title: Transient activation of c-MYC expression is critical for efficient platelet generation from human induced pluripotent stem cells.
abstract: Human (h) induced pluripotent stem cells (iPSCs) are a potentially abundant source of blood cells, but how best to select iPSC clones suitable for this purpose from among the many clones that can be simultaneously established from an identical source is not clear. Using an in vitro culture system yielding a hematopoietic niche that concentrates hematopoietic progenitors, we show that the pattern of c-MYC reactivation after reprogramming influences platelet generation from hiPSCs. During differentiation, reduction of c-MYC expression after initial reactivation of c-MYC expression in selected hiPSC clones was associated with more efficient in vitro generation of CD41a(+)CD42b(+) platelets. This effect was recapitulated in virus integration-free hiPSCs using a doxycycline-controlled c-MYC expression vector. In vivo imaging revealed that these CD42b(+) platelets were present in thrombi after laser-induced vessel wall injury. In contrast, sustained and excessive c-MYC expression in megakaryocytes was accompanied by increased p14 (ARF) and p16 (INK4A) expression, decreased GATA1 expression, and impaired production of functional platelets. These findings suggest that the pattern of c-MYC expression, particularly its later decline, is key to producing functional platelets from selected iPSC clones.
authors.affiliation: Stem Cell Bank, Center for Stem Cell Biology and Regenerative Medicine, the Institute of Medical Science, Tokyo, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Takayama
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nishimura
authors.forename: Sou
authors.initials: S
authors.lastname: Nakamura
authors.forename: Takafumi
authors.initials: T
authors.lastname: Shimizu
authors.forename: Ryoko
authors.initials: R
authors.lastname: Ohnishi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Endo
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Makoto
authors.initials: M
authors.lastname: Otsu
authors.forename: Ken
authors.initials: K
authors.lastname: Nishimura
authors.forename: Mahito
authors.initials: M
authors.lastname: Nakanishi
authors.forename: Akira
authors.initials: A
authors.lastname: Sawaguchi
authors.forename: Ryozo
authors.initials: R
authors.lastname: Nagai
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamanaka
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Nakauchi
authors.forename: Koji
authors.initials: K
authors.lastname: Eto
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D046529:Microscopy, Electron, Transmission; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D017713:Platelet Transfusion; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013921:Thrombocytopenia / Q000628:therapy; D025803:Tumor Suppressor Protein p14ARF / Q000235:genetics / Q000378:metabolism
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D019038:Platelet Glycoprotein GPIb-IX Complex; D040201:Platelet Membrane Glycoprotein IIb; D016271:Proto-Oncogene Proteins c-myc; D025803:Tumor Suppressor Protein p14ARF