Proteomic Analysis of Extracellular Vesicles for Cancer Diagnostics

Wu, Anthony Yan-Tang; Ueda, Koji; Lai, Charles P.

doi:10.1002/pmic.201800162

Cited by 35 publications

(37 citation statements)

References 150 publications

(140 reference statements)

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“…Label‐free quantitation is most commonly used for global EV profiling due to its capacity for high throughput at minimal cost; however, this method relies heavily on accurate peptide calling . Meanwhile, metabolic or isotopic labeling methodologies can aid in relative quantitation and sample comparison but are expensive and are limited to small cohorts . It is important that researchers adopt a methodology that is appropriate for the study aim and sample type in question.…”

Section: Limitationsmentioning

confidence: 99%

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Tumor‐Derived Extracellular Vesicles as a Novel Source of Protein Biomarkers for Cancer Diagnosis and Monitoring

Ruhen

Meehan

2019

Proteomics

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“Liquid biopsies” have received attention as a complementary tool for traditional tissue biopsies that may enhance the spectrum of analysis for tumor‐derived factors. One such factor gaining prominence in the liquid biopsy field is extracellular vesicles (EVs), membrane‐bound nanovesicles which are secreted by cells into biofluids such as blood, urine, and saliva. EVs are released in both physiological and pathological conditions and can transport a variety of molecules, including proteins, metabolites, RNA, microRNAs, and DNA, to distant sites throughout the body. As such, they are emerging as a promising source of tumor biomarkers for the noninvasive diagnosis, prognosis, and monitoring of cancer patients. In particular, the wealth of tumor‐related information that can be gleaned from the EV proteomic cargo has become apparent through mass spectrometric analysis, which has provided new benchmarks for clinically focused biomarker research. In this review, the current achievements in the use of MS for identifying potential EV‐derived protein biomarkers of cancer are explored, and the techniques and challenges involved in this pursuit are summarized.

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Section: Limitationsmentioning

confidence: 99%

“…[13] Meanwhile, metabolic or isotopic labeling methodologies can aid in relative quantitation and sample comparison but are expensive and are limited to small cohorts. [53] It is important that researchers adopt a methodology that is appropriate for the study aim and sample type in question.…”

Section: Limitationsmentioning

confidence: 99%

Tumor‐Derived Extracellular Vesicles as a Novel Source of Protein Biomarkers for Cancer Diagnosis and Monitoring

Ruhen

Meehan

2019

Proteomics

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“…Extracellular vesicles (EVs) are nanosized bioparticles (30–10,000 nm) released by cells to deliver bioactive cargoes to enable communications over short and long distances, both within and between organisms 1,2 . Subtypes include exosomes (30–100 nm in diameter), shed microvesicles (or ectosomes, microparticles; 100-1,000 nm) and apoptotic bodies (50–4,000 nm) 3 . Using asymmetric flow field-flow fractionation (AF4), exosomes can be further subcategorized as small (Exo-S; 60–80 nm) or large (Exo-L; 90–120 nm) exosomes or bionanoparticle exomeres (< 50 nm) 4 .…”

Section: Introductionmentioning

confidence: 99%

“…mRNA, miRNA and lncRNA) and proteins ( e.g . ligands and receptors), which have been associated with development, immune response, neurodegenerative diseases, developmental disorders and cancer progression 3, 6–10 . Tumour exosomes have been shown to be organotropic in facilitating subsequent metastasis to the tropic organs 11 .…”

Section: Introductionmentioning

confidence: 99%

Multi-resolution imaging using bioluminescence resonance energy transfer identifies distinct biodistribution profiles of extracellular vesicles and exomeres with redirected tropism

