研究者総覧

口丸 高弘 (クチマル タカヒロ)

  • 分子病態研究部 講師
Last Updated :2020/09/28

研究者情報

学位

  • 博士(工学)(大阪大学)

ホームページURL

科研費研究者番号

  • 10570591

J-Global ID

研究分野

  • ライフサイエンス / 腫瘍生物学 / 光イメージング / 生体化学

研究活動情報

論文

  • Wanaporn Yimchuen, Tetsuya Kadonosono, Yumi Ota, Shinichi Sato, Maika Kitazawa, Tadashi Shiozawa, Takahiro Kuchimaru, Masumi Taki, Yuji Ito, Hiroyuki Nakamura, Shinae Kizaka-Kondoh
    RSC Advances 10 26 15154 - 15162 2020年04月 [査読有り][通常論文]
     
    This journal is © 2020 The Royal Society of Chemistry. Tumor-binding peptides such as human epidermal growth factor receptor 2 (HER2)-binding peptides are attractive therapeutic and diagnostic options for cancer. However, the HER2-binding peptides (HBPs) developed thus far are susceptible to proteolysis and lose their affinity to HER2 in vivo. In this report, a method to create a HER2-binding fluctuation-regulated affinity protein (HBP-FLAP) consisting of a fibronectin type III domain (FN3) scaffold with a structurally immobilized HBP is presented. HBPs were selected by phage-library screening and grafted onto FN3 to create FN3-HBPs, and the HBP-FLAP with the highest affinity (HBP sequence: YCAHNM) was identified after affinity maturation of the grafted HBP. HBP-FLAP containing the YCAHNM peptide showed increased proteolysis-resistance, binding to HER2 with a dissociation constant (KD) of 58 nM in ELISA and 287 nM in biolayer interferometry and specifically detects HER2-expressing cancer cells. In addition, HBP-FLAP clearly delineated HER2-expressing tumors with a half-life of 6 h after intravenous injection into tumor-bearing mice. FN3-based FLAP is an excellent platform for developing target-binding small proteins for clinical applications.
  • Tetsuya Kadonosono, Wanaporn Yimchuen, Yumi Ota, Kyra See, Tadaomi Furuta, Tadashi Shiozawa, Maika Kitazawa, Yu Goto, Akash Patil, Takahiro Kuchimaru, Shinae Kizaka-Kondoh
    Scientific reports 10 1 891 - 891 2020年01月 [査読有り][通常論文]
     
    Monoclonal antibodies (mAbs) are attractive therapeutics for treating a wide range of human disorders, and bind to the antigen through their complementarity-determining regions (CDRs). Small stable proteins containing structurally retained CDRs are promising alternatives to mAbs. In this report, we present a method to create such proteins, named fluctuation-regulated affinity proteins (FLAPs). Thirteen graft acceptor (GA) sites that efficiently immobilise the grafted peptide structure were initially selected from six small protein scaffolds by computational identification. Five CDR peptides extracted by binding energy calculations from mAbs against breast cancer marker human epithelial growth factor receptor type 2 (HER2) were then grafted to the selected scaffolds. The combination of five CDR peptides and 13 GA sites in six scaffolds revealed that three of the 65 combinations showed specific binding to HER2 with dissociation constants (KD) of 270-350 nM in biolayer interferometry and 24-65 nM in ELISA. The FLAPs specifically detected HER2-overexpressing cancer cells. Thus, the present strategy is a promising and practical method for developing small antibody mimetics.
  • Shinichiro Fuse, Kensuke Suzuki, Takahiro Kuchimaru, Tetsuya Kadonosono, Hiroki Ueda, Shinichi Sato, Shinae Kizaka-Kondoh, Hiroyuki Nakamura
    Bioorganic & medicinal chemistry 28 1 115207 - 115207 2020年01月 [査読有り][通常論文]
     
    HIF-1 is regarded as a promising target for the drugs used in cancer chemotherapy, and creating readily accessible templates for the development of synthetic drug candidates that could inhibit HIF-1 transcriptional activity is an important pursuit. In this study, indeno[2,1-c]pyrazolones were designed as readily available synthetic inhibitors of HIF-1 transcriptional activity. Nine compounds were synthesized in 4-5 steps from commercially available starting materials. In evaluations of the ability to inhibit the hypoxia-induced transcriptional activity of HIF-1, compound 3c showed a higher level compared with that of known inhibitor, YC-1. The compound 3c suppressed HIF-1α protein accumulation without affecting the levels of HIF-1α mRNA.
  • Emi Nomura, Yasuyuki Ohta, Koh Tadokoro, Jingwei Shang, Tian Feng, Xia Liu, Xiaowen Shi, Namiko Matsumoto, Ryo Sasaki, Keiichiro Tsunoda, Kota Sato, Mami Takemoto, Nozomi Hishikawa, Toru Yamashita, Takahiro Kuchimaru, Shinae Kizaka-Kondoh, Koji Abe
    Neuroscience 415 31 - 43 2019年09月 [査読有り][通常論文]
     
