Organismal Biosystems Laboratories
Laboratory of Immunology and Cell Biology
Imaging, Immunity, Bone tissue, Bone marrow, Blood cell
Living cells are constantly moving and working in our living bodies
The main theme of our laboratory is to reveal the cellular dynamics in various kinds of tissues and organs in vivo, by using advanced imaging techniques, especially focusing on the dynamic phenomenon in immune and inflammatory systems. By exploiting advanced multiphoton microscopy, we have originally developed original intravital imaging systems that enable the visualization of the movements of the diverse cell types resident in various immune tissues and organs in situ. We first established a live imaging system for visualizing living phenomena in intact bone marrow cavities. The dynamic nature of different cell types can be visualized in a time-dependent manner, in addition to the spatial and structural information generated by conventional histological analyses, resulting in a paradigm shift in research on immunology and cell biology.
Dynamic immune network wiring different tissues and organs and constituting integrity of biosystems
|Masaru Ishii (Professor)||mishii[at]icb.med.osaka-u.ac.jp|
|Nishikawa keizo (Guest Associate Professor, Doshisha Univ.)|
|Junichi Kikuta (Associate Professor)||jkikuta[at]icb.med.osaka-u.ac.jp|
|Takao Sudo (Assistant Professor)||sudo[at]icb.med.osaka-u.ac.jp|
|Yutaka Uchida (Specially Appointed Assistant Professor, Graduate School of Medicine)|
|Lok Laurence (Specially Appointed Researcher, IFReC)|
|Erika Yamashita (Specially Appointed Researcher, Graduate School of Medicine)|
|Akito Morimoto (Specially Appointed Researcher, Graduate School of Medicine, Division of health sciences)|
|Mayu Fujita (Guest Researcher)|
|Tetsuo Hasegawa (Guest Researcher)|
|Tomoka Awao (Guest Researcher, NIBIOHN)|
|Ayumi Sakai (Technical Staff)|
|Mami Kawasaki (Technical Staff)|
|Maiko Yorioka (Technical Staff)|
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- What is your hot research topic?
- Organisms, such as animals, are shaped by organizational dynamics, and the spatiotemporal control of dynamic cellular movement in the body is crucial for life activity. In our lab, we intend to comprehensively investigate dynamic behaviors and their molecular basis in vivo using combinatory cutting-edge technologies, such as intravital imaging and single-cell sequencing analyses that were originally developed by the proposer of this study. Drawing on novel cross-disciplinary approaches based on mathematics, and bioinformatics, we seek to understand the essence of life by identifying the invisible hand ruling cellular dynamics in vivo.
- What is your recent breakthrough or research progress?
- 1. Osteoclasts have a unique bone-destroying capacity, playing key roles in steady-state bone remodeling and arthritic bone erosion. Whether the osteoclasts in these different tissue settings arise from the same precursor states of monocytoid cells is presently unknown. Here, we show that osteoclasts in pannus originate exclusively from circulating bone marrow-derived cells and not from locally resident macrophages. We identify murine CX3CR1hiLy6CintF4/80+I-A+/I-E+ macrophages (termed here arthritis-associated osteoclastogenic macrophages (AtoMs)) as the osteoclast precursor-containing population in the inflamed synovium, comprising a subset distinct from conventional osteoclast precursors in homeostatic bone remodeling. Tamoxifen-inducible Foxm1 deletion suppressed the capacity of AtoMs to differentiate into osteoclasts in vitro and in vivo. Furthermore, synovial samples from human patients with rheumatoid arthritis contained CX3CR1+HLA-DRhiCD11c+CD80-CD86+ cells that corresponded to mouse AtoMs, and human osteoclastogenesis was inhibited by the FoxM1 inhibitor thiostrepton, constituting a potential target for rheumatoid arthritis treatment.（Hasegawa et al., Nat Immunol, 2019）
