Researchers Database

uosaki hideki

    DivisionofRegenerativeMedicine,CenterformolecularMedicine Associate Professor
Last Updated :2021/12/08

Researcher Information

J-Global ID


  • Research interests: Heart Development, Stem Cells (ES/iPS cells)

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Research Interests

  • 遺伝子治療   ゲノム編集   Regenerative Medicine   Stem Cell Biology   Transcriptome Analysis   Maturation   Differentiation   Cardiomyocytes   Pluripotent Stem Cells   

Research Areas

  • Life sciences / Developmental biology
  • Life sciences / Anatomy
  • Life sciences / Cardiology

Academic & Professional Experience

  • 2017/12 - Today  Jichi Medical UniversityCenter for Molecular Medicine, Division of Regenerative MedicineAssociate Professor
  • 2016/09 - 2017/11  Jichi Medical UniversityCenter for Molecular Medicine, Division of Regenerative MedicineLecturer
  • 2011/06 - 2016/08  Johns Hopkins UniversitySchool of Medicine, Division of CardiologyPostdoctoral Fellow
  • 2011/04 - 2011/05  Kyoto UniversityCenter for iPS research and applicationResearch Fellow


  • 2006/04 - 2011/03  Kyoto University  Graduate School of Medicine  医学専攻
  • 1998/04 - 2004/03  Hokkaido University  School of Medicine  医学科