    Hypoxia inducible factor-1α (HIF-1α) is a key transcription factor that maintains oxygen homeostasis. Hypoxic stress is related to the pathogenesis of amyotrophic lateral sclerosis (ALS), and impaired HIF-1α induces motor neuron degeneration in ALS. Dimethyloxalylglycine (DMOG) upregulates the stability of HIF-1α expression and shows neuroprotective effects, but has not been used in ALS as an anti-hypoxic stress treatment. In the present study, we investigated hypoxic stress in ALS model mice bearing G93A-human Cu/Zn superoxide dismutase by in vivo HIF-1α imaging, and treated the ALS mice with DMOG. In vivo HIF-1α imaging analysis showed enhanced hypoxic stress in both the spinal cord and muscles of lower limbs of ALS mice, even at the pre-symptomatic stage. HIF-1α expression decreased as the disease progressed until 126 days of age. DMOG treatment significantly ameliorated the decrease in HIF-1α expression, the degeneration of both spinal motor neurons and myofibers in lower limbs, gliosis and apoptosis in the spinal cord. This was accompanied by prolonged survival. The present study suggests that in vivo bioluminescence resonance energy transfer (BRET) HIF-1α imaging is useful for evaluating hypoxic stress in ALS, and that the enhancement of HIF-1α is a therapeutic target for ALS patients.
  • Tadashi Ishida, Takuya Shimamoto, Maho Kaminaga, Takahiro Kuchimaru, Shinae Kizaka-Kondoh, Toru Omata
    Micromachines 10 2 2019年02月 [査読有り][通常論文]
     
    The small number of high-migratory cancer cells in a cell population make studies on high-migratory cancer cells difficult. For the development of migration assays for such cancer cells, several microfluidic devices have been developed. However, they measure migration that is influenced by microstructures and they collect not only high-migratory cells, but also surrounding cells. In order to find high-migratory cells in cell populations while suppressing artifacts and to collect these cells while minimizing damages, we developed a microfluidic high-migratory cell collector with the ability to sort cancer cells according to cellular migration and mechanical detachment. High-migratory cancer cells travel further from the starting line when all of the cells are seeded on the same starting line. The high-migratory cells are detached using a stretch of cell adhesive surface using a water-driven balloon actuator. Using this cell collector, we selected high-migratory HeLa cells that migrated about 100m in 12 h and collected the cells.
  • Ryohei Saito, Takahiro Kuchimaru, Shoko Higashi, Shijia W Lu, Masahiro Kiyama, Satoshi Iwano, Rika Obata, Takashi Hirano, Shinae Kizaka-Kondoh, Shojiro A Maki
    Bulletin of the Chemical Society of Japan 92 3 608 - 618 2018年12月 [査読有り][通常論文]
  • Pongsuchart Mongkol, Kuchimaru Takahiro, Yonezawa Sakiko, Tran Diem Thi Phuong, Kha Nguyen The, Hoang Ngoc, Thi Hong, Kadonosono Tetsuya, Kizaka-Kondoh Shinae
    Cancer science 109 9 2746 - 2756 2018年09月 [査読有り][通常論文]
     
    Lung metastasis is a major cause of mortality in patients with osteosarcoma (OS). A better understanding of the molecular mechanism of OS lung metastasis may facilitate development of new therapeutic strategies to prevent the metastasis. We have established high- and low-metastatic sublines (LM8-H and LM8-L, respectively) from Dunn OS cell line LM8 by using in vivo image-guided screening. Among the genes whose expression was significantly increased in LM8-H compared to LM8-L, the transcription factor lymphoid enhancer-binding factor 1 (LEF1) was identified as a factor that promotes LM8-H cell extravasation into the lungs. To identify downstream effectors of LEF1 that are involved in OS lung metastasis, 13 genes were selected based on LM8 microarray data and genomewide meta-analysis of a public database for OS patients. Among them, the cytoglobin (Cygb) gene was identified as a key effector in promoting OS extravasation into the lungs. CYGB overexpression increased the extravasation ability of LM8-L cells, whereas knocking out the Cygb gene in LM8-H cells reduced this ability. Our results showed a novel LEF1-CYGB axis in OS lung metastasis and may provide a new way of developing therapeutic strategies to prevent OS lung metastasis.
  • Takahiro Kuchimaru, Naoya Kataoka, Kenji Nakagawa, Tatsuhiro Isozaki, Hitomi Miyabara, Misa Minegishi, Tetsuya Kadonosono, Shinae Kizaka-Kondoh
    Nature Communications 9 1 2981 - 2981 2018年07月 [査読有り][通常論文]
     
    Although the current murine model of bone metastasis using intracardiac (IC) injection successfully recapitulates the process of bone metastasis, further progress in the study of bone metastasis requires a new model to circumvent some limitations of this model. Here, we present a new murine model of bone metastasis achieved by injecting cancer cells through the intra-caudal arterial (CA). This model does not require high technical proficiency, predominantly delivers cancer cells to bone marrow of hind limbs with much higher efficiency than IC injection, and greatly shortens the period of overt bone metastasis development. Moreover, CA injection barely causes acute death of mice, enabling us to inject a larger number of cancer cells to further accelerate the development of bone metastasis with a wide variety of cell lines. Our model may open a new avenue for understanding the bone metastatic processes and development of drugs preventing bone metastasis and recurrence.
  • Mahiro Iizuka-Ohashi, Motoki Watanabe, Mamiko Sukeno, Mie Morita, Ngoc Thi Hong Hoang, Takahiro Kuchimaru, Shinae Kizaka-Kondoh, Yoshihiro Sowa, Koichi Sakaguchi, Tetsuya Taguchi, Toshiyuki Sakai
    Oncotarget 9 28 19597 - 19612 2018年04月 [査読有り][通常論文]
     
