Single-Vesicle Landscape of Ti-EVs in Age-related and High
Myopic Cataracts
Using the ExoDisc, we isolated Ti-EVs from the lens capsules of 9
patients with age-related cataracts and 7 patients with high myopic
cataracts. This analysis identified a total of 260 proteins, and
unsupervised clustering of their expression profiles indicated
significant pathological heterogeneity between the two types of
cataracts (Figure 2A). Among these proteins, 21 were differentially
expressed between the groups (Figure 2B; Table S2); specifically, 13
proteins, including ABCG2, ERBB2, and CD86, were significantly enriched
in high myopic cataracts, while 8 proteins were more abundant in the
Ti-EVs of age-related cataracts (Figure 2C, D). Further analysis using
Harmony to adjust for batch effects identified 65 subclusters comprising
a total of 102,156 Ti-EVs, which were evenly distributed across both
types of cataracts (Figure 2E, F; Table S3). Based on protein markers
within each subcluster, we classified the Ti-EVs into seven clusters:
Adhesion, AQP1, Immunity, Lysosome, Metabolize, Signal transduction, and
SLC12A1&SLC12A3 (Figure 2G). Notably, each subcluster displayed
distinct and clearly distinguishable protein profiles (Figure 2H).
Our data indicated that the Immunity cluster was the predominant Ti-EVs
type in cataracts. Additionally, despite having two fewer samples, the
Lysosome cluster was more prevalent in pathological cataracts (Figure
2I). There were no significant differences in Ti-EV between samples, and
each cluster was well represented across samples, suggesting that the
observed patterns are consistent and not significantly affected by
individual sample variability (Figure 2J). The Ro/e values for the
Lysosome, AQP1, Immunity, and SLC12A1&SLC12A3 clusters were higher in
high myopic cataract samples, indicating these ones may potential
pathogenic factors (Figure 2K). Notably enriched proteins in the
Lysosome cluster included CDH17, LAMP1, HLA-DRA, and IL6 (Figure S1A),
while AQP1 and CDCP1 were enriched in the AQP1 cluster (Figure S1B),
with a high correlation observed between these two groups (Figure S1C).
Interestingly, ABCG2, ERBB2, and CD86 were primarily located in the
Immunity cluster, suggesting a possible role in the progression of high
myopic cataracts.