Published Papers

  • Matthew Miyamoto, Peter Andersen, Edrick Sulistio, Xihe Liu, Sean Murphy, Suraj Kannan, Lucy Nam, William Miyamoto, Emmanouil Tampakakis, Narutoshi Hibino, Hideki Uosaki, Chulan Kwon
    Biochemical and Biophysical Research Communications 577 12 - 16 0006-291X 2021/11 [Refereed][Not invited]
  • Takeshi Tokuyama, Razan Elfadil Ahmed, Nawin Chanthra, Tatsuya Anzai, Hideki Uosaki
    Biology 2021/09
  • Takafumi Miyamoto, Hideki Uosaki, Yuhei Mizunoe, Song-Iee Han, Satoi Goto, Daisuke Yamanaka, Masato Masuda, Yosuke Yoneyama, Hideki Nakamura, Naoko Hattori, Yoshinori Takeuchi, Hiroshi Ohno, Motohiro Sekiya, Takashi Matsuzaka, Fumihiko Hakuno, Shin-Ichiro Takahashi, Naoya Yahagi, Koichi Ito, Hitoshi Shimano
    Cell Reports Methods 100052 - 100052 2667-2375 2021/08
  • Tatsuya Anzai, Hiromasa Hara, Nawin Chanthra, Taketaro Sadahiro, Masaki Ieda, Yutaka Hanazono, Hideki Uosaki
    Methods in molecular biology (Clifton, N.J.) 2320 247 - 259 2021/07 [Not refereed][Invited]
    A knock-in can generate fluorescent or Cre-reporter under the control of an endogenous promoter. It also generates knock-out or tagged-protein with fluorescent protein and short tags for tracking and purification. Recent advances in genome editing with clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9) significantly increased the efficiencies of making knock-in cells. Here we describe the detailed protocols of generating knock-in mouse and human pluripotent stem cells (PSCs) by electroporation and lipofection, respectively.
  • Azumi Noguchi, Kenji Ito, Yuichi Uosaki, Maky Ideta-Otsuka, Katsuhide Igarashi, Hideyuki Nakashima, Toshikazu Kakizaki, Ruri Kaneda, Hideki Uosaki, Yuchio Yanagawa, Kinichi Nakashima, Hirokazu Arakawa, Takumi Takizawa
    Neuroscience Research 0168-0102 2021/05 [Refereed][Not invited]
  • Suvd Byambaa, Hideki Uosaki, Tsukasa Ohmori, Hiromasa Hara, Hitoshi Endo, Osamu Nureki, Yutaka Hanazono
    Molecular therapy. Methods & clinical development 20 451 - 462 2021/03 
    We conducted two lines of genome-editing experiments of mouse hematopoietic stem cells (HSCs) with the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9). First, to evaluate the genome-editing efficiency in mouse bona fide HSCs, we knocked out integrin alpha 2b (Itga2b) with Cas9 ribonucleoprotein (Cas9/RNP) and performed serial transplantation in mice. The knockout efficiency was estimated at approximately 15%. Second, giving an example of X-linked severe combined immunodeficiency (X-SCID) as a target genetic disease, we showed a proof-of-concept of universal gene correction, allowing rescue of most of X-SCID mutations, in a completely non-viral setting. We inserted partial cDNA of interleukin-2 receptor gamma chain (Il2rg) into intron 1 of Il2rg via non-homologous end-joining (NHEJ) with Cas9/RNP and a homology-independent targeted integration (HITI)-based construct. Repaired HSCs reconstituted T lymphocytes and thymuses in SCID mice. Our results show that a non-viral genome-editing of HSCs with CRISPR/Cas9 will help cure genetic diseases.
  • Sean A Murphy, Matthew Miyamoto, Anaïs Kervadec, Suraj Kannan, Emmanouil Tampakakis, Sandeep Kambhampati, Brian Leei Lin, Sam Paek, Peter Andersen, Dong-Ik Lee, Renjun Zhu, Steven S An, David A Kass, Hideki Uosaki, Alexandre R Colas, Chulan Kwon
    Nature communications 12 (1) 1648 - 1648 2021/03 
    Cardiomyocytes undergo significant structural and functional changes after birth, and these fundamental processes are essential for the heart to pump blood to the growing body. However, due to the challenges of isolating single postnatal/adult myocytes, how individual newborn cardiomyocytes acquire multiple aspects of the mature phenotype remains poorly understood. Here we implement large-particle sorting and analyze single myocytes from neonatal to adult hearts. Early myocytes exhibit wide-ranging transcriptomic and size heterogeneity that is maintained until adulthood with a continuous transcriptomic shift. Gene regulatory network analysis followed by mosaic gene deletion reveals that peroxisome proliferator-activated receptor coactivator-1 signaling, which is active in vivo but inactive in pluripotent stem cell-derived cardiomyocytes, mediates the shift. This signaling simultaneously regulates key aspects of cardiomyocyte maturation through previously unrecognized proteins, including YAP1 and SF3B2. Our study provides a single-cell roadmap of heterogeneous transitions coupled to cellular features and identifies a multifaceted regulator controlling cardiomyocyte maturation.
  • Razan E. Ahmed, Nawin Chanthra, Tatsuya Anzai, Keiichiro Koiwai, Tomoki Murakami, Hiroaki Suzuki, Yutaka Hanazono, Hideki Uosaki
    Journal of Visualized Experiments (169) 2021/03 [Refereed]
  • Tomoyuki Abe, Hideki Uosaki, Hiroaki Shibata, Hiromasa Hara, Borjigin Sarentonglaga, Yoshikazu Nagao, Yutaka Hanazono
    Experimental Hematology 2021/03
  • Suvd Byambaa, Hideki Uosaki, Hiromasa Hara, Yasumitsu Nagao, Tomoyuki Abe, Hiroaki Shibata, Osamu Nureki, Tsukasa Ohmori, Yutaka Hanazono
    Experimental animals 69 (2) 189 - 198 1341-1357 2020/04 [Refereed][Not invited]
    X-linked severe combined immunodeficiency (X-SCID) is an inherited genetic disorder. A majority of X-SCID subjects carries point mutations in the Interleukin-2 receptor gamma chain (IL2RG) gene. In contrast, Il2rg-knockout mice recapitulating X-SCID phenotype lack a large part of Il2rg instead of point mutations. In this study, we generated novel X-SCID mouse strains with small insertion and deletion (InDel) mutations in Il2rg by using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9. To this end, we injected Streptococcus pyogenes Cas9 (SpCas9) mRNA and single guide RNA targeting the exon 2, 3 or 4 of Il2rg into mouse zygotes. In the F0 generation, we obtained 35 pups and 25 out of them were positive for Surveyor assay, and most of mutants displayed dramatic reductions of T and B lymphocytes in the peripheral blood. By amplicon sequencing, 15 out of 31 founder mice were determined as monoallelic mutants with possible minor mosaicisms while 10 mice were mosaic. Finally, we established new strains with 7-nucleotide deletion and 1-nucleotide insertions in the exon 2 and the exons 3 and 4, respectively. Although no IL2RG protein was detected on T cells of exons 3 and 4 mutants, IL2RG protein was unexpectedly detected in the exon 2 mutants. These data indicated that CRISPR/Cas9 targeting Il2rg causes InDel mutations effectively and generates genetically X-SCID mice. Genetic mutations, however, did not necessarily grant phenotypical alteration, which requires an intensive analysis after establishing a strain to confirm their phenotypes.
  • Tatsuya Anzai, Takanori Yamagata, Hideki Uosaki
    Frontiers in cell and developmental biology 8 268 - 268 2020/04 [Refereed][Not invited]
    Transcriptome landscape of organs from mice and humans offers perspectives on the process of how organs develop and the similarity and diversity in each organ between the species. Among multi-species and multi-organ dataset, which was previously generated, we focused on the mouse and human dataset and performed a reanalysis to provide a more specific perspective on the maturation of human cardiomyocytes. First, we examined how organs diversify their transcriptome during development across and within two species. We unexpectedly identified that ribosomal genes were differentially expressed between mice and humans. Second, we examined the corresponding ages of organs in mice and humans and found that the corresponding developmental ages did not match throughout organs. Mouse hearts at P0-3 and human hearts at 18-19 wpc showed the most proximity in the regard of the transcriptome. Third, we identified a novel set of maturation marker genes that are more consistent between mice and humans. In contrast, conventionally used maturation marker genes only work well with mouse hearts. Finally, we compared human pluripotent stem cell-derived cardiomyocytes (PSC-CMs) in maturation-enhanced conditions to human fetal and adult hearts and revealed that human PSC-CMs only expressed low levels of the potential maturation marker genes. Our findings provide a novel foundation to study cardiomyocyte maturation and highlight the importance of studying human samples rather than relying on a mouse time-series dataset.
  • Hirofumi Nishizono, Mohamed Darwish, Hideki Uosaki, Nanami Masuyama, Motoaki Seki, Hiroyuki Abe, Nozomu Yachie, Ryohei Yasuda
    Journal of visualized experiments : JoVE (158) 2020/04 [Refereed][Not invited]
    The use of genetically modified (GM) mice has become crucial for understanding gene function and deciphering the underlying mechanisms of human diseases. The CRISPR/Cas9 system allows researchers to modify the genome with unprecedented efficiency, fidelity, and simplicity. Harnessing this technology, researchers are seeking a rapid, efficient, and easy protocol for generating GM mice. Here we introduce an improved method for cryopreservation of one-cell embryos that leads to a higher developmental rate of the freeze-thawed embryos. By combining it with optimized electroporation conditions, this protocol allows for the generation of knockout and knock-in mice with high efficiency and low mosaic rates within a short time. Furthermore, we show a step-by-step explanation of our optimized protocol, covering CRISPR reagent preparation, in vitro fertilization, cryopreservation and thawing of one-cell embryos, electroporation of CRISPR reagents, mouse generation, and genotyping of the founders. Using this protocol, researchers should be able to prepare GM mice with unparalleled ease, speed, and efficiency.
  • Razan Elfadil Ahmed, Tatsuya Anzai, Nawin Chanthra, Hideki Uosaki
    Frontiers in cell and developmental biology 8 178 - 178 2020/03 [Refereed][Not invited]
