Статті в журналах: "Urinary exosome"

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Huang, Kun, Sudha Garimella, Alyssa Clay-Gilmour, Lucia Vojtech, Bridget Armstrong, Madison Bessonny, and Alexis Stamatikos. "Comparison of Human Urinary Exosomes Isolated via Ultracentrifugation Alone versus Ultracentrifugation Followed by SEC Column-Purification." Journal of Personalized Medicine 12, no. 3 (February 24, 2022): 340. http://dx.doi.org/10.3390/jpm12030340.

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Chronic kidney disease is a progressive, incurable condition that involves a gradual loss of kidney function. While there are no non-invasive biomarkers available to determine whether individuals are susceptible to developing chronic kidney disease, small RNAs within urinary exosomes have recently emerged as a potential candidate to use for assessing renal function. Ultracentrifugation is the gold standard for urinary exosome isolation. However, extravesicular small RNA contamination can occur when isolating exosomes from biological fluids using ultracentrifugation, which may lead to misidentifying the presence of certain small RNA species in human urinary exosomes. Therefore, we characterized human urinary exosomal preparations isolated by ultracentrifugation alone, or via ultracentrifugation followed by size exclusion chromatography (SEC) column-purification. Using nanoparticle tracking analysis, we identified SEC fractions containing robust amounts of exosome-sized particles, that we further characterized using immunoblotting. When compared to exosomal preparations isolated by ultracentrifugation only, SEC fractionated exosomal preparations showed higher levels of the exosome-positive marker CD81. Moreover, while the exosome-negative marker calnexin was undetectable in SEC fractionated exosomal preparations, we did observe calnexin detection in the exosomal preparations isolated by ultracentrifugation alone, which implies contamination in these preparations. Lastly, we imaged SEC fractionated exosomal preparations using transmission electron microscopy to confirm these preparations contained human urinary exosomes. Our results indicate that combining ultracentrifugation and SEC column-purification exosome isolation strategies is a powerful approach for collecting contaminant-free human urinary exosomes and should be considered when exosomes devoid of contamination are needed for downstream applications.

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Abdeen, Ahmed, Hiroko Sonoda, Ragab El-Shawarby, Saki Takahashi, and Masahiro Ikeda. "Urinary excretion pattern of exosomal aquaporin-2 in rats that received gentamicin." American Journal of Physiology-Renal Physiology 307, no. 11 (December 1, 2014): F1227—F1237. http://dx.doi.org/10.1152/ajprenal.00140.2014.

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Urinary exosomes are nano-sized vesicles secreted into urine from all types of renal epithelial cells and are known to contain possible biomarker proteins for renal diseases. Gentamicin has been reported to decrease the level of renal aquaporin (AQP)2, which is known to be mainly expressed in renal collecting ducts and excreted into the urine via exosomes. In the present study, we investigated whether urinary exosomal AQP2 could serve as a potential biomarker for gentamicin-induced nephrotoxicity, especially collecting duct cell dysfunction. Gentamicin was given to rats intraperitoneally once every day starting on day 0. Gentamicin significantly increased the plasma creatinine concentration from day 5 and beyond. Also, gentamicin induced polyuria and a defective urine concentration mechanism on day 7, suggesting gentamicin-induced collecting duct cell dysfunction. Immunoblot analysis showed that gentamicin significantly increased urinary exosomal AQP2 excretion on day 1 but decreased it on day 7 compared with the control group. Similarly, increased excretion of exosomal tumor susceptibility gene 101 protein, frequently used as an exosome marker protein, was observed on day 1. However, gentamicin did not significantly affect the urinary excretion of exosomal tumor susceptibility gene 101 on day 7. Gentamicin slightly decreased renal AQP2 expression on day 2 and markedly decreased it on day 8. These data strongly suggest that the use of urinary exosomal AQP2 as a biomarker may allow detection of gentamicin-induced collecting duct cell dysfunction. Furthermore, urinary exosomal AQP2 might also be useful for the early detection of gentamicin-induced renal injury in addition to collecting duct injury.

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Liu, Yu-Ru, and Yi-Fen Lee. "Urinary exosome and beyond." Translational Cancer Research 5, S2 (August 2016): S321—S324. http://dx.doi.org/10.21037/tcr.2016.07.16.

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Xiang, Xiaochao, Fulin Guan, Fenglong Jiao, Hang Li, Wanjun Zhang, Yangjun Zhang, and Weijie Qin. "A new urinary exosome enrichment method by a combination of ultrafiltration and TiO2 nanoparticles." Analytical Methods 13, no. 13 (2021): 1591–600. http://dx.doi.org/10.1039/d1ay00102g.

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The workflow of separation and enrichment of exosomes by ultrafiltration–TiO₂. We proposed a new strategy for facile exosome isolation from human urine by utilizing the ultrafiltration technique and TiO₂, which can significantly reduce urine volumes, increase exposure of the material to exosomes in urine and obtain high purity exosomes.

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Lv, Lin-Li, Yu-Han Cao, Hai-Feng Ni, Min Xu, Dan Liu, Hong Liu, Ping-Sheng Chen, and Bi-Cheng Liu. "MicroRNA-29c in urinary exosome/microvesicle as a biomarker of renal fibrosis." American Journal of Physiology-Renal Physiology 305, no. 8 (October 15, 2013): F1220—F1227. http://dx.doi.org/10.1152/ajprenal.00148.2013.

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Micro (mi)RNAs are frequently dysregulated in the development of renal fibrosis. Exosomes are small membrane vesicles that could be isolated from urine secreted from all nephron segments. Here we sought to observe for the first time whether miRNA in urine exosome could serve as a potential biomarker of renal fibrosis. Urine samples were collected from 32 chronic kidney disease (CKD) patients who underwent kidney biopsy and 7 controls. Exosome was isolated and confirmed by immunogold staining of exosome marker. Members of miR-29, miR-200, and RNU6B as endogenous control were detected by RT quantitative PCR. Electronic microscopy verified a typical shape of exosome with average size of 65.1 nm and labeled it with anti-CD9 and anti-aquaporin 2 antibody. Members of miR-29 and miR-200 are readily measured with reduced levels compared with controls ( P < 0.05) and can robustly distinguish CKD from controls [area under the curve (AUC) varied from 0.902 to 1 by receiver operating characteristics analysis]. miR-29c correlated with both estimated glomerular filtration rate ( r = 0.362; P < 0.05) and degree of tubulointerstitial fibrosis ( r = −0.359; P < 0.05) for CKD patients. Moreover, miRNA in exosome was decreased in mild fibrosis group compared with moderated to severe group. miR-29a and miR-29c could predict degree of tubulointerstitial fibrosis with AUC of 0.883 and 0.738 ( P < 0.05). The sensitivity and specificity for distinguishing mild from moderate to severe fibrosis were 93.8 and 81.3% with the use of miR-29a and 68.8 and 81.3% for miR-29c. Overall, miR-29c in urinary exosome correlates with both renal function and degree of histological fibrosis, suggesting it as a novel, noninvasive marker for renal fibrosis.

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Garcia-Vives, Eloi, Cristina Solé, Teresa Moliné, Marta Vidal, Irene Agraz, Josep Ordi-Ros, and Josefina Cortés-Hernández. "The Urinary Exosomal miRNA Expression Profile is Predictive of Clinical Response in Lupus Nephritis." International Journal of Molecular Sciences 21, no. 4 (February 18, 2020): 1372. http://dx.doi.org/10.3390/ijms21041372.

