Inaugurating High‐Throughput Profiling of Extracellular Vesicles for Earlier Ovarian Cancer Detection

Abstract

Detecting early cancer through liquid biopsy is challenging due to the lack of specific biomarkers for early lesions and potentially low levels of these markers. The current study systematically develops an extracellular‐vesicle (EV)‐based test for early detection, specifically focusing on high‐grade serous ovarian carcinoma (HGSOC). The marker selection is based on emerging insights into HGSOC pathogenesis, notably that it arises from precursor lesions within the fallopian tube. This work thus establishes murine fallopian tube (mFT) cells with oncogenic mutations and performs proteomic analyses on mFT‐derived EVs. The identified markers are then evaluated with an orthotopic HGSOC animal model. In serially‐drawn blood of tumor‐bearing mice, mFT‐EV markers increase with tumor initiation, supporting their potential use in early cancer detection. A pilot clinical study (n = 51) further narrows EV markers to five candidates, EpCAM, CD24, VCAN, HE4, and TNC. The combined expression of these markers distinguishes HGSOC from non‐cancer with 89% sensitivity and 93% specificity. The same markers are also effective in classifying three groups (non‐cancer, early‐stage HGSOC, and late‐stage HGSOC). The developed approach, for the first time inaugurated in fallopian tube‐derived EVs, could be a minimally invasive tool to monitor women at high risk of ovarian cancer for timely intervention.

Proteomic analysis of ascitic extracellular vesicles describes tumour microenvironment and predicts patient survival in ovarian cancer

“…Higher levels of EpCAM-positive EVs are thus interpreted as reflecting increased tumour burden, and consequently, negatively associated with patient survival across various cancers. Notably, EpCAM is often utilized as one of markers for EVs for the early diagnosis of HGSC (Jo et al, 2023;Li et al, 2023). Similarly, CKB (Creatine Kinase B), expressed in malignant cells, exhibited a negative correlation with patient survival.…”

Recent advances in chemical biology tools for protein and RNA profiling of extracellular vesicles

“…The Lee and Weissleder group developed the SAViA and DEST assay using HRP based radical reaction for tyramide polymerization mediated signal amplification. 70,82 The Shao group applied HRP based radical reaction for the deposition of metal 154 or insoluble optical products 143 to improve the sensitivity of EV analysis. The Shao group also achieved excellent EV detection sensitivity by mechanical transition of a metamaterial using a HRP based radical reaction.…”

“…3(A) ). 82 The SAViA assay, integrating EV physisorption and tyramide-assisted signal amplification, increased analytical sensitivity over conventional immunoassays by more than 10 000-fold, enabling the detection of a small amount of EVs (limit of detection (LOD): 600 vesicles) within a practical microwell-plate format (384 wells). Notably, this SAViA assay represents the first reported approach for targeting EVs derived from fallopian tubes.…”

Thermophoretic glycan profiling of extracellular vesicles for triple-negative breast cancer management

“…EVs (10 6 μL − 1 , 100 μL) were immobilized onto 96-well ELISA plates with polystyrene substrate (437112, Thermo Scientific, USA) through passive adsorption for 2 h at room temperature 60 , 61 . After EV immobilization, the supernatant was discarded, and the wells were blocked using 100 μL of 1× PBS containing 1% BSA for 1 h at room temperature.…”