Coupling Lipid Labeling and Click Chemistry Enables Isolation of Extracellular Vesicles for Noninvasive Detection of Oncogenic Gene Alterations

Sun, Na; Tran, Benjamin; Peng, Zishan; Wang, Jing; Zhang, Ceng; Yang, Peng; Zhang, Tiffany X.; Widjaja, Josephine; Zhang, Ryan Y.; Xia, Weiguo; Keir, Alexandra; She, Jia‐Wei; Yu, Hsiao‐hua; Shyue, Jing‐Jong; Zhu, Hongguang; Agopian, Vatche G.; Pei, Renjun; Tomlinson, James S.; Toretsky, Jeffrey A.; Jonas, Steven J.; Federman, Noah; Lu, Shiqi; Tseng, Hsian‐Rong; Zhu, Yazhen

doi:10.1002/advs.202105853

Cited by 25 publications

(28 citation statements)

References 45 publications

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“…To conduct HCC EV SPA, we (i) produced Click Beads [18] (i.e., methyltetrazine [mTz]-modified microbeads) according to a chemical modification procedure (Figure S1) and optimized them as described in Supplementary Methods and Figures S2-S4, (ii) prepared the four transcyclooctene (TCO)-conjugated HCC-associated antibodies using the copper-free click chemistry coupling method (Figure S5), and (iii) conjugated DNA barcode onto anti-CD63 and anti-CD9 to target EV markers, followed by validation (Figure S6, the primers and probes for the DNA barcodes were purchased from Integrated DNA Technologies and the sequences are listed in Table S1).…”

Section: General Information For Hcc Ev Spamentioning

confidence: 99%

HCC EV ECG score: An extracellular vesicle‐based protein assay for detection of early‐stage hepatocellular carcinoma

et al. 2023

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Background and Aims: The sensitivity of current surveillance methods for detecting early-stage hepatocellular carcinoma (HCC) is suboptimal. Extracellular vesicles (EVs) are promising circulating biomarkers for early cancer detection. In this study, we aim to develop an HCC EV-based surface protein assay for early detection of HCC.Approach and Results: Tissue microarray was used to evaluate four potential HCC-associated protein markers. An HCC EV surface protein assay, composed of covalent chemistry-mediated HCC EV purification and real-time immuno-polymerase chain reaction readouts, was developed and optimized for quantifying subpopulations of EVs. An HCC EV ECG score, calculated from the readouts of three HCC EV subpopulations (EpCAM + CD63 + , CD147 + CD63 + , and GPC3 + CD63 + HCC EVs), was established for detecting early-stage HCC. A phase 2 biomarker study was conducted to evaluate the performance of ECG score in a training cohort (n = 106) and an independent validation cohort (n = 72). Overall, 99.7% of tissue microarray stained positive for at least one of the four HCC-associated protein markers (EpCAM, CD147, GPC3, and ASGPR1) that were subsequently validated in HCC EVs. In the training cohort, HCC EV ECG score demonstrated an area under the receiver operating curve (AUROC) of 0.95 (95% confidence interval [CI], 0.90-0.99) for distinguishing early-stage HCC from cirrhosis with a sensitivity of 91% and a specificity of 90%. The AUROCs of the HCC EV ECG score remained excellent in the validation cohort (0.93; 95% CI, 0.87-0.99) and in the subgroups by etiology (viral: 0.95; 95% CI, 0.90--1.00; nonviral: 0.94; 95% CI, 0.88-0.99). Conclusion: HCC EV ECG score demonstrated great potential for detecting early-stage HCC. It could augment current surveillance methods and improve patients' outcomes.

