Extracellular vesicle isolation and characterization: toward clinical application

Xu, Rong; Greening, David W.; Zhu, Hong; Takahashi, Nobuhiro; Simpson, Richard J.

doi:10.1172/jci81129

Cited by 726 publications

(637 citation statements)

References 147 publications

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“…To confirm that the purified exosomes also met the standards in this field, we analyzed the features of exosomes by transmission electron microcopy. As shown in Figure 1D, the exosomes presented as cup-shaped membrane vesicles of 50 to 90 nm, as previously reported (32,33). Approximately 42% of TPO, 42% of ANGPTL2, and 62% of ANGPTL3 were located in exosomes marked by TSG101 and flotillin 1 (FLOT1) ( Figure 1E), and purified exosomes from human plasma contained a large amount of TPO, ANGPTL2, and ANGPTL3 proteins ( Figure 1F).…”

Section: Introductionsupporting

confidence: 81%

Sorting protein VPS33B regulates exosomal autocrine signaling to mediate hematopoiesis and leukemogenesis

Chen

Hao

et al. 2016

Journal of Clinical Investigation

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Section: Introductionsupporting

confidence: 81%

Sorting protein VPS33B regulates exosomal autocrine signaling to mediate hematopoiesis and leukemogenesis

Chen

Hao

et al. 2016

Journal of Clinical Investigation

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“…Importantly, this data cannot distinguish whether increased alternative pathway activation is a cause of disease or simply a marker of inflammation in CKD. Another potential limitation is the absence of standardized methodology for the measurement of microparticles in humans 51. Notwithstanding these limitations, data such as ours may lead to greater interest in microparticles, and future studies should focus on evaluating the methodology and whether in fact microparticles may be a valid marker of CVD.…”

Section: Discussionmentioning

confidence: 94%

Endothelial Microparticles and Systemic Complement Activation in Patients With Chronic Kidney Disease

Jalal

Renner

Laskowski

et al. 2018

JAHA

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BackgroundEndothelial microparticles are associated with chronic kidney disease (CKD) and complement activation. We hypothesized that the complement pathway is activated in patients with CKD via endothelial microparticles and that complement activation correlates with endothelial dysfunction in CKD.Methods and ResultsWe analyzed complement data of 30 healthy subjects, 30 patients with stage III/IV CKD, and 30 renal transplant recipients with stage III/IV CKD, evaluating the potential correlation of complement fragments with brachial artery flow–mediated dilation, Chronic Kidney Disease Epidemiology Collaboration glomerular filtration rate, and urinary albumin/creatinine ratio. Endothelial microparticles were characterized via proteomic analysis and compared between study groups. Complement fragment Ba was significantly increased in CKD and post–kidney transplant CKD. Plasma Ba levels correlated significantly with lower brachial artery flow–mediated dilation, lower Chronic Kidney Disease Epidemiology Collaboration glomerular filtration rate, and higher urinary albumin/creatinine ratio. Factor D levels were significantly higher in the plasma microparticles of patients with CKD versus healthy controls. Plasma microparticles isolated from patients with CKD and containing factor D activated the alternative pathway in vitro.ConclusionThe alternative complement pathway is activated in CKD and correlates with endothelial dysfunction and markers of CKD. Future studies are needed to evaluate whether endothelial microparticles with increased factor D play a pathologic role in CKD‐associated vascular disease.Clinical Trial Registration URL: http://www.clinicaltrials.gov. Unique identifier: NCT02230202.

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“…For example, development of a pre-amplification of EV mRNA or a more sensitive quantification method. In addition, although there will be some difficulties and cautionary notes, [18][19][20] in order to not only make Ct-OATP1B3 mRNA detection more easily, but also to specify its origin, consideration should be given to collect and concentrate CRC-specific EVs from whole serum in future studies.…”

Section: Discussionmentioning

confidence: 99%

Cancer-Type OATP1B3 mRNA in Extracellular Vesicles as a Promising Candidate for a Serum-Based Colorectal Cancer Biomarker

Morio

Sun

Harada

et al. 2018

Biological & Pharmaceutical Bulletin

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Cancer-type organic anion transporting polypeptide 1B3 (Ct-OATP1B3) mRNA is a variant isoform of the liver-type OATP1B3. Because Ct-OATP1B3 mRNA shows an excellent cancer-specific expression profile in colorectal cancer (CRC), and that its expression levels are associated with CRC prognosis, it holds the potential to become a useful CRC detection and diagnosis biomarker. While the potential is currently justified only at the tissue level, if existence of Ct-OATP1B3 mRNA in CRC-derived extracellular vesicles (EVs) is validated, the findings could enhance its translational potential as a CRC detection and diagnosis biomarker. Therefore, this study aims at proving that Ct-OATP1B3 mRNA exists in CRC-derived EVs, and can be detected using serum specimens. To examine the possibility of Ct-OATP1B3 mRNA being existed in extracellular milieu, we isolated EVs from the human CRC (HCT116, HT-29, and SW480) cell lines, and prepared their cDNAs. The RT-PCR results showed that Ct-OATP1B3 mRNA was clearly present in EVs derived from the human CRC cell lines. Then, in order to further explore the possibility that Ct-OATP1B3 mRNA in CRCderived EVs can be detected in serum, we isolated serum EVs derived from human CRC xenograft mice, and then performed RT-PCR. The results showed that Ct-OATP1B3 mRNA could be found in all serum EV and CRC tissue samples of the mice examined. Collectively, our findings, which show that Ct-OATP1B3 mRNA exists in EVs and can be detected in (at least) mouse serum, strengthen the potential use of Ct-OATP1B3 mRNA as a serum-based CRC biomarker.Key words organic anion transporting polypeptide 1B3; SLCO1B3; colorectal cancer; extracellular vesicle; cancer biomarker Worldwide, colorectal cancer (CRC) is one of the most frequently encountered malignant tumors and is a major cause of related deaths.

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Extracellular vesicle isolation and characterization: toward clinical application

Cited by 726 publications

References 147 publications

Sorting protein VPS33B regulates exosomal autocrine signaling to mediate hematopoiesis and leukemogenesis

Sorting protein VPS33B regulates exosomal autocrine signaling to mediate hematopoiesis and leukemogenesis

Endothelial Microparticles and Systemic Complement Activation in Patients With Chronic Kidney Disease

Cancer-Type OATP1B3 mRNA in Extracellular Vesicles as a Promising Candidate for a Serum-Based Colorectal Cancer Biomarker

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