Machine intelligence-driven classification of cancer patients-derived extracellular vesicles using fluorescence correlation spectroscopy: results from a pilot study

Patient-derived extracellular vesicles (EVs) that contains a complex biological cargo is a valuable source of liquid-biopsy diagnostics to aid in early detection, cancer screening, and precision nanotherapeutics. In this study, we predicted that coupling cancer patient blood-derived EVs to time-resolved spectroscopy and artificial intelligence (AI) could provide a robust cancer screening and follow-up tools. In our pilot study, fluorescence correlation spectroscopy (FCS) measurements were taken on 24 blood samples-derived EVs. Blood samples were obtained from 15 cancer patients (presenting five different types of cancers), and nine healthy controls (including patients with benign lesions). EVs samples were labeled with PKH67 dye. The obtained FCS autocorrelation spectra were processed into power spectra using the fast Fourier transform algorithm. The processed power spectra were subjected to various machine learning algorithms to distinguish cancer spectra from healthy control spectra. The performance of AdaBoost Random Forest (RF) classifier, support vector machine, and multilayer perceptron were tested on selected frequencies in the N = 118 power spectra. The RF classifier exhibited the highest classification accuracy and performance metrics in distinguishing the FCS power spectra of cancer patients from those of healthy controls. Further, neural computing via an image convolutional neural network (CNN), ResNet network, and a quantum CNN were assessed on the power spectral images as additional validation tools. All image-based CNNs exhibited a nearly equal classification performance and reasonably high sensitivity and specificity scores. Our pilot study demonstrates that AI-algorithms coupled to time-resolved FCS power spectra can accurately and differentially classify the complex patient-derived EVs from different cancer samples of distinct tissue subtypes. As such, our findings hold promise in the diagnostic and prognostic screening in clinical medicine.