Cytotoxic T-cell infiltration contributes to RPE-choroidal hyperpermeability under stress revealed by single-cell RNA sequencing

Abstract

Purpose : Psychological stress associates with various ocular diseases, especially central serous chorioretinopathy, featuring RPE-choroidal hyperpermeability and subretinal fluid accumulation. We aimed to reveal the alterations in RPE-choroidal complexes under stress and the underlying mechanisms.

Methods : C57BL/6J mice went through modified chronic social defect stress procedure. Swept-source optical coherence tomography and transmission electron microscopy were used for phenotype observation, then the RPE-choroidal complexes were subjected to single-cell RNA sequencing.

Results : Endothelial dysfunctions, more choriocapillary fenestrations and interrupted RPE tight junctions were observed in stressed mice. The cell-cell interaction analysis identified enhanced cytotoxic T-cell activities under stress and revealed that choriocapillaris is the most susceptible site. However, the alterations in corticosterone-treated mice were much weaker than those in stressed mice.

Conclusions : Psychological stress could induce RPE-choroidal hyperpermeability, which is greatly attributed to cytotoxic T-cell infiltration rather than elevated glucocorticoids alone.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.

 

Figure 1. Choroidal phenotype changes under stress. (A) Reduced choroidal thickness under stress (Control=20, stress=21). (B) Swept-source optical coherence tomography angiography showing vortex vein congestion, blood vessel constriction and venous anastomosis in stressed mice. (C-D) Increased choriocapillary fenestration density (Control=8, stress=8). (E) Impaired RPE tight junctions under stress. (F) Undefined cell infiltration in stressed mice. Scale bars, 1 mm (B), 200 nm (C, E), 500 nm (F). Values in A,D: mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001. CEC, choroidal endothelial cells; BM, Bruch’s membrane; BI, basal infoldings.

 

Figure 2. T-cell heterogeneity in RPE-choroidal complexes under stress. (A-B) Enhanced cell-cell interactions and increased T-cell ratio under stress. (C-E) T-cell subclustering. (F-H) Pseudotime analysis indicates the tendency of T cells to enter cell fate 2 (where subcluster T2 is located) under stress. (I) Enhanced cell-cell interactions sourced from T cells and targeting endothelial cells. (J) Upregulated cytotoxic ligand-receptor pairs in stress group that sourced from subcluster T2 and targeting at endothelial cells. A, arterial; V, venous; C, capillary.