title: LIN28A expression reduces sickling of cultured human erythrocytes.
abstract: 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.
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Jaira F
authors.initials: JF
authors.lastname: de Vasconcellos
authors.affiliation: Center for Cancer and Blood Disorders, Children's National Medical Center, Washington, District of Columbia, United States of America.
authors.forename: Ross M
authors.initials: RM
authors.lastname: Fasano
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Y Terry
authors.initials: YT
authors.lastname: Lee
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America; Center for Cancer and Blood Disorders, Children's National Medical Center, Washington, District of Columbia, United States of America.
authors.forename: Megha
authors.initials: M
authors.lastname: Kaushal
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Colleen
authors.initials: C
authors.lastname: Byrnes
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America; Center for Cancer and Blood Disorders, Children's National Medical Center, Washington, District of Columbia, United States of America.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Meier
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Molly
authors.initials: M
authors.lastname: Anderson
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Antoinette
authors.initials: A
authors.lastname: Rabel
authors.affiliation: Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Raul
authors.initials: R
authors.lastname: Braylan
authors.affiliation: Cell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: David F
authors.initials: DF
authors.lastname: Stroncek
authors.affiliation: Molecular Genomics and Therapeutics Section, Molecular Medicine Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Jeffery L
authors.initials: JL
authors.lastname: Miller
mesh_terms: D000293:Adolescent; D000755:Anemia, Sickle Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D002454:Cell Differentiation; D048430:Cell Shape; D002648:Child; D004900:Erythroblasts / Q000378:metabolism / Q000473:pathology; D017707:Erythrocyte Transfusion; D004913:Erythrocytes, Abnormal / Q000378:metabolism* / Q000473:pathology; D005319:Fetal Hemoglobin / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D061251:Primary Cell Culture; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection; D055544:beta-Globins / Q000235:genetics* / Q000378:metabolism
chemical_list: C516326:LIN28B protein, human; D035683:MicroRNAs; D016601:RNA-Binding Proteins; D055544:beta-Globins; C487931:mirnlet7 microRNA, human; D005319:Fetal Hemoglobin