Abstract
Extracellular vesicles (EVs) secreted by cancer cells provide an important insight into cancer biology and could be leveraged to enhance diagnostics and disease monitoring. This paper details a high-throughput label-free extracellular vesicle analysis approach to study fundamental EV biology, toward diagnosis and monitoring of cancer in a minimally invasive manner and with the elimination of interpreter bias. We present the next generation of our single particle automated Raman trapping analysis—SPARTA—system through the development of a dedicated standalone device optimized for single particle analysis of EVs. Our visualization approach, dubbed dimensional reduction analysis (DRA), presents a convenient and comprehensive method of comparing multiple EV spectra. We demonstrate that the dedicated SPARTA system can differentiate between cancer and noncancer EVs with a high degree of sensitivity and specificity (>95% for both). We further show that the predictive ability of our approach is consistent across multiple EV isolations from the same cell types. Detailed modeling reveals accurate classification between EVs derived from various closely related breast cancer subtypes, further supporting the utility of our SPARTA-based approach for detailed EV profiling.
Highlights
Extracellular vesicles (EVs) secreted by cancer cells provide an important insight into cancer biology and could be leveraged to enhance diagnostics and disease monitoring
This study investigates the potential of EVs as biomarkers of breast cancer by applying advances to our recently developed high-throughput single particle automated Raman trapping analysis (SPARTA) platform.[45]
A label-free, nondestructive analysis of EVs in their hydrated state is performed to identify the compositional differences between EVs of cancerous and noncancerous origin in an unbiased manner
Summary
Extracellular vesicles (EVs) secreted by cancer cells provide an important insight into cancer biology and could be leveraged to enhance diagnostics and disease monitoring. This paper details a high-throughput label-free extracellular vesicle analysis approach to study fundamental EV biology, toward diagnosis and monitoring of cancer in a minimally invasive manner and with the elimination of interpreter bias. The primary concerns regarding this approach relate to the invasive nature and the sampling error that can occur with inaccurate biopsy positioning Both X-ray screening and histopathological assessment depend strongly on the individual judgment of trained specialists, with variations between individuals and laboratories frequently reported.[3] These limitations demonstrate the need for approaches that focus on noninvasive and minimally invasive techniques and reduce interpretation variability and bias. Most frequently blood samples, have shown increasing promise as a minimally invasive alternative to biopsy.[14] Blood-borne entities such as circulating tumor cells, circulating tumor DNA, and of key interest in this study tumor-derived extracellular vesicles (EVs) are all potential biomarkers.[15,16]. Addressing all of these extrinsic and intrinsic study parameters through the use of the SPARTA platform
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