    Cardiovascular diseases are the leading cause of death worldwide. Therefore, the discovery of induced pluripotent stem cells (iPSCs) and the subsequent generation of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) was a pivotal point in regenerative medicine and cardiovascular research. They constituted an appealing tool for replacing dead and dysfunctional cardiac tissue, screening cardiac drugs and toxins, and studying inherited cardiac diseases. The problem is that these cells remain largely immature, and in order to utilize them, they must reach a functional degree of maturity. To attempt to mimic in vivo environment, various methods including prolonging culture time, co-culture and modulations of chemical, electrical, mechanical culture conditions have been tried. In addition to that, changing the topology of the culture made huge progress with the introduction of the 3D culture that closely resembles the in vivo cardiac topology and overcomes many of the limitations of the conventionally used 2D models. Nonetheless, 3D culture alone is not enough, and using a combination of these methods is being explored. In this review, we summarize the main differences between immature, fetal-like hiPSC-CMs and adult cardiomyocytes, then glance at the current approaches used to promote hiPSC-CMs maturation. In the second part, we focus on the evolving 3D culture model - it's structure, the effect on hiPSC-CMs maturation, incorporation with different maturation methods, limitations and future prospects.
  • Nawin Chanthra, Tomoyuki Abe, Matthew Miyamoto, Kiyotoshi Sekiguchi, Chulan Kwon, Yutaka Hanazono, Hideki Uosaki
    Scientific reports 10 (1) 4249 - 4249 2020/03 [Refereed][Not invited]
    Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) hold great promise for disease modeling and drug discovery. However, PSC-CMs exhibit immature phenotypes in culture, and the lack of maturity limits their broad applications. While physical and functional analyses are generally used to determine the status of cardiomyocyte maturation, they could be time-consuming and often present challenges in comparing maturation-enhancing strategies. Therefore, there is a demand for a method to assess cardiomyocyte maturation rapidly and reproducibly. In this study, we found that Myomesin-2 (Myom2), encoding M-protein, is upregulated postnatally, and based on this, we targeted TagRFP to the Myom2 locus in mouse embryonic stem cells. Myom2-RFP+ PSC-CMs exhibited more mature phenotypes than RFP- cells in morphology, function and transcriptionally, conductive to sarcomere shortening assays. Using this system, we screened extracellular matrices (ECMs) and identified laminin-511/521 as potent enhancers of cardiomyocyte maturation. Together, we developed and characterized a novel fluorescent reporter system for the assessment of cardiomyocyte maturation and identified potent maturation-enhancing ECMs through this simple and rapid assay. This system is expected to facilitate use of PSC-CMs in a variety of scientific and medical investigations.
  • 阿部 朋行, 柴田 宏昭, 魚崎 英毅, 原 弘真, ボラジギン・サラントラガ, 長尾 慶和, 花園 豊
    Organ Biology (一社)日本臓器保存生物医学会 26 (3) 87 - 87 1340-5152 2019/10
  • 原 弘真, 伊藤 拓哉, 菱川 修司, 中野 和明, 阿部 朋行, 柴田 宏昭, 魚崎 英毅, 渡邊 將人, 國田 智, 長嶋 比呂志, 花園 豊
    Organ Biology (一社)日本臓器保存生物医学会 26 (3) 89 - 89 1340-5152 2019/10
  • 原 弘真, 菱川 修司, 伊藤 拓哉, 阿部 朋行, 柴田 宏昭, 魚崎 英毅, 國田 智, 花園 豊
    Organ Biology (一社)日本臓器保存生物医学会 26 (3) 121 - 121 1340-5152 2019/10
  • Mohamed Darwish, Hirofumi Nishizono, Hideki Uosaki, Hitomi Sawada, Taketaro Sadahiro, Masaki Ieda, Keizo Takao
    Journal of Neuroscience Methods 317 149 - 156 0165-0270 2019/04 [Refereed][Not invited]
  • 和食の健康維持・増進効果の機序解明を目指した研究
    原 弘真, 永山 学, 須田 亙, 柴田 宏昭, 菱川 修司, 國田 智, 阿部 朋行, 魚崎 英毅, 新 幸二, 本田 賢也, 花園 豊
    自治医科大学紀要 (学)自治医科大学 41 92 - 92 1881-252X 2019/03
  • Taketaro Sadahiro, Mari Isomi, Naoto Muraoka, Hidenori Kojima, Sho Haginiwa, Shota Kurotsu, Fumiya Tamura, Hidenori Tani, Shugo Tohyama, Jun Fujita, Hiroyuki Miyoshi, Yoshifumi Kawamura, Naoki Goshima, Yuka W Iwasaki, Kensaku Murano, Kuniaki Saito, Mayumi Oda, Peter Andersen, Chulan Kwon, Hideki Uosaki, Hirofumi Nishizono, Keiichi Fukuda, Masaki Ieda
    Cell stem cell 23 (3) 382 - 395 1934-5909 2018/09 [Refereed][Not invited]
    The mesoderm arises from pluripotent epiblasts and differentiates into multiple lineages; however, the underlying molecular mechanisms are unclear. Tbx6 is enriched in the paraxial mesoderm and is implicated in somite formation, but its function in other mesoderms remains elusive. Here, using direct reprogramming-based screening, single-cell RNA-seq in mouse embryos, and directed cardiac differentiation in pluripotent stem cells (PSCs), we demonstrated that Tbx6 induces nascent mesoderm from PSCs and determines cardiovascular and somite lineage specification via its temporal expression. Tbx6 knockout in mouse PSCs using CRISPR/Cas9 technology inhibited mesoderm and cardiovascular differentiation, whereas transient Tbx6 expression induced mesoderm and cardiovascular specification from mouse and human PSCs via direct upregulation of Mesp1, repression of Sox2, and activation of BMP/Nodal/Wnt signaling. Notably, prolonged Tbx6 expression suppressed cardiac differentiation and induced somite lineages, including skeletal muscle and chondrocytes. Thus, Tbx6 is critical for mesoderm induction and subsequent lineage diversification.
  • Hiromasa Hara, Hiroaki Shibata, Kazuaki Nakano, Tomoyuki Abe, Hideki Uosaki, Takahiro Ohnuki, Shuji Hishikawa, Satoshi Kunita, Masahito Watanabe, Osamu Nureki, Hiroshi Nagashima, Yutaka Hanazono
    Experimental Animals 67 (2) 139 - 146 1341-1357 2018 [Refereed][Not invited]
    Pigs with X-linked severe combined immunodeficiency (X-SCID) caused by a mutation of the interleukin-2 receptor gamma chain gene (IL2RG) are of value for a wide range of studies. However, they do not survive longer than 8 weeks because of their susceptibility to infections. To allow longer survival of X-SCID pigs, the animals must be born and reared under germ-free conditions. Here, we established an efficient system for piglet derivation by hysterectomy and used it to obtain and maintain a germ-free X-SCID pig. In four trials using pregnant wild-type pigs, 66% of piglets after hysterectomy started spontaneous breathing (range of 20–100% per litter). The resuscitation rate was found to negatively correlate with elapsed time from the uterus excision to piglet derivation (r=−0.97, P< 0.05). Therefore, it is critical to deliver piglets within 5 min to achieve a high resuscitation rate (82% estimated from regression analysis). In a fifth trial with an IL2RG+/− pig, four piglets were delivered within 4.2 min of uterus excision and three were alive (75%). One of the live born piglets was genotypically and phenotypically determined to be X-SCID and was reared for 12 weeks. The X-SCID piglet was free from both bacteria and fungi at all time points tested by microbial culture and grew without any abnormal signs or symptoms. This study showed successful production and rearing of germ-free pigs, enabling experiments involving long-term follow-up of X-SCID pigs.
  • Gun-Sik Cho, Dong I. Lee, Emmanouil Tampakakis, Sean Murphy, Peter Andersen, Hideki Uosaki, Stephen Chelko, Khalid Chakir, Ingie Hong, Kinya Seo, Huei-Sheng Vincent Chen, Xiongwen Chen, Cristina Basso, Steven R. Houser, Gordon F. Tomaselli, Brian O'Rourke, Daniel P. Judge, David A. Kass, Chulan Kwon
    CELL REPORTS 18 (2) 571 - 582 2211-1247 2017/01 [Refereed][Not invited]
    Pluripotent stem cells (PSCs) offer unprecedented opportunities for disease modeling and personalized medicine. However, PSC-derived cells exhibit fetal-like characteristics and remain immature in a dish. This has emerged as a major obstacle for their application for late-onset diseases. We previously showed that there is a neonatal arrest of long-term cultured PSC-derived cardiomyocytes (PSC-CMs). Here, we demonstrate that PSC-CMs mature into adult CMs when transplanted into neonatal hearts. PSC-CMs became similar to adult CMs in morphology, structure, and function within a month of transplantation into rats. The similarity was further supported by single-cell RNA-sequencing analysis. Moreover, this in vivo maturation allowed patient-derived PSC-CMs to reveal the disease phenotype of arrhythmogenic right ventricular cardiomyopathy, which manifests predominantly in adults. This study lays a foundation for understanding human CM maturation and pathogenesis and can be instrumental in PSC-based modeling of adult heart diseases.
  • Hideki Uosaki, Y-h Taguchi
    GENOMICS PROTEOMICS & BIOINFORMATICS 14 (4) 207 - 215 1672-0229 2016/08 [Refereed][Not invited]
    Understanding how human cardiomyocytes mature is crucial to realizing stem cell-based heart regeneration, modeling adult heart diseases, and facilitating drug discovery. However, it is not feasible to analyze human samples for maturation due to inaccessibility to samples while cardiomyocytes mature during fetal development and childhood, as well as difficulty in avoiding variations among individuals. Using model animals such as mice can be a useful strategy; nonetheless, it is not well-understood whether and to what degree gene expression profiles during maturation are shared between humans and mice. Therefore, we performed a comparative gene expression analysis of mice and human samples. First, we examined two distinct mice microarray platforms for shared gene expression profiles, aiming to increase reliability of the analysis. We identified a set of genes displaying progressive changes during maturation based on principal component analysis. Second, we demonstrated that the genes identified had a differential expression pattern between adult and earlier stages (e.g., fetus) common in mice and humans. Our findings provide a foundation for further genetic studies of cardiomyocyte maturation.
  • Hideki Uosaki, Patrick Cahan, Dong I. Lee, Songnan Wang, Matthew Miyamoto, Laviel Fernandez, David A. Kass, Chulan Kwon
    CELL REPORTS 13 (8) 1705 - 1716 2211-1247 2015/11 [Refereed][Not invited]
    Decades of progress in developmental cardiology has advanced our understanding of the early aspects of heart development, including cardiomyocyte (CM) differentiation. However, control of the CM maturation that is subsequently required to generate adult myocytes remains elusive. Here, we analyzed over 200 microarray datasets from early embryonic to adult hearts and identified a large number of genes whose expression shifts gradually and continuously during maturation. We generated an atlas of integrated gene expression, biological pathways, transcriptional regulators, and gene regulatory networks (GRNs), which show discrete sets of key transcriptional regulators and pathways activated or suppressed during CM maturation. We developed a GRN-based program named MatStat CM that indexes CM maturation status. MatStat CM reveals that pluripotent-stem-cell-derived CMs mature early in culture but are arrested at the late embryonic stage with aberrant regulation of key transcription factors. Our study provides a foundation for understanding CM maturation.
  • Lincoln T. Shenje, Peter Andersen, Hideki Uosaki, Laviel Fernandez, Peter P. Rainer, Gun-Sik Cho, Dong-ik Lee, Weimin Zhong, Richard P. Harvey, David A. Kass, Chulan Kwon