    With increasing clinical demands for MEK inhibitors in cancer treatment, overcoming the resistance to MEK inhibitors is an urgent problem to be solved. Numerous reports have shown that MEK inhibition results in the activation of PI3K-Akt signaling, which may confer apoptotic resistance to MEK inhibitors. We here demonstrate that the blockade of the mevalonate pathway using the antilipidemic drug statins represses Akt activation following MEK inhibition and induces significant apoptosis when co-treated with CH5126766 or trametinib. These events were clearly negated by the addition of mevalonate or geranylgeranyl pyrophosphate, indicating that the protein geranylgeranylation is implicated in the apoptotic resistance to MEK inhibitors. Furthermore, mechanistically, the combined treatment of CH5126766 with statins upregulated TNF-related apoptosis-inducing ligand (TRAIL), which was dependent on inhibition of the mevalonate pathway and is involved in apoptosis induction in human breast cancer MDA-MB-231 cells. The present study not only revealed that the mevalonate pathway could be targetable to enhance the efficacy of MEK inhibitors, but also proposes that combinatorial treatment of MEK inhibitors with statins may be a promising therapeutic strategy to sensitize cancer cells to apoptosis.
  • Satoshi Iwano, Mayu Sugiyama, Hiroshi Hama, Akiya Watakabe, Naomi Hasegawa, Takahiro Kuchimaru, Kazumasa Z Tanaka, Megumu Takahashi, Yoko Ishida, Junichi Hata, Satoshi Shimozono, Kana Namiki, Takashi Fukano, Masahiro Kiyama, Hideyuki Okano, Shinae Kizaka-Kondoh, Thomas J McHugh, Tetsuo Yamamori, Hiroyuki Hioki, Shojiro Maki, Atsushi Miyawaki
    Science (New York, N.Y.) 359 6378 935 - 939 2018年02月 [査読有り][通常論文]
     
    Bioluminescence is a natural light source based on luciferase catalysis of its substrate luciferin. We performed directed evolution on firefly luciferase using a red-shifted and highly deliverable luciferin analog to establish AkaBLI, an all-engineered bioluminescence in vivo imaging system. AkaBLI produced emissions in vivo that were brighter by a factor of 100 to 1000 than conventional systems, allowing noninvasive visualization of single cells deep inside freely moving animals. Single tumorigenic cells trapped in the mouse lung vasculature could be visualized. In the mouse brain, genetic labeling with neural activity sensors allowed tracking of small clusters of hippocampal neurons activated by novel environments. In a marmoset, we recorded video-rate bioluminescence from neurons in the striatum, a deep brain area, for more than 1 year. AkaBLI is therefore a bioengineered light source to spur unprecedented scientific, medical, and industrial applications.
  • Tsubaki Takuya, Kadonosono Tetsuya, Sakurai Shimon, Shiozawa Tadashi, Goto Toshiki, Sakai Shiori, Kuchimaru Takahiro, Sakamoto Takeharu, Watanabe Hitomi, Kondoh Gen, Kizaka-Kondoh Shinae
    Oncotarget 9 13 11209 - 11226 2018年02月 [査読有り][通常論文]
     
    The immunosuppressive tumor microenvironment is a hallmark of cancer. Myeloid-derived suppressor cells (MDSCs) are CD11b+ Gr-1+ tumor-infiltrating immature myeloid cells that strongly mediate tumor immunosuppression. The CD11b+ Gr-1+ cells are a heterogeneous cell population, and the impacts of each subpopulation on tumor progression are not yet completely understood. In the present study, we identified a novel subpopulation of CD11b+ Gr-1+ cells from murine lung carcinoma tumors according to their strongly adherent abilities. Although strong adherent activity is a unique property of macrophages, their marker expression patterns are similar to those of MDSCs; thus, we named this novel subpopulation MDSC-like adherent cells (MLACs). Unlike known MDSCs, MLACs lack the ability to suppress cytotoxic T lymphocytes and differentiate into tumor-associated macrophages (TAMs), but could still directly facilitate tumor growth and angiogenesis through secreting CCL2, CXCL1/2/5, PAI-1, MMPs, and VEGFA. Furthermore, MLACs recruited MDSCs via the secretion of CCL2/5 and CXCL1/2/5, thereby enhancing the immunosuppressive tumor microenvironment and promoting TAMs-mediated tumor progression. Our findings suggest that MLACs may function as an initiator of the immunosuppressive tumor microenvironment and highlight a new therapeutic target to prevent the onset or delay malignant progression.
  • Satoshi Iwano, Mayu Sugiyama, Hiroshi Hama, Akiya Watakabe, Naomi Hasegawa, Takahiro Kuchimaru, Kazumasa Z. Tanaka, Megumu Takahashi, Yoko Ishida, Junichi Hata, Satoshi Shimozono, Kana Namiki, Takashi Fukano, Masahiro Kiyama, Hideyuki Okano, Shinae Kizaka-Kondoh, Thomas J. McHugh, Tetsuo Yamamori, Hiroyuki Hioki, Shojiro Maki, Atsushi Miyawaki
    Science 359 6378 935  American Association for the Advancement of Science ({AAAS}) 2018年02月 [査読有り][通常論文]
  • Kadonosono Tetsuya, Yimchuen Wanaporn, Tsubaki Takuya, Shiozawa Tadashi, Suzuki Yasuhiro, Kuchimaru Takahiro, Sato Yasufumi, Kizaka-Kondoh Shinae
    Protein Science 26 3 452 - 463 2017年03月 [査読有り][通常論文]
     
