Single Cell RNA-Seq Analysis of Human Red Cells.

Area of Science:

• Hematology
• Genomics
• Molecular Biology

Background:

• Human red blood cells (RBCs) are crucial for gas exchange, and RBC diseases impact millions globally.
• Phenotypic heterogeneity within RBC populations is recognized but poorly understood.
• Previous analyses of RBC RNA transcripts did not capture sub-population heterogeneity.

Purpose of the Study:

• To analyze transcriptional heterogeneity in human RBCs using single-cell RNA sequencing.
• To identify distinct RBC subpopulations and their characteristics.
• To investigate the impact of blood bank storage on RBC heterogeneity.

Main Methods:

• Single-cell RNA sequencing (scRNA-Seq) was performed on RBCs from three healthy adult donors.
• RBCs were assayed at day 1 and day 15 of storage under blood bank conditions.
• Bioinformatic analysis was used to cluster cells based on gene expression patterns.

Main Results:

• scRNA-Seq identified seven distinct RBC clusters.
• A cluster expressing fetal hemoglobin (HbF) was identified and annotated as F cells.
• Clusters were annotated based on canonical gene expression, including HBG2, HBB, NIX, ACVR2B, and HEMGN.
• RBC storage was associated with an increase in ACVR2B-expressing and F-cell clusters.

Conclusions:

• Single-cell RNA sequencing is a powerful tool for discovering RBC population heterogeneity.
• Distinct RBC subpopulations, including F cells, can be identified and characterized.
• Blood storage conditions can alter RBC transcriptional profiles, affecting specific subpopulations.