Abstract
Purpose:
Immune cells play a critical role in pathogenesis of glaucoma. However, their ontogeny, gene expression profiles, spatial distribution, and functional changes in the retina and optic nerve in glaucoma remain poorly understood. We performed single-cell RNA sequencing to investigate the changes in immune cell populations in the microbead mouse model of glaucoma.
Methods:
Elevated intraocular pressure was induced by injecting microbeads into the anterior chamber of 3-month-old C57BL/6J mice, while control mice received sham injections. 30 days after injection, retinas and optic nerves were harvested, and CD45+ immune cells were isolated using flow cytometry and sequenced using the 10X Genomics platform (n = 3-4 samples per group, each sample pooled from 8-10 eyes). Immunohistochemistry was used to validate the spatial distribution of specific immune cell subsets.
Results:
28,277 immune cells were successfully sequenced from microbead- and sham-injected retinas and optic nerves, identifying many different immune cell subsets. In control mice, optic nerve microglia were less homeostatic than retinal microglia and were characterized by elevated expression of genes such as Spp1 and Gpnmb, which are classically upregulated by microglia in neurodegeneration. In glaucomatous retinas, there was a significant increase in disease-associated microglia (DAM), macrophages, and T cells compared to sham-injected retinas (DAM, p = 0.0039; macrophages, p = 0.0004; T cells, p = 0.024). Moreover, macrophages in the glaucomatous retina shifted towards a state characterized by high MHCII expression, suggesting a potential for crosstalk between retinal macrophages and peripheral immune cells through enhanced antigen presentation and immune response. Interestingly, the immune cell proportions in glaucomatous optic nerves were similar to those in sham optic nerves, with only subtle changes noted in differentially expressed genes between the two conditions.
Conclusions:
Our findings reveal the extensive and dynamic changes in immune cell subsets within glaucomatous retinas, in contrast to the relatively minor changes detected in glaucomatous optic nerves. These findings provide novel insights into the immune cell landscape in glaucoma and may help identify new therapeutic targets for this common blinding disease.
Immune cells play a critical role in pathogenesis of glaucoma. However, their ontogeny, gene expression profiles, spatial distribution, and functional changes in the retina and optic nerve in glaucoma remain poorly understood. We performed single-cell RNA sequencing to investigate the changes in immune cell populations in the microbead mouse model of glaucoma.
Methods:
Elevated intraocular pressure was induced by injecting microbeads into the anterior chamber of 3-month-old C57BL/6J mice, while control mice received sham injections. 30 days after injection, retinas and optic nerves were harvested, and CD45+ immune cells were isolated using flow cytometry and sequenced using the 10X Genomics platform (n = 3-4 samples per group, each sample pooled from 8-10 eyes). Immunohistochemistry was used to validate the spatial distribution of specific immune cell subsets.
Results:
28,277 immune cells were successfully sequenced from microbead- and sham-injected retinas and optic nerves, identifying many different immune cell subsets. In control mice, optic nerve microglia were less homeostatic than retinal microglia and were characterized by elevated expression of genes such as Spp1 and Gpnmb, which are classically upregulated by microglia in neurodegeneration. In glaucomatous retinas, there was a significant increase in disease-associated microglia (DAM), macrophages, and T cells compared to sham-injected retinas (DAM, p = 0.0039; macrophages, p = 0.0004; T cells, p = 0.024). Moreover, macrophages in the glaucomatous retina shifted towards a state characterized by high MHCII expression, suggesting a potential for crosstalk between retinal macrophages and peripheral immune cells through enhanced antigen presentation and immune response. Interestingly, the immune cell proportions in glaucomatous optic nerves were similar to those in sham optic nerves, with only subtle changes noted in differentially expressed genes between the two conditions.
Conclusions:
Our findings reveal the extensive and dynamic changes in immune cell subsets within glaucomatous retinas, in contrast to the relatively minor changes detected in glaucomatous optic nerves. These findings provide novel insights into the immune cell landscape in glaucoma and may help identify new therapeutic targets for this common blinding disease.
| Original language | English |
|---|---|
| Journal | Investigative ophthalmology & visual science (iovs) |
| Volume | 66 |
| Issue number | 8 |
| Publication status | Published - Jun 2025 |
| Externally published | Yes |