    Vasohibins (VASH1 and VASH2) are recently identified regulators of angiogenesis and cancer cell functions. They are secreted proteins without any classical secretion signal sequences, and are thought to be secreted instead via an unconventional protein secretion (UPS) pathway in a small vasohibin-binding protein (SVBP)-dependent manner. However, the precise mechanism of SVBP-dependent UPS is poorly understood. In this study, we identified a novel UPS regulatory system in which essential domain architecture (VASH-PS) of VASHs, comprising regions VASH191-180 and VASH280-169 , regulate the cytosolic punctate structure formation in the absence of SVBP. We also demonstrate that SVBP form a complex with VASH1 through the VASH1274-282 (SIa), VASH1139-144 (SIb), and VASH1133-137 (SIc), leading to the dispersion in the cytosol and extracellular release of VASH1. The amino acid sequences of VASH-SIa and VASH-PS, containing SIb and SIc, are highly conserved among VASH family members in vertebrates, suggesting that SVBP-dependent UPS may be common within the VASH family. This novel UPS regulatory system may open up new avenues for understanding fundamental protein secretion in vertebrates.
  • Kuchimaru Takahiro, Suka Tomoya, Hirota Keisuke, Kadonosono Tetsuya, Kizaka-Kondoh Shinae
    Scientific Reports 6 34311 - 34311 2016年10月 [査読有り][通常論文]
     
    The ubiquitin-proteasome system (UPS) is a selective protein degradation system that plays a critical role in many essential biological processes by regulating the existence of various cellular proteins. The target proteins of UPS are recognized and tagged with polyubiquitin chains by E3 ubiquitin ligases, which have high substrate-specific activities. Here we present a novel injectable imaging probe POL-N that can detect the UPS-regulated hypoxia-inducible factor (HIF) activity in vivo. Because the luciferase is fused to the E3 ligase-recognition domain of the HIF-1α, POL-N is intact only in the HIFα-overexpressing cells, that is, HIF-active cells, generating signals via an intramolecular bioluminescence resonance energy transfer (BRET) between luciferase and a near-infrared (NIR) fluorescent dye at the C-terminal end of the probe. Off-target signals of the NIR-BRET were so low that we could achieve highly sensitive and fast detection of intratumoral HIF-activity. Notably, we successfully detected hypoxic liver metastasis, which is extremely difficult to detect by injectable imaging probes due to strong off-target signals, as early as 1 h after systemic injection of POL-N. Our probe design can be widely adapted to UPS-target proteins and may contribute to the exploration of their roles in animal disease models.
  • Tadashi Ishida, Takuya Shimamoto, Nobuya Ozaki, Satoshi Takaki, Takahiro Kuchimaru, Sinae Kizaka-Kondoh, Toru Omata
    Micromachines 7 9 2016年09月 [査読有り][通常論文]
     
    A microfluidic device capable of precise chemical control is helpful to mimic tumor microenvironments in vitro, which are closely associated with malignant progression, including metastasis. Cancer cells under a concentration gradient of oxygen and other sustenance materials inside a tumor in vivo have recently been reported to increase the probability of metastasis. The influence of glucose concentration on cancer cells has not been measured well, whereas that of oxygen concentration has been thoroughly examined using microfluidic devices. This is because glucose concentrations can be controlled using microfluidic concentration gradient generators, which trade off temporal stability of the glucose concentration and shear stress on the cells; by contrast, oxygen concentration can be easily controlled without microfluidic device-induced shear stresses. To study cell division and migration responses as a function of glucose concentration, we developed a microfluidic device to observe cell behaviors under various chemical conditions. The device has small-cross-section microchannels for generating a concentration gradient and a large-cross-section chamber for cell culture. With this design, the device can achieve both a cell culture with sufficiently low shear stress on cell activity and a stable glucose concentration gradient. Experiments revealed that a low glucose concentration increased the total migration length of HeLa cells and that HeLa cells under a glucose concentration gradient exhibit random motion rather than chemotaxis.
  • Ngoc Thi Hong Hoang, Tetsuya Kadonosono, Takahiro Kuchimaru, Shinae Kizaka-Kondoh
    Cancer science 107 8 1151 - 8 2016年08月 [査読有り][通常論文]
     
