title: Selective Inhibition of ADAM17 Efficiently Mediates Glycoprotein Ibα Retention During Ex Vivo Generation of Human Induced Pluripotent Stem Cell-Derived Platelets.
abstract: Donor-independent platelet concentrates for transfusion can be produced in vitro from induced pluripotent stem cells (iPSCs). However, culture at 37°C induces ectodomain shedding on platelets of glycoprotein Ibα (GPIbα), the von Willebrand factor receptor critical for adhesive function and platelet lifetime in vivo, through temperature-dependent activation of a disintegrin and metalloproteinase 17 (ADAM17). The shedding can be suppressed by using inhibitors of panmetalloproteinases and possibly of the upstream regulator p38 mitogen-activated protein kinase (p38 MAPK), but residues of these inhibitors in the final platelet products may be accompanied by harmful risks that prevent clinical application. Here, we optimized the culture conditions for generating human iPSC-derived GPIbα+platelets, focusing on culture temperature and additives, by comparing a new and safe selective ADAM17 inhibitor, KP-457, with previous inhibitors. Because cultivation at 24°C (at which conventional platelet concentrates are stored) markedly diminished the yield of platelets with high expression of platelet receptors, 37°C was requisite for normal platelet production from iPSCs. KP-457 blocked GPIbα shedding from iPSC platelets at a lower half-maximal inhibitory concentration than panmetalloproteinase inhibitor GM-6001, whereas p38 MAPK inhibitors did not. iPSC platelets generated in the presence of KP-457 exhibited improved GPIbα-dependent aggregation not inferior to human fresh platelets. A thrombus formation model using immunodeficient mice after platelet transfusion revealed that iPSC platelets generated with KP-457 exerted better hemostatic function in vivo. Our findings suggest that KP-457, unlike GM-6001 or p38 MAPK inhibitors, effectively enhances the production of functional human iPSC-derived platelets at 37°C, which is an important step toward their clinical application. Stem Cells Translational Medicine 2017;6:720-730.
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.|Kaken Pharmaceutical Co., Ltd., Tokyo, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Hirata
authors.affiliation: Kaken Pharmaceutical Co., Ltd., Tokyo, Japan.
authors.forename: Takahiko
authors.initials: T
authors.lastname: Murata
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Suzuki
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
authors.forename: Sou
authors.initials: S
authors.lastname: Nakamura
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Jono-Ohnishi
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.|Kyoto Development Center, Megakaryon Co., Ltd., Kyoto, Japan.
authors.forename: Hidenori
authors.initials: H
authors.lastname: Hirose
authors.affiliation: Department of Anatomy, Faculty of Medicine, University of Miyazaki, Miyazaki, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Sawaguchi
authors.affiliation: Center for Molecular Medicine, Jichi Medical University, Tochigi, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nishimura
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
authors.forename: Naoshi
authors.initials: N
authors.lastname: Sugimoto
authors.affiliation: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.|Department of Innovation Stem Cell Therapy, Chiba University Graduate School of Medicine, Chiba, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Eto
mesh_terms: D000072198:ADAM17 Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000375:Aging / Q000378:metabolism; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002478:Cells, Cultured; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006487:Hemostasis / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D008533:Megakaryocytes / Q000187:drug effects / Q000378:metabolism; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism*; D013696:Temperature; D034061:Thrombopoiesis / Q000187:drug effects
chemical_list: D019038:Platelet Glycoprotein GPIb-IX Complex; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000072198:ADAM17 Protein
keywords: Cell culture; Cell transplantation; Induced pluripotent stem cell; Megakaryocyte; Mitogen-activated protein kinase; Thrombopoiesis