Original Research: Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells

Christina M. Ward, Biaoru Li, Betty S. Pace

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Sickle cell anemia is a common genetic disorder caused by a point mutation in the sixth codon of the β-globin gene affecting people of African descent worldwide. A wide variety of clinical phenotypes ranging from mild to severe symptoms and complications occur due to hemoglobin S polymerization, red blood cell sickling, and vaso-occlusion. Research efforts are ongoing to develop strategies of fetal hemoglobin (HbF; α2γ2) induction to inhibit sickle hemoglobin polymerization and improve clinical outcomes. Insights have been gained from investigating mutations in the β-globin locus or transcription factors involved in the mechanisms of hemoglobin switching. Recent efforts to expand molecular targets that modulate γ-globin expression involve microRNAs that work through posttranscriptional gene regulation. Therefore, the goal of our study was to identify novel microRNA genes involved in fetal hemoglobin expression. Using in silico analysis, we identified a miR-34a binding site in the γ-globin mRNA which was tested for functional relevance. Stable expression of the shMIMIC miR-34a lentivirus vector increased fetal hemoglobin levels in single cell K562 clones consistent with silencing of a γ-globin gene repressor. Furthermore, miR-34a promoted cell differentiation supported by increased expression of KLF1, glycophorin A, and the erythropoietin receptor. Western blot analysis of known negative regulators of γ-globin including YY1, histone deacetylase 1, and STAT3, which are regulated by miR-34a showed no change in YY1 and histone deacetylase 1 levels; however, total- and phosphorylated-STAT3 levels were decreased in single cell miR-34a K562 clones. These data support a mechanism of fetal hemoglobin activation by miR-34a involving STAT3 gene silencing.

Original languageEnglish (US)
Pages (from-to)719-729
Number of pages11
JournalExperimental Biology and Medicine
Volume241
Issue number7
DOIs
StatePublished - Jan 1 2016

Fingerprint

Fetal Hemoglobin
Globins
K562 Cells
Genes
Histone Deacetylase 1
Research
Sickle Hemoglobin
MicroRNAs
Polymerization
Clone cells
Clone Cells
Erythropoietin Receptors
Glycophorin
Lentivirus
Inborn Genetic Diseases
Sickle Cell Anemia
Gene Silencing
Point Mutation
Gene expression
Codon

Keywords

  • STAT3
  • fetal hemoglobin
  • gamma-globin
  • miR-34a

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Original Research : Stable expression of miR-34a mediates fetal hemoglobin induction in K562 cells. / Ward, Christina M.; Li, Biaoru; Pace, Betty S.

In: Experimental Biology and Medicine, Vol. 241, No. 7, 01.01.2016, p. 719-729.

Research output: Contribution to journalArticle

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