    ELIFE 3 e02164  2050-084X 2014/04 [Refereed][Not invited]
    Cardiac progenitor cells (CPCs) must control their number and fate to sustain the rapid heart growth during development, yet the intrinsic factors and environment governing these processes remain unclear. Here, we show that deletion of the conserved cell-fate regulator Numb and its homologue Numblike (Numbl) depletes CPCs in second pharyngeal arches (PA2s) and is associated with an atrophic heart. With histological, flow cytometric and functional analyses, we find that CPCs remain undifferentiated and expansive in the PA2, but differentiate into cardiac cells as they exit the arch. Tracing of Numb-and Numbl-deficient CPCs by lineage-specific mosaicism reveals that the CPCs normally populate in the PA2, but lose their expansion potential in the PA2. These findings demonstrate that Numb and Numbl are intrinsic factors crucial for the renewal of CPCs in the PA2 and that the PA2 serves as a microenvironment for their expansion.
  • Hideki Uosaki, Ajit Magadum, Kinya Seo, Hiroyuki Fukushima, Ayako Takeuchi, Yasuaki Nakagawa, Kara White Moyes, Genta Narazaki, Koichiro Kuwahara, Michael Laflamme, Satoshi Matsuoka, Norio Nakatsuji, Kazuwa Nakao, Chulan Kwon, David A. Kass, Felix B. Engel, Jun K. Yamashita
    Circulation: Cardiovascular Genetics 6 (6) 624 - 633 1942-325X 2013/12 [Refereed][Not invited]
    Background-The proliferation of cardiomyocytes is highly restricted after postnatal maturation, limiting heart regeneration. Elucidation of the regulatory machineries for the proliferation and growth arrest of cardiomyocytes is imperative. Chemical biology is efficient to dissect molecular mechanisms of various cellular events and often provides therapeutic potentials. We have been investigating cardiovascular differentiation with pluripotent stem cells. The combination of stem cell and chemical biology can provide novel approaches to investigate the molecular mechanisms and manipulation of cardiomyocyte proliferation. Methods and Results-To identify chemicals that regulate cardiomyocyte proliferation, we performed a screening of a defined chemical library based on proliferation of mouse pluripotent stem cell-derived cardiomyocytes and identified 4 chemical compound groups: inhibitors of glycogen synthase kinase-3, p38 mitogen-activated protein kinase, and Ca 2+/calmodulin-dependent protein kinase II, and activators of extracellular signal-regulated kinase. Several appropriate combinations of chemicals synergistically enhanced proliferation of cardiomyocytes derived from both mouse and human pluripotent stem cells, notably up to a 14-fold increase in mouse cardiomyocytes. We also examined the effects of identified chemicals on cardiomyocytes in various developmental stages and species. Whereas extracellular signal-regulated kinase activators and Ca2+/calmodulin-dependent protein kinase II inhibitors showed proliferative effects only on cardiomyocytes in early developmental stages, glycogen synthase kinase-3 and p38 mitogen-activated protein kinase inhibitors substantially and synergistically induced re-entry and progression of cell cycle in neonatal but also as well as adult cardiomyocytes. Conclusions-Our approach successfully uncovered novel molecular targets and mechanisms controlling cardiomyocyte proliferation in distinct developmental stages and offered pluripotent stem cell-derived cardiomyocytes as a potent tool to explore chemical-based cardiac regenerative strategies. © 2013 American Heart Association, Inc.
  • Hideki Uosaki, Peter Andersen, Lincoln T. Shenje, Laviel Fernandez, Sofie Lindgren Christiansen, Chulan Kwon
    PLOS ONE 7 (10) e46413  1932-6203 2012/10 [Refereed][Not invited]
    Rationale: Pluripotent stem cell-derived cardiac progenitor cells (CPCs) have emerged as a powerful tool to study cardiogenesis in vitro and a potential cell source for cardiac regenerative medicine. However, available methods to induce CPCs are not efficient or require high-cost cytokines with extensive optimization due to cell line variations. Objective: Based on our in-vivo observation that early endodermal cells maintain contact with nascent pre-cardiac mesoderm, we hypothesized that direct physical contact with endoderm promotes induction of CPCs from pluripotent cells. Method and Result: To test the hypothesis, we cocultured mouse embryonic stem (ES) cells with the endodermal cell line End2 by co-aggregation or End2-conditioned medium. Co-aggregation resulted in strong induction of Flk1(+) PDGFRa(+) CPCs in a dose-dependent manner, but the conditioned medium did not, indicating that direct contact is necessary for this process. To determine if direct contact with End2 cells also promotes the induction of committed cardiac progenitors, we utilized several mouse ES and induced pluripotent (iPS) cell lines expressing fluorescent proteins under regulation of the CPC lineage markers Nkx2.5 or Isl1. In agreement with earlier data, co-aggregation with End2 cells potently induces both Nkx2.5(+) and Isl1(+) CPCs, leading to a sheet of beating cardiomyocytes. Furthermore, co-aggregation with End2 cells greatly promotes the induction of KDR+ PDGFRa(+) CPCs from human ES cells. Conclusions: Our co-aggregation method provides an efficient, simple and cost-effective way to induce CPCs from mouse and human pluripotent cells.
  • Hidetoshi Masumoto, Takehiko Matsuo, Kohei Yamamizu, Hideki Uosaki, Genta Narazaki, Shiori Katayama, Akira Marui, Tatsuya Shimizu, Tadashi Ikeda, Teruo Okano, Ryuzo Sakata, Jun K. Yamashita
    STEM CELLS 30 (6) 1196 - 1205 1066-5099 2012/06 [Refereed][Not invited]
    Although stem cell therapy is a promising strategy for cardiac restoration, the heterogeneity of transplanted cells has been hampering the precise understanding of the cellular and molecular mechanisms. Previously, we established a cardiovascular cell differentiation system from mouse pluripotent stem cells, in which cardiomyocytes (CMs), endothelial cells (ECs), and mural cells (MCs) can be systematically induced and purified. Combining this with cell sheet technology, we generated cardiac tissue sheets reassembled with defined cardiovascular populations. Here, we show the potentials and mechanisms of cardiac tissue sheet transplantation in cardiac function after myocardial infarction (MI). Transplantation of the cardiac tissue sheet to a rat MI model showed significant and sustained improvement of systolic function accompanied by neovascularization. Reduction of the infarct wall thinning and fibrotic length indicated the attenuation of left ventricular remodeling. Cell tracing with species-specific fluorescent in situ hybridization after transplantation revealed a relatively early loss of transplanted cells and an increase in endogenous neovascularization in the proximity of the graft, suggesting an indirect angiogenic effect of cardiac tissue sheets rather than direct CM contributions. We prospectively dissected the functional mechanisms with cell type-controlled sheet analyses. Sheet CMs were the main source of vascular endothelial growth factor. Transplantation of sheets lacking CMs resulted in the disappearance of neovascularization and subsequent functional improvement, indicating that the beneficial effects of the sheet were achieved by sheet CMs. ECs and MCs enhanced the sheet functions and structural integration. Supplying CMs to ischemic regions with cellular interaction could be a strategic key in future cardiac cell therapy. STEM CELLS2012;30:11961205
  • Peter Andersen, Hideki Uosaki, Lincoln T. Shenje, Chulan Kwon
    TRENDS IN CELL BIOLOGY 22 (5) 257 - 265 0962-8924 2012/05 [Refereed][Not invited]
    Notch is an ancient transmembrane receptor with crucial roles in cell-fate choices. Although the 'canonical' Notch pathway and its core members are well established involving ligand-induced cleavage of Notch for transcriptional regulation - it has been unclear whether Notch can also function independently of ligand and transcription ('non-canonically') through a common mechanism. Recent studies suggest that Notch can non-canonically exert its biological functions by post-translationally targeting Wnt/beta-catenin signaling, an important cellular and developmental regulator. The non-canonical Notch pathway appears to be highly conserved from flies to mammals. Here, we discuss the emerging conserved mechanism and role of ligand/transcription-independent Notch signaling in cell and developmental biology.
  • Hideki Uosaki, Hiroyuki Fukushima, Ayako Takeuchi, Satoshi Matsuoka, Norio Nakatsuji, Shinya Yamanaka, Jun K. Yamashita
    PLOS ONE 6 (8) e23657  1932-6203 2011/08 [Refereed][Not invited]
    Rationale: Human embryonic and induced pluripotent stem cells (hESCs/hiPSCs) are promising cell sources for cardiac regenerative medicine. To realize hESC/hiPSC-based cardiac cell therapy, efficient induction, purification, and transplantation methods for cardiomyocytes are required. Though marker gene transduction or fluorescent-based purification methods have been reported, fast, efficient and scalable purification methods with no genetic modification are essential for clinical purpose but have not yet been established. In this study, we attempted to identify cell surface markers for cardiomyocytes derived from hESC/hiPSCs. Method and Result: We adopted a previously reported differentiation protocol for hESCs based on high density monolayer culture to hiPSCs with some modification. Cardiac troponin-T (TNNT2)-positive cardiomyocytes appeared robustly with 30-70% efficiency. Using this differentiation method, we screened 242 antibodies for human cell surface molecules to isolate cardiomyocytes derived from hiPSCs and identified anti-VCAM1 (Vascular cell adhesion molecule 1) antibody specifically marked cardiomyocytes. TNNT2-positive cells were detected at day 7-8 after induction and 80% of them became VCAM1-positive by day 11. Approximately 95-98% of VCAM1-positive cells at day 11 were positive for TNNT2. VCAM1 was exclusive with CD144 (endothelium), CD140b (pericytes) and TRA-1-60 (undifferentiated hESCs/hiPSCs). 95% of MACS-purified cells were positive for TNNT2. MACS purification yielded 5-10x10(5) VCAM1-positive cells from a single well of a six-well culture plate. Purified VCAM1-positive cells displayed molecular and functional features of cardiomyocytes. VCAM1 also specifically marked cardiomyocytes derived from other hESC or hiPSC lines. Conclusion: We succeeded in efficiently inducing cardiomyocytes from hESCs/hiPSCs and identifying VCAM1 as a potent cell surface marker for robust, efficient and scalable purification of cardiomyocytes from hESC/hiPSCs. These findings would offer a valuable technological basis for hESC/hiPSC-based cell therapy.
