LIN28A Expression Reduces Sickling of Cultured Human Erythrocytes

Induction of fetal hemoglobin (HbF) has therapeutic importance for patients with sickle cell disease (SCD) and the beta-thalassemias. It was recently reported that increased expression of LIN28 proteins or decreased expression of its target let-7 miRNAs enhances HbF levels in cultured primary human erythroblasts from adult healthy donors. Here LIN28A effects were studied further using erythrocytes cultured from peripheral blood progenitor cells of pediatric subjects with SCD. Transgenic expression of LIN28A was accomplished by lentiviral transduction in CD34(+) sickle cells cultivated ex vivo in serum-free medium. LIN28A over-expression (LIN28A-OE) increased HbF, reduced beta (sickle)-globin, and strongly suppressed all members of the let-7 family of miRNAs. LIN28A-OE did not affect erythroblast differentiation or prevent enucleation, but it significantly reduced or ameliorated the sickling morphologies of the enucleated erythrocytes.

[1]  Sergei A. Vinogradov,et al.  Direct measurement of local oxygen concentration in the bone marrow of live animals , 2014, Nature.

[2]  D. Weatherall,et al.  The role of the inherited disorders of hemoglobin, the first "molecular diseases," in the future of human genetics. , 2013, Annual review of genomics and human genetics.

[3]  Y. T. Lee,et al.  LIN28B-mediated expression of fetal hemoglobin and production of fetal-like erythrocytes from adult human erythroblasts ex vivo. , 2013, Blood.

[4]  K. Neville,et al.  Candidate Sequence Variants and Fetal Hemoglobin in Children with Sickle Cell Disease Treated with Hydroxyurea , 2013, PloS one.

[5]  S. Orkin,et al.  The switch from fetal to adult hemoglobin. , 2013, Cold Spring Harbor perspectives in medicine.

[6]  G. Neale,et al.  Hydroxycarbamide alters erythroid gene expression in children with sickle cell anaemia , 2012, British journal of haematology.

[7]  Cong Peng,et al.  Correction of Sickle Cell Disease in Adult Mice by Interference with Fetal Hemoglobin Silencing , 2011, Science.

[8]  Jeffery L. Miller,et al.  Expression patterns of fetal hemoglobin in sickle cell erythrocytes are both patient‐ and treatment‐specific during childhood , 2011, Pediatric blood & cancer.

[9]  M. Platt,et al.  Sickle cell biomechanics. , 2010, Annual review of biomedical engineering.

[10]  S. Orkin,et al.  Transcriptional silencing of {gamma}-globin by BCL11A involves long-range interactions and cooperation with SOX6. , 2010, Genes & development.

[11]  G. Daley,et al.  Lin28: A MicroRNA Regulator with a Macro Role , 2010, Cell.

[12]  Ingo Müller,et al.  Low physiologic oxygen tensions reduce proliferation and differentiation of human multipotent mesenchymal stromal cells , 2010, BMC Cell Biology.

[13]  F. Marincola,et al.  Let-7 microRNAs are developmentally regulated in circulating human erythroid cells , 2009, Journal of Translational Medicine.

[14]  S. Goh,et al.  Cytokine-mediated increases in fetal hemoglobin are associated with globin gene histone modification and transcription factor reprogramming. , 2009, Blood.

[15]  Y. T. Lee,et al.  Identification of TWSG1 as a second novel erythroid regulator of hepcidin expression in murine and human cells. , 2009, Blood.

[16]  D. Strachan,et al.  Genetic variation in LIN28B is associated with the timing of puberty. , 2009, Nature genetics.

[17]  P. Ridker,et al.  Genome-wide association studies identify novel loci associated with age at menarche and age at natural menopause , 2009, Nature Genetics.

[18]  Y. T. Lee,et al.  Identification of TWSG 1 as a second novel erythroid regulator of hepcidin expression in murine and human cells , 2009 .

[19]  J. Hirschhorn,et al.  Supporting Online Material Materials and Methods Figs. S1 to S10 Tables S1 to S7 References Human Fetal Hemoglobin Expression Is Regulated by the Developmental Stage-specific Repressor Bcl11a , 2022 .

[20]  A. Schechter,et al.  Hemoglobin research and the origins of molecular medicine. , 2008, Blood.

[21]  P. Sebastiani,et al.  BCL11A is a major HbF quantitative trait locus in three different populations with beta-hemoglobinopathies. , 2008, Blood cells, molecules & diseases.

[22]  C. Gieger,et al.  Identification of ten loci associated with height highlights new biological pathways in human growth , 2008, Nature Genetics.

[23]  Gonçalo R. Abecasis,et al.  Genome-wide association study shows BCL11A associated with persistent fetal hemoglobin and amelioration of the phenotype of β-thalassemia , 2008, Proceedings of the National Academy of Sciences.

[24]  Shulan Tian,et al.  Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells , 2007, Science.

[25]  Simon Heath,et al.  A QTL influencing F cell production maps to a gene encoding a zinc-finger protein on chromosome 2p15 , 2007, Nature Genetics.

[26]  T. Spector,et al.  Intergenic variants of HBS1L-MYB are responsible for a major quantitative trait locus on chromosome 6q23 influencing fetal hemoglobin levels in adults , 2007, Proceedings of the National Academy of Sciences.

[27]  A. Harel-Bellan,et al.  Lin-28 binds IGF-2 mRNA and participates in skeletal myogenesis by increasing translation efficiency. , 2007, Genes & development.

[28]  V. Ambros,et al.  The Cold Shock Domain Protein LIN-28 Controls Developmental Timing in C. elegans and Is Regulated by the lin-4 RNA , 1997, Cell.

[29]  T Asakura,et al.  Morphologic studies of sickle erythrocytes by image analysis. , 1990, The Journal of laboratory and clinical medicine.

[30]  A. Schechter,et al.  Levels of fetal hemoglobin necessary for treatment of sickle cell disease. , 1988, The New England journal of medicine.

[31]  A. Schechter,et al.  Hemoglobin S polymerization: primary determinant of the hemolytic and clinical severity of the sickling syndromes. , 1985, Blood.

[32]  S. Orkin,et al.  Hydroxyurea enhances fetal hemoglobin production in sickle cell anemia. , 1984, The Journal of clinical investigation.

[33]  T. Ley,et al.  5-azacytidine selectively increases gamma-globin synthesis in a patient with beta+ thalassemia. , 1982, The New England journal of medicine.

[34]  A. Schechter,et al.  The intracellular polymerization of sickle hemoglobin and its relevance to sickle cell disease. , 1981, Blood.

[35]  S. Edelstein,et al.  Diameter of haemoglobin S fibres in sickled cells , 1978, Nature.