Single-cell transcriptomics reveals transcriptional and metabolic alterations associated with disease severity in β-thalassemia/Hb E

Single-cell RNA sequencing was used to analyze transcriptional profiles during erythropoiesis in erythroblasts from normal subjects, mild, and severe β-thalassemia/Hb E patients, revealing disease severity- and differentiation stage-associated alterations in erythroid transcriptional states. Erythroblasts from each group were collected on days 8 and 10 of differentiation phase and labeled with lipid-modified oligonucleotide (LMO)-based MULTI-seq barcodes. GEM generation and library preparation were performed according to the protocol provided with the Chromium GEM-X Single Cell 3′ Reagent Kits v4 (10x Genomics). In this study, two libraries were generated: library 1 contained 16,673 cells from erythroblasts on day 8, whereas library 2 contained 12,754 cells from erythroblasts on day 10. R script used for integration, clustering, differential expression analysis, functional enrichment analysis, and visualization of single-cell RNA sequencing data is available in “Rscript_CS07_integ” file. All sequencing raw and processed data have been uploaded to NCBI’s Gene Expression Omnibus (accession no. GSE348715).