  • Masataka Fujiwara, Peishi Yan, Tomomi G. Otsuji, Genta Narazaki, Hideki Uosaki, Hiroyuki Fukushima, Koichiro Kuwahara, Masaki Harada, Hiroyuki Matsuda, Satoshi Matsuoka, Keisuke Okita, Kazutoshi Takahashi, Masato Nakagawa, Tadashi Ikeda, Ryuzo Sakata, Christine L. Mummery, Norio Nakatsuji, Shinya Yamanaka, Kazuwa Nakao, Jun K. Yamashita
    PLOS ONE 6 (2) e16734  1932-6203 2011/02 [Refereed][Not invited]
    Induced pluripotent stem cells (iPSCs) are novel stem cells derived from adult mouse and human tissues by reprogramming. Elucidation of mechanisms and exploration of efficient methods for their differentiation to functional cardiomyocytes are essential for developing cardiac cell models and future regenerative therapies. We previously established a novel mouse embryonic stem cell (ESC) and iPSC differentiation system in which cardiovascular cells can be systematically induced from Flk1(+) common progenitor cells, and identified highly cardiogenic progenitors as Flk1(+)/CXCR4(+)/VE-cadherin(-) (FCV) cells. We have also reported that cyclosporin-A (CSA) drastically increases FCV progenitor and cardiomyocyte induction from mouse ESCs. Here, we combined these technologies and extended them to mouse and human iPSCs. Co-culture of purified mouse iPSC-derived Flk1(+) cells with OP9 stroma cells induced cardiomyocyte differentiation whilst addition of CSA to Flk1(+) cells dramatically increased both cardiomyocyte and FCV progenitor cell differentiation. Spontaneously beating colonies were obtained from human iPSCs by co-culture with END-2 visceral endoderm-like cells. Appearance of beating colonies from human iPSCs was increased approximately 4.3 times by addition of CSA at mesoderm stage. CSA-expanded human iPSC-derived cardiomyocytes showed various cardiac marker expressions, synchronized calcium transients, cardiomyocyte-like action potentials, pharmacological reactions, and ultra-structural features as cardiomyocytes. These results provide a technological basis to obtain functional cardiomyocytes from iPSCs.
  • Kohei Yamamizu, Taichi Matsunaga, Hideki Uosaki, Hiroyuki Fukushima, Shiori Katayama, Mina Hiraoka-Kanie, Kohnosuke Mitani, Jun K. Yamashita
    JOURNAL OF CELL BIOLOGY 189 (2) 325 - 338 0021-9525 2010/04 [Refereed][Not invited]
    Molecular mechanisms controlling arterial-venous specification have not been fully elucidated. Previously, we established an embryonic stem cell differentiation system and demonstrated that activation of cAMP signaling together with VEGF induces arterial endothelial cells (ECs) from Flk1(+) vascular progenitor cells. Here, we show novel arterial specification machinery regulated by Notch and beta-catenin signaling. Notch and GSK3 beta-mediated beta-catenin signaling were activated downstream of cAMP through phosphatidylinositol-3 kinase. Forced activation of Notch and beta-catenin with VEGF completely reconstituted cAMP-elicited arterial EC induction, and synergistically enhanced target gene promoter activity in vitro and arterial gene expression during in vivo angiogenesis. A protein complex with RBP-J, the intracellular domain of Notch, and beta-catenin was formed on RBP-J binding sites of arterial genes in arterial, but not venous ECs. This molecular machinery for arterial specification leads to an integrated and more comprehensive understanding of vascular signaling.
  • Shinobu Kuratomi, Yoko Ohmori, Masayuki Ito, Kuniko Shimazaki, Shin-ichi Muramatsu, Hiroaki Mizukami, Hideki Uosaki, Jun K. Yamashita, Yuji Arai, Koichiro Kuwahara, Makoto Takano
    CARDIOVASCULAR RESEARCH 83 (4) 682 - 687 0008-6363 2009/09 [Refereed][Not invited]
    Hcn4, which encodes the hyperpolarization-activated, cyclic nucleotide-sensitive channel (I(h)), is a well-established marker of the cardiac sino-atrial node. We aimed to identify cis-elements in the genomic locus of the Hcn4 gene that regulate the transcription of Hcn4. We screened evolutionarily conserved non-coding sequences (CNSs) that are often involved in the regulation of gene expression. The VISTA Enhancer Browser identified 16 regions, termed CNS 1-16, within the Hcn4 locus. Using the luciferase reporter assay in primary neonatal rat cardiomyocytes, we found that CNS13 conferred a prominent enhancer activity (more than 30-fold) on the Hcn4 promoter. Subsequent mutation analysis revealed that the Hcn4 enhancer function was dependent on myocyte enhancer factor-2 (MEF2) and activator protein-1 (AP1) binding sequences located in CNS13. Electrophoretic mobility shift assay and chromatin immunoprecipitation confirmed that MEF2 and AP1 proteins bound CNS13. Furthermore, overexpression of a dominant negative MEF2 mutant inhibited the enhancer activity of CNS13, decreased Hcn4 mRNA expression and also decreased the amplitude of I(h) current in myocytes isolated from the inflow tract of embryonic heart. These results suggest that the novel enhancer CNS13 and MEF2 may play a critical role in the transcription of Hcn4 in the heart.
  • Peishi Yan, Atsushi Nagasawa, Hideki Uosaki, Akihiro Sugimoto, Kohei Yamamizu, Mizue Teranishi, Hiroyuki Matsuda, Satoshi Matsuoka, Tadashi Ikeda, Masashi Komeda, Ryuzo Sakata, Jun K. Yamashita
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 379 (1) 115 - 120 0006-291X 2009/01 [Refereed][Not invited]
    Though cardiac progenitor cells should be a suitable material for cardiac regeneration, efficient ways to induce cardiac progenitors from embryonic stein (ES) cells have not been established. Extending our systematic cardiovascular differentiation method of ES cells, here we show efficient and specific expansion of cardiomyocytes and highly cardiogenic progenitors from ES cells. An immunosuppressant, cyclosporin A (CSA), showed a novel effect specifically acting on mesoderm cells to drastically increase cardiac progenitors as well as cardiomyocytes by 10-20 times. Approximately 200 cardiomyocytes could be induced from one mouse ES cell using this method. Expanded Progenitors Successfully integrated into scar tissue of infracted heart as cardiomyocytes after cell transplantation to rat myocardial infarction model. CSA elicited specific induction of cardiac lineage from mesoderm in a novel mesoderm-specific, NFAT independent fashion. This simple but efficient differentiation technology Would be extended to induce Pluripotent stein (iPS) cells and broadly contribute to cardiac regeneration. (C) 2008 Elsevier Inc. All rights reserved.
  • Genta Narazaki, Hideki Uosaki, Mizue Teranishi, Keisuke Okita, Bongju Kim, Satoshi Matsuoka, Shinya Yamanaka, Jun K. Yamashita
    CIRCULATION 118 (5) 498 - 506 0009-7322 2008/07 [Refereed][Not invited]
    Background - Induced pluripotent stem ( iPS) cells are a novel stem cell population induced from mouse and human adult somatic cells through reprogramming by transduction of defined transcription factors. However, detailed differentiation properties and the directional differentiation system of iPS cells have not been demonstrated. Methods and Results - Previously, we established a novel mouse embryonic stem ( ES) cell differentiation system that can reproduce the early differentiation processes of cardiovascular cells. We applied our ES cell system to iPS cells and examined directional differentiation of mouse iPS cells to cardiovascular cells. Flk1 ( also designated as vascular endothelial growth factor receptor-2)-expressing mesoderm cells were induced from iPS cells after approximate to 4- day culture for differentiation. Purified Flk1(+) cells gave rise to endothelial cells and mural cells by addition of vascular endothelial growth factor and serum. Arterial, venous, and lymphatic endothelial cells were also successfully induced. Self-beating cardiomyocytes could be induced from Flk1(+) cells by culture on OP9 stroma cells. Time course and efficiency of the differentiation were comparable to those of mouse ES cells. Occasionally, reexpression of transgene mRNAs, including c-myc, was observed in long-term differentiation cultures. Conclusions - Various cardiovascular cells can be systematically induced from iPS cells. The differentiation properties of iPS cells are almost completely identical to those of ES cells. This system would greatly contribute to a novel understanding of iPS cell biology and the development of novel cardiovascular regenerative medicine.
  • Kentoku Yanagi, Makoto Takano, Genta Narazaki, Hideki Uosaki, Takuhiro Hoshino, Takahiro Ishii, Takurou Misaki, Jun K. Yamashita
    STEM CELLS 25 (11) 2712 - 2719 1066-5099 2007/11 [Refereed][Not invited]
    Regeneration of cardiac pacemakers is an important target of cardiac regeneration. Previously, we developed a novel embryonic stem (ES) cell differentiation system that could trace cardiovascular differentiation processes at the cellular level. In the present study, we examine expressions and functions of ion channels in ES cell-derived cardiomyocytes during their differentiation and identify ion channels that confer their automaticity. ES cell-derived Flk1(+) mesoderm cells give rise to spontaneously beating cardiomyocytes on OP9 stroma cells. Spontaneously beating colonies observed at day 9.5 of Flk1(+) cell culture (Flk-d9.5) were significantly decreased at Flk-d23.5. Expressions of ion channels in pacemaker cells hyperpolarization-activated cyclic nucleotidegated (HCN)l and -4 and voltage-gated calcium channel (Cav)3.1 and -3.2 were significantly decreased in purified cardiomyocytes at Flk-d23.5 compared with at Flk-d9.5, whereas expression of an atrial and ventricular ion channel, inward rectifier potassium channel (Kir)2.1, did not change. Blockade of HCNs and Cav ion channels significantly inhibited beating rates of cardiomyocyte colonies. Electrophysiological studies demonstrated that spontaneously beating cardiomyocytes at Flk-d9.5 showed almost similar features to those of the native mouse sinoatrial node except for relatively deep maximal diastolic potential and faster maximal upstroke velocity. Although similar to 60% of myocytes at Flk-d23.5 revealed almost the same properties as those at Flk-d9.5, similar to 40% of myocytes showed loss of HCN and decreased Cav3 currents and ceased spontaneous beating, with no remarkable increase of Kir2.1. Thus, HCN and Cav3 ion channels should be responsible for the maintenance of automaticity in ES cell-derived cardiomyocytes. Controlled regulation of these ion channels should be required to generate complete biological pacemakers.