    Pancreatic cancer is one of the most lethal digestive system cancers with a 5-year survival rate of 4-7%. Despite extensive efforts, recent chemotherapeutic regimens have provided only limited benefits to pancreatic cancer patients. Gemcitabine and TS-1, the current standard-of-care chemotherapeutic drugs for treatment of this severe cancer, have a low response rate. Hypoxia is one of the factors contributing to treatment resistance. Specifically, overexpression of hypoxia-inducible factor, a master transcriptional regulator of cell adaption to hypoxia, is strongly correlated with poor prognosis in many human cancers. TAT-ODD-procaspase-3 (TOP3) is a protein prodrug that is specifically processed and activated in hypoxia-inducible factor-active cells in cancers, leading to cell death. Here, we report combination therapies in which TOP3 was combined with gemcitabine or TS-1. As monotherapy, gemcitabine and TS-1 showed a limited effect on hypoxic and starved pancreatic cancer cells, whereas co-treatment with TOP3 successfully overcame this limitation in vitro. Furthermore, combination therapies of TOP3 with these drugs resulted in a significant improvement in survival of orthotopic pancreatic cancer models involving the human pancreatic cancer cell line SUIT-2. Overall, our study indicates that the combination of TOP3 with current chemotherapeutic drugs can significantly improve treatment outcome, offering a promising new therapeutic option for patients with pancreatic cancer.
  • Kuchimaru Takahiro, Iwano Satoshi, Kiyama Masahiro, Mitsumata Shun, Kadonosono Tetsuya, Niwa Haruki, Maki Shojiro, Kizaka-Kondoh Shinae
    Nature Communications 7 11856 - 11856 2016年06月 [査読有り][通常論文]
     
    In preclinical cancer research, bioluminescence imaging with firefly luciferase and D-luciferin has become a standard to monitor biological processes both in vitro and in vivo. However, the emission maximum (λmax) of bioluminescence produced by D-luciferin is 562 nm where light is not highly penetrable in biological tissues. This emphasizes a need for developing a red-shifted bioluminescence imaging system to improve detection sensitivity of targets in deep tissue. Here we characterize the bioluminescent properties of the newly synthesized luciferin analogue, AkaLumine-HCl. The bioluminescence produced by AkaLumine-HCl in reactions with native firefly luciferase is in the near-infrared wavelength ranges (λmax=677 nm), and yields significantly increased target-detection sensitivity from deep tissues with maximal signals attained at very low concentrations, as compared with D-luciferin and emerging synthetic luciferin CycLuc1. These characteristics offer a more sensitive and accurate method for non-invasive bioluminescence imaging with native firefly luciferase in various animal models.
  • Takahiro Kuchimaru, Satoshi Iwano, Masahiro Kiyama, Shun Mitsumata, Tetsuya Kadonosono, Haruki Niwa, Shojiro Maki, Shinae Kizaka-Kondoh
    NATURE COMMUNICATIONS 7 2016年06月 [査読有り][通常論文]
     
    In preclinical cancer research, bioluminescence imaging with firefly luciferase and D-luciferin has become a standard to monitor biological processes both in vitro and in vivo. However, the emission maximum (lambda(max)) of bioluminescence produced by D-luciferin is 562 nm where light is not highly penetrable in biological tissues. This emphasizes a need for developing a red-shifted bioluminescence imaging system to improve detection sensitivity of targets in deep tissue. Here we characterize the bioluminescent properties of the newly synthesized luciferin analogue, AkaLumine-HCl. The bioluminescence produced by AkaLumine-HCl in reactions with native firefly luciferase is in the near-infrared wavelength ranges (lambda(max) = 677 nm), and yields significantly increased target-detection sensitivity from deep tissues with maximal signals attained at very low concentrations, as compared with D-luciferin and emerging synthetic luciferin CycLuc1. These characteristics offer a more sensitive and accurate method for non-invasive bioluminescence imaging with native firefly luciferase in various animal models.
  • Hiroshi Inaba, Nusrat J M Sanghamitra, Kenta Fujita, Takeya Sho, Takahiro Kuchimaru, Susumu Kitagawa, Shinae Kizaka-Kondoh, Takafumi Ueno
    Molecular bioSystems 11 11 3111 - 8 2015年11月 [査読有り][通常論文]
     
    Carbon monoxide (CO) has been recognized as a messenger for signal transduction in living cells and tissues. For intracellular CO delivery, several metal carbonyl complexes have been used as CO-releasing molecules (CO-RMs). To improve the properties of CO-RMs, such as the stability and the CO release rate, ligands and carriers of the metal complexes have been exploited. Here we report the development of an efficient intracellular CO delivery system using a protein scaffold. We used a protein needle reconstructed from gene product 5 of bacteriophage T4, which has high cellular permeability and stability. When ruthenium carbonyl complexes are conjugated to the needle using a His-tag triad at the C-terminus, the resulting composite has a significantly higher cellular uptake efficiency of Ru carbonyl and a 12-fold prolonged CO release rate relative to Ru(CO)3Cl(glycinate), a widely used CO-RM. We demonstrate that CO delivered by the composite activates the transcriptional factor nuclear factor-kappaB (NF-κB), which in turn leads to significant induction of expression of its target genes, HO1, NQO1, and IL6, through generation of reactive oxygen species (ROS). The signaling pathway is distinct from that of tumor necrosis factor (TNF)-α-induced activation of NF-κB. The protein needle-based CO-RM can be exploited to elucidate the biological functions of CO and used in the development of protein-based organometallic tools for modulation of cellular signaling.
  • Shinsuke Takata, Tomomi Masuda, Shinsuke Nakamura, Takahiro Kuchimaru, Kazuhiro Tsuruma, Masamitsu Shimazawa, Hideko Nagasawa, Shinae Kizaka-Kondoh, Hideaki Hara
    Scientific reports 5 9898 - 9898 2015年04月 [査読有り][通常論文]
     
