Domain-focused CRISPR screen identifies HRI as a fetal hemoglobin regulator in human erythroid cells

Science. 2018 Jul 20;361(6399):285-290. doi: 10.1126/science.aao0932.

Abstract

Increasing fetal hemoglobin (HbF) levels in adult red blood cells provides clinical benefit to patients with sickle cell disease and some forms of β-thalassemia. To identify potentially druggable HbF regulators in adult human erythroid cells, we employed a protein kinase domain-focused CRISPR-Cas9-based genetic screen with a newly optimized single-guide RNA scaffold. The screen uncovered the heme-regulated inhibitor HRI (also known as EIF2AK1), an erythroid-specific kinase that controls protein translation, as an HbF repressor. HRI depletion markedly increased HbF production in a specific manner and reduced sickling in cultured erythroid cells. Diminished expression of the HbF repressor BCL11A accounted in large part for the effects of HRI depletion. Taken together, these results suggest HRI as a potential therapeutic target for hemoglobinopathies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anemia, Sickle Cell / drug therapy
  • Anemia, Sickle Cell / genetics*
  • CRISPR-Cas Systems
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism
  • Cell Line
  • Erythroid Cells / metabolism*
  • Fetal Hemoglobin / genetics*
  • Gene Expression Regulation*
  • Genetic Testing
  • Humans
  • Molecular Targeted Therapy
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / metabolism
  • RNA, Guide, CRISPR-Cas Systems
  • Repressor Proteins
  • eIF-2 Kinase / genetics*
  • eIF-2 Kinase / metabolism

Substances

  • BCL11A protein, human
  • Carrier Proteins
  • Nuclear Proteins
  • RNA, Guide, CRISPR-Cas Systems
  • Repressor Proteins
  • Fetal Hemoglobin
  • EIF2AK1 protein, human
  • eIF-2 Kinase