2. Bone homeostasis is regulated by communication between bone-forming mature osteoblasts (mOBs) and bone-resorptive mature osteoclasts (mOCs). However, the spatial-temporal relationship and mode of interaction in vivo remain elusive. Here we show, by using an intravital imaging technique, that mOB and mOC functions are regulated via direct cell-cell contact between these cell types. The mOBs and mOCs mainly occupy discrete territories in the steady state, although direct cell-cell contact is detected in spatiotemporally limited areas. In addition, a pH-sensing fluorescence probe reveals that mOCs secrete protons for bone resorption when they are not in contact with mOBs, whereas mOCs contacting mOBs are non-resorptive, suggesting that mOBs can inhibit bone resorption by direct contact. Intermittent administration of parathyroid hormone causes bone anabolic effects, which lead to a mixed distribution of mOBs and mOCs, and increase cell-cell contact. This study reveals spatiotemporal intercellular interactions between mOBs and mOCs affecting bone homeostasis in vivo.（Furuya et al., Nat Commun, 2018）。
3. Metabolic reprogramming occurs in response to the cellular environment to mediate differentiation, but the fundamental mechanisms linking metabolic processes to differentiation programs remain to be elucidated. During osteoclast differentiation, a shift toward more oxidative metabolic processes occurs. In this study we identified the de novo DNA methyltransferase 3a (Dnmt3a) as a transcription factor that couples these metabolic changes to osteoclast differentiation. We also found that receptor activator of nuclear factor-κB ligand (RANKL), an essential cytokine for osteoclastogenesis, induces this metabolic shift towards oxidative metabolism, which is accompanied by an increase in S-adenosylmethionine (SAM) production. We found that SAM-mediated DNA methylation by Dnmt3a regulates osteoclastogenesis via epigenetic repression of anti-osteoclastogenic genes. The importance of Dnmt3a in bone homeostasis was underscored by the observations that Dnmt3a-deficient osteoclast precursor cells do not differentiate efficiently into osteoclasts and that mice with an osteoclast-specific deficiency in Dnmt3a have elevated bone mass due to a smaller number of osteoclasts. Furthermore, inhibition of DNA methylation by theaflavin-3,3'-digallate abrogated bone loss in models of osteoporosis. Thus, this study reveals the role of epigenetic processes in the regulation of cellular metabolism and differentiation, which may provide the molecular basis for a new therapeutic strategy for a variety of bone disorders.(Nishikawa et al., Nat Med, 2015)
- What kind of background do your lab members have？
- We have a variety of members, including graduates from the areas of science and medicine.
- Do you collaborate with other institutions and universities?
- We are in collaboration with the Graduate School of Information Science and Technology and the Graduate School of Engineering at Osaka University. We are also studying with a variety of research institutions, such as the University of Tokyo, Kyoto University, and RIKEN.
- What kind of careers do your Lab's alumni go on to?
- Academic positions (Osaka University), pharmaceutical companies, etc
- How do you develop your research?
- The ultimate aim of the proposed research is to expand the concept of morphology, which is based on the form of cells and tissue, and to establish a novel scientific field, which posits cellular movement as the universal principle of life.
Publications (Research Articles, Reviews, Books)
SLPI is a critical mediator that controls PTH-induced bone formation
Nature Communications 12(1):2136 2021
Group 2 innate lymphoid cells support hematopoietic recovery under stress conditions
Journal of Experimental Medicine 218(5):e20200817 2021
In vivo dynamic analysis of BMP-2-induced ectopic bone formation
Sci Rep 10(1):4751 2020
Intravital multiphoton microscopy as a novel tool in the field of immunopharmacology.
Pharmacol. Ther. 206:107429 2020
Thrombomodulin induces anti-inflammatory effects by inhibiting the rolling adhesion of leukocytes in vivo.
Pharmacol. Sci. 143(1):17-22 2020
Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by FoxM1
Nat. Immunol. 20(12):1631-1643 2019
Mesenchymal stem cells cultured under hypoxic conditions had a greater therapeutic effect on mice with liver cirrhosis compared to those cultured under normal oxygen conditions.