Books etc

  • モデル動物の作製と利用-循環器疾患2021
    西園 啓文, 魚崎 英毅 (ContributorCRISPR-CAS9による遺伝子編集マウスを用いた心血管疾患研究)
    エル・アイ・シー 2021/09
  • Pluripotent Stem-Cell Derived Cardiomyocytes. (In: Yoshida Y. (eds), Methods in Molecular Biology, vol 2320.)
    Tatsuya Anzai, Hiromasa Hara, Nawin Chanthra, Taketaro Sadahiro, Masaki Ieda, Yutaka Hanazono, Hideki Uosaki (Generation of Efficient Knock-in Mouse and Human Pluripotent Stem Cells Using CRISPR-Cas9.)
    Humana, New York, NY 2021/07
  • Cardiac Regeneration using Stem Cells (eds., Fukuda K. & Yuasa S.)
    Hideki Uosaki, Jun K Yamashita (Joint workDirected and Systematic Differentiation of Cardiovascular Cells from Mouse and Human Pluripotent Stem Cells)
    CRC press 2013
  • Embryonic Stem Cells: The Hormonal Regulation of Pluripotency and Embryogenesis (eds., Atwood C.)
    Hideki Uosaki, Jun K Yamashita (Joint workChemicals Regulating Cardiomyoyte Differentiation)
    InTech 2011

