Fig. 5: RCA-ExM-based cancer diagnosis and evlaution of immunotherapy response.

A Heatmap indicating relative miR-21 expression levels detected in EpCAM⁺ PD-L1⁺ EVs from healthy donors (HD, n = 13) and cancer patients with stable disease (SD, n = 15), partial response (PR, n = 17), or progressive disease (PD, n = 41) following immunotherapy. B miR-21 expression levels in EpCAM⁺ PD-L1⁺ EVs and (C) PD-L1 expression levels in EpCAM⁺ EVs from each subgroup. Open circles indicate the individual data points, solid dot indicate the mean value, box indicate the interquartile range spanning the 25th to 75th percentiles, vertical line within box indicate the mean ± SD. P values were calculated by one-way ANOVA (two-sided) with Tukey’s multiple comparisons test. **** indicates P < 0.0001, ns indicates no significant difference. For miR-21 expression levels in EpCAM⁺ PD-L1⁺ EVs assay, P (PR vs. SD) < 0.0001, q (82) = 7. 140, d = −1.788 [95% CI: −17.37, −5.494]; P (SD vs. PD) < 0.0001, q (82) = 15.50, d = −3.311 [95% CI: −26.21, −16.09]; P (PR vs. PD) < 0.0001, q (82) = 24.99, d = −5.099 [95% CI: −37.41, −27.74]; P (HD vs. PD) < 0.0001, q (82) = 35.26, d = −7.954 [95% CI: −56.06, −45.39]. For PD-L1 expression levels in EpCAM⁺ EVs assay, P (PR vs. SD) = 0.5470, q (82) = 1.880, d = −0.471 [95% CI: −24.10,7.883]; P (SD vs. PD) < 0.0001, q (82) = 9.584, d = −2.045 [95% CI: −48.82, −21.58]; P (PR vs. PD) < 0.0001, q (82) = 12.33, d = −2.516 [95% CI: −56.32, −30.28]; P (HD vs. PD) < 0.0001, q (82) = 20.30, d = −4.570 [95% CI: -93.01, −64.28]. Source data are provided as a Source Data file. D ROC curves indicating for the ability to distinguish specific immunotherapy responses (PR vs. SD, SD vs. PD, and PD vs. PR) in 73 cancer patients, when analyzing miR-21 expression in EpCAM⁺ PD-L1⁺ EVs (left) and PD-L1 expression in EpCAM⁺ EVs (right). Source data are provided as a Source Data file.