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Data on exosomal-derived urinary miRNAs have identified several miRNAs associated with disease activity and fibrosis formation, but studies on prognosis are lacking. We conducted a qPCR array screening on urinary exosomes from 14 patients with biopsy-proven proliferative lupus glomerulonephritis with a renal outcome of clinical response (n = 7) and non-response (n = 7) following therapy. Validation studies were performed by qRT-PCR in a new lupus nephritis (LN) cohort (responders = 22 and non-responders = 21). Responder patients expressed significantly increased levels of miR-31, miR-107, and miR-135b-5p in urine and renal tissue compared to non-responders. MiR-135b exhibited the best predictive value to discriminate responder patients (area under the curve = 0.783). In vitro studies showed exosome-derived miR-31, miR-107, and miR-135b-5p expression to be mainly produced by tubular renal cells stimulated with inflammatory cytokines (e.g IL1, TNFα, IFNα and IL6). Uptake of urinary exosomes from responders by mesangial cells was superior compared to that from non-responders (90% vs. 50%, p < 0.0001). HIF1A was identified as a potential common target, and low protein levels were found in non-responder renal biopsies. HIF1A inhibition reduced mesangial proliferation and IL-8, CCL2, CCL3, and CXCL1 mesangial cell production and IL-6/VCAM-1 in endothelial cells. Urinary exosomal miR-135b-5p, miR-107, and miR-31 are promising novel markers for clinical outcomes, regulating LN renal recovery by HIF1A inhibition.

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Ledet, Elisa M., Patrick J. Miller, Ratish Gambhira, Aryeneesh Dotiwala, and A. Oliver Sartor. "Characterization of plasma-derived and urinary exosomal microRNA from metastatic CRPC patients." Journal of Clinical Oncology 34, no. 2_suppl (January 10, 2016): 248. http://dx.doi.org/10.1200/jco.2016.34.2_suppl.248.

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248 Background: Exosomes are nano-sized (50-100nm) vesicles derived from normal and tumor cells that function in cell-cell communication. These vesicles and their nucleic acid cargo may potentially serve as biomarkers for assessment of risk stratification and therapeutic response. The goal of this study was to characterize exosome derived microRNA (miRNA) isolated from plasma (pExos) and urine (uExos) of metastatic CRPC patients. Methods: Plasma samples were obtained from 18 mCRPC patients and 1 normal control. Following exosome isolation, RNA extraction and library prep, paired-end sequencing was performed using Illumina Hi-Seq 2000. A bioinformatics pipeline was used for data processing including alignment, duplicate removal, normalization, and variant calling. Visualization and differential analyses were performed with SNP & Variation Suite v8.x. RNA derived from uExos was amplified using whole transcriptome amplification and interrogated with Prostate Cancer (PCa) miScript miRNA PCR Array. Results: Exosomes from both plasma and urine had similar amounts of miRNA/total RNA with average 34% miRNA (range 19%-51%). pExos had larger RNA fragments (range 10-333 nt) while uExos were more highly fragmented (range 10-60 nt). The amount of miRNA and fragmentation pattern was highly variable amongst patients. In pExos, RNA from PDPK1, USP9X, MAGI2, HMGA2 and PTGFR were present and previously shown in PCa. Also in pExos, miR941-2, miR4454, miR1302-2, miR143HG and miR22HG were annotated in prostate cancer patients; these miRNA have previously been identified in cancer. In uExos miR-16-5p and miR-375 were present and are shown to be differentially regulated in prostate cancer. Expression analyses will be presented. PCR validation is ongoing. Conclusions: The identification of cancer associated miRNA in pExos and uExos may potentially serve as biomarkers in mCRPC patients. The abundance and stability of miRNA contained in exosomes may provide insight into tumor evolution and disease progression. Additional studies evaluating the clinical relevance and prognostic value of exosomal miRNA are warranted.

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Shirley, James Forrest, Joshua Drourr, W. Taylor Edwards, Kubra Tuna, Lisa K. Ryan, Abdel Alli, Ying Tang та Sarah C. Glover. "Mast Cells in Patients with Hereditary α-Tryptasemia Promote HLA-DR Expression and a Th2-Polarizing Microenvironment in the Gastrointestinal Tract". Journal of Immunology 202, № 1_Supplement (1 травня 2019): 192.13. http://dx.doi.org/10.4049/jimmunol.202.supp.192.13.

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Abstract Hereditary α-tryptasemia (HαT) is a recently identified, genetic disorder characterized by multisystem comorbidities resulting from increased monoallelic α-tryptase copy number (CN) at TPSAB1 (α-TPSAB1). Affected individuals frequently exhibit heterogeneous multisystem symptoms and comorbidities which negatively impact daily functioning and quality of life. In the cohorts examined thus far, functional dyspepsia and irritable bowel syndrome, were most frequently reported. However, the mechanism by which increased α-TPSAB1 CN produces GI disease is unknown. Tryptase is almost exclusively produced by mast cells (MC). In the gut, MC play important roles in immune responses, leukocyte recruitment, neuroimmune inflammation, and tissue repair. MC can also promote T-cell activation and polarization via cytokine secretion, co-stimulatory marker expression and MHC II antigen presentation. MCs also release exosomes, nanosized vesicles secreted by most cell types present in biological fluids such as blood, cerebrospinal fluid, and urine, which have a reported influence on T cell differentiation. In early experiments, we found increased numbers of GI MCs, activated T cells, and urinary exosomes in HαT patients compared to healthy donors. Thus, we evaluated MC surface antigen expression, exosome proteomics, and cytokine expression of exosome-treated T cells. We found increased HLA-DR and FcɛR1α expression in HαT-patient GI MCs, increased Th2 associated signaling proteins in HαT urinary exosomes, and elevated Th2 cytokine expression in HαT-exosome treated T cells. These results suggest that HαT GI morbidity involves aggravated MC hyperplasia and antigen presentation which promotes T cell activation and Th2 polarization.

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El Fekih, Rania, James Hurley, Vasisht Tadigotla, Areej Alghamdi, Anand Srivastava, Christine Coticchia, John Choi, et al. "Discovery and Validation of a Urinary Exosome mRNA Signature for the Diagnosis of Human Kidney Transplant Rejection." Journal of the American Society of Nephrology 32, no. 4 (March 3, 2021): 994–1004. http://dx.doi.org/10.1681/asn.2020060850.

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BackgroundDeveloping a noninvasive clinical test to accurately diagnose kidney allograft rejection is critical to improve allograft outcomes. Urinary exosomes, tiny vesicles released into the urine that carry parent cells’ proteins and nucleic acids, reflect the biologic function of the parent cells within the kidney, including immune cells. Their stability in urine makes them a potentially powerful tool for liquid biopsy and a noninvasive diagnostic biomarker for kidney-transplant rejection.MethodsUsing 192 of 220 urine samples with matched biopsy samples from 175 patients who underwent a clinically indicated kidney-transplant biopsy, we isolated urinary exosomal mRNAs and developed rejection signatures on the basis of differential gene expression. We used crossvalidation to assess the performance of the signatures on multiple data subsets.ResultsAn exosomal mRNA signature discriminated between biopsy samples from patients with all-cause rejection and those with no rejection, yielding an area under the curve (AUC) of 0.93 (95% CI, 0.87 to 0.98), which is significantly better than the current standard of care (increase in eGFR AUC of 0.57; 95% CI, 0.49 to 0.65). The exosome-based signature’s negative predictive value was 93.3% and its positive predictive value was 86.2%. Using the same approach, we identified an additional gene signature that discriminated patients with T cell–mediated rejection from those with antibody-mediated rejection (with an AUC of 0.87; 95% CI, 0.76 to 0.97). This signature’s negative predictive value was 90.6% and its positive predictive value was 77.8%.ConclusionsOur findings show that mRNA signatures derived from urinary exosomes represent a powerful and noninvasive tool to screen for kidney allograft rejection. This finding has the potential to assist clinicians in therapeutic decision making.

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Cao, Yuhan, Yuanhui Shi, Yuwei Wang, Yanlang Yang, Wenjun Guo, Cuifeng Zhang, Wenjun Pei, and Cong Fu. "Exosomal hsa_circ_0008925 from Urine Is Related to Chronic Renal Fibrosis." Disease Markers 2022 (February 18, 2022): 1–10. http://dx.doi.org/10.1155/2022/1899282.