Conference Activities & Talks

  • Implication of Hormonal and Mechanical Regulation in Maturation of Cardiomyocytes Derived from Pluripotent Stem Cells  [Invited]
    Hideki Uosaki, Razan Elfadil Ahmed, Nawin Chanthra
    JCS-Council Forum on Basic CardioVascular Research  2021/09
  • Disease Modeling with Cardiomyocytes derived from iPS cells  [Invited]
    Hideki Uosaki
    CVMW2020  2021/03
  • 疾患iPS細胞を用いた病態解明に向けた基盤構築  [Invited]
    魚崎 英毅
    第20回日本再生医療学会総会  2021/03
  • 蛍光標識サルコメアを利用したiPS細胞由来心筋細胞のサルコメアショートニング  [Not invited]
    魚崎英毅, 安済達也, Nawin Chanthra, Razan E Ahmed, 花園 豊
    第20回再生医療学会  2021/03
  • Disease Modeling of Cardiomyopathy Using Pluripotent Stem Cells.  [Invited]
    Hideki Uosaki
    MIS-Korea 2021  2021/03
  • Disease modeling of cardiomyopathy using iPS cells  [Invited]
    Hideki Uosaki
    CATs – Cardiovasucular Thrombosis Seminar -  2021/01
  • Identifying Cross-Organ Similarities During Development and Maturation Arrest of Pluripotent Stem Cell Differentiation  [Not invited]
    Sandeep Kambhampati, Sean Murphy, Hideki Uosaki, Chulan Kwon
    BMES 2020 Virtual Annual Meeting
  • In vitro maturation of cardiomyocytes derived from stem cells.  [Invited]
    Hideki Uosaki
    MIS-Korea 2020  2020/08
  • Systematic Profiling of Cardiomyocyte Maturation Enhancing Factors.  [Not invited]
    Hideki Uosaki, Nawin Chanthra, Yutaka Hanazono
    IRG Meeting  2020/01
  • Visualizations of Cardiomyocyte Maturation on Extracellular Matrices Using a Novel Myom2-RFP Reporter Line  [Not invited]
    Nawin Chanthra, Yutaka Hanazono, Hideki Uosaki
    CVMW 2019  2019/12
  • Transcriptional Network Regulating Cardiomyocyte Maturation  [Invited]
    The 42nd Annual Meeting of the Molecular Biology Society of Japan  2019/12
    Yutaka Hanazono, Hiromasa Hara, Hideki Uosaki, Osamu Nureki
    Frontiers in Genome Engineering 2019  2019/11
    Hiromasa Hara, Hideki Uosaki, Tomoyuki Abe, Hiroaki Shibata, Osamu Nureki, Yutaka Hanazono
    Frontiers in Genome Engineering 2019  2019/11
    Suvd Byambaa, Hideki Uosaki, Hiromasa Hara, Hiroaki Shibata, Tomoyuki Abe, Yasumitsu Nagao, Osamu Nureki, Tsukasa Ohmori, Yutaka Hanazono
    Frontiers in Genome Engineering 2019  2019/11
  • 大型動物胎仔を用いたヒト臓器再生技術:ヒツジで作るヒト造血幹細胞  [Not invited]
    阿部朋行, 柴田宏昭, 魚崎英毅, 原 弘真, ボラジギン・サラントラガ, 長尾慶和, 花園 豊
    第46回日本臓器保存生物医学会学術集会  2019/11
  • ピッグ同種輸血の試み  [Not invited]
    原弘真, 菱川修司, 伊藤拓哉, 阿部朋行, 柴田宏昭, 魚崎英毅, 國田智, 花園豊
    第46回日本臓器保存生物医学会学術集会  2019/11
  • SCIDピッグの長期飼育:再生医療・細胞治療評価系の確立をめざして  [Not invited]
    原弘真, 伊藤拓哉, 菱川修司, 中野和明, 阿部朋行, 柴田宏昭, 魚崎英毅, 渡邊將人, 國田智, 長嶋比呂志, 花園豊
    第46回日本臓器保存生物医学会学術集会  2019/11
  • 汎血球減少ピッグへの同種輸血  [Not invited]
    原弘真, 菱川修司, 伊藤拓哉, 阿部朋行, 柴田宏昭, 魚崎英毅, 國田智, 花園豊
    日本先進医工学ブタ研究会  2019/10
  • Hormonal and Transcriptional Regulation of Cardiomyocyte Maturation  [Not invited]
    Uosaki H, Chanthra N, Hanazono Y
    16th Nikko Symposium  2019/10
  • Decoding Molecular Responses to Electrical and Mechanical Stimuli Enhancing Cardiomyocyte Maturation  [Invited]
    Hideki Uosaki
    3rd JCS Council Forum on Basic CardioVascular Research  2019/09
  • Hormonal Regulation of Cardiomyocyte Maturation  [Not invited]
    3rd JCS Council Forum on Basic CardioVascular Research  2019/09
  • Phenotypical Correction of X-SCID Mice After CRISPR/Cas9-mediated Genome-Editing Therapy of Hematopoietic Stem Cells  [Not invited]
    Byambaa S, Uosaki H, Hara H, Shibata H, Abe T, Nagao Y, Nureki O, Ohmori T, Hanazono Y
    The 25th Annual Meeting of Japan Society of Gene and Cell Therapy  2019/07
    Chanthra N, Hanazono Y, Uosaki H
    International Society for Stem Cell Research 2019 Annual Meeting  2019/06
  • Genome Editing of Murine Hematopoietic Stem Cells by Cas9/RNP Targeting Integrin-alpha IIb Gene  [Not invited]
    Byambaa S, Uosaki H, Ohmori T, Hanazono Y
    The 4th Annual Meeting of the Japanese Society for Genome Editing  2019/06
  • 培養心筋細胞の成熟におけるDNA脱メチル化代謝胴体の心筋細胞特異的エピジェネティックドメイン形成との関係  [Not invited]
    小田 真由美, 魚崎 英毅
    第13回日本エピジェネティクス研究会年会  2019/05
  • Cure of X-SCID Mice After Ex Vivo Non-viral Genome-Editing Therapy of Hematopoietic Stem Cells  [Not invited]
    Byambaa S, Uosaki H, Hara H, Shibata H, Abe T, Hayakawa H, Nagao Y, Nureki O, Ohmori T, Hanazono Y
    the 21st American Society of Gene and Cell therapy  2019/05
  • The relationship between DNA methylation dynamics and cardiomyocyte-specific epigenetic domain formation in cardiomyocyte maturation.  [Not invited]
    Oda M, Uosaki H
    BDR SYmposium 2019  2019/03
  • 多能性幹細胞由来心筋細胞の定量的成熟度評価法の開発  [Not invited]
    魚崎英毅, Nawin Chanthra, 花園豊
    第18回日本再生医療学会総会  2019/03
  • Hormones and Nuclear Receptors Regulating Cardiomyocyte Maturation  [Not invited]
    Uosaki H, Chanthra N, Kannan S, Murphy S, Kwon C, Hanazono Y
    CVMW2018  2018/12
  • Targeted Gene Integration in Hematopoietic Stem and Progenitor Cells with Cas9-RNP Method to Universally Correct X-SCID Mutations.  [Not invited]
    Byambaa S, Uosaki H, Hara H, Shibata H, Abe T, Nagao Y, Nureki O, Ohmori T, Hanazono Y
    the 41st Annual Meeting of MBSJ  2018/11
  • 試験管内心筋細胞分化における細胞成熟と細胞種特異的エピジェネティック・ドメイン形成  [Not invited]
    小田 真由美, 魚崎 英毅
    第41回日本分子生物学会年会  2018/11
  • 腸内細菌研究におけるピッグのモデル動物としての可能性  [Not invited]
    原弘真, 永山学, 須田亙, 魚崎英毅, 阿部朋行, 柴田宏昭, 菱川修司, 國田智, 新幸二, 本田賢也, 花園豊
    第6回先進医工学ブタ研究会  2018/10
  • Targeted Genome Editing of Murine Hematopoietic Stem Cells by CRISPR/Cas9  [Not invited]
    The 24th Annual Meeting of Japan Society of Gene and Cell Therapy  2018/06
    Hideki Uosaki
    International Society for Stem Cell Research 2018 Annual Meeting  2018/06
  • Novel SCID mice generation by CRISPR/Cas9  [Not invited]
    The 65th Annual Meeting of Japanese Association for Laboratory Animal Science  2018/05
  • ヒト腸内細菌定着ブタとヒトの腸内細菌叢の比較  [Not invited]
    原弘真, 永山学, 須田亙, 大貫貴広, 菱川修司, 柴田宏昭, 阿部朋行, 魚崎英毅, 國田智, 新幸二, 本田賢也, 花園豊
    第65回日本実験動物学会  2018/05
  • Noncanonical Notch Signaling Plays a Biphasic Role during Heart Development  [Not invited]
    Miyamoto M, Andersen P, Sulistio E, Kannan S, Murphy S, Uosaki H, Kwon C
    Weinstein Conference  2018/05
  • Activation of the PPAR/PGC1a pathway promotes pluripotent stem cell-derived cardiomyocyte maturation  [Not invited]
    Hideki Uosaki
    Weinstein Conference  2018/05
  • 新生ブタに対する麻酔管理  [Not invited]
    伊藤拓哉, 芝順太郎, 五十嵐孝, 五十嵐かほり, 半田紀子, 菱川修司, 柴田宏明, 魚崎英毅, 原弘真, 和久井亨, 宮澤和志, 栗田瑞希, 伊藤昌紀, 國田智, 竹内護, 花園豊
    第65回日本実験動物学会  2018/05
  • Qualitative and Quantitative Measurements of Maturity Revealed Novel Regulators of Cardiomyocyte Maturation  [Not invited]
    Chanthra N, Miyamoto M, Kwon C, Hanazono Y, Uosaki H
    Weinstein Conference  2018/05
  • 異種移植系における同一ドナー由来リンパ球の影響  [Not invited]
    阿部朋行, 柴田宏昭, 原 弘真, 魚崎英毅, 大貫貴広, 竹内絢香, 原明日香, ボラジギン・サラントラガ, 長尾慶和, 花園 豊
    第20回日本異種移植研究会  2018/03
  • 無菌環境下におけるSCIDブタ長期飼育の試み  [Not invited]
    原弘真, 柴田宏昭, 中野和明, 阿部朋行, 魚崎英毅, 大貫貴広, 菱川修司, 國田智, 渡邊將人, 濡木理, 長嶋比呂志, 花園豊
    第5回先進医工学ブタ研究会  2017/10
  • Generation of CD45-positive Hematopoietic Cells from Human iPS Cells in Vivo in Ovine Fetal Liver.  [Not invited]
    Abe T, Shibata H, Uosaki H, Hara H, Ohnuki T, Byambaa S, Chanthra N, Sarentonglaga B, Fukumori R, Nagao Y, Hanazono Y
    International Society for Stem Cell Research 2017 Annual Meeting  2017/06
  • Long-term Hematopoietic Engraftment of Human iPS Cell-Derivatives After Sheep in Utero Transplantation  [Not invited]
    Tomoyuki Abe, Hiroaki Shibata, Hideki Uosaki, Hiromasa Hara, Takahiro Ohnuki, Suvd Byambaa, Nawin Chanthra, Borjigin Sarentonglaga, Rika Fukumori, Yoshikazu Nagao, Yutaka Hanazono
    JALAS  2017/05
  • Generation of Adult-like Cardiomyocytes from Pluripotent Stem Cells in Vivo  [Not invited]
    Hideki Uosaki
    15th Stem Cell Symposium  2017/05
  • Successful derivation/rearing of germfree pigs and a trial to rear a SCID pig  [Not invited]
    Hiromasa Hara, Hiroaki Shibata, Kazuaki Nakano, Tomoyuki Abe, Hideki Uosaki, Takahiro Ohnuki, Suvd Byambaa, Nawin Chanthra, Shuji Hishikawa, Satoshi Kunita, Hiroshi Nagashima, Osamu Nureki, Yutaka Hanazono
    JALAS  2017/05
  • Resistance of Mouse Hematopoietic Stem and Progenitor Cells to the Genome-Editing with Cas9  [Not invited]
    Byambaa S, Uosaki H, Hara H, Abe T, Nagao Y, Nureki O, Ohmori T, Hanazono Y
    ASGCT  2017/05
  • Prediction of Cardiomyocyte Maturity with Transcriptome  [Invited]
    Hideki Uosaki
    ANALYTIX 2017  2017/03
  • GRNs of Cardiomyocyte Maturation Revealed Maturation Arrest of  [Not invited]
    Hideki Uosaki, Patrick Cahan, Chulan Kwon
    The 13th Nikko International Symposium  2016/10
  • 多能性幹細胞からの心筋細胞分化誘導と将来展望  [Invited]
    魚崎 英毅
    第14回 日本再生医療学会  2015
  • Signaling Pathways Involved in the Differentiation of PSC to Cardiomyocytes: Lessens from Development  [Invited]
    Hideki Uosaki
    FAES workshop (BioTech 54: Making Cardiomyocytes from iPSCs)  2015
  • Identification of Chemicals Inducing Cardiomyocyte Proliferation in Developmental Stage-Specific Manner with Pluripotent Stem Cells  [Not invited]
    Hideki Uosaki
    Japanese Circulation Society  2014
  • 低分子化合物による ES/iPS 細胞由来心筋細胞の分化および増殖制御-再生医療へのケミカルハイオロジー-  [Invited]
    Hideki Uosaki
    第10回 日本再生医療学会  2011/03
  • ハイコンテンツスクリーニングシステムとES 細胞を用いた新規心筋分化・増 殖化合物の探索  [Invited]
    魚崎英毅, 福島弘之, 山下潤
    第11 回モレキュラーデバイステクニカルセミナー.2010. 6. 2 東京,6. 3.大阪  2010
  • 低分子化合物を用いた ES/iPS 細胞由来心筋細胞の増殖制御  [Not invited]
    魚崎 英毅, 藤原 正隆, 山下 潤
    第31回 日本炎症再生医学会  2010
  • サイクロスポリンAを用いたES細胞及びiPS細胞からの心筋及び心筋前駆細胞 の効率的誘導法の確立  [Not invited]
    魚崎英毅, 顔培実, 山下潤
    第8回再生医療学会総会,2009.3.5-6,東京  2009
  • ES 細胞を用いた生物学的ペースメーカー樹立の試み  [Not invited]
    魚崎英毅, 楢崎元太, 柳堅徳, 鷹野誠, 山下潤
    第7回再生医療学会(2008.3.13-14.名古屋)  2008
  • ES 細胞およびiPS 細胞からの心筋前駆細胞の効率的誘導法  [Not invited]
    魚崎英毅, 顔培実, 藤原正隆, 山下潤
    12th Molecular Cardiovascular Conference(2008.9.5-7.小樽)  2008

