Takushi Miyoshi, MD, PhD

Takushi Miyoshi, MD, PhD

Assistant Professor
Molecular and Integrative Physiology

    About me

    Research Interest

    Stereocilia are a bundle of cylindrical F-actin protrusions developed on the apical surface of inner ear hair cells and function as biological mechanosensors to detect sound, acceleration and gravity. The long-term goal of my research is to elucidate the molecular mechanisms that permit stereocilia to be developed from tiny microvilli, equipped with mechanotransduction channel complexes and structurally stable for years. There are currently no clinically effective treatments to regenerate or restore stereocilia although degeneration of stereocilia is a process that often accompanies sensorineural hearing loss caused by genetic variations, aging, drugs and noise exposure, which is one of the reasons why sensorineural hearing loss is often irreversible. Understanding the protein dynamics in stereocilia will be a basis to (1) formulate therapeutic strategies for sensorineural hearing loss and (2) provide experimental data necessary for genetic diagnosis of patients suspected of having hereditary hearing loss.

    My research carrier initially focused on the dynamic regulation of actin cytoskeleton. Single-molecule microscopy, which I learned at Kyoto University, has been a powerful tool to elucidate the dynamics of actin regulatory proteins, such as CapZ, the Arp2/3 complex and DIAPH1. Single-molecule microscopy poses two challenges of interest to me: (1) solving a clinical problem and (2) elucidating the in vivo protein dynamics, especially in stereocilia. To accomplish the first challenge, I have been collaborating in an international research effort with Japanese and Korean researchers. I have worked on the pathophysiology of DIAPH1 variants associated with human deafness DFNA1 and used single-molecule microscopy to visualize abnormal actin elongation activity of mutant DIAPH1. The second challenge started at NIDCD/NIH. Under the mentorship of Dr. Thomas B. Friedman (NIDCD/NIH) and Dr. Hari Shroff (formerly NIBIB/NIH, currently Janelia Research Campus), I developed a workflow for single-molecule microscopy in stereocilia of live hair cells and started analyses of active cargo transport driven by unconventional myosins and the molecular turnover of stereocilia components.

    At SIU, I am utilizing my single-molecule microscopy technique to visualize trafficking of MYO7A, which are essential for localizing the mechanotransduction complex at stereocilia tips. I am also expanding the imaging target to other unconventional myosins whose variants are associated with human hearing loss and to actin and regulatory proteins, which are replenished maintaining the entire architecture of stereocilia. My single-molecule microscopy will be a novel tool for analyzing the normal and aberrant behavior of proteins in stereocilia, improving the efficacy and reliability of genetic diagnoses based on experimental data and formulating therapeutic strategies for sensorineural hearing loss. In addition, approaches developed in my study will be generally useful outside the field of hearing research to analyze the pathophysiology of mutant human genes affecting other organ systems.
     

    Please visit the Miyoshi lab website for more information: https://takushim.github.io/

    Gender

    Male

    Education & training

    Board Certifications
    Board Certified Otolaryngologist in Japan
    Medical School
    Kyoto University Faculty of Medicine, Kyoto, Japan
    Doctorate Degree
    Kyoto University Graduate School of Medicine, Kyoto, Japan
    Residency
    Kitano Hospital, Osaka, Japan
    Japanese Red Cross Society Wakayama Medical Center, Wakayama, Japan

