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Extracellular Vesicles for Clinical Diagnostics: From Bulk Measurements to Single-Vesicle Analysis

  • Hai Linh Tran
  • , Wenshu Zheng
  • , David A. Issadore
  • , Hyungsoon Im
  • , Yoon Kyoung Cho
  • , Yuanqing Zhang
  • , Dingbin Liu
  • , Yang Liu
  • , Bo Li
  • , Fei Liu
  • , David Tai Wai Wong
  • , Jiashu Sun
  • , Kun Qian
  • , Mei He
  • , Meihua Wan
  • , Yong Zeng
  • , Ke Cheng
  • , Tony Jun Huang
  • , Daniel T. Chiu
  • , Luke P. Lee
  • Lei Zheng, Andrew K. Godwin, Raghu Kalluri, Steven A. Soper, Tony Y. Hu
  • Tulane University
  • University of Pennsylvania
  • Harvard University
  • Ulsan National Institute of Science and Technology
  • Sun Yat-Sen University
  • Nankai University
  • Tianjin Medical University
  • Nanfang Hospital
  • University of California at Los Angeles
  • National Center for Nanoscience and Technology
  • University of Chinese Academy of Sciences
  • Shanghai Jiao Tong University
  • University of Florida
  • Sichuan University
  • Columbia University
  • Duke University
  • University of Washington
  • Guangzhou University of Chinese Medicine
  • University of Kansas
  • University of Texas MD Anderson Cancer Center
  • Rice University
  • Baylor College of Medicine

Research output: Contribution to journalReview articlepeer-review

Abstract

Extracellular vesicles (EVs) play a crucial role in intercellular communication, signaling pathways, and disease pathogenesis by transporting biomolecules such as DNA, RNA, proteins, and lipids derived from their cells of origin, and they have demonstrated substantial potential in clinical applications. Their clinical significance underscores the need for sensitive methods to fully harness their diagnostic potential. In this comprehensive review, we explore EV heterogeneity related to biogenesis, structure, content, origin, sample type, and function roles; the use of EVs as disease biomarkers; and the evolving landscape of EV measurement for clinical diagnostics, highlighting the progression from bulk measurement to single vesicle analysis. This review covers emerging technologies such as single-particle tracking microscopy, single-vesicle RNA sequencing, and various nanopore-, nanoplasmonic-, immuno-digital droplet–, microfluidic-, and nanomaterial-based techniques. Unlike traditional bulk analysis methods, these methods contribute uniquely to EV characterization. Techniques like droplet-based single EV-counting enzyme-linked immunosorbent assays (ELISA), proximity-dependent barcoding assays, and surface-enhanced Raman spectroscopy further enhance our ability to precisely identify biomarkers, detect diseases earlier, and significantly improve clinical outcomes. These innovations provide access to intricate molecular details that expand our understanding of EV composition, with profound diagnostic implications. This review also examines key research challenges in the field, including the complexities of sample analysis, technique sensitivity and specificity, the level of detail provided by analytical methods, and practical applications, and we identify directions for future research. This review underscores the value of advanced EV analysis methods, which contribute to deep insights into EV-mediated pathological diversity and enhanced clinical diagnostics.

Original languageEnglish
Pages (from-to)28021-28109
Number of pages89
JournalACS Nano
Volume19
Issue number31
DOIs
StatePublished - 12 Aug 2025

Keywords

  • analytical techniques
  • biomarkers
  • diagnostics
  • extracellular vesicles (EVs)
  • single EV analysis

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