Ishida, Hanako

写真a

Affiliation

Faculty of Pharmacy, Department of Pharmaceutical Sciences (Shiba-Kyoritsu)

Position

Research Associate/Assistant Professor/Instructor

Related Websites
Page Top ▲

Career 【 Display / hide 】

  • 2022.04
    -
    Present

    Faculty of Pharmacy, Keio University, Assistant professor

  • 2014.04
    -
    2022.03

    Graduate School of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, The University of Tokyo, project researcher

  • 2011.10
    -
    2014.03

    Graduate School of Engineering, Osaka University, technical assistant

  • 2009.02
    -
    2011.09

    Chiome Bioscience Inc.

  • 2006.04
    -
    2009.01

    International College of Arts and Sciences, Yokohama City University, technical assistant

Page Top ▲

Academic Background 【 Display / hide 】

  • 2004.04
    -
    2006.03

    Tottori University, Faculty of Agriculture, Department of Agriculture

    Graduate School, Completed, Master's course

  • 2000.04
    -
    2004.03

    Tokyo University of Agriculture, Faculty of Agriculture, Department of Agriculture

    University, Graduated

Page Top ▲

Academic Degrees 【 Display / hide 】

  • 博士（薬科学）, The University of Tokyo, Dissertation, 2019.03

    Structural biology of activation mechanism of innate immune receptor TLR9

Page Top ▲
 

Papers 【 Display / hide 】

  • KMN003 activates Nrf2 via disruption of the Keap1-Nrf2 interaction and p38-dependent transcriptional regulation

    Komeda K., Toyoshima K., Yasuda D., Ishida H., Kojima K., Osawa M., Hirano T., Ohe T., Tago K., Funakoshi-Tago M.

    Cellular Signalling 137 2026.01

    ISSN  08986568

    　View Summary

    Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that plays a crucial role in cellular defenses against oxidative stress and inflammation. Under normal conditions, Kelch-like ECH-associated protein 1 (Keap1), a ubiquitin ligase adaptor, binds to Nrf2, facilitating its ubiquitination and subsequent degradation via the proteasome. In this study, we investigated the properties of KMN003, a novel Nrf2 activator specifically designed to stabilize Nrf2 by disrupting its interaction with Keap1. X-ray crystallographic analysis revealed that KMN003 binds to the DGR-Cul3 (DC) domain of Keap1, occupying the Nrf2 interaction site. An AlphaScreen assay further showed that KMN003 effectively inhibits the binding between the Keap1 DC domain and the DLG motif of Nrf2 (IC₅₀ = 300 nM). We also investigated the mechanism of Nrf2 activation by KMN003 and its anti-inflammatory properties using murine macrophage-like RAW264.7 cells. KMN003 significantly reduced the lipopolysaccharide (LPS)-induced production of nitric oxide, CCL2, and tumor necrosis factor-alpha (TNFα) as well as the mRNA expression of inducible nitric oxide synthase, CCL2, and TNFα, which are essential inflammatory markers. KMN003 strongly inhibited nuclear translocation and transcriptional activation of nuclear factor-kappa B (NF-κB), a central regulator of inflammatory gene expression. KMN003 did not affect the LPS-induced phosphorylation of ERK or JNK, but strongly induced p38 phosphorylation in the absence of the LPS stimulation. The inhibition of p38 with SB203580 blocked KMN003-induced Nrf2 transcriptional activation despite promoting Nrf2 accumulation. These results highlight KMN003 as a promising anti-inflammatory drug that selectively stabilizes and activates Nrf2 via the p38 pathway.

  • Development of Keap1-Nrf2 Protein–Protein Interaction Inhibitor Activating Intracellular Nrf2 Based on the Naphthalene-2-acetamide Scaffold, and its Anti-Inflammatory Effects

    Yasuda D., Toyoshima K., Kojima K., Ishida H., Kaitoh K., Imamura R., Kanamitsu K., Kojima H., Funakoshi-Tago M., Osawa M., Ohe T., Hirano T.

    Chemmedchem 20 ( 21 ) e202500474 2025.11

    ISSN  18607179

    　View Summary

    Nuclear factor erythroid 2-related factor 2 (Nrf2) and Kelch-like ECH-associated protein 1 (Keap1) axis is an attractive therapeutic target for various intractable diseases. Although protein–protein interaction inhibitors against Keap1-Nrf2 have been developed over the past decade, more structural expansion is needed to improve efficacy. In this article, several candidate compounds are designed and synthesized as novel Nrf2 activators and their intracellular Nrf2-activating effects are evaluated. Among the synthesized compounds, a novel naphthalene-1,4-(4-ethoxybenzensulfonamide) bearing a tertiary acetamide side chain at the 2-position strongly activated intracellular Nrf2. Particularly, the pyrrolidine-type acetamide compound showed the strongest intracellular Nrf2 activation. X-ray cocrystallography revealed that this compound can bind to the DC domain of Keap1. Additionally, the pyrrolidine-type acetamide compound induced the mRNA expression of the representative Nrf2 target genes heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1. Moreover, the compound exhibited anti-inflammatory effects in a lipopolysaccharide-stimulated macrophage cell line. Conclusively, these results suggest that the pyrrolidine-type naphthalene-2-acetamide is a promising compound for the development of Nrf2 activators that can be applied to treat inflammatory diseases.

  • Cryo-EM structures of the zinc transporters ZnT3 and ZnT4 provide insights into their transport mechanisms.