Industrial Property Rights

Awards & Honors

  • 2018/03 Hokkaido University School of Medicine Alumni Association Furate Young Investigator Award
    受賞者: Hideki Uosaki
  • 2015/12 BMB Travel Grants for Early Career Researchers in Abroad
    受賞者: Hideki Uosaki
  • 2015/11 American Heart Association BCVS Abstract Travel Grant
    受賞者: Hideki Uosaki
  • 2015/05 Weinstein Conference Travel Award
    受賞者: Hideki Uosaki
  • 2014/05 Weinstein Conference Travel Award
    受賞者: Hideki Uosaki
  • 2014/03 The Japanese Circulation Society Young Investigator's Award
     Identification of Chemicals Inducing Cardiomyocyte Proliferation in Developmental Stage-Specific Manner with Pluripotent Stem Cells 
    受賞者: Hideki Uosaki
  • 2010 Global COE, Kyoto University Travel Award
    受賞者: Hideki Uosaki
  • 2010 American Heart Association BCVS International Travel Grant
    受賞者: Hideki Uosaki
  • 2010 日本炎症再生医学会 優秀発表演題
    受賞者: 魚崎 英毅
  • 2010 Molecular Cardiovascular Conference Poster Award
    受賞者: Hideki Uosaki

Research Grants & Projects

  • 哺乳類の出生の異時性に基づく心筋再生機構の解明
    日本学術振興会:科学研究費助成事業 挑戦的研究(開拓)
    Date (from‐to) : 2021/07 -2024/03 
    Author : 木村 航
  • 心筋症の塩基編集治療に関する研究開発
    Date (from‐to) : 2021/11 -2024/03 
    Author : 魚崎 英毅, 小板橋紀通
  • 心筋症に対する遺伝子治療効果の予測手法の開発と実証
    日本医療研究開発機構:創薬基盤推進研究事業 医薬品創出に資する革新的技術の研究
    Date (from‐to) : 2021 -2024/03 
    Author : 魚崎英毅, 小板橋 紀通, 西園 啓文
  • Comprehensive Platform for Studying Molecular Mechanisms Regulating Cardiomyocyte Maturation
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
    Date (from‐to) : 2019/10 -2023/03 
    Author : Hideki Uosaki
  • The Acceleration Program for Intractable Diseases Research utilizing Disease-specific iPS cells
    Date (from‐to) : 2020/08 -2023/03 
    Author : Hideki Uosaki, Hitoshi Osaka, Kei Murayama, Tomoyuki Abe
  • 電気的・機械的刺激に対する多能性幹細胞由来心筋細胞の成熟適応応答メカニズムの解明
    Date (from‐to) : 2019/09 -2021/08 
    Author : 魚崎 英毅
  • 足場の硬化による心筋細胞の機能低下メカニズムの解明
    SENSHIN Medical Research Foundation:Research Grant for Young Investigator
    Date (from‐to) : 2018/12 -2019/11 
    Author : Hideki Uosaki
  • 足場の硬さによる心筋細胞の成熟メカニズムの解明
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Early-Career Scientists
    Date (from‐to) : 2019/04 -2019/10 
    Author : Hideki Uosaki
  • 心筋細胞の成熟を促進する外因性シグナル
    Uehara Memorial Foundation:Research Incentive Grant
    Date (from‐to) : 2018/04 -2019/03 
    Author : Hideki Uosaki
  • 心筋細胞の成熟促進因子の同定と分子メカニズムの解明
    Takeda Science Foundation:Research Grant
    Date (from‐to) : 2017 -2019/03 
    Author : Hideki Uosaki
  • ホルモン受容体が多能性幹細胞由来心筋細胞の成熟に果たす機能の解明
    Date (from‐to) : 2017 -2019/03 
    Author : 魚崎 英毅
  • 未成熟心筋細胞の成熟心筋細胞へのリプログラミングとその分子メカニズムの解明
    Date (from‐to) : 2016/11 -2019/03 
    Author : 魚崎 英毅
  • 遺伝性心筋症モデルの作出とゲノム治療法の開発
    Date (from‐to) : 2017/04 -2018/03 
    Author : 魚崎 英毅
  • 疾患iPS細胞と生体内成熟法を用いた心筋症発症早期の病態解明
    Date (from‐to) : 2017 -2018 
    Author : 魚崎 英毅
  • ファロー四徴症を持つGATA4変異ブタの開発
    Date (from‐to) : 2017/01 -2017/12 
    Author : 魚崎 英毅
  • MicroRNA-based Maturation of Cardiomyocytes Derived from Human Pluripotent Stem Cells
    Maryland Stem Cell Research Fund:Post-Doctoral Fellowship Programs
    Date (from‐to) : 2015/07 -2017/06 
    Author : Hideki Uosaki
    2016/8, Terminated in favor of a Lecturer position at Jichi Medical University
  • 心臓発生におけるnicheとしての外胚葉機能の解明と心筋分化誘導系への応用
    Date (from‐to) : 2013/04 -2015/03 
    Author : 魚崎 英毅
  • 治療応用を目指したヒトES・iPS細胞からの効率的心筋分化誘導法の確立
    JSPS:Research Fellowships for Young Scientists (DC2)
    Date (from‐to) : 2009/04 -2011/03 
    Author : Hideki Uosaki
  • 心筋細胞の多核化機構の解明による新規心臓再生療法の開発
    Date (from‐to) : 2011 
    Author : 魚崎 英毅
  • 治療応用を目指したヒトES・iPS細胞からの効率的心筋分化誘導法の確立
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for JSPS Fellows
    Date (from‐to) : 2009 -2010 
    Author : 魚崎 英毅
    Chemical Biologyを用い、ES細胞から心筋を効率的に分化誘導する化合物、誘導心筋細胞を増殖させる化合物を同定した。心筋増殖化合物は、4つの異なるシグナル伝達系に作用する7化合物を同定し、synergistic effectにより、ほぼ増殖することのないマウスES細胞由来心筋細胞を最大で14倍まで増殖させることを見出した(論文投稿準備中、特許申請中)。また、ヒトiPS細胞からの効率的な心筋分化誘導法を開発し、同分化系を用いることで心筋細胞を30-70%の効率で分化誘導することが可能となった。さらに細胞表面マーカーを同定し、FACSあるいはMACSを用いて95%以上まで純化することに成功した(論文投稿中、特許申請中)。本研究では、治療応用を目指したヒトES・iPS細胞からの効率的心筋分化誘導法の確立を目指した研究を実施し、治療応用を目指す上でのハードルであった、低い分化誘導効率、純度の高い心筋を得られないという問題に対して、誘導効率の向上と純化法の開発という両面からアプローチし、問題の解決を果たした。本研究成果により、ヒトiPS細胞由来心筋細胞の移植といった、治療応用研究が実施可能となり、本研究室において引き続き同研究に取り組んでいる。

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