Regen. Ther. 11:269-281 2019
Single-cell Transcriptome Analysis of Mouse Leukocytes in Inflammatory Stimulation
2019 IEEE International Conference on Bioinformatics and Biomedicine (BIBM) 2019
Analyzing Leukocyte Migration Trajectories by Deformable Image Matching
2019 IEEE 19th International Conference on Bioinformatics and Bioengineering (BIBE) 2019
In Vivo Multicolor Imaging with Fluorescent Probes Revealed the Dynamics and Function of Osteoclast Proton Pumps.
ACS Central Sci. 5(6):1059-1066 2019
GPR31-dependent dendrite protrusion of intestinal CX3CR1+ cells by bacterial metabolites
Nature 566(7742):110-114 2019
High-endothelial cell-derived S1P regulates dendritic cell localization and vascular integrity in the lymph node.
eLife 8:e41239 2019
Mesenchymal stem cells and induced bone marrow-derived macrophages synergistically improve liver fibrosis in mice.
Stem Cells Transl. Med. 8(3):271-284 2019
Imaging the Bone-Immune Cell Interaction in Bone Destruction.
Front. Immunol. 0.830555556 2019
Folliculin Regulates Osteoclastogenesis Through Metabolic Regulation.
J. Bone Miner. Res. 33(10):1785-98 2018
Presenilin 1 regulates NF-κB activation via association with breakpoint cluster region and casein kinase II.
J. Immunol. 201(8):2256-2263 2018
Intravital imaging with two-photon microscopy reveals cellular dynamics in the ischeamia-reperfused rat heart.
Sci Rep 8(1):15991 2018
Dynamic analyses of the short-term effects of different bisphosphonates using intravital two-photon microscopy.
JBMR Plus. 2(6):362-366 2018
Roles of Enhancer RNAs in RANKL-induced Osteoclast Differentiation Identified by Genome-wide Cap-analysis of Gene Expression using CRISPR/Cas9.
Sci Rep 14(1):7504 2018
Histone methylation regulator PTIP is required to maintain normal and leukemic bone marrow niches.
Proc. Natl. Acad. Sci. U. S. A. 115(43):E10137-E10146 2018
In vivo visualisation of different modes of action of biological DMARDs inhibiting osteoclastic bone resorption.
Ann. Rheum. Dis. 77(8):1219-1225 2018
Chiral cell sliding drives left-right asymmetric organ twisting.
eLife 7:e32506 2018
Variable SATB1 Levels Regulate Hematopoietic Stem Cell Heterogeneity with Distinct Lineage Fate.
Cell Reports 23(11):3223-3235 2018
In vivo imaging of T cell lymphoma infiltration process at the colon.
Sci Rep 8(1):3978. 2018
Intravital Imaging of the Heart at the Cellular Level Using Two-Photon Microscopy.
Methods Mol Biol. 1763:145-151 2018
Intravital Imaging of Liver Cell Dynamics.
Methods Mol Biol. 1763:137-143 2018
Bone Imaging: Osteoclast and Osteoblast Dynamics.
Methods Mol Biol. 1763:1-9 2018
Dynamic analysis of hematopoietic stem cells in the bone marrow by intravital imaging.
Clin Calcium. 28(2):231-236 2018
In vivo imaging of osteoblasts.
Biol. Reprod. 28(2):217-222 2018
Development of methodology for living bone imaging.
Biomaterials 28(2):181-185 2018
In vivo live imaging of bone cells.
Histochem. Cell Biol. 149(4):417-422 2018
Direct cell-cell contact between mature osteoblasts and osteoclasts dynamically controls their functions in vivo.
Biomolecules 9(1):300 2018
Identification of MS4A3 as a reliable marker for early myeloid differentiation in human hematopoiesis.
Biophys. J. 495(3):2338-2343 2018
The 17,18-epoxyeicosatetraenoic acid-G protein-coupled receptor 40 axis ameliorates contact hypersensitivity by inhibiting neutrophil mobility in mice and cynomolgus macaques.