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At present, there is no noninvasive biomarker of renal fibrosis. The potential diagnostic value of urinary exosome-derived circRNAs from glomerular disease patients for renal fibrosis is still uncertain. Here, we first detected the expression of hsa_circ_0008925 in TGF-β1-cultured HK-2 cell-derived exosomes. Secondly, we collected urine samples from 95 biopsy-proven glomerular disease patients and 34 healthy controls. The expression of hsa_circ_0008925 was analyzed, and the correlation with renal function and pathological changes was calculated. The receiver operating characteristic (ROC) curve for the diagnosis of renal fibrosis was performed. The results showed that in exosomes derived from TGF-β1-cultured HK-2 cells, the expression of hsa_circ_0008925 was increased compared with normal cultured. Further, the expression level of hsa_circ_0008925 was increased in urinary exosomes from renal fibrosis patients and correlated with serum creatinine, blood urea nitrogen (BUN), estimated glomerular filtration rate, and cystatin C. The level of hsa_circ_0008925 was furthermore correlated with the score of tubulointerstitial fibrosis (TIF) and the score of glomerular sclerosis. The ROC curve showed that hsa_circ_0008925 can diagnose renal fibrosis at a cut-off value of 0.093 with a sensitivity of 52.2% and specificity of 96.4%. In summary, we indicated that urinary exosomal hsa_circ_0008925 could be acted as a noninvasive biomarker for renal fibrosis in glomerular diseases patients.

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Fenner, Annette. "Urinary exosome biomarkers of radiation exposure." Nature Reviews Urology 13, no. 8 (June 28, 2016): 437. http://dx.doi.org/10.1038/nrurol.2016.122.

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Bielopolski, Dana, Andrea Ronning, Dacia Vasquez, Glenis George-Alexander, Jeanne Walker, Jonathan N. Tobin, and Rhonda G. Kost. "4393 Translational Characterization of Blood Pressure Changes Following the DASH Diet– from Nutrition to Electrolytes to Exosomes." Journal of Clinical and Translational Science 4, s1 (June 2020): 41. http://dx.doi.org/10.1017/cts.2020.157.

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OBJECTIVES/GOALS: 1.analyze urinary protein exosome content pattern before and during DASH diet.2.characterize urine electrolyte changes associated with changes in protein profiles, and hormonal changes before/after DASH diet.3.analyze the association of these changes to the DASH-related BP response.METHODS/STUDY POPULATION: In this proof of concept study, hypertension stage 1 volunteers will receive a DASH based menu during 14 consecutive days of elective admission to the RU research hospital. Participants will complete a food frequency questionnaire (VioScreen) with a bionutritionist. Throughout the intervention period, participants will be assessed for blood pressure, plasma renin and aldosterone, and 24 hour urines for electrolytes, creatinine, protein, albumin and first morning urine collected for exosomes. Exosome analysis will be performed by a commercial lab. Proteome analysis will be conducted in the RU Mass-spectrometry service. RESULTS/ANTICIPATED RESULTS: The causal pathway we will elucidate hypothesizes that: 1) changes in diet affect blood electrolytes, and through these, aldosterone. 2) Aldosterone alters the expression of specific transporter proteins in the renal tubule; protein expression will be reflected in the urine exosome. 3) These transporters affect the excretion of electrolytes, as reflected by urinary ratio of sodium (Na) to Potassium (K). During consumption of the Western diet, the Na/K ratio is approximately 2-2.5, whereas we expect the urinary sodium/potassium ratio to be <1, when the participant is eating a DASH based diet. DISCUSSION/SIGNIFICANCE OF IMPACT: This assay provides a clinical tool to assess dietary adherence, and the project will provide insights into the mechanism whereby DASH reduces blood pressure.

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Roberts, Douglas, Emily Mitsock, Olubode Ogunlusi, Seth Yu, and Johan Skog. "Biomarker screening for the early detection of prostate cancer using an exosomal-enrichment RNA liquid biopsy test." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): e15553-e15553. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e15553.

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e15553 Background: Prostate cancer is one of the most common cancers in men, with approximately 10% of all new cancer cases and ~5% of all cancer deaths in 2019. The standard test for prostate cancer is the Prostate Serum Antigen (PSA) test. The PSA test suffers from low specificity (20-40%) in patients including ‘grey zone’ levels (4-10 ng/mL); moreover, the PSA test fails to identify patients with high-risk cancers. Previously we developed ExoDx Prostate Intelliscore (EPI), a urine-based exosome prostate cancer test optimized to rule out the need for a biopsy (risk stratification for high-grade prostate cancer). This study utilized a next generation exosome-based test that specifically enriches a subtype of prostate cancer exosomes from urine. Early detection of prostate cancer via a non-invasive method is desirable and the identification of patients with high-risk cancer is critical. Here we describe the development of a prostate-specific urinary exosome test for the identification of patients with prostate cancer. Methods: We have developed a prostate-specific enrichment method to isolate exosomes of prostate origin from urine samples. Using an affinity-based method against surface marker proteins found on prostate cells, we were able to selectively enrich for exosomes shed by the prostate gland with demonstrated specificity. Subsequent analysis of exosomal nucleic acids enables a promising panel of gene expression biomarkers capable of distinguishing patients with prostate cancer from healthy individuals. Results: RNA from prostate cancer enriched exosomes was compared to total exosomes from urine. Enrichment of prostate cancer specific exosomes significantly enhanced the RNA signature compared to total urine exosomes. Conclusions: Prostate cancer tests have recently been developed for RNA signatures in urine. Exosomes provide a source of nucleic acids as they are actively shed continuously from living cells as part of their normal life cycle. The urine exosomes can be used for total RNA transcriptome analysis and are therefore a very rich source of biomarkers for prostate cancer that can be tailored to different clinical indications. An affinity-based enrichment for tissue-specific exosomes allow for better resolution of the gene expression profile from the tissue of interest and reduces the RNA targets from non-relevant processes of the bladder and kidneys. The gene signature identified in this ongoing study could potentially provide a non-invasive molecular means for the early diagnosis of prostate cancer.

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Riffo-Campos, Angela L., Javier Perez-Hernandez, Olga Martinez-Arroyo, Ana Ortega, Ana Flores-Chova, Josep Redon, and Raquel Cortes. "Biofluid Specificity of Long Non-Coding RNA Profile in Hypertension: Relevance of Exosomal Fraction." International Journal of Molecular Sciences 23, no. 9 (May 6, 2022): 5199. http://dx.doi.org/10.3390/ijms23095199.

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Non-coding RNA (ncRNA)-mediated targeting of various genes regulates the molecular mechanisms of the pathogenesis of hypertension (HTN). However, very few circulating long ncRNAs (lncRNAs) have been reported to be altered in essential HTN. The aim of our study was to identify a lncRNA profile in plasma and plasma exosomes associated with urinary albumin excretion in HTN by next-generation sequencing and to assess biological functions enriched in response to albuminuria using GO and KEGG analysis. Plasma exosomes showed higher diversity and fold change of lncRNAs than plasma, and low transcript overlapping was found between the two biofluids. Enrichment analysis identified different biological pathways regulated in plasma or exosome fraction, which were implicated in fatty acid metabolism, extracellular matrix, and mechanisms of sorting ncRNAs into exosomes, while plasma pathways were implicated in genome reorganization, interference with RNA polymerase, and as scaffolds for assembling transcriptional regulators. Our study found a biofluid specific lncRNA profile associated with albuminuria, with higher diversity in exosomal fraction, which identifies several potential targets that may be utilized to study mechanisms of albuminuria and cardiovascular damage.

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Li, Guorong, Nora Mallouk, Pascale Flandrin, Arnauld Garcin, Claude Lambert, Sid Ali Berremila, Hocine Habchi, and Nicolas Mottet. "Presence of Urinary Exosomes for Liquid Biopsy of Clear Cell Renal Cell Carcinoma: Protocol for a Pilot Feasibility Study." JMIR Research Protocols 10, no. 7 (July 20, 2021): e24423. http://dx.doi.org/10.2196/24423.

