Single Cell Transcriptomics and Surface Protein Expression Reveal Distinct Cellular and Molecular Phenotypes in Human...

Area of Science:

• Ophthalmology
• Regenerative Medicine
• Cell Biology

Background:

• Current age-related macular degeneration (AMD) treatments utilize retinal pigment epithelium (RPE) cell replacement strategies.
• These strategies involve either pluripotent stem cell (PSC)-derived RPE or adult RPE stem cell (RPESC)-derived RPE.

Purpose of the Study:

• To compare the transcriptomic and surface protein profiles of RPESC-RPE and PSC-RPE.
• To identify potential differences impacting transplantation outcomes for AMD.

Main Methods:

• Utilized Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq).
• Simultaneously assessed single-cell transcriptomic and surface protein expression for both RPE sources.
• Identified key RPE markers and cell-specific surface proteins.

Main Results:

• Both RPESC-RPE and PSC-RPE expressed key RPE markers and showed cellular heterogeneity.
• RPESC-RPE exhibited higher expression of genes related to mature retinal functions.
• PSC-RPE showed greater expression of genes involved in stem cell development and differentiation.
• CD24 was robustly detected on RPESC-RPE, while CD57 was detected on PSC-RPE.

Conclusions:

• Significant differences exist in gene and surface protein expression between RPESC-RPE and PSC-RPE.
• These molecular differences suggest variations in functional, adhesion, and immunomodulatory properties.
• Such differences may influence the success of RPE cell transplantation in AMD patients.