J. Allergy Clin. Immunol. 142(2):470-484 2017
Imaging of inflammation and regeneration: a novel trend dissecting dynamic features of biological phenomena in vivo.
Inflamm Regen. 37:26:00 2017
Claudin-3 Loss Causes Leakage of Sweat from the Sweat Gland to Contribute to the Pathogenesis of Atopic Dermatitis.
J. Invest. Dermatol. 138(6):1279-1287 2017
Update on recent progress in vitamin D research. The effects of vitamin D in autoinflammatory diseases.
Clin Calcium. 27(11):1551-1559 2017
Soluble APP functions as a vascular niche signal that controls adult neural stem cell number.
Development 144:2730-2736 2017
Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes.
Nat. Cell Biol. 19(5):530-541 2017
Non-labeling multiphoton excitation microscopy as a novel diagnostic tool for discriminating normal tissue and colorectal cancer lesions.
Sci Rep 7(1):6959 2017
Bone marrow cavity segmentation using graph-cuts with wavelet-based texture feature.
J. Bioinform. Comput. Biol. 15(5):1740004 2017
Roles of cell-cell adhesion and contact in obesity-induced hepatic myeloid cell accumulation and glucose intolerance.
Cell Reports 18(11):2766-2779 2017
Immunology provides a great success for treating systemic autoimmune diseases - a perspective on immunopharmacology - IUPHAR Review X.
Br. J. Pharmacol. 174(13):1875-1880 2017
Intravital imaging technology reveals immune system dynamics in vivo.
Allergol. Int. 65(3):225-227 2016
ESAM is a novel human hematopoietic stem cell marker associated with a subset of human leukemias.
Exp. Hematol. 44(4):269-281 2016
Endothelial Cell-Selective Adhesion Molecule Expression in Hematopoietic Stem/Progenitor Cells Is Essential for Erythropoiesis Recovery after Bone Marrow Injury.
PLoS One 11(4):e0154189. 2016
Prickle1 promotes focal adhesion disassembly in cooperation with the CLASP-LL5β complex in migrating cells.
J. Cell Sci. 129(16):3115-29. 2016
Analysis of bone tissues by intravital imaging
Clin Calcium. 26(5):729-34 2016
Intercellular communication between keratinocytes and fibroblasts induces local osteoclast differentiation: a mechanism underlying cholesteatoma-induced bone destruction.
Mol. Cell. Biol. 36(11):1610-20. 2016
Cell-cycle-controlled radiation therapy was effective for treating a murine malignant melanoma cell line in vitro and in vivo.
Sci Rep 6:30689 2016
Fibroblast Growth Factor-2 facilitates the growth and chemo-resistance of leukemia cells in the bone marrow by modulating osteoblast functions.
Sci Rep 6:30779 2016
Real-time intravital imaging of pH variation associated with cell osteoclast activity and motility using designed small molecular probe.
Nat. Chem. Biol. 12(8):579-85 2016
Intravital imaging of Ca(2+) signals in lymphocytes of Ca(2+) biosensor transgenic mice: indication of autoimmune diseases before the pathological onset.
Sci Rep 6:18738 2016
Fibroblastic reticular cell-derived lysophosphatidic acid regulates confined intranodal T-cell motility.
eLife 5:e10561 2016
Trafficking of Osteoclast Precursors
Osteoimmunology (Second Edition) 25-40 2016
Our ideal candidate (as a graduate student)
We are looking for a highly motivated person to work on our research topics as our lab member. Our lab welcomes the person who loves taking care of creatures, hand working and handcraft too. Any kind of background (such as your expertise or major) is available.
Laboratory of Immunology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University,
2-2 Yamadaoka, Suita, Osaka 565-0871 Japan.
E-mail: mishii[at]icb.med.osaka-u.ac.jp (Prof. Masaru Ishii)
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