    Publications

    https://www.ncbi.nlm.nih.gov/myncbi/takushi.miyoshi.1/bibliography/public/

    • Miyoshi T, Belyantseva IA, Sajeevadathan M, Friedman TB. Pathophysiology of human hearing loss associated with variants in myosins. Front Physiol. 2024;15:1374901. doi: 10.3389/fphys.2024.1374901. eCollection 2024. Review. PubMed PMID: 38562617; PubMed Central PMCID: PMC10982375.
    • Kim BJ, Miyoshi T, Chaudhry T, Friedman TB, Choi BY, Ueyama T. Late-onset hearing loss case associated with a heterozygous truncating variant of DIAPH1. Clin Genet. 2022 Apr;101(4):466-471. doi: 10.1111/cge.14115. Epub 2022 Feb 2. PubMed PMID: 35060117; PubMed Central PMCID: PMC8981108.
    • Miyoshi T, Belyantseva IA, Kitajiri SI, Miyajima H, Nishio SY, Usami SI, Kim BJ, Choi BY, Omori K, Shroff H, Friedman TB. Human deafness-associated variants alter the dynamics of key molecules in hair cell stereocilia F-actin cores. Hum Genet. 2022 Apr;141(3-4):363-382. doi: 10.1007/s00439-021-02304-0. Epub 2021 Jul 7. Review. PubMed PMID: 34232383.
    • Miyoshi T, Friedman TB, Watanabe N. Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy. STAR Protoc. 2021 Dec 17;2(4):100967. doi: 10.1016/j.xpro.2021.100967. eCollection 2021 Dec 17. PubMed PMID: 34841279; PubMed Central PMCID: PMC8605432.
    • Miyoshi T, Zhang Q, Miyake T, Watanabe S, Ohnishi H, Chen J, Vishwasrao HD, Chakraborty O, Belyantseva IA, Perrin BJ, Shroff H, Friedman TB, Omori K, Watanabe N. Semi-automated single-molecule microscopy screening of fast-dissociating specific antibodies directly from hybridoma cultures. Cell Rep. 2021 Feb 2;34(5):108708. doi: 10.1016/j.celrep.2021.108708. PubMed PMID: 33535030; PubMed Central PMCID: PMC7904085.
    • Kim BJ, Ueyama T, Miyoshi T, Lee S, Han JH, Park HR, Kim AR, Oh J, Kim MY, Kang YS, Oh DY, Yun J, Hwang SM, Kim NKD, Park WY, Kitajiri SI, Choi BY. Differential disruption of autoinhibition and defect in assembly of cytoskeleton during cell division decide the fate of human DIAPH1-related cytoskeletopathy. J Med Genet. 2019 Dec;56(12):818-827. doi: 10.1136/jmedgenet-2019-106282. Epub 2019 Aug 31. PubMed PMID: 31473629.
    • Tona R, Chen W, Nakano Y, Reyes LD, Petralia RS, Wang YX, Starost MF, Wafa TT, Morell RJ, Cravedi KD, du Hoffmann J, Miyoshi T, Munasinghe JP, Fitzgerald TS, Chudasama Y, Omori K, Pierpaoli C, Banfi B, Dong L, Belyantseva IA, Friedman TB. The phenotypic landscape of a Tbc1d24 mutant mouse includes convulsive seizures resembling human early infantile epileptic encephalopathy. Hum Mol Genet. 2019 May 1;28(9):1530-1547. doi: 10.1093/hmg/ddy445. PubMed PMID: 30602030; PubMed Central PMCID: PMC6466106.
    • Miyoshi T, Yamaguchi T, Ogita K, Tanaka Y, Ishibashi KI, Ito H, Kobayashi T, Nakagawa T, Ito J, Omori K, Yamamoto N. Quantitative Analysis of Aquaporin Expression Levels during the Development and Maturation of the Inner Ear. J Assoc Res Otolaryngol. 2017 Apr;18(2):247-261. doi: 10.1007/s10162-016-0607-3. Epub 2016 Dec 21. PubMed PMID: 28004290; PubMed Central PMCID: PMC5352615.
    • Ueyama T, Ninoyu Y, Nishio SY, Miyoshi T, Torii H, Nishimura K, Sugahara K, Sakata H, Thumkeo D, Sakaguchi H, Watanabe N, Usami SI, Saito N, Kitajiri SI. Constitutive activation of DIA1 (DIAPH1) via C-terminal truncation causes human sensorineural hearing loss. EMBO Mol Med. 2016 Nov;8(11):1310-1324. doi: 10.15252/emmm.201606609. Print 2016 Nov. PubMed PMID: 27707755; PubMed Central PMCID: PMC5090661.
    • Tona Y, Hamaguchi K, Ishikawa M, Miyoshi T, Yamamoto N, Yamahara K, Ito J, Nakagawa T. Therapeutic potential of a gamma-secretase inhibitor for hearing restoration in a guinea pig model with noise-induced hearing loss. BMC Neurosci. 2014 May 22;15:66. doi: 10.1186/1471-2202-15-66. PubMed PMID: 24884926; PubMed Central PMCID: PMC4051152.
    • Miyoshi T, Watanabe N. Can filament treadmilling alone account for the F-actin turnover in lamellipodia?. Cytoskeleton (Hoboken). 2013 Apr;70(4):179-90. doi: 10.1002/cm.21098. Epub 2013 Mar 5. PubMed PMID: 23341338.
    • Miyoshi T, Takebayashi S, Suzuki C, Hiwatashi N, Ikeda H, Ono K, Miura M. Early-onset postirradiation sarcoma of the tongue after pseudotumor phase. ORL J Otorhinolaryngol Relat Spec. 2011;73(4):201-5. doi: 10.1159/000328977. Epub 2011 Jun 23. PubMed PMID: 21701246.
    • Tsuji T, Miyoshi T, Higashida C, Narumiya S, Watanabe N. An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia. PLoS One. 2009;4(3):e4921. doi: 10.1371/journal.pone.0004921. Epub 2009 Mar 17. PubMed PMID: 19290054; PubMed Central PMCID: PMC2654150.
    • Miyoshi T, Tsuji T, Higashida C, Hertzog M, Fujita A, Narumiya S, Scita G, Watanabe N. Actin turnover-dependent fast dissociation of capping protein in the dendritic nucleation actin network: evidence of frequent filament severing. J Cell Biol. 2006 Dec 18;175(6):947-55. doi: 10.1083/jcb.200604176. PubMed PMID: 17178911; PubMed Central PMCID: PMC2064704.
    • Higashida C, Miyoshi T, Watanabe N. [Probing actin biochemistry in living cells using single-molecule speckle microscopy]. Tanpakushitsu Kakusan Koso. 2005 Sep;50(11):1436-42. Review. PubMed PMID: 16146192.
    • Higashida C, Miyoshi T, Fujita A, Oceguera-Yanez F, Monypenny J, Andou Y, Narumiya S, Watanabe N. Actin polymerization-driven molecular movement of mDia1 in living cells. Science. 2004 Mar 26;303(5666):2007-10. doi: 10.1126/science.1093923. PubMed PMID: 15044801.

    Grants

    2016-2018 JSPS KAKENHI, Grant-in-Aid for JSPS fellows (DC2)
    2018-2018 JSPS KAKENHI, Grant-in-Aid for Early-Career Scientists (ended due to the transfer to NIH)
    2024-2026 NIH Pathway to Independence Award (R00, 1R00DC019949)

    Awards

    2019-2021 JSPS Overseas Research Fellowships
    2022-2023 NIH Pathway to Independence Award (K99, 1K99DC019949)