    Hypoxic stress is a risk factor of ocular neovascularization. Hypoxia visualization may provide clues regarding the underlying cause of angiogenesis. Recently, we developed a hypoxia-specific probe, protein transduction domain-oxygen-dependent degradation domain-HaloTag-Rhodamine (POH-Rhodamine). In this study, we observed the localization of HIF-1α proteins by immunohistochemistry and the fluorescence of POH-Rhodamine on RPE-choroid flat mounts. Moreover, we compared the localization of POH-Rhodamine with pimonidazole which is a standard reagent for detecting hypoxia. Next, we investigated the effects of triamcinolone acetonide (TAAC) against visual function that was evaluated by recording electroretinogram (ERG) and choroidal neovascularization (CNV) development. Mice were given laser-induced CNV using a diode laser and treated with intravitreal injection of TAAC. Finally, we investigated POH-Rhodamine on CNV treated with TAAC. In this study, the fluorescence of POH-Rhodamine and HIF-1α were co-localized in laser-irradiated sites, and both the POH-Rhodamine and pimonidazole fluorescent areas were almost the same. Intravitreal injection of TAAC restored the reduced ERG b-wave but not the a-wave and decreased the mean CNV area. Furthermore, the area of the POH-Rhodamine-positive cells decreased. These findings indicate that POH-Rhodamine is useful for evaluating tissue hypoxia in a laser-induced CNV model, suggesting that TAAC suppressed CNV through tissue hypoxia improvement.
  • Tetsuya Kadonosono, Akihiro Yamano, Toshiki Goto, Takuya Tsubaki, Mizuho Niibori, Takahiro Kuchimaru, Shinae Kizaka-Kondoh
    Journal of controlled release : official journal of the Controlled Release Society 201 14 - 21 2015年03月 [査読有り][通常論文]
     
    Cell-penetrating peptides (CPPs), also referred to as protein transduction domains (PTDs), can mediate the cellular uptake of a wide range of macromolecules including peptides, proteins, oligonucleotides, and nanoparticles, and thus have received considerable attention as a promising method for drug delivery in vivo. Here, we report that CPP/PTDs facilitate the extravasation of fused proteins by binding to neuropilin-1 (NRP1), a vascular endothelial growth factor (VEGF) co-receptor expressed on the surface of endothelial and some tumor cells. In this study, we examined the capacity of the amphipathic and cationic CPP/PTDs, PTD-3 and TAT-PTD, respectively, to bind cells in vitro and accumulate in xenograft tumors in vivo. Notably, these functions were significantly suppressed by pre-treatment with NRP1-neutralizing Ab. Furthermore, co-injection of iRGD, a cyclic peptide known to increase NRP1-dependent vascular permeability, significantly reduced CPP/PTD tumor delivery. This data demonstrates a mechanism by which NRP1 promotes the extravasation of CPP/PTDs that may open new avenues for the development of more efficient CPP/PTD delivery systems.
  • Hiroyasu Tabe, Takuya Shimoi, Kenta Fujita, Satoshi Abe, Hiroshi Ijiri, Masahiko Tsujimoto, Takahiro Kuchimaru, Shinae Kizaka-Kondo, Hajime Mori, Susumu Kitagawa, Takafumi Ueno
    CHEMISTRY LETTERS 44 3 342 - 344 2015年03月 [査読有り][通常論文]
     
    A crystalline protein assembly of cypovirus polyhedra was engineered to develop a carbon monoxide (CO) releasing extracellular scaffold by immobilizing ruthenium carbonyls. The molecular design includes introduction of a hexahistidine tag to the C-terminus and provides immobilization of about 2-fold more Ru carbonyls per protein monomer and effectively releases three times more CO for activation of nuclear factor kappa B (NF-kappa B) in living cells relative to wild-type polyhedra with Ru carbonyls.
  • Hiroyasu Tabe, Kenta Fujita, Satoshi Abe, Masahiko Tsujimoto, Takahiro Kuchimaru, Shinae Kizaka-Kondoh, Mikio Takano, Susumu Kitagawa, Takafumi Ueno
    Inorganic chemistry 54 1 215 - 20 2015年01月 [査読有り][通常論文]
     
    Protein crystals generally are stable solid protein assemblies. Certain protein crystals are suitable for use as nanovessels for immobilizing metal complexes. Here we report the preparation of ruthenium carbonyl-incorporated cross-linked hen egg white lysozyme crystals (Ru·CL-HEWL). Ru·CL-HEWL retains a Ru carbonyl moiety that can release CO, although a composite of Ru carbonyl-HEWL dissolved in buffer solution (Ru·HEWL) does not release CO. We found that treatment of cells with Ru·CL-HEWL significantly increased nuclear factor kappa B (NF-κB) activity as a cellular response to CO. These results demonstrate that Ru·CL-HEWL has potential for use as an artificial extracellular scaffold suitable for transport and release of a gas molecule.
  • Kenta Fujita, Yuya Tanaka, Takeya Sho, Shuichi Ozeki, Satoshi Abe, Tatsuo Hikage, Takahiro Kuchimaru, Shinae Kizaka-Kondoh, Takafumi Ueno
    Journal of the American Chemical Society 136 48 16902 - 8 2014年12月 [査読有り][通常論文]
     
