Discrimination of Dormant and Active Hematopoietic Stem Cells by G0 Marker Reveals Dormancy Regulation by Cytoplasmic Calcium
Tsuyoshi Fukushima,
Yosuke Tanaka,
Fiona K. Hamey,
Chih-Hsiang Chang,
Toshihiko Oki,
Shuhei Asada,
Yasutaka Hayashi,
Takeshi Fujino,
Taishi Yonezawa,
Reina Takeda,
Kimihito Cojin Kawabata,
Tomofusa Fukuyama,
Terumasa Umemoto,
Keiyo Takubo,
Hitoshi Takizawa,
Susumu Goyama,
Yasushi Ishihama,
Hiroaki Honda,
Berthold Göttgens,
Toshio Kitamura
Affiliations
Tsuyoshi Fukushima
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Yosuke Tanaka
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan; Corresponding author
Fiona K. Hamey
Department of Haematology, Wellcome Trust and MRC Cambridge Stem Cell Institute and Cambridge Institute for Medical Research, Cambridge University, Cambridge CB2 0XY, UK
Chih-Hsiang Chang
Department of Molecular and Cellular BioAnalysis, Graduate School of Pharmaceutical Sciences, Sakyo-ku, Kyoto University, Kyoto 606-8501, Japan
Toshihiko Oki
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA
Shuhei Asada
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Yasutaka Hayashi
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Takeshi Fujino
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Taishi Yonezawa
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Reina Takeda
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Kimihito Cojin Kawabata
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan; Department of Hematology/Oncology, Weill Cornell Medical College, New York, NY 10021, USA
Tomofusa Fukuyama
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Terumasa Umemoto
International Research Center for Medical Sciences, Chuo-ku, Kumamoto University, Kumamoto 860-0811, Japan
Keiyo Takubo
Department of Stem Cell Biology, Research Institute, National Center for Global Health and Medicine, Shinjuku-ku, Tokyo 162-8655, Japan
Hitoshi Takizawa
International Research Center for Medical Sciences, Chuo-ku, Kumamoto University, Kumamoto 860-0811, Japan
Susumu Goyama
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Yasushi Ishihama
Department of Molecular and Cellular BioAnalysis, Graduate School of Pharmaceutical Sciences, Sakyo-ku, Kyoto University, Kyoto 606-8501, Japan
Hiroaki Honda
Field of Human Disease Models, Major in Advanced Life Sciences and Medicine, Tokyo Women’s Medical University, Shinjuku-ku, Tokyo 162-8666, Japan
Berthold Göttgens
Department of Haematology, Wellcome Trust and MRC Cambridge Stem Cell Institute and Cambridge Institute for Medical Research, Cambridge University, Cambridge CB2 0XY, UK
Toshio Kitamura
Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan
Summary: Quiescent hematopoietic stem cells (HSCs) are typically dormant, and only a few quiescent HSCs are active. The relationship between “dormant” and “active” HSCs remains unresolved. Here we generate a G0 marker (G0M) mouse line that visualizes quiescent cells and identify a small population of active HSCs (G0Mlow), which are distinct from dormant HSCs (G0Mhigh), within the conventional quiescent HSC fraction. Single-cell RNA-seq analyses show that the gene expression profiles of these populations are nearly identical but differ in their Cdk4/6 activity. Furthermore, high-throughput small-molecule screening reveals that high concentrations of cytoplasmic calcium ([Ca2+]c) are linked to dormancy of HSCs. These findings indicate that G0M separates dormant and active adult HSCs, which are regulated by Cdk4/6 and [Ca2+]c. This G0M mouse line represents a useful resource for investigating physiologically important stem cell subpopulations. : Fukushima et al. show that G0 marker (G0M) discriminates between dormant and active HSCs within the conventional quiescent HSC fraction. Small-molecule screening reveals that high [Ca2+]c is linked to dormancy of HSCs. Moreover, upregulation of [Ca2+]c by thapsigargin enhances the bone marrow reconstitution ability of HSCs. Keywords: hematopoietic stem cell, HSC, cell cycle, G0 phase, p27, dormancy, CDK, calcium