Sung

Chen

et al. 2020

Preprint

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Extracellular particles (EP) including extracellular vesicles (EVs) and exomeres have been shown to play significant roles in diseases and therapeutic applications. However, their spatiotemporal dynamics in vivo have remained largely unresolved in detail due to the lack of a suitable method. We therefore created a bioluminescence resonance energy transfer (BRET)-based reporter, PalmGRET, to enable pan-EP labelling ranging from exomeres (< 50 nm) to small (< 200 nm) and medium and large (> 200 nm) EVs. PalmGRET emits robust, sustained signals and allows the visualization, tracking and quantification of the EPs from whole-animal to nanoscopic resolutions under different imaging modalities, including bioluminescence, BRET and fluorescence. Using PalmGRET, we show that EPs released by lung metastatic hepatocellular carcinoma (HCC) exhibit lung tropism with varying distributions to other major organs in immunocompetent mice. We further demonstrate that gene knockdown of lung-tropic membrane proteins, solute carrier organic anion transporter family member 2A1 (Slco2a1), alanine aminopeptidase (Cd13) and chloride intracellular channel (Clic1) decreases HCC-EP distribution to the lungs and yields distinct biodistribution profiles. We anticipate that EP-specific imaging, quantitative assays and detailed in vivo characterization to be a starting point for more accurate and comprehensive in vivo models of EP biology and therapeutic design.

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“…EVs are released by almost all cell types and provide an effective and ubiquitous path for intercellular communication and the transmission of pathogenic and signaling molecules among cells [3][4][5] . Their potentially important cellular functions in disease onset and progression make them intriguing sources for biomarker discovery and disease diagnosis [6][7][8][9] . In particular, these EV-based disease markers can be identified well before the onset of symptoms or physiological detection of an ailment, making them promising candidates for early-stage disease detection [9][10][11][12][13][14] .…”

Section: Introductionmentioning

confidence: 99%

Sequential phosphoproteomics and N-glycoproteomics of plasma-derived extracellular vesicles

et al. 2019

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Extracellular vesicles (EVs) are increasingly being recognized as important vehicles for intercellular communication and as promising sources for biomarker discovery. Because the state of protein post-translational modifications (PTMs) such as phosphorylation and glycosylation can be a key determinant of cellular physiology, comprehensive characterization of protein PTMs in EVs can be particularly valuable for early-stage diagnostics and monitoring of disease status. However, the analysis of PTMs in EVs has been complicated by limited amounts of purified EVs, low-abundance PTM proteins, and interference from proteins and metabolites in biofluids. Recently, we developed an approach to isolate phosphoproteins and glycoproteins in EVs from small volumes of human plasma that enabled us to identify nearly 10,000 unique phosphopeptides and 1,500 unique N-glycopeptides. The approach demonstrated the feasibility of using these data to identify potential markers to differentiate disease from healthy states. Here we present an updated workflow to sequentially isolate phosphopeptides and N-glycopeptides, enabling multiple PTM analyses of the same clinical samples. In this updated workflow, we have improved the reproducibility and efficiency of EV isolation, protein extraction, and phosphopeptide/Nglycopeptide enrichment to achieve sensitive analyses of low-abundance PTMs in EVs isolated from 1 mL of plasma. The modularity of the workflow also allows for the characterization of phospho-or glycopeptides only and enables additional analysis of total proteomes and other PTMs of interest. After blood collection, the protocol takes 2 d, including EV isolation, PTM/peptide enrichment, mass spectrometry analysis, and data quantification. Reprints and permissions information is available at www.nature.com/reprints.

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Proteomic Analysis of Extracellular Vesicles for Cancer Diagnostics

Cited by 35 publications

References 150 publications

Tumor‐Derived Extracellular Vesicles as a Novel Source of Protein Biomarkers for Cancer Diagnosis and Monitoring

Tumor‐Derived Extracellular Vesicles as a Novel Source of Protein Biomarkers for Cancer Diagnosis and Monitoring

Multi-resolution imaging using bioluminescence resonance energy transfer identifies distinct biodistribution profiles of extracellular vesicles and exomeres with redirected tropism

Sequential phosphoproteomics and N-glycoproteomics of plasma-derived extracellular vesicles

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