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

• Published in: Experimental hematology Vol. 43; no. 10; pp. 849 - 857
• Main Authors: Iizuka, Hiromitsu, Kagoya, Yuki, Kataoka, Keisuke, Yoshimi, Akihide, Miyauchi, Masashi, Taoka, Kazuki, Kumano, Keiki, Yamamoto, Takashi, Hotta, Akitsu, Arai, Shunya, Kurokawa, Mineo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.10.2015
• Subjects: Adult; Advanced Basic Science; 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; Hematology, Oncology and Palliative Medicine; 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
• ISSN: 0301-472X; 1873-2399
• DOI: 10.1016/j.exphem.2015.05.004

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.