    Protein cages have been utilized as templates in the development of biomaterials. Here we report protein engineering of the ferritin (Fr) cage for encapsulating carbon monoxide releasing molecules (CORMs) and release of CO gas which serves as a cell signaling molecule. The protein cages enable us to increase the half-life for CO release, providing a release rate that is 18-fold slower than the rate of a typical CORM, Ru(CO)3Cl(glycinate) (CORM-3). Moreover, the uptake ratio of the composite is about 4-fold greater than that of CORM-3. We found that these effects enhance the activation of nuclear factor κB 10-fold higher than CORM-3. The protein cage of Fr thus provides the basis for new CORMs that can be used for in vitro cell research.
  • Takahiro Kuchimaru, Takuya Hoshino, Tomoya Aikawa, Hisataka Yasuda, Tatsuya Kobayashi, Tetsuya Kadonosono, Shinae Kizaka-Kondoh
    CANCER SCIENCE 105 5 553 - 559 2014年05月 [査読有り][通常論文]
     
    Bone metastasis is a multistep process that includes cancer cell dissemination, colonization, and metastatic growth. Furthermore, this process involves complex, reciprocal interactions between cancer cells and the bone microenvironment. Bone resorption is known to be involved in both osteolytic and osteoblastic bone metastasis. However, the precise roles of the bone resorption in the multistep process of osteoblastic bone metastasis remain unidentified. In this study, we show that bone resorption plays important roles in cancer cell colonization during the initial stage of osteoblastic bone metastasis. We applied bioluminescence/X-ray computed tomography multimodal imaging that allows us to spatiotemporally analyze metastasized cancer cells and bone status in osteoblastic bone metastasis models. We found that treatment with receptor activator of factor-B ligand (RANKL) increased osteoblastic bone metastasis when given at the same time as intracardiac injection of cancer cells, but failed to increase metastasis when given 4days after cancer cell injection, suggesting that RANKL-induced bone resorption facilitates growth of cancer cells colonized in the bone. We show that insulin-like growth factor-1 released from the bone during bone resorption and hypoxia-inducible factor activity in cancer cells cooperatively promoted survival and proliferation of cancer cells in bone marrow. These results suggest a mechanism that bone resorption and hypoxic stress in the bone microenvironment cooperatively play an important role in establishing osteoblastic metastasis.
  • Takahiro Kuchimaru, Takuya Hoshino, Tomoya Aikawa, Hisataka Yasuda, Tatsuya Kobayashi, Tetsuya Kadonosono, Shinae Kizaka-Kondoh
    Cancer Science 105 5 553 - 559 2014年 [査読有り][通常論文]
     
    Bone metastasis is a multistep process that includes cancer cell dissemination, colonization, and metastatic growth. Furthermore, this process involves complex, reciprocal interactions between cancer cells and the bone microenvironment. Bone resorption is known to be involved in both osteolytic and osteoblastic bone metastasis. However, the precise roles of the bone resorption in the multistep process of osteoblastic bone metastasis remain unidentified. In this study, we show that bone resorption plays important roles in cancer cell colonization during the initial stage of osteoblastic bone metastasis. We applied bioluminescence/X-ray computed tomography multimodal imaging that allows us to spatiotemporally analyze metastasized cancer cells and bone status in osteoblastic bone metastasis models. We found that treatment with receptor activator of factor-κB ligand (RANKL) increased osteoblastic bone metastasis when given at the same time as intracardiac injection of cancer cells, but failed to increase metastasis when given 4 days after cancer cell injection, suggesting that RANKL-induced bone resorption facilitates growth of cancer cells colonized in the bone. We show that insulin-like growth factor-1 released from the bone during bone resorption and hypoxia-inducible factor activity in cancer cells cooperatively promoted survival and proliferation of cancer cells in bone marrow. These results suggest a mechanism that bone resorption and hypoxic stress in the bone microenvironment cooperatively play an important role in establishing osteoblastic metastasis. © 2014 The Authors.
  • Tetsuya Kadonosono, Etsuri Yabe, Tadaomi Furuta, Akihiro Yamano, Takuya Tsubaki, Takuya Sekine, Takahiro Kuchimaru, Minoru Sakurai, Shinae Kizaka-Kondoh
    PloS one 9 8 e103397  2014年 [査読有り][通常論文]
     
    Peptides that have high affinity for target molecules on the surface of cancer cells are crucial for the development of targeted cancer therapies. However, unstructured peptides often fail to bind their target molecules with high affinity. To efficiently identify high-affinity target-binding peptides, we have constructed a fluorescent protein scaffold, designated gFPS, in which structurally constrained peptides are integrated at residues K131-L137 of superfolder green fluorescent protein. Molecular dynamics simulation supported the suitability of this site for presentation of exogenous peptides with a constrained structure. gFPS can present 4 to 12 exogenous amino acids without a loss of fluorescence. When gFPSs presenting human epidermal growth factor receptor type 2 (HER2)-targeting peptides were added to the culture medium of HER2-expressing cells, we could easily identify the peptides with high HER2-affinity and -specificity based on gFPS fluorescence. In addition, gFPS could be expressed on the yeast cell surface and applied for a high-throughput screening. These results demonstrate that gFPS has the potential to serve as a powerful tool to improve screening of structurally constrained peptides that have a high target affinity, and suggest that it could expedite the one-step identification of clinically applicable cancer cell-binding peptides.
  • Ando H, Natsume A, Iwami K, Ohka F, Kuchimaru T, Kizaka-Kondoh S, Ito K, Saito K, Sugita S, Hoshino T, Wakabayashi T
    Biochemical and biophysical research communications 433 1 139 - 144 2013年03月 [査読有り][通常論文]
  • Ayano Takeuchi, Makihito Hori, Shinichi Sato, Hyun Seung Ban, Takahiro Kuchimaru, Shinae Kizaka-Kondoh, Takao Yamori, Hiroyuki Nakamura
    MedChemComm 3 11 1455 - 1461 2012年11月 [査読有り][通常論文]
     
    New YC-1 derivatives were synthesized and evaluated with HIF-1 transcriptional activity, and a potent derivative shows activity in vivo. Furthermore, for the mechanistic study of the YC-1 derivative, we evaluate the antiproliferative activity against 39 human cancer cell lines and the localization of active probes in cells. © The Royal Society of Chemistry.
  • Nishida C, Kusubata K, Tashiro Y, Gritli I, Sato A, Ohki-Koizumi M, Morita Y, Nagano M, Sakamoto T, Koshikawa N, Kuchimaru T, Kizaka-Kondoh S, Seiki M, Nakauchi H, Heissig B, Hattori K
    Blood 119 23 5405 - 5416 2012年06月 [査読有り][通常論文]
  • Kadonosono T, Kuchimaru T, Yamada S, Takahashi Y, Murakami A, Tani T, Watanabe H, Tanaka T, Hirota K, Inoue M, Tsukamoto T, Toyoda T, Urano K, Machida K, Eto T, Ogura T, Tsutsumi H, Ito M, Hiraoka M, Kondoh G, Kizaka-Kondoh S
    PloS one 6 11 e26640  2011年11月 [査読有り][通常論文]
  • Shinae Kizaka-Kondoh, Takahiro Kuchimaru, Tetsuya Kadonosono
    JOURNAL OF PHARMACOLOGICAL SCIENCES 115 4 440 - 445 2011年04月 [査読有り][通常論文]
     
    The microenvironment of solid tumors is characterized by low pO(2) that is well below physiological levels. Intratumoral hypoxia is a major factor contributing to cancer progression and is exacerbated as a result of oxygen consumption by rapidly proliferating tumor cells near blood vessels, poor lymphatic drainage resulting in high interstitial pressure, and irregular blood supply through immature tumor vasculature. Hypoxia-inducible factor-1 (HIF-1) is the main transcription factor that regulates cellular responses to hypoxia. Cellular changes induced by HIF-1 are extremely important targets for cancer therapy. Therefore, targeting strategies to counteract HIF-1-active cells are essential for cancer therapy. In this study, we introduce a novel strategy for targeting HIF-1-active cells.
  • Takahiro Kuchimaru, Fuminobu Sato, Souichi Tanaka, Isao Murata, Yushi Kato, Toshiyuki Iida
    Journal of Nuclear Science and Technology 47 12 1206 - 1210 2010年12月 [査読有り][通常論文]
     
    A cell chip was developed for the examination of biological damage of cells irradiated by high-energy alpha particles. A CR-39 track detector was employed as a chip substrate to identify high-energy charged particles traversing target cells. Moreover, the patterning of a photopolymer layer spatially controlled the cellular adhesiveness on the chip substrate. HeLa cancer cells were cultured on a micropatterned photopolymer layer. In this way, all the cells on the chip were individually addressed through the block number in the photopolymer pattern. The biocompatibility of the cell chip was examined through a viability test with fluophor reagent and measurement of the cell proliferation rate. HeLa cancer cells on the chip were irradiated with alpha particles and stained with a fluorescent probe molecule for DNA damage detection. The CR-39 substrate was etched by means of an alkali solution during cell incubation. The HeLa cells and alpha tracks were successfully observed by microscopy at once. It was confirmed that fluorescent spots corresponding to DNA damage were located in the direction of the major axis of oval alpha tracks. © Atomic Energy Society of Japan.
  • Takahiro Kuchimaru, Tetsuya Kadonosono, Shotaro Tanaka, Takashi Ushiki, Masahiro Hiraoka, Shinae Kizaka-Kondoh
    PLOS ONE 5 12 e15736  2010年12月 [査読有り][通常論文]
     
    Hypoxia-inducible factor (HIF) functions as a master transcriptional regulator for adaptation to hypoxia by inducing adaptive changes in gene expression for regulation of proliferation, angiogenesis, apoptosis and energy metabolism. Cancers with high expression of the alpha subunit of HIF (HIF alpha) are often malignant and treatment-resistant. Therefore, the development of a molecular probe that can detect HIF activity has great potential value for monitoring tumor hypoxia. HIF prolyl hydroxylases (HPHDs) act as oxygen sensors that regulate the fate of HIF alpha protein through its oxygen-dependent degradation (ODