Erythroid-Specific Expression of LIN28A Is Sufficient for Robust Gamma-Globin Gene and Protein Expression in Adult Erythroblasts

• Published in: PloS one Vol. 10; no. 12; p. e0144977
• Main Authors: Lee, Y Terry, de Vasconcellos, Jaira F, Byrnes, Colleen, Kaushal, Megha, Rabel, Antoinette, Tumburu, Laxminath, Allwardt, Joshua M, Miller, Jeffery L
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 16-12-2015; Public Library of Science (PLoS)
• Subjects: Adult; Cell Differentiation; Cell Proliferation; Cloning; Colonies; Diabetes; Enzymes; Erythroblasts; Erythroblasts - cytology; Erythroblasts - metabolism; Expression vectors; Fetal Hemoglobin - genetics; Fetal Hemoglobin - metabolism; Fetuses; gamma-Globins - genetics; gamma-Globins - metabolism; Gene expression; Gene Expression Regulation; Genes; Genomics; Hemoglobin; Hemoglobins; Humans; Kidney diseases; Kruppel-Like Transcription Factors - genetics; Medicine; MicroRNA; MicroRNAs; MicroRNAs - genetics; miRNA; Organ Specificity - genetics; Overexpression; Promoter Regions, Genetic; Proteins; Puromycin; Ribonucleic acid; RNA; RNA-Binding Proteins - genetics; Stem cells; Transcription, Genetic; United States > US; Maryland
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0144977

Increasing fetal hemoglobin (HbF) levels in adult humans remains an active area in hematologic research. Here we explored erythroid-specific LIN28A expression for its effect in regulating gamma-globin gene expression and HbF levels in cultured adult erythroblasts. For this purpose, lentiviral transduction vectors were produced with LIN28A expression driven by erythroid-specific gene promoter regions of the human KLF1 or SPTA1 genes. Transgene expression of LIN28A with a linked puromycin resistance marker was restricted to the erythroid lineage as demonstrated by selective survival of erythroid colonies (greater than 95% of all colonies). Erythroblast LIN28A over-expression (LIN28A-OE) did not significantly affect proliferation or inhibit differentiation. Greater than 70% suppression of total let-7 microRNA levels was confirmed in LIN28A-OE cells. Increases in gamma-globin mRNA and protein expression with HbF levels reaching 30-40% were achieved. These data suggest that erythroblast targeting of LIN28A expression is sufficient for increasing fetal hemoglobin expression in adult human erythroblasts.