Single-cell transcriptomics combined with spatial proteomics defines phagocytes type-specific immune regulation in...

Area of Science:

• Ophthalmology
• Immunology
• Genomics

Background:

• Diabetic cataract (DC) is a major cause of vision loss in diabetes, driven by complex immune mechanisms.
• Understanding the immune cell characteristics and molecular pathways in DC is crucial for developing effective treatments.

Purpose of the Study:

• To investigate immune cell profiles and molecular mechanisms in diabetic rat lenses using multiomic data.
• To identify potential therapeutic targets for diabetic cataract.

Main Methods:

• Single-cell RNA sequencing (scRNA-seq) to identify lens cell types and proportions.
• Quantitative proteomics to screen immune targets and pathological mechanisms.
• Gene Ontology (GO) and KEGG pathway analyses for differential protein analysis.
• Anterior segment optical coherence tomography (AS-OCT) and immunofluorescence staining for immune cell detection.

Main Results:

• scRNA-seq revealed increased mononuclear phagocytes, specifically pro-inflammatory M1 macrophages, in diabetic rat lenses.
• Immunofluorescence and AS-OCT confirmed a higher immune cell presence in DC patients compared to age-related cataract (ARC) patients.
• Proteomics identified upregulated macrophage migration inhibitory factor (MIF) and pathways related to immune regulation and protein ubiquitination (K63 ubiquitination).

Conclusions:

• Immune mechanisms, particularly mononuclear phagocytes, play a significant role in diabetic cataract development.
• Upregulated immune regulation and altered protein ubiquitination pathways present potential therapeutic targets for DC.
• Further research is needed to elucidate specific immune cell functions and develop targeted therapies.