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Background Approximately 70%-80% of kidney cancers are clear cell renal cell carcinomas (CCRCCs). Patient management is based on imaging (abdominal ultrasound and computerized tomography), surgical excision of the tumor, and pathological analysis. A tissue biopsy is therefore necessary to confirm the diagnosis and avoid unnecessary nephrectomy. For metastatic cancers, a tissue biopsy is essential for establishing the targeted therapy. This biopsy of tumor material is invasive and painful. Other techniques such as liquid biopsy would help reduce the need for tissue biopsy. The development of a simple biological test for diagnosis is essential. CA9 is a powerful marker for the diagnosis of CCRCC. Exosomes have become a major source of liquid biopsy because they carry tumor proteins, RNA, and lipids. Urine is the most convenient biological liquid for exosome sampling. Objective The aim of this study (PEP-C study) is mainly to determine whether it is possible to detect urinary exosomal CA9 for the molecular diagnosis of CCRCC. Methods This study will include 60 patients with CCRCC and 40 noncancer patients. Exosomes will be isolated from urine samples and exosomal CA9 will be detected by transmission electron microscopy, flow cytometry, and reverse transcription-quantitative polymerase chain reaction. Results This study is currently underway with funding support from the CHU Saint-Etienne of France. Conclusions We expect to demonstrate that urinary tumor exosomes could be a novel liquid biopsy to diagnose CCRCC and to guide clinicians in treatment decision-making. Trial Registration ClinicalTrials.gov NCT04053855; https://clinicaltrials.gov/ct2/show/NCT04053855 International Registered Report Identifier (IRRID) DERR1-10.2196/24423

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Fernández-Llama, Patricia, Sookkasem Khositseth, Patricia A. Gonzales, Robert A. Star, Trairak Pisitkun, and Mark A. Knepper. "Tamm-Horsfall protein and urinary exosome isolation." Kidney International 77, no. 8 (April 2010): 736–42. http://dx.doi.org/10.1038/ki.2009.550.

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Ni, Jianshu, Hongchao Li, Yiwen Zhou, Baojun Gu, Yuemin Xu, Qiang Fu, Xufeng Peng, et al. "Therapeutic Potential of Human Adipose-Derived Stem Cell Exosomes in Stress Urinary Incontinence – An in Vitro and in Vivo Study." Cellular Physiology and Biochemistry 48, no. 4 (2018): 1710–22. http://dx.doi.org/10.1159/000492298.

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Background/Aims: To evaluate whether local injection of exosomes derived from human adipose-derived stem cells (hADSCs) facilitates recovery of stress urinary incontinence (SUI) in a rat model. Methods: For the in vitro study, a Cell Counting Kit-8 (CCK-8) array and proteomic analysis were performed. For the in vivo study, female rats were divided into four groups: sham, SUI, adipose-derived stem cell (ADSC), and exosomes (n = 12 each). The SUI model was generated by pudendal nerve transection and vaginal dilation. Vehicle, hADSCs, or exosomes were injected into the peripheral urethra. After 2, 4, and 8 weeks, the rats underwent cystometrography and leak point pressure (LPP) testing, and tissues were harvested for histochemical analyses. Results: The CCK-8 experiment demonstrated that ADSC-derived exosomes could enhance the growth of skeletal muscle and Schwann cell lines in a dose-dependent manner. Proteomic analysis revealed that ADSC-derived exosomes contained various proteins of different signaling pathways. Some of these proteins are associated with the PI3K-Akt, Jak-STAT, and Wnt pathways, which are related to skeletal muscle and nerve regeneration and proliferation. In vivo experiments illustrated that rats of the exosome group had higher bladder capacity and LPP, and had more striated muscle fibers and peripheral nerve fibers in the urethra than rats of the SUI group. Both urethral function and histology of rats in the exosome group were slightly better than those in the ADSC group. Conclusions: Local injection of hADSC-derived exosomes improved functional and histological recovery after SUI.

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Bijnsdorp, I., L. A. Erozenci, J. Koetsier, F. Feenstra, S. R. Piersma, R. J. Van Moorselaar, A. Vis, et al. "The prostate cancer urinary exosome protein biomarker landscape." European Urology Open Science 19 (July 2020): e517-e518. http://dx.doi.org/10.1016/s2666-1683(20)32911-6.

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Lv, Lin-Li, Ye Feng, Yi Wen, Wei-Jun Wu, Hai-Feng Ni, Zuo-Lin Li, Le-Ting Zhou, et al. "Exosomal CCL2 from Tubular Epithelial Cells Is Critical for Albumin-Induced Tubulointerstitial Inflammation." Journal of the American Society of Nephrology 29, no. 3 (January 2, 2018): 919–35. http://dx.doi.org/10.1681/asn.2017050523.

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Albuminuria is a key instigator of tubulointerstitial inflammation associated with CKD, but the mechanism through which filtered albumin propagates renal injury remains unclear. In this study, we explored the role in this process of exosome mRNA released from tubular epithelial cells (TECs). Compared with control mice, acute and chronic kidney injury models had more exosomes containing inflammatory cytokine mRNA, particularly the chemokine CCL2, in kidneys and urine. In vitro stimulation of TECs with BSA recapitulated this finding. Notably, the internalization of purified TEC exosomes by cultured macrophages increased if TECs were exposed to BSA. Macrophage internalization of exosomes from BSA-treated TECs led to an enhanced inflammatory response and macrophage migration, but CCL2 silencing in TECs prevented these effects. Using a GFP-CCL2 fusion mRNA construct, we observed direct transfer of CCL2 mRNA from TEC exosomes to macrophages. Mice subjected to tail vein injection of purified BSA-treated TEC exosomes developed tubular injury with renal inflammatory cell infiltration. However, injection of exosomes from BSA-treated CCL2-deficient TECs induced less severe kidney inflammation. Finally, in patients with IgA nephropathy, the increase of proteinuria correlated with augmented urinary excretion of exosomes with exaggerated expression of CCL2 mRNA. Moreover, the level of CCL2 mRNA in urinary exosomes correlated closely with levels of renal interstitial macrophage infiltration in these patients. Our studies demonstrate that the increasing release of exosomes that transfer CCL2 mRNA from TECs to macrophages constitutes a critical mechanism of albumin-induced tubulointerstitial inflammation.

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Fernandez-Llama, P., S. Khositseth, PA Gonzales, RA Star, T. Pisitkun, and MA Knepper. "TAMM-HORSFALL PROTEIN AND URINARY EXOSOME ISOLATION: PP.9.380." Journal of Hypertension 28 (June 2010): e164. http://dx.doi.org/10.1097/01.hjh.0000378704.37967.40.

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Bruschi, Maurizio, Laura Santucci, Silvia Ravera, Giovanni Candiano, Martina Bartolucci, Daniela Calzia, Chiara Lavarello, et al. "Human urinary exosome proteome unveils its aerobic respiratory ability." Journal of Proteomics 136 (March 2016): 25–34. http://dx.doi.org/10.1016/j.jprot.2016.02.001.

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Zhao, Yao, Kaiyong Chen, Haining Li, and Huiyi Wu. "Effect of pH on the isolation of urinary exosome." International Urology and Nephrology 49, no. 1 (September 27, 2016): 165–69. http://dx.doi.org/10.1007/s11255-016-1408-7.

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Ramirez-Alvarado, Marina, Christopher J. Ward, Bing Q. Huang, Xun Gong, Marie C. Hogan, Benjamin J. Madden, Cristine Charlesworth, Angela Dispenzieri, Morie A. Gertz, and Nelson Leung. "Detection of High Molecular Weight Light Chain Oligomers in Urinary Exosomes of Patients with AL Amyloidosis." Blood 114, no. 22 (November 20, 2009): 4886. http://dx.doi.org/10.1182/blood.v114.22.4886.4886.

