title: Targeted gene correction of RUNX1 in induced pluripotent stem cells derived from familial platelet disorder with propensity to myeloid malignancy restores normal megakaryopoiesis.
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.
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Iizuka
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kagoya
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Kataoka
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Akihide
authors.initials: A
authors.lastname: Yoshimi
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Miyauchi
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Taoka
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Keiki
authors.initials: K
authors.lastname: Kumano
authors.affiliation: Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: Department of Reprogramming Science, Center for iPS cell Research and Applications (CiRA), and Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University, Kyoto, Japan; PRESTO, Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Akitsu
authors.initials: A
authors.lastname: Hotta
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors.forename: Shunya
authors.initials: S
authors.lastname: Arai
authors.affiliation: Department of Hematology & Oncology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan. Electronic address: kurokawa-tky@umin.ac.jp.
authors.forename: Mineo
authors.initials: M
authors.lastname: Kurokawa
mesh_terms: D000328:Adult; D025861:Blood Coagulation Disorders, Inherited / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D001791:Blood Platelet Disorders / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D018389:Codon, Nonsense; D050676:Core Binding Factor Alpha 2 Subunit / Q000096:biosynthesis / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000473:pathology; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D034061:Thrombopoiesis / Q000235:genetics*
chemical_list: D018389:Codon, Nonsense; D050676:Core Binding Factor Alpha 2 Subunit; D018076:DNA, Complementary; C493728:RUNX1 protein, human