Targeted gene correction of RUNX1 in induced pluripotent stem cells derived from familial platelet disorder with propensity to myeloid malignancy restores normal megakaryopoiesis

Exp Hematol. 2015 Oct;43(10):849-57. doi: 10.1016/j.exphem.2015.05.004. Epub 2015 Jun 11.

Abstract

Familial platelet disorder with propensity to acute myeloid leukemia (FPD/AML) is an autosomal dominant disease associated with a germline mutation in the RUNX1 gene and is characterized by thrombocytopenia and an increased risk of developing myeloid malignancies. We generated induced pluripotent stem cells (iPSCs) from dermal fibroblasts of a patient with FPD/AML possessing a nonsense mutation R174X in the RUNX1 gene. Consistent with the clinical characteristics of the disease, FPD iPSC-derived hematopoietic progenitor cells were significantly impaired in undergoing megakaryocytic differentiation and subsequent maturation, as determined by colony-forming cell assay and surface marker analysis. Notably, when we corrected the RUNX1 mutation using transcription activator-like effector nucleases in conjunction with a donor plasmid containing normal RUNX1 cDNA sequences, megakaryopoiesis and subsequent maturation were restored in FPD iPSC-derived hematopoietic cells. These findings clearly indicate that the RUNX1 mutation is robustly associated with thrombocytopenia in patients with FPD/AML, and transcription activator-like effector nuclease-mediated gene correction in iPSCs generated from patient-derived cells could provide a promising clinical application for treatment of the disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blood Coagulation Disorders, Inherited* / genetics
  • Blood Coagulation Disorders, Inherited* / metabolism
  • Blood Coagulation Disorders, Inherited* / pathology
  • Blood Coagulation Disorders, Inherited* / therapy
  • Blood Platelet Disorders* / genetics
  • Blood Platelet Disorders* / metabolism
  • Blood Platelet Disorders* / pathology
  • Blood Platelet Disorders* / therapy
  • Codon, Nonsense*
  • Core Binding Factor Alpha 2 Subunit* / biosynthesis
  • Core Binding Factor Alpha 2 Subunit* / genetics
  • DNA, Complementary / genetics
  • Female
  • Genetic Therapy / methods*
  • Humans
  • Induced Pluripotent Stem Cells / metabolism*
  • Induced Pluripotent Stem Cells / pathology
  • Leukemia, Myeloid, Acute* / genetics
  • Leukemia, Myeloid, Acute* / metabolism
  • Leukemia, Myeloid, Acute* / pathology
  • Leukemia, Myeloid, Acute* / therapy
  • Thrombopoiesis / genetics*

Substances

  • Codon, Nonsense
  • Core Binding Factor Alpha 2 Subunit
  • DNA, Complementary
  • RUNX1 protein, human

Supplementary concepts

  • Platelet Disorder, Familial, with Associated Myeloid Malignancy