MoSERS profiles
of blood-borne EVs from GBM patients harboring
distinct molecular alterations. (a) Schematic illustrating the classification
of patient-derived EVs using MoSERS single EV fingerprints analyzed
by machine learning. Each class represents a different GBM marker.
(b) RT-PCR agarose gel of EGFR and EGFRvIII cDNA in control and patient-derived
circulating EV samples. (c) The probability distribution of belonging
to each of the classes based on CNN output, EGFR amplification (blue),
EGFRvIII (green), and MGMT methylation (orange). (d) The ROC curve
of assessing the single EV spectra prediction accuracy over clinical
annotation demonstrates an overall AUC of 0.85. The probability that
EVs are positive for (e) EGFR amplification, (f) EGFRvIII, and (g)
MGMT partitioned into classes based on clinical read-out: healthy
(gray), negative-variant patients (light color), and individual positive-variant
patients (dark color). (h–j) ANOVA analysis of all spectra
partitioned based on clinical annotations, demonstrating the ability
to distinguish samples from negative and positive variant patients
as well as healthy patients. (k) Samples with positive variants of
EGFR amplification, EGFRvIII and MGMT methylation were pooled and
classified by the probability distribution of each sample. (l) The
ROC curve of assessing the overall MoSERS prediction accuracy of individual
patients carrying one of the three molecular GBM-associated alterations
over clinical annotation demonstrates an overall AUC of 0.91.