Distribution and Function of Immunity Cluster from all Ti-EVs in Cataracts
We subset 42,199 Ti-EVs from Immunity cluster, including Macrophage, T, HAVCR1, and other clusters (Figure 3A). The Macrophage-derived cluster exhibited high expression of CD68, MRC1, CD80, CD86, and CD14, while the T cell-derived cluster was marked by high expression of CD3D and CD3E (Figure 3B). Functionally, the T cell-derived cluster was primarily involved in pathways related to cell adhesion mediated by integrins and the positive regulation of fibroblast migration, whereas the increased presence of Macrophage-derived cluster appeared to respond to bacterial invasion (Figure 3C). It is worth noting that the other immune-related Ti-EV cluster comprised the majority of the cataract immunity landscape (Figure 3D, E), and further division was deemed unnecessary as their protein contents showed no significant differences.
Notably, the Ro/e value for Macrophage-derived Ti-EVs cluster was higher in high myopic cataracts, while the value for T cell-derived Ti-EVs cluster was higher in age-related cataracts (Figure 3F). Additionally, the total expression of the 264 proteins in each Ti-EV correlated highly with overall protein expression (R=0.908; Figure 3G). Using BayesPrism to deconvolute individual Ti-EV and project this information onto total proteins, we observed that the Macrophage-derived cluster was more abundant in high myopic cataracts and showed a strong correlation with the AQP1 cluster (R=0.680, P=0.004; Figure 3I, J). Interestingly, among proteins linked to cataract progression, AQP1, ITGAV, and ITGB1 were significantly upregulated in Ti-EVs, with AQP1 more prominent in high myopic cataracts (Figure S1D). These findings suggest that Macrophage-derived Ti-EVs and AQP1 in Ti-EVs may synergistically contribute to drive the progression of high myopic cataracts.