LIN28B-mediated expression of fetal hemoglobin and production of fetal-like erythrocytes from adult human erythroblasts ex vivo.

Abstract

Reactivation of fetal hemoglobin (HbF) holds therapeutic potential for sickle cell disease and β-thalassemias. In human erythroid cells and hematopoietic organs, LIN28B and its targeted let-7 microRNA family, demonstrate regulated expression during the fetal-to-adult developmental transition. To explore the effects of LIN28B in human erythroid cell development, lentiviral transduction was used to knockdown LIN28B expression in erythroblasts cultured from human umbilical cord CD34+ cells. The subsequent reduction in LIN28B expression caused increased expression of let-7 and significantly reduced HbF expression. Conversely, LIN28B overexpression in cultured adult erythroblasts reduced the expression of let-7 and significantly increased HbF expression. Cellular maturation was maintained including enucleation. LIN28B expression in adult erythroblasts increased the expression of γ-globin, and the HbF content of the cells rose to levels >30% of their hemoglobin. Expression of carbonic anhydrase I, glucosaminyl (N-acetyl) transferase 2, and miR-96 (three additional genes marking the transition from fetal-to-adult erythropoiesis) were reduced by LIN28B expression. The transcription factor BCL11A, a well-characterized repressor of γ-globin expression, was significantly down-regulated. Independent of LIN28B, experimental suppression of let-7 also reduced BCL11A expression and significantly increased HbF expression. LIN28B expression regulates HbF levels and causes adult human erythroblasts to differentiate with a more fetal-like phenotype.

• Authors: Y Terry Lee, Jaira F de Vasconcellos, Joan Yuan, Colleen Byrnes, Seung-Jae Noh, Emily R Meier, Ki Soon Kim, Antoinette Rabel, Megha Kaushal, Stefan A Muljo, Jeffery L Miller
• Journal: Blood (Blood) Vol. 122 Issue 6 Pg. 1034-41 (Aug 08 2013) ISSN: 1528-0020 [Electronic] United States
• PMID: 23798711 (Publication Type: Journal Article, Research Support, N.I.H., Intramural)

Chemical References

• Antigens, CD34
• DNA-Binding Proteins
• LIN28B protein, human
• MIRN96 microRNA, human
• MicroRNAs
• RNA-Binding Proteins
• mirnlet7 microRNA, human
• Hemoglobin A
• Fetal Hemoglobin
• N-Acetylglucosaminyltransferases
• Carbonic Anhydrase I

Topics

• Antigens, CD34 (metabolism)
• Carbonic Anhydrase I (metabolism)
• Cell Culture Techniques
• DNA-Binding Proteins (metabolism)
• Erythroblasts (cytology)
• Erythrocytes (cytology)
• Fetal Blood (cytology)
• Fetal Hemoglobin (metabolism)
• Gene Expression Regulation
• Hemoglobin A (metabolism)
• Humans
• MicroRNAs (metabolism)
• N-Acetylglucosaminyltransferases (metabolism)
• Phenotype
• RNA-Binding Proteins