    Ishida H, Yo R, Zhang Z, Shimizu T, Ohto U

    FEBS letters 599 ( 1 ) 41 - 52 2024.10

    Research paper (scientific journal), Lead author, Accepted,  ISSN  0014-5793

    　View Summary

    Zinc transporters (ZnTs) act as H+/Zn2+ antiporters, crucial for zinc homeostasis. Brain-specific ZnT3 expressed in synaptic vesicles transports Zn2+ from the cytosol into vesicles and is essential for neurotransmission, with ZnT3 dysfunction associated with neurological disorders. Ubiquitously expressed ZnT4 localized to lysosomes facilitates the Zn2+ efflux from the cytosol to lysosomes, mitigating the cell injury risk. Despite their importance, the structures and Zn2+ transport mechanisms remain unclear. We characterized the three-dimensional structures of human ZnT3 (inward-facing) and ZnT4 (outward-facing) using cryo-electron microscopy. By combining these structures, we assessed the conformational changes that could occur within the transmembrane domain during Zn2+ transport. Our results provide a structural basis for a more comprehensive understanding of the H+/Zn2+ exchange mechanisms exhibited by ZnTs.

  • Structure of the bile acid transporter and HBV receptor NTCP

    Asami J., Kimura K.T., Fujita-Fujiharu Y., Ishida H., Zhang Z., Nomura Y., Liu K., Uemura T., Sato Y., Ono M., Yamamoto M., Noda T., Shigematsu H., Drew D., Iwata S., Shimizu T., Nomura N., Ohto U.

    Nature (Nature)  606 ( 7916 ) 1021 - 1026 2022.06

    Research paper (scientific journal), Accepted,  ISSN  00280836

    　View Summary

    Chronic infection with hepatitis B virus (HBV) affects more than 290 million people worldwide, is a major cause of cirrhosis and hepatocellular carcinoma, and results in an estimated 820,000 deaths annually1,2. For HBV infection to be established, a molecular interaction is required between the large glycoproteins of the virus envelope (known as LHBs) and the host entry receptor sodium taurocholate co-transporting polypeptide (NTCP), a sodium-dependent bile acid transporter from the blood to hepatocytes3. However, the molecular basis for the virus–transporter interaction is poorly understood. Here we report the cryo-electron microscopy structures of human, bovine and rat NTCPs in the apo state, which reveal the presence of a tunnel across the membrane and a possible transport route for the substrate. Moreover, the cryo-electron microscopy structure of human NTCP in the presence of the myristoylated preS1 domain of LHBs, together with mutation and transport assays, suggest a binding mode in which preS1 and the substrate compete for the extracellular opening of the tunnel in NTCP. Our preS1 domain interaction analysis enables a mechanistic interpretation of naturally occurring HBV-insusceptible mutations in human NTCP. Together, our findings provide a structural framework for HBV recognition and a mechanistic understanding of sodium-dependent bile acid translocation by mammalian NTCPs.

  • Structural basis for the oligomerization-mediated regulation of NLRP3 inflammasome activation

    Ohto U., Kamitsukasa Y., Ishida H., Zhangami K., Hirama C., Maekawa S., Shimizu T. Z., Murak

    Proceedings of the National Academy of Sciences of the United States of America (Proceedings of the National Academy of Sciences of the United States of America)  119 ( 11 ) e2121353119 2022.03

    Research paper (scientific journal), Accepted,  ISSN  00278424

    　View Summary

    The nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain containing 3 (NLRP3) responds to a vast variety of stimuli, and activated NLRP3 forms an inflammasome, which in turn is associated with conditions such as atherosclerosis, Alzheimer's disease, and diabetes. A multilayered regulatory mechanism ensures proper NLRP3 inflammasome activation, although the structural basis for this process remains unclear. This study aimed to investigate the cryo-electron microscopy structure of the dodecameric form of full-length NLRP3 bound to the clinically relevant NLRP3-specific inhibitor MCC950. The inhibitor binds to the cavity distinct from the nucleotide binding site in the NACHT domain and stabilizes the closed conformation of NLRP3. The barrel-shaped dodecamer composed of the inactive form of NLRP3 is formed mainly through LRR-LRR interactions on the lateral side, and the highly positively charged top and bottom sides composed of NACHT domains provide a scaffold for membrane association. The cryo-electron microscopy structure suggests that oligomerization of NLRP3 is necessary for its membrane association; it is subsequently disrupted for activation, hence serving as a key player in controlling the spatiotemporal NLRP3 inflammasome activation. These findings are expected to contribute to the development of drugs targeting NLRP3 in future.

display all >>

Page Top ▲

Papers, etc., Registered in KOARA 【 Display / hide 】

Page Top ▲

Reviews, Commentaries, etc. 【 Display / hide 】

Page Top ▲

Presentations 【 Display / hide 】

  • Structural basis of the activation and trafficking mechanisms of nucleic acid-sensing TLR

    Hanako Ishida

    [Domestic presentation]  Molecular Biology Society of Japan (神戸ポートアイランド) , 

    2023.12
    , 

    Oral presentation (invited, special), 日本分子生物学会年会事務局

Page Top ▲
 

Courses Taught 【 Display / hide 】

  • STUDY OF MAJOR FIELD:(PHYSICS FOR LIFE FUNCTIONS)SPRING

    2025

  • STUDY OF MAJOR FIELD:(PHYSICS FOR LIFE FUNCTIONS)AUTUMN

    2025

  • STUDY OF MAJOR FIELD:(PHYSICS FOR LIFE FUNCTIONS)

    2025

  • SEMINAR:(PHYSICS FOR LIFE FUNCTIONS)SPRING

    2025

  • SEMINAR:(PHYSICS FOR LIFE FUNCTIONS)AUTUMN

    2025

display all >>

Page Top ▲