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Section: General Information For Hcc Ev Spamentioning

confidence: 99%

HCC EV ECG score: An extracellular vesicle‐based protein assay for detection of early‐stage hepatocellular carcinoma

et al. 2023

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“…Sun et al ( 78 ) also developed a click chemistry-based approach for the purification of EV ( Tables 2 , 3 ). They first optimized this approach in conditioned medium from an Ewing sarcoma cell line, and then validated its in vivo potential in plasma from Ewing sarcoma patients and even patients with pancreatic cancer, coupled to a cohort of healthy controls.…”

Section: Resultsmentioning

confidence: 99%

Extracellular Vesicles: A New Source of Biomarkers in Pediatric Solid Tumors? A Systematic Review

et al. 2022

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Virtually every cell in the body releases extracellular vesicles (EVs), the contents of which can provide a “fingerprint” of their cellular origin. EVs are present in all bodily fluids and can be obtained using minimally invasive techniques. Thus, EVs can provide a promising source of diagnostic, prognostic, and predictive biomarkers, particularly in the context of cancer. Despite advances using EVs as biomarkers in adult cancers, little is known regarding their use in pediatric cancers. In this review, we provide an overview of published clinical and in vitro studies in order to assess the potential of using EV-derived biomarkers in pediatric solid tumors. We performed a systematic literature search, which yielded studies regarding desmoplastic small round cell tumor, hepatoblastoma, neuroblastoma, osteosarcoma, and rhabdomyosarcoma. We then determined the extent to which the in vivo findings are supported by in vitro data, and vice versa. We also critically evaluated the clinical studies using the GRADE (Grading of Recommendations Assessment, Development, and Evaluation) system, and we evaluated the purification and characterization of EVs in both the in vivo and in vitro studies in accordance with MISEV guidelines, yielding EV-TRACK and PedEV scores. We found that several studies identified similar miRNAs in overlapping and distinct tumor entities, indicating the potential for EV-derived biomarkers. However, most studies regarding EV-based biomarkers in pediatric solid tumors lack a standardized system of reporting their EV purification and characterization methods, as well as validation in an independent cohort, which are needed in order to bring EV-based biomarkers to the clinic.

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“…In order to achieve rapid capture of EVs, Sun et al (2022) proposed the concept of Click Beads. TCO-PEG-NHS was co-incubated with DSPE-PEG 1000 -NH 2 to form DSPE-PEG 1000 -TCO conjugates.…”

Section: Capturing Exosomes With Covalent Chemistrymentioning

confidence: 99%

Recent developments in isolating methods for exosomes

Gao

et al. 2023

Front. Bioeng. Biotechnol.

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Exosomes are the smallest extracellular vesicles that can be released by practically all cell types, and range in size from 30 nm to 150 nm. As the major marker of liquid biopsies, exosomes have great potential for disease diagnosis, therapy, and prognosis. However, their inherent heterogeneity, the complexity of biological fluids, and the presence of nanoscale contaminants make the isolation of exosomes a great challenge. Traditional isolation methods of exosomes are cumbersome and challenging with complex and time-consuming operations. In recent years, the emergence of microfluidic chips, nanolithography, electro-deposition, and other technologies has promoted the combination and innovation of the isolation methods. The application of these methods has brought very considerable benefits to the isolation of exosomes such as ultra-fast, portable integration, and low loss. There are significant functional improvements in isolation yield, isolation purity, and clinical applications. In this review, a series of methods for the isolation of exosomes are summarized, with emphasis on the emerging methods, and in-depth comparison and analysis of each method are provided, including their principles, merits, and demerits.

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Coupling Lipid Labeling and Click Chemistry Enables Isolation of Extracellular Vesicles for Noninvasive Detection of Oncogenic Gene Alterations

Cited by 25 publications

References 45 publications

HCC EV ECG score: An extracellular vesicle‐based protein assay for detection of early‐stage hepatocellular carcinoma

HCC EV ECG score: An extracellular vesicle‐based protein assay for detection of early‐stage hepatocellular carcinoma

Extracellular Vesicles: A New Source of Biomarkers in Pediatric Solid Tumors? A Systematic Review

Recent developments in isolating methods for exosomes

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