Exosomes In Urine Research Articles

LincRNA-p21/AIF-1/CMPK2/NLRP3 pathway promoted inflammation, autophagy and apoptosis of human tubular epithelial cell induced by urate via exosomes

Urate nephropathy, a common complication of hyperuricemia, has garnered increasing attention worldwide. However, the exact pathogenesis of this condition remains unclear. Currently, inflammation is widely accepted as the key factor in urate nephropathy. Therefore, the aim of this study was to elucidate the interaction of lincRNA-p21/AIF-1/CMPK2/NLRP3 via exosomes in urate nephropathy. This study evaluated the effect of lincRNA-p21/AIF-1/CMPK2/NLRP3 using clinical data collected from patients with urate nephropathy and human renal tubular epithelial cells (HK2) cultured with different concentrations of urate. In clinical research section, the level of lincRNA-p21/AIF-1 in exosomes of urine in patients with hyperuricemia or urate nephropathy was found to be increased, particularly in patients with urate nephropathy. In vitro study section, the level of exosomes, inflammation, autophagy, and apoptosis was increased in HK2 cells induced by urate. Additionally, the expression of lincRNA-p21, AIF-1, CMPK2, and NLRP3 was upregulated in exosomes and HK2 cells. Furthermore, manipulating the activity of lincRNA-p21, AIF-1, CMPK2, and NLRP3 through overexpression or interference vectors regulated the level of inflammation, autophagy, and apoptosis in HK2 cells. In conclusion, the pathway of lincRNA-p21/AIF-1/CMPK2/NLRP3 contributed to inflammation, autophagy, and apoptosis of human renal tubular epithelial cell induced by urate via exosomes. Additionally, the specific exosomes in urine might serve as novel biomarkers for urate nephropathy.

3D-Nanostructured microfluidic device arranged in a herringbone pattern for the highly effective capture of HER2-Positive cancer-derived exosomes in urine

Exosomes in body fluids are important in liquid biopsy as they reflect the information of their originating cells. Thus, exosome analysis can provide valuable insights into diseases such as cancer. However, the low concentration of exosomes in body fluids is accompanied by background noise, making exosome analysis challenging. Here, a microfluidic chip in which three-dimensional nanostructures were arranged in a herringbone pattern (nanochip) that could efficiently capture specific exosomes was developed. Nanostructures were prepared by stacking silica nanoparticles to enhance the contact and interaction between the exosomes and structures, which were then arranged in a herringbone pattern to improve mass transfer through micromixing. To analyze exosomes derived from human epidermal growth factor receptor 2 (HER2, an important marker for cancer progression and patient survival)-positive cancer, anti-HER2 antibody was introduced into the nanostructures in the nanochip and approximately 97.7% of exosome capture efficiency was confirmed. The nanochip performed better than chips with a solid herringbone structure or without a structure (solid and flat chips). The feasibility of capturing multiple exosomes was demonstrated using both in vitro and in vivo samples by employing a dual nanochip configuration in which nanochips with different antibodies were interconnected in a series. This nanochip can effectively capture HER2-positive exosomes and has potential for multiple exosome isolations. Additionally, this chip can capture and detect various disease-related exosomes because various antibodies can be applied; this nanochip will be useful for exosome-based disease diagnosis and monitoring in liquid biopsies.

Characterization of exosomal microRNAs in preterm infants fed with breast milk and infant formula.

Breastfeeding not only reduces infection-related morbidity, but also increases growth of preterm infants. Advantages of breast milk (BM) for preterm infants are significant. They continue to be studied. However, because not all preterm infants can receive breastfeeding, bovine-based infant formula (IF) is used as an alternative, which may increase the risk of several preterm complications. Exosomes isolated from biofluids are emerging as biomarkers in research of various diseases. Here, we characterized miRNA contents of exosomes in urine and serum samples of preterm infants who were BM and IF fed and performed transcriptomic analysis of small RNA libraries. We identified significantly up-regulated 6 miRNAs and 10 miRNAs, respectively. Gene Ontology (GO) analysis revealed that target genes of these miRNAs might participate in neuronal development, immunity modulation, detoxification of reactive oxygen species, and transmembrane exchange. Our data suggest that exosome-based systemic screening for preterm infants with breastfeeding might be a screening tool for identifying target molecules involved in therapy for preterm infants in neonatal intensive care unit (NICU) and for future application as nutraceutical formulations or pharmaceuticals.

Colorimetric Detection of HER2-Overexpressing-Cancer-Derived Exosomes in Mouse Urine Using Magnetic-Polydiacetylene Nanoparticles.

Breast cancer (BC) is a major global health problem, with ≈20-25% of patients overexpressing human epidermal growth factor receptor 2 (HER2), an aggressive marker, yet access to early detection and treatment varies across countries. A low-cost, equipment-free, and easy-to-use polydiacetylene (PDA)-based colorimetric sensor is developed for HER2-overexpressing cancer detection, designed for use in low- and middle-income countries (LMICs). PDA nanoparticles are first prepared through thin-film hydration. Subsequently, hydrophilic magnetic nanoparticles and HER2 antibodies are sequentially conjugated to them. The synthesized HER2-MPDA can be concentrated and separated by a magnetic field while inheriting the optical characteristics of PDA. The specific binding of HER2 antibody in HER2-MPDA to HER2 receptor in HER2-overexpressing exosomes causes a blue-to-red color change by altering the molecular structure of the PDA backbone. This colorimetric sensor can simultaneously separate and detect HER2-overexpressing exosomes. HER2-MPDA can detect HER2-overexpressingexosomes in the culture medium of HER2-overexpressingBCcellsandinmouse urine samples from a HER2-overexpressing BCmousemodel.Itcanselectively isolate and detect only HER2-overexpressingexosomesthroughmagneticseparation, and its detectionlimitisfoundtobe8.5×108particlesmL-1.Thiscolorimetric sensorcanbeusedforpoint-of-carediagnosisofHER2-overexpressing BC in LMICs.

Role of tumor-derived exosomes in metastasis, drug resistance and diagnosis of clear cell renal cell carcinoma.

Renal cancer is one of the most extensively studied human tumors today, with clear cell renal cell carcinoma accounting for approximately 80% of all cases. Despite recent advances in research on clear cell renal cell carcinoma, advanced distant metastasis of the disease, delay in diagnosis, as well as drug resistance remain major problems. In recent years, as an important mediator of material and information exchange between cells in the tumor microenvironment, exosomes have attracted widespread attention for their role in tumor development. It has been reported that tumor-derived exosomes may act as regulators and have an important effect on the metastasis, drug resistance formation, and providing targets for early diagnosis of clear cell renal cell carcinoma. Therefore, the extensive study of tumour-derived exosomes will provide a meaningful reference for the development of the diagnostic and therapeutic field of clear cell renal cell carcinoma. This article reviews the biological role and research progress of tumor-derived exosomes in different aspects of premetastatic niche formation, tumor angiogenesis, and epithelial-mesenchymal transition during the progression of clear cell renal cell carcinoma. In addition, the role of tumor-derived exosomes in the development of drug resistance in clear cell renal cell carcinoma is also addressed in this review. Furthermore, recent studies have found that cargoes of exosomes in serum and urine, for example, a series of miRNAs, have the potential to be biological markers of clear cell renal cell carcinoma and provide meaningful targets for early diagnosis and monitoring of tumors, which is also covered in this article.

Tumor-derived exosomal non-coding RNAs as diagnostic biomarkers in cancer.

Almost all clinical oncologists agree that the discovery of reliable, accessible, and non-invasive biomarkers is necessary to decrease cancer mortality. It is possible to employ reliable biomarkers to diagnose cancer in the early stages, predict the patient prognosis, follow up the response to treatment, and estimate the risk of disease recurrence with high sensitivity and specificity. Extracellular vesicles (EVs), especially exosomes, have been the focus of translational research to develop such biomarkers over the past decade. The abundance and distribution of exosomes in bodily fluids, including serum, saliva, and urine, as well as their ability to transport various biomolecules (nucleic acids, proteins, and lipids) derived from their parent cells, make exosomes reliable, accessible, and potent biomarkers for diagnosis and follow-up of solid and hematopoietic tumors. In addition, exosomes play a vital role in various cellular processes, including tumor progression, by participating in intercellular communication. Although these advantages underline the high potential of tumor-derived exosomes as diagnostic biomarkers, the lack of standardized effective methods for their isolation, identification, and precise characterization makes their application challenging in clinical settings. We discuss the importance of non-coding RNAs (ncRNAs) in cellular processes, and the role of tumor-derived exosomes containing ncRNAs as potential biomarkers in several types of cancer. In addition, the advantages and challenges of these studies for translation into clinical applications are covered.

Design and application of hydrophilic bimetallic metal-organic framework magnetic nanoparticles for rapid capture of exosomes

Functional materials with good biocompatibility have been widely used in the study of genomics, proteomics and disease diagnosis, which has improved the progress of life science. In this paper, the material not only exhibited a strong affinity to the phosphate groups on the exosomal membrane due to the coexistence of Zr–O clusters and Ti4+, but also owned great hydrophilicity to reduce non-specific adsorption of contaminated proteins, achieving the separation and purification of exosomes from complex biosamples. The model exosomes extracted by ultracentrifugation (UC) were used to evaluate the feasibility of Fe3O4@UiO-66-NH2@PA-Ti4+ capturing exosomes. The process of Fe3O4@UiO-66-NH2@PA-Ti4+ capturing exosomes was simple to operate with a high recovery rate (97.3%) within a short time (5 min). Then Fe3O4@UiO-66-NH2@PA-Ti4+ was further applied to capture exosomes in media and urine followed by the downstream proteomics analysis. 348 and 284 exosomal proteins were identified for cell medium and urine, respectively. This work shows great potential of the material for subsequent function research of disease-related exosomes by separating exosomes rapidly and efficiently.

Rapid isolation and proteome analysis of urinary exosome based on double interactions of Fe3O4@TiO2-DNA aptamer

There are accumulating evidence that proteins carried by exosomes in urine are most possibly used as biomarkers or therapeutic carriers for certain diseases. The isolation of exosomes is therefore highly desirable for aiding the downstream protein analysis. Particularly, urine is a dynamic biological fluid changing within a short time, resulting in that the separation of urinary exosome requires more efficient technology. Here, a new biocompatible material (denoted as Fe3O4@TiO2-CD63 aptamer) is designed and synthesized for rapid exosome isolation from human urine, depending on the double interactions of TiO2 with phosphate groups as well as aptamers with specific exosome proteins. Moreover, within 10 min, 92.6% exosomes with intact structure are captured from urine by Fe3O4@TiO2-CD63 aptamers, from which 999 proteins are detected through LC-MS/MS.

Urine and Serum Exosomes as Novel Biomarkers in Detection of Bladder Cancer.

Background:The gold standard for initial clinical diagnosis of bladder cancer involves cystoscopic examination of bladder and histological evaluation of tissues. There is a critical need to identify non-invasive and sensitive biomarkers. Early detection is essential challenge in diagnosis and surveillance of bladder carcinoma. Exosomes are nano- sized vesicles present in many biological fluids and have significant role in cancer. Thus, quantification of exosomes in different stages of bladder cancer may be of critical concern for clinical diagnosis and prognosis. Methods:Tumor derived exosomes levels in urine and serum samples of 70 bladder cancer Egyptian patients from stages T0-T3 and 12 healthy control people were measured using ELISA technique. Results:When compared to health subjects, exosomes levels in bladder cancer patients were increased in urine and serum samples at different stages of the disease. A gradual increase in tumor derived exosomes in serum (1.21, 3.31, 4.71, 6.47µg/ml) and urine (1.59, 2.84, 4.75, 6.67µg/ml) was observed comparative to invasiveness of tumor (T0-T3). Serum was more specific (100%) sample for detection of exosomes in bladder cancer. Conclusion: our findings suggest that tumor derived exosomes may offer a convenient tool for early diagnosis and monitoring of bladder cancer.

A phase II study for prostate cancer monitoring using 18F-DCFPyL and blood-based biomarkers.

TPS3154 Background: Assessing treatment response in castrate resistant prostate cancer (CRPC), remains a challenge due to the limited sensitivity and specificity of existing imaging modalities. Understanding prostate cancer biology with tumor biopsies does not address the issue of tumor heterogeneity or cellular degradation during the decalcification process of bone biopsies. Next generation positron emission tomography (PET) imaging and circulating biomarkers might provide additional insights on treatment responses and inform clinical decision-making earlier in therapy. 18F-DCFPyL (PyL) is a second-generation fluorinated PSMA PET tracer that has superior sensitivity and specificity to detect prostate cancer compared to standard imaging. Its role in assessing tumor response to therapy has not been evaluated. Circulating tumor DNA (ctDNA) in blood can provide tumor genomic information, while exosomes in serum and urine may provide data on the proteomic landscape of tumors. Methods: We are conducting a prospective study of 15 men with metastatic CRPC who are scheduled to start a new systemic therapy for their disease. Upon enrollment, subjects will have baseline assessments with standard cross-sectional imaging, 99mTc bone scan, and blood work. Standard scans will be performed every 8-12 weeks until progression of disease. PyL PET/CT scans and liquid biopsies (ctDNA and exosomes) will occur at baseline, 6 weeks after starting their new therapy, and at disease progression. Lesions seen on PET/CT images will be identified by a certified reader. The maximum standardized uptake value (SUVmax) will be measured and recorded in up to the five hottest lesions and normalized to a background SUVmean measured in the liver, spleen, kidney, mediastinum, and parotid glands. Changes in the normalized SUV from baseline to the 6-week PyL PET scan will be correlated to PSA response by the Pearson’s correlation coefficient. The Kaplan-Meier method will be used to evaluate progression free survival and overall survival dichotomized by the median value of SUV change. Blood for ctDNA and exosomes will be stored for future analysis. The study is open with two patients enrolled at the time of submission. One patient has completed his initial PyL PET/CT scan. Clinical trial information: NCT03585114.

Ultrasensitive and Reversible Nanoplatform of Urinary Exosomes for Prostate Cancer Diagnosis.

Prostate cancer cell-derived exosomes in urine have been extensively studied recently and regarded as novel biomarkers for cancer diagnosis and prognosis, which presents wide prospects in clinical applications. Sensitive detection and specific capture methods are essential for exosomes analysis. Herein, a dual functional platform composed of superparamagnetic conjunctions and molecular beacons (SMC-MB) is reported. The SMC-MB platform is designed based on aptamer immunoaffinity with ultrasensitive detection efficiency and reversible isolation capacity, which, respectively, profit from nonenzymatic amplification methods and magnetic separation along with restriction cleavage. It is noteworthy that exosomes quantification was exactly amplified and transformed into single strand DNA detection. Correlated measurements evidence that the limit of detection of SMC-MB is as low as ∼100 particles/μL in urine, and a linear relationship meets between the logarithmic concentration of exosomes and fluorescence intensity of the molecular beacon. Furthermore, employing prostate specific membrane antigen (PSMA) aptamer, the platform adapted to detect and capture PMSA-positive exosomes from urine samples provides excellent diagnostic efficiency for prostate cancer (PCa). The expression of typical biomarkers of PCa, i.e., PSA and PCA3 mRNA, is significantly higher in PSMA-positive exosomes. Altogether, the platform and strategy described in this paper are promising in urinary exosomes analysis and prostate cancer detection.

Urinary exosome as a potential biomarker for urinary tract infection.

Unlike urinary tract infection (UTI), asymptomatic bacteriuria (ABU) should not be treated, with some exceptions such as pregnant women and patients who will undergo traumatic urologic interventions. However, there has been no clinically available marker for their differential diagnosis. Exosomes or small extracellular vesicles carry proteins contained in cells from which they are derived, thus having the potential as a biomarker of several diseases. On the basis of the hypothesis that the molecular signature of exosomes in urine may differ between UTI and ABU patients, we examined if urinary exosomes could serve as a marker for their differential diagnosis. Exosomes were isolated by ultracentrifugation or affinity-based method from cell culture medium of monocytic THP-1 and uroepithelial SV-HUC-1 cells and human urine. Protein expression was examined by Western blot analysis, ELISA, and CLEIA. The results showed that the levels of intracellular signalling molecules Akt and ERK and transcription factor NF-κB increased in exosomes isolated from THP-1 and SV-HUC-1 cells cocultured with Escherichia coli and/or treated with lipopolysaccharide. In urinary exosomes of UTI patients, Akt significantly diminished, and an exosomal marker CD9 showed a trend to decrease after treatment with antimicrobial agents. More importantly, Akt and CD9 levels in urinary exosomes were higher in UTI patients than in ABU patients, which was also observed after correction by urine creatinine. Collectively, these results suggest that Akt and CD9 in urinary exosomes could be useful markers for differential diagnosis of UTI and ABU.

Increased Podocyte Exosome Release in Glomerular Injury induced by NLRP3 Inflammasome Activation during Hyperhomocysteinemia

The NOD‐like receptor containing pyrin domain 3 (NLRP3) inflammasome has been implicated in podocyte injury and glomerular sclerosis to elevated homocysteine (Hcys) levels in vitro and in vivo. However, it remains unknown how the products of NRLP3 inflammasome activated in cytoplasm are secreted out of podocytes and whether exosome release serves as a critical mechanism to mediate the action of NLRP3 inflammasome activation in Hcys‐induced kidney injury. In the present study, we tried to answer these questions using Smpd1trg/Podocre (a podocye‐specific sphingomyelin phosphodiesterase 1 (Smpd1) transgenic mouse strain) because Smpd1 encodes acid sphingomyelinase that produces ceramide in lysosomes and regulates lysosome function and potentially controls exosome release. It was found that Smpd1trg/Podocre mice had obvious overexpression of Smpd1 gene and remarkable increase in ceramide concentrations in podocytes. These Smpd1trg/Podocre mice were fed a folate‐free (FF) diet to induce hyperhomocysteinemia (hHcys), and ~ 3 folds increase in colocalization of inflammasome proteins, NLRP3 with ASC or with caspase‐1 and ~ 4 folds of IL‐1β production were found in the glomeruli of these FF‐treated Smpd1trg/Podocre mice, suggesting NLRP3 inflammasome formation and activation. Increased IL‐1β production in the glomeruli was accompanied by decreased podocin levels, but enhanced desmin expression in podocytes with glomerular injury as shown by morphological examinations. We also observed significantly increased colocalization of Annexin‐2 and alkaline phosphatase (exosome markers) in glomeruli of FF‐treated Smpd1trg/Podocre mice than their control littermates, indicating the increased exosome release in mice with Smpd1 gene overexpression in podocytes. By Nanoparticle Tracking Analysis, we found that urinary exosomes (60–140 nm in size) were significantly increased especially from 90nm to 140nm in FF‐treated Smpd1trg/Podocre mice, but not in their control littermates. These increased exosomes in urine from FF‐treated Smpd1trg/Podocre mice contained more detectable podocin protein than control littermates, meaning enrichment of podocyte‐derived exosomes in urine from the podocyte‐specific Smpd1 transgenic mice. Taken together, our results suggest that increased ceramide in podocytes due to Smpd1 gene overexpression may increase exosome release and thereby enhance excretion of inflammasome products to induce podocyte dysfunction and glomerular injury.Support or Funding Informationsupported by NIH grants DK54927 to P.L. and, DK107991 to N.L.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effective enrichment of urinary exosomes by polyethylene glycol for RNA detection

Urinary exosome is a kind of extracellular vesicles secreted by the cells of kidney and urinary tract. It is an ideal non-invasive biomarker for obtaining information about the state of renal cells. There are few exosomes in urine which are affected by many factors. Enriching exosomes in urine with inexpensive and efficient method is a major challenge. Because exosomes are virus-sized minuscule membrane vesicles or sacs and people used to enrich virus by Polyethylene Glycol (PEG), here we use PEG to enrich the exosomes in urine of 17 volunteers. To confirm the harvest of exosomes, the morphology of exosomes were observed by Transmission Electron Microscopy (TEM), the common exosomal protein markers (CD63, CD9, tumor susceptibility gene 101, Flotillin-1 and β-actin) were detected by Western blot, and the size distribution was analysed by Malvin Nano-ZS. Next, the exosomal housekeeping gene of miRNA (SnRNA-U6) and mRNA (β-actin) were tested by qRT-PCR to prove that this method could be used in subsequent experiments.

Serum and urinary levels of CD222 in cancer: origin and diagnostic value.

The mannose 6-phosphate/insulin-like growth factor 2 receptor (CD222, M6P/IGF2R) is a multifunctional transmembrane type I receptor, mostly localized intracellularly, less on the surface of all types of mammalian cells. It is known both to transport lysosomal enzymes through their mannose 6-phosphate moieties and to internalize extracellular ligands like insulin-like growth factor 2 or plasminogen. CD222 is involved in regulation of cell proliferation, migration, T cell activation, and apoptosis. Soluble CD222 has been found in higher concentrations in sera of liver disease patients. In this study, we analysed the level of CD222 present in body fluids, namely in serum and urine, of cancer patients. We found significantly elevated levels of soluble CD222 in sera of cancer patients compared to healthy controls irrespective of the type of disease. The urine CD222 levels were increased specifically in breast cancer and multiple myeloma. In contrast to serum, CD222 was present within CD222-positive exosomes in urine pointing to different origins of CD222 present in various human body fluids. Based on this work, we propose serum soluble CD222 as a general biomarker for tumorigenesis.

Cancer exosomes in urine: a promising biomarker source

Cancer exosomes in urine: a promising biomarker source

Urinary exosomes as a novel biomarker for evaluation of α-lipoic acid's protective effect in early diabetic nephropathy.

Long-term administration of α-lipoic acid (α-LA) is proved to ameliorate renal impairment. Herein we assessed serum, urinary biomarkers and vascular endothelium function to evaluate its short-period therapeutic effect and identify novel biomarkers for diabetic nephropathy (DN). Sixty-two microalbuminuria-stage DN patients were randomly divided into two groups and received the following treatment for 8weeks: (1) routine treatment(DM group); (2) routine treatment with 600mg/d α-lipoic acid intravenously (α-LA group). Another total of 21 patients were recruited for the second-stage study and randomly divided into two groups: normoalbuminuria (UAER <30mg/24h) and microalbuminuria (UAER from 30-300mg/24h). With α-LA treatment, urinary albumin excretion rates (UAER), serum creatinine (SCr) and malonaldehyde (MDA) declined significantly, whereas plasma superoxide dismutase (SOD)activity increased and endothelium-dependent flow mediated vasodilation (FMD) flexibility improved dramatically. Furthermore, the improvement of FMD showed positive correlation with the variation in MDA and SOD as well (r values are .516 and .435, P<.01 and P<.05, respectively). In contrast, these markers have no significant difference in the DM group with routine treatment. Notably, the CD63 expressing of exosomes in urine was found higher in the normoalbuminuria patients compared with those in microalbuminuria, parallelly only declined markedly after α-LA administration in normoalbuminuria patients. In summary, we emphasize short-term α-LA could protect the kidney in the early DN against general oxidative stress, particularly the urinary CD63-positive exosome could be a potential sensitive and therapeutic indicator.

Extracellular Vesicle miRNA Detection Using Molecular Beacons.

Many studies have recently focused on extracellular vesicles. Extracellular vesicles including exosomes are nanosized vesicles that are produced and secreted by cells. Extracellular vesicles contain various types of nucleic acid biomarkers, including microRNAs (miRNAs ) from their parent cells. As high concentrations of extracellular vesicles are found in most body fluids, miRNA detection of exosomes in body fluids, including blood and urine, enables early and easy disease diagnosis. In this context, the in situ detection method of extracellular vesicles miRNA using molecular beacons is described here.

Rodent models of chronic kidney disease for studies of early renal tubulointerstitial fibrosis

Chronic kidney disease (CKD) is the progressive decline of renal function resulting from renal fibrosis because of several causes [1]. CKD is an important health care problem that impacts upon the global economy because of the need for long-term renal replacement therapy at the final stage, referred to as “end stage renal disease” or “ESRD”. Interestingly, similar clinical manifestations, such as anemia, nausea and vomiting, are present in patients with ESRD regardless of the initial causes of renal injury. CKD could be unofficially differentiated into chronic glomerular disease (CGD) and chronic tubulointerstitial disease (CTID). Because of the progressive natural history of CKD, biomarkers for the early diagnosis and monitoring of CKD are essential. The diagnosis of CKD is currently based on the levels of serum creatinine (Scr) and proteinuria (urine albumin or urine total proteins), which has several limitations in the early detection of CKD. Scr levels in CKD are confounded by several nonrenal factors (e.g. muscle mass and liver function) [2] and often increase after the presence of proteinuria. By contrast, proteinuria levels are influenced by diet and the diurnal variation [3]. Therefore, new sensitive and reliable biomarkers for CKD are needed. Exosomes, nano-sized vesicles containing cytosolic and membrane proteins secreted from a wide variety of cell types, are new and interesting sources of urine biomarkers, because exosomes can be detected in urine and the analysis of exosomes resembles a random biopsy of the cell [4]. Thus, a new noninvasive method of so-called “liquid biopsy” through isolation of exosomes in urine should open up more opportunity for biomarker discovery.

807 - Discovery of high-Gleason prostate cancer biomarker candidates from exosomes in urine after massage by proteomic analysis and subsequent verification using selected reaction monitoring

807 - Discovery of high-Gleason prostate cancer biomarker candidates from exosomes in urine after massage by proteomic analysis and subsequent verification using selected reaction monitoring