drug delivery (2021, background): “Exosomes are now widely found in all bodily fluids and tissues, including blood [6], urine [7], breast milk [8], amniotic/synovial/ascites fluid [9], saliva [10], and adipose tissue [11].”
Tumor-derived exosomes and their role in cancer progression (2016, background): “Exosomes are small extracellular vesicles (EVs) produced by all cells and present in all body fluids [1].”
Standardization of sample collection, isolation and analysis methods in extracellular vesicle research (2013, neutral): “…humor (36), breast milk (35,37), ascites (38,39), amniotic fluid (40,41), semen (‘‘prostasomes’’ and ‘‘epididymosomes’’) (42 (cid:2) 50), saliva (35,41,51,52), nasal secretions (53), cerebrospinal fluid (CSF) (54), bronchoalveolar lavage (BAL) (55 (cid:2) 57), synovial fluid (58 (cid:2) 65), bile…”
Deep sequencing of RNA from immune cell-derived vesicles uncovers the selective incorporation of small non-coding RNA biotypes with potential regulatory functions (2012, background): “Since the molecular make-up and release of cell-derived vesicles is regulated by the producing cell, these vesicles are of interest for disease-related biomarkers (14).”