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Abstract Abstract 4886 Detection of high molecular weight light chain oligomers in urinary exosomes of patients with AL amyloidosis. Background Exosomes are microvesicles that are part of the multivesicular body (MVB) pathway. They are created by the inward budding of the cell surface membrane and contain both surface bound membrane proteins and cytosolic proteins which can be used to identify the cell of origin. Immunoglobulin light chain amyloidosis (AL) occurs as the result of amyloid formation by the misfolding of monoclonal light chains (LC) and deposition of these amyloid fibrils in various soft tissues. This reaction requires the organization of the monoclonal LC's into protofibrils which are then weave into amyloid fibrils. This study was undertaken to determine whether urinary exosomes of glomerular origin contain intermediate species of amyloid formation. Method Urine samples from patients with AL, light chain deposition disease (LCDD), multiple myeloma (MM) and monoclonal clonal gammopathy of undetermined significance (MGUS) were collected. Urinary exosomes were isolated and separated into fractions by gradient centrifugation. Western blots were performed on the urinary exosome fractions using anti-kappa or anti-lambda antibodies. Glomerular fractions were identified using antibodies directed toward podocin. Results Urine samples were collected from 5 patients with AL, 2 from LCDD, 1 from MM and 1 MGUS. On the Western blot, immunoglobulin LC were seen in all exosomal fractions in patients with AL amyloidosis, LCDD, MM but not MGUS which is similar to normal controls (not shown). In patients with AL, oligomeric species were found in the highest concentrations in fraction 4 and 5 (Figure 1). Fraction 4 and 5 were also stained for podocin, a glomerular protein (not shown). The highest molecular weight species was ∼250 kd which corresponds to a LC decamer. High molecular weight species were also identified in 1 of 2 LCDD patients corresponding to a tetramer. The band was identified in fraction 10 which had polycystin-1 expression suggesting a tubular origin. No high molecular weight LC species was found in patients with MM or MGUS. Conclusion Our study found high molecular weight LC species corresponding to the intermediates involved in protofibril formation in urinary exosomes of patients with AL. Smaller (tetramer) high molecular weight LC species were also found in a patient with LCDD but not in patients with MM and MGUS. Not only were the high molecular weight LC species found exclusively in the diseases characterized by deposition of LC aggregates, they were also found in the segments of the nephron where the deposits were expected: glomerulus for AL and tubular epithelium for LCDD. This is consistent with our current understanding of the pathogenic mechanisms of these diseases. We believe urinary exosomes are a powerful tool in the study of diseases involving self-aggregation of monoclonal proteins. It has tremendous potential in both diagnostic and scientific research in this area. Disclosures Gertz: celgene: Honoraria; millenium: Honoraria, Membership on an entity's Board of Directors or advisory committees.

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Bruschi, Maurizio, Silvia Ravera, Laura Santucci, Giovanni Candiano, Martina Bartolucci, Daniela Calzia, Chiara Lavarello, et al. "The human urinary exosome as a potential metabolic effector cargo." Expert Review of Proteomics 12, no. 4 (July 4, 2015): 425–32. http://dx.doi.org/10.1586/14789450.2015.1055324.

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Lv, Lin-Li, Yu-Han Cao, Ming-Ming Pan, Hong Liu, Ri-Ning Tang, Kun-Ling Ma, Ping-Sheng Chen, and Bi-Cheng Liu. "CD2AP mRNA in urinary exosome as biomarker of kidney disease." Clinica Chimica Acta 428 (January 2014): 26–31. http://dx.doi.org/10.1016/j.cca.2013.10.003.

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Awdishu, Linda, Shirley Tsunoda, Michelle Pearlman, Chanthel Kokoy-Mondragon, Majid Ghassemian, Robert K. Naviaux, Heather M. Patton, Ravindra L. Mehta, Bhavya Vijay, and Satish P. RamachandraRao. "Identification of Maltase Glucoamylase as a Biomarker of Acute Kidney Injury in Patients with Cirrhosis." Critical Care Research and Practice 2019 (April 16, 2019): 1–8. http://dx.doi.org/10.1155/2019/5912804.

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Background. Acute kidney injury (AKI) is a frequent complication of decompensated cirrhosis with increased mortality. Traditional biomarkers such as serum creatinine are not sensitive for detecting injury without functional change. We hypothesize that urinary exosomes potentially carry markers that differentiate the type of kidney injury in cirrhotic patients. Methods. This is a prospective, single-center, and observational study of adult patients with cirrhosis. The patient groups included healthy normal controls, compensated cirrhosis with normal kidney function, decompensated cirrhosis with normal kidney function, and decompensated cirrhosis with AKI. Data were extracted from the electronic health record including etiology of liver disease, MELD score, history of decompensation, Child-Turcotte-Pugh score, history of AKI, and medication exposures. Urine samples were collected at the time of consent. Urine exosome protein content was analyzed, and proteomic data were validated by immunoblotting. Statistical analysis included partial least squares-discriminant analysis coupled with variable importance in projection identification. Results. Eighteen cirrhotic subjects were enrolled, and six healthy control subjects were extracted from our biorepository. Urine exosomes were isolated, and 1572 proteins were identified. Maltase-glucoamylase was the top discriminating protein confirmed by western blotting. Conclusions. Patients with cirrhosis and AKI have upregulation of renal brush border disaccharidase, MGAM, in urinary exosomes which may differentiate the type of kidney injury in cirrhosis; however, the clinical significance of this requires further validation.

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Valera, Vladimir, Beatriz Walter, and Maria J. Merino. "Abstract 5831: Urinary exosome analysis as a marker of treatment response in bladder cancer patients." Cancer Research 82, no. 12_Supplement (June 15, 2022): 5831. http://dx.doi.org/10.1158/1538-7445.am2022-5831.

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Abstract Introduction: Intravesical Bacillus Calmette-Guerin (BCG) is an effective immunotherapy for non-muscle invasive bladder cancer (NMIBC). However, BCG treatment failure will lead to recurrence and tumor progression. In this study, urinary exosomes content (miRNA profile) was evaluated as a possible marker of BCG treatment response in NMIBC patients. Methods: Urine samples from patients with bladder cancer were collected at the time of surgery and during patient follow up to 1 year. Urine from healthy volunteers were also included. Clinical and pathological information such as tumor grade (High Grade (HG), Low Grade (LG)), depth of invasion (Ta, T1, CIS), and response to BCG treatment was also obtained. Exosome Isolation and total RNA extraction including microRNAs from cell-free urine after centrifugation were obtained. Library preparation for miRNA expression was done (QIAseq® miRNA Library) for Next-Generation Sequencing (NGS) analysis in a NextSeq 2000 single read platform, 75 bp with 15-20 million reads per sample. Reads were then queried against miRDeep2 software for identification. Only miRNAs having at least 20 counts considering all samples were included. After normalization, significantly and deferentially expressed microRNAs (>2-Fold) were selected for analysis. Bioinformatic analysis including sequence alignment was performed under the STAR-based approach. Identified microRNAs were then used to classify/predict the response to treatment and its relationship with other clinicopathologic variables. Results: A total of 56 urine samples from 13 patients were available/used for analysis including 10 High Grade Ta and 3 High Grade T1 patients. Urine from normal healthy donors (N=3) was also included. Clinicopathological features were patients with HGTa=10, HGT1=3 and 3 control samples. Regarding treatment response 9 patients were BCG responders and 4 BCG unresponsive. When compared to BCG unresponsive patients, BCG responders showed 45 differentially expressed miRNAs. Statistically significant differentially expressed miRNAs (Fold-change >2, p value <0.05) were 12 miRNAs, upregulated were miR132-3p (p=0.042); miR-187-3p (p=0.021); miR-409-3p (p=0.043) and miR1301-3p (p=0.048). Downregulated miRNAs were miR-let7-5p (p=0.007), miR-3605-3p (p=0.047), miR-140-5p (p=0.031), miR-500a-5p (p=0.051), miR-629-5p (p=0.039), miR-454-3p (p=0.05), miR-2110 (p=0.049) and miR-30c-5p (p=0.03). Interestingly, miRPathDABv2.0 analysis predicted those microRNAs be related to cancer, including bladder cancer. They showed targeting important pathways as PI3K.Akt, Wnt/B catenin and P53 signaling related with disease progression and treatment response. Conclusion: Our study supports the value of urinary exosomal microRNAs as non-invasive biomarkers to predict BCG treatment response in nonmuscle-invasive bladder cancer. Citation Format: Vladimir Valera, Beatriz Walter, Maria J. Merino. Urinary exosome analysis as a marker of treatment response in bladder cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5831.

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Chun-yan, Lv, Zhao Zi-yi, Yang Tian-lin, Wang Yi-li, Li Bao, Lv Jiao, and Ding Wei-jun. "Liquid biopsy biomarkers of renal interstitial fibrosis based on urinary exosome." Experimental and Molecular Pathology 105, no. 2 (October 2018): 223–28. http://dx.doi.org/10.1016/j.yexmp.2018.08.004.

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Choi, Hojun, Myung-Yoon Kim, Dae-Hwan Kim, Hanoul Yun, Byung-Koo Oh, Su-Bin Kim, In-Ho Song, et al. "Quantitative Biodistribution and Pharmacokinetics Study of GMP-Grade Exosomes Labeled with 89Zr Radioisotope in Mice and Rats." Pharmaceutics 14, no. 6 (May 24, 2022): 1118. http://dx.doi.org/10.3390/pharmaceutics14061118.

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For the successful clinical advancement of exosome therapeutics, the biodistribution and pharmacokinetic profile of exogenous exosomes in various animal models must be determined. Compared with fluorescence or bioluminescence imaging, radionuclide imaging confers multiple advantages for the in vivo tracking of biomolecular therapeutics because of its excellent sensitivity for deep tissue imaging and potential for quantitative measurement. Herein, we assessed the quantitative biodistribution and pharmacokinetics of good manufacturing practice-grade therapeutic exosomes labeled with zirconium-89 (89Zr) after systemic intravenous administration in mice and rats. Quantitative biodistribution analysis by positron emission tomography/computed tomography and gamma counting in mice and rats revealed that the total 89Zr signals in the organs were lower in rats than in mice, suggesting a higher excretion rate of exosomes in rats. A prolonged 89Zr signal for up to 7 days in most organs indicated that substantial amounts of exosomes were taken up by the parenchymal cells in those organs, highlighting the therapeutic potential of exosomes for the intracellular delivery of therapeutics. Exosomes were mainly distributed in the liver and to a lesser extent in the spleen, while a moderately distributed in the kidney, lung, stomach, intestine, urinary bladder, brain, and heart. Exosomes were rapidly cleared from the blood circulation, with a rate greater than that of free 89Zr, indicating that exosomes might be rapidly taken up by cells and tissues.

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Bruschi, Granata, Candiano, Fabris, Petretto, Ghiggeri, Gambaro, and Zaza. "Proteomic Analysis of Urinary Extracellular Vesicles Reveals a Role for the Complement System in Medullary Sponge Kidney Disease." International Journal of Molecular Sciences 20, no. 21 (November 5, 2019): 5517. http://dx.doi.org/10.3390/ijms20215517.

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Medullary sponge kidney (MSK) disease is a rare and neglected kidney condition often associated with nephrocalcinosis/nephrolithiasis and cystic anomalies in the precalyceal ducts. Little is known about the pathogenesis of this disease, so we addressed the knowledge gap using a proteomics approach. The protein content of microvesicles/exosomes isolated from urine of 15 MSK and 15 idiopathic calcium nephrolithiasis (ICN) patients was investigated by mass spectrometry, followed by weighted gene coexpression network analysis, support vector machine (SVM) learning, and partial least squares discriminant analysis (PLS-DA) to select the most discriminative proteins. Proteomic data were verified by ELISA. We identified 2998 proteins in total, 1764 (58.9%) of which were present in both vesicle types in both diseases. Among the MSK samples, only 65 (2.2%) and 137 (4.6%) proteins were exclusively found in the microvesicles and exosomes, respectively. Similarly, among the ICN samples, only 75 (2.5%) and 94 (3.1%) proteins were exclusively found in the microvesicles and exosomes, respectively. SVM learning and PLS-DA revealed a core panel of 20 proteins that distinguished extracellular vesicles representing each clinical condition with an accuracy of 100%. Among them, three exosome proteins involved in the lectin complement pathway maximized the discrimination between MSK and ICN: Ficolin 1, Mannan-binding lectin serine protease 2, and Complement component 4-binding protein β. ELISA confirmed the proteomic results. Our data show that the complement pathway is involved in the MSK, revealing a new range of potential therapeutic targets and early diagnostic biomarkers.

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Kwon, H. J., C. E. Yoon, D. Shin, S. Shin, H. W. Moon, H. J. Cho, U. Ha, et al. "Urinary exosome microRNA signatures as a noninvasive prognostic biomarker for prostate cancer." European Urology 81 (February 2022): S1371—S1372. http://dx.doi.org/10.1016/s0302-2838(22)01010-7.

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Nolte, Adam, George Wayne*, Elizabeth Nagoda, Vivian Wong, Juan Cedeno, Alejandra Perez, and Alan Nieder. "MP27-08 URINARY EXOSOME TEST AND MP-MRI FOR PROSTATE CANCER SCREENING." Journal of Urology 203 (April 2020): e412-e413. http://dx.doi.org/10.1097/ju.0000000000000866.08.

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Silvers*, Christopher, Hiroshi Miyamoto, Edward M. Messing, and Yi-Fen Lee. "PD47-08 URINARY EXOSOME CYTOKINE LEVELS CORRELATE WITH PELVIC LYMPH NODE INFLAMMATION." Journal of Urology 203 (April 2020): e928. http://dx.doi.org/10.1097/ju.0000000000000934.08.

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Walker, Jillian Marie, Padraic O’Malley, and Mei He. "Applications of Exosomes in Diagnosing Muscle Invasive Bladder Cancer." Pharmaceutics 14, no. 10 (September 23, 2022): 2027. http://dx.doi.org/10.3390/pharmaceutics14102027.

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Muscle Invasive Bladder Cancer (MIBC) is a subset of bladder cancer with a significant risk for metastases and death. It accounts for nearly 25% of bladder cancer diagnoses. A diagnostic work-up for MIBC is inclusive of urologic evaluation, radiographic imaging with a CT scan, urinalysis, and cystoscopy. These evaluations, especially cystoscopy, are invasive and carry the risk of secondary health concerns. Non-invasive diagnostics such as urine cytology are an attractive alternative currently being investigated to mitigate the requirement for cystoscopy. A pitfall in urine cytology is the lack of available options with high reliability, specificity, and sensitivity to malignant bladder cells. Exosomes are a novel biomarker source which could resolve some of the concerns with urine cytology, due to the high specificity as the surrogates of tumor cells. This review serves to define muscle invasive bladder cancer, current urine cytology methods, the role of exosomes in MIBC, and exosomes application as a diagnostic tool in MIBC. Urinary exosomes as the specific populations of extracellular vesicles could provide additional biomarkers with specificity and sensitivity to bladder malignancies, which are a consistent source of cellular information to direct clinicians for developing treatment strategies. Given its strong presence and differentiation ability between normal and cancerous cells, exosome-based urine cytology is highly promising in providing a perspective of a patient’s bladder cancer.

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Earle, Angel, Madison Bessonny, Josh Benito, Kun Huang, Hannah Parker, Emily Tyler, Brittany Crawford, et al. "Urinary Exosomal MicroRNAs as Biomarkers for Obesity-Associated Chronic Kidney Disease." Journal of Clinical Medicine 11, no. 18 (September 7, 2022): 5271. http://dx.doi.org/10.3390/jcm11185271.

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The early detection of chronic kidney disease (CKD) is key to reducing the burden of disease and rising costs of care. This need has spurred interest in finding new biomarkers for CKD. Ideal bi-omarkers for CKD should be: easy to measure; stable; reliably detected, even when interfering substances are present; site-specific based on the type of injury (tubules vs. glomeruli); and its changes in concentration should correlate with disease risk or outcome. Currently, no single can-didate biomarker fulfills these criteria effectively, and the mechanisms underlying kidney fibrosis are not fully understood; however, there is growing evidence in support of microRNA-mediated pro-cesses. Specifically, urinary exosomal microRNAs may serve as biomarkers for kidney fibrosis. In-creasing incidences of obesity and the recognition of obesity-associated CKD have increased interest in the interplay of obesity and CKD. In this review, we provide: (1) an overview of the current scope of CKD biomarkers within obese individuals to elucidate the genetic pathways unique to obesi-ty-related CKD; (2) a review of microRNA expression in obese individuals with kidney fibrosis in the presence of comorbidities, such as diabetes mellitus and hypertension; (3) a review of thera-peutic processes, such as diet and exercise, that may influence miR-expression in obesity-associated CKD; (4) a review of the technical aspects of urinary exosome isolation; and (5) future areas of research.

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Xie, Yijun, Yijie Jia, Xie Cuihua, Fang Hu, Meng Xue, and Yaoming Xue. "Urinary Exosomal MicroRNA Profiling in Incipient Type 2 Diabetic Kidney Disease." Journal of Diabetes Research 2017 (September 5, 2017): 1–10. http://dx.doi.org/10.1155/2017/6978984.

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Background. Albuminuria is an early sign but not a strong predictor of diabetic kidney disease (DKD). Owing to their high stability, urinary exosomal miRNAs can be useful predictors of the progression of early-stage DKD to renal failure; fluid biopsies are ideal for detecting abnormalities in these miRNAs. The aim of this study was to identify novel differentially expressed miRNAs as urine biomarkers for type 2 DKD by comparing between patients of type 2 diabetes (T2D) with and without macroalbuminuria. Methods. Ten patients with T2D, including five who had no renal disease and five with macroalbuminuria (DKD G1-2A3), were selected for this study. Exosome- (UExo-) derived miRNA profiles were used to identify candidate biomarkers, a subset of which was verified using quantitative reverse transcription PCR. Results. A total of 496 UExo-derived miRNA species were found to be differentially expressed (>2-fold) in patients with DKD, compared to those with T2D. A validation analysis revealed that three miRNAs (miR-362-3p, miR-877-3p, and miR-150-5p) were upregulated and one (miR-15a-5p) was downregulated. These miRNAs might regulate DKD through p53, mTOR, and AMPK pathways. Conclusions. In conclusion, UExo-derived miRNAs were altered in type 2 DKD. MiR-362-3p, miR-877-3p, miR-150-5p, and miR-15a-5p might be novel biomarkers for incipient DKD.

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Li, Yun, Jin Ji, Ji Lyu, Xin Jin, Xing He, Shaojia Mo, Huan Xu, et al. "A Novel Urine Exosomal lncRNA Assay to Improve the Detection of Prostate Cancer at Initial Biopsy: A Retrospective Multicenter Diagnostic Feasibility Study." Cancers 13, no. 16 (August 13, 2021): 4075. http://dx.doi.org/10.3390/cancers13164075.

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Purpose: This study aimed at developing and validating a novel noninvasive urinary exosome-based post-DRE (digital rectal examination) lncRNA assay to diagnose PCa (prostate cancer) and clinically significant PCa (Gleason score ≥ 7) from the initial prostate biopsy. Methods: A total of 602 urine samples from eligible participants were collected. The expression levels of urinary exosomal PCA3 (prostate cancer antigen 3) and MALAT1 (metastasis-associated lung adenocarcinoma transcript 1) were detected by qPCR (quantitative real-time PCR). Receiver operating characteristic (ROC) analysis was applied to evaluate the diagnostic performance of PCA3, MALAT1 and the lncRNA assay. A decision curve analysis (DCA) and waterfall plots were used to assess the clinical value of the lncRNA assay. Results: Urinary exosomal PCA3 and MALAT1 were overexpressed in PCa and clinically significant PCa (p < 0.001). The lncRNA assay combining PCA3 and MALAT1 had a better diagnostic performance (AUC 0.828) than the current clinical parameters in detecting PCa. More importantly, the lncRNA assay yielded an AUC of 0.831 to detect clinically significant PCa, which is much higher than that of the current clinical parameters. The lncRNA assay was superior to PSA, f/tPSA and the base model for detecting PCa and clinically significant PCa, with a higher net benefit for almost all threshold probabilities. At the cutoff value of 95% sensitivity, the lncRNA assay could avoid 24.2% unnecessary biopsies while only missing 1.2% of the cases of clinically significant PCa. Conclusion: We developed and validated a novel noninvasive post-DRE urine-based lncRNA assay that presented good diagnostic power and clinical utility for the early diagnosis of PCa and high-grade PCa.

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Ortega, Ana, Olga Martínez-Arroyo, Javier Perez-Hernandez, Felipe J. Chaves, Josep Redon, and Raquel Cortes. "URINARY EXOSOME MIR-146A IS A POTENTIAL MARKER OF ALBUMINURIA IN ESSENTIAL HYPERTENSION." Journal of Hypertension 39, Supplement 1 (April 2021): e296. http://dx.doi.org/10.1097/01.hjh.0000747872.49377.26.

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Gildea, John J., Julia M. Carlson, Cynthia D. Schoeffel, Robert M. Carey, and Robin A. Felder. "Urinary exosome miRNome analysis and its applications to salt sensitivity of blood pressure." Clinical Biochemistry 46, no. 12 (August 2013): 1131–34. http://dx.doi.org/10.1016/j.clinbiochem.2013.05.052.

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Zatz, Roberto, and Clarice Kazue Fujihara. "Discarding the haystack to examine the needles: the potential role of urinary exosome analysis." American Journal of Physiology-Renal Physiology 305, no. 11 (December 1, 2013): F1544—F1545. http://dx.doi.org/10.1152/ajprenal.00504.2013.

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LEE, KIYOUNG, DAEHO LEE, and IE BYUNG PARK. "500-P: The Performance of Four Different Exosome Isolation Methods for Urinary Exosomal microRNA Profiling in Patients with Diabetic Kidney Disease." Diabetes 68, Supplement 1 (June 2019): 500—P. http://dx.doi.org/10.2337/db19-500-p.

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Hinrichs, Gitte R., Jannie S. Michelsen, Rikke Zachar, Ulla G. Friis, Per Svenningsen, Henrik Birn, Claus Bistrup, and Boye L. Jensen. "Albuminuria in kidney transplant recipients is associated with increased urinary serine proteases and activation of the epithelial sodium channel." American Journal of Physiology-Renal Physiology 315, no. 1 (July 1, 2018): F151—F160. http://dx.doi.org/10.1152/ajprenal.00545.2017.

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Albuminuria predicts adverse renal outcome in kidney transplant recipients. The present study addressed the hypothesis that albuminuria is associated with increased urine serine proteases with the ability to activate the epithelial sodium channel (ENaC) and with greater extracellular volume and higher blood pressure. In a cross-sectional design, kidney transplant recipients with ( n = 18) and without ( n = 19) albuminuria were included for office blood pressure measurements, estimation of volume status by bioimpedance, and collection of spot urine and plasma samples. Urine was analyzed for serine proteases and for the ability to activate ENaC current in vitro. Urine exosome protein was immunoblotted for prostasin and γ-ENaC protein. In the present study, it was found that, compared with nonalbuminuria (8.8 mg/g creatinine), albuminuric (1,722 mg/g creatinine) kidney transplant recipients had a higher systolic and diastolic blood pressure, despite receiving significantly more antihypertensives, and a greater urinary total plasminogen, active plasmin, active urokinase-type plasminogen activator, and prostasin protein abundance, which correlated significantly with u-albumin. Fluid overload correlated with systolic blood pressure, urinary albumin/creatinine, and plasminogen/creatinine. Urine from albuminuric kidney transplant recipients evoked a greater amiloride- and aprotinin-sensitive inward current in single collecting duct cells (murine cell line M1). γENaC subunits at 50 and 75 kDa showed increased abundance in urine exosomes from albuminuric kidney transplant recipients when compared with controls. These findings show that albuminuria in kidney transplant recipients is associated with hypertension, ability of urine to proteolytically activate ENaC current, and increased abundance of γENaC. ENaC activity could contribute to hypertension and adverse outcome in posttransplant proteinuria.

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Wang, Chao, Xiang Liu, Hongyan Li, Libo Zhao, Guanyi Kong, Jing Chen, Zhi Li, Jianfei Qi, Ye Tian, and Fengbo Zhang. "Urinary exosome-based androgen receptor-variant 7 detection in metastatic castration-resistant prostate cancer patients." Translational Andrology and Urology 11, no. 2 (February 2022): 202–12. http://dx.doi.org/10.21037/tau-21-1136.

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DAI, Yi, Yuan ZHANG, Feng QIU, Yan-Yan LI, and Zong-Yin QIU. "Analysis of Differentially Expressed Proteome in Urinary Exosome from Non-small Cell lung Cancer Patients." CHINESE JOURNAL OF ANALYTICAL CHEMISTRY (CHINESE VERSION) 38, no. 3 (May 20, 2010): 325–31. http://dx.doi.org/10.3724/sp.j.1096.2010.00325.

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DAI, Yi, Yuan ZHANG, Feng QIU, Yan-Yan LI, and Zong-Yin QIU. "Analysis of Differentially Expressed Proteome in Urinary Exosome from Non-small Cell Lung Cancer Patients." Chinese Journal of Analytical Chemistry 38, no. 3 (March 2010): 325–31. http://dx.doi.org/10.1016/s1872-2040(09)60030-x.

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Channavajjhala, Sarath, Wenjing Jia, Mahli Jalland, Kevin O'Shaughnessy, Ian Hall, and Mark Glover. "OS 08-05 NOVEL MOLECULAR INSIGHTS IN URINARY EXOSOME PROTEIN PROFILING IN THIAZIDE INDUCED HYPONATREMIA." Journal of Hypertension 34, Supplement 1 (September 2016): e69. http://dx.doi.org/10.1097/01.hjh.0000500030.16778.dd.

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Costa de Freitas, Renata Caroline, Raul Hernandes Bortolin, Fabiana Dalla Vecchia Genvigir, Vivian Bonezi, Thiago Dominguez Crespo Hirata, Claudia Rosso Felipe, Helio Tedesco-Silva, et al. "Differentially expressed urinary exo-miRs and clinical outcomes in kidney recipients on short-term tacrolimus therapy: a pilot study." Epigenomics 12, no. 22 (November 2020): 2019–34. http://dx.doi.org/10.2217/epi-2020-0160.

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Aim: To analyze the expression of urinary exosome-derived miRNAs (exo-miRs) in kidney recipients on tacrolimus-based therapy. Patients & methods: Clinical and drug monitoring data were recorded from 23 kidney recipients. Expression of 93 exo-miRs was measured by quantitative PCR array and mRNA targets were explored. Results: 16 exo-miRs were differentially expressed, including marked upregulation of miR-155-5p, and downregulation of miR-223-3p and miR-1228-3p. Expression of miR-155-5p and miR-223-3p correlated with tacrolimus dose (p < 0.05), miR-223-3p with serum creatinine (p < 0.05), and miR-223-3p and miR-1228-3p with blood leukocytes (p < 0.05). 12 miRNAs have predicted targets involved in cell proliferation, apoptosis, stress response, PIK3/AKT/mTOR and TGF-β signaling pathways. Conclusion: Differentially expressed urinary exo-miRs may be useful markers to monitor tacrolimus therapy and graft function in kidney transplantation.

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Cheruvanky, Anita, Hua Zhou, Trairak Pisitkun, Jeffrey B. Kopp, Mark A. Knepper, Peter S. T. Yuen, and Robert A. Star. "Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator." American Journal of Physiology-Renal Physiology 292, no. 5 (May 2007): F1657—F1661. http://dx.doi.org/10.1152/ajprenal.00434.2006.

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Urinary exosomes are excreted from all nephron segments and may serve as biomarkers for classifying renal diseases. Isolation of urinary exosomes by the established ultracentrifugation method has some limitations for use in a clinical laboratory. We sought a rapid and simple way to obtain urinary exosomes. We used a commercially available nanomembrane concentrator to enrich exosomes from urine by centrifugation at 3,000 g for 10–30 min. Urinary exosomal markers tumor susceptibility gene 101, aquaporin-2, neuron-specific enolase, annexin V, angiotensin-converting enzyme, and podocalyxin (PODXL) were recovered from the nanomembrane concentrator and detected by Western blotting, and typical features of urinary vesicles were found by electron microscopy. Exosomal markers were detected in as little as 0.5 ml of urine. By the nanomembrane method, exosomal proteins could be recovered from urine samples frozen at −80°C or refrigerated overnight at 4°C then stored at −80°C. By enriching exosomes we could detect PODXL, a podocyte marker, which decreased by 71% in five male patients with focal segmental glomerulosclerosis and abundant proteinuria. We conclude that 1) use of a nanomembrane concentrator simplifies and accelerates the enrichment of urinary exosomes; and 2) the nanomembrane concentrator can concentrate exosomal proteins from clinical urine samples. This enhanced method may accelerate the translation of urinary exosomal biomarkers from bench to bedside for the diagnosis, classification, and prognostication of renal diseases.

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Miranda, Kevin C., Daniel T. Bond, Joshua Z. Levin, Xian Adiconis, Andrey Sivachenko, Carsten Russ, Dennis Brown, Chad Nusbaum, and Leileata M. Russo. "Massively Parallel Sequencing of Human Urinary Exosome/Microvesicle RNA Reveals a Predominance of Non-Coding RNA." PLoS ONE 9, no. 5 (May 9, 2014): e96094. http://dx.doi.org/10.1371/journal.pone.0096094.

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Huang, Zhibin, Yong Zhang, Jianhua Zhou, and Yu Zhang. "Urinary Exosomal miR-193a Can Be a Potential Biomarker for the Diagnosis of Primary Focal Segmental Glomerulosclerosis in Children." BioMed Research International 2017 (2017): 1–6. http://dx.doi.org/10.1155/2017/7298160.

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Background. Glomerular upregulation of miR-193a has been detected in primary focal segmental glomerulosclerosis (FSGS) but not in other glomerular diseases. We aimed to isolate exosomes from urine of children with primary FSGS and to assess the diagnostic potential of urinary exosomal miR-193a for primary FSGS. Methods. The first morning urine samples were collected from children with primary FSGS (n=8) and minimal change disease (MCD, n=5). Isolated urinary exosomes were confirmed by electron microscopy and Western blotting. Urinary exosomal microRNA was extracted, and the expression levels of exosomal miR-193a were quantified by real-time PCR. The diagnosis value of urinary exosomal miR-193a levels for primary FSGS was evaluated by ROC analysis. Results. The isolated vesicles were qualitatively compatible with exosomes. The levels of urinary exosomal miR-193a were significantly higher in children with primary FSGS than those in children with MCD. Moreover, the area under the ROC for the diagnosis of primary FSGS using urinary exosomal miR-193a was 0.85. Conclusions. A significant increase in the levels of urinary exosomal miR-193a in primary FSGS patients compared to those in MCD ones was observed. This study suggests that urinary exosomal miR-193a may be a new noninvasive biomarker for the diagnosis of primary FSGS.

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