High-level transgene expression in induced pluripotent stem cell-derived megakaryocytes: correction of Glanzmann thrombasthenia

Blood. 2014 Jan 30;123(5):753-7. doi: 10.1182/blood-2013-10-530725. Epub 2013 Dec 13.

Abstract

Megakaryocyte-specific transgene expression in patient-derived induced pluripotent stem cells (iPSCs) offers a new approach to study and potentially treat disorders affecting megakaryocytes and platelets. By using a Gp1ba promoter, we developed a strategy for achieving a high level of protein expression in human megakaryocytes. The feasibility of this approach was demonstrated in iPSCs derived from two patients with Glanzmann thrombasthenia (GT), an inherited platelet disorder caused by mutations in integrin αIIbβ3. Hemizygous insertion of Gp1ba promoter-driven human αIIb complementary DNA into the AAVS1 locus of iPSCs led to high αIIb messenger RNA and protein expression and correction of surface αIIbβ3 in megakaryocytes. Agonist stimulation of these cells displayed recovery of integrin αIIbβ3 activation. Our findings demonstrate a novel approach to studying human megakaryocyte biology as well as functional correction of the GT defect, offering a potential therapeutic strategy for patients with diseases that affect platelet function.

Publication types

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

MeSH terms

  • Gene Expression
  • Humans
  • Induced Pluripotent Stem Cells / metabolism*
  • Megakaryocytes / metabolism*
  • Membrane Glycoproteins / genetics*
  • Platelet Glycoprotein GPIIb-IIIa Complex / genetics*
  • Platelet Glycoprotein GPIb-IX Complex
  • Promoter Regions, Genetic
  • Thrombasthenia / genetics*
  • Transgenes*

Substances

  • Membrane Glycoproteins
  • Platelet Glycoprotein GPIIb-IIIa Complex
  • Platelet Glycoprotein GPIb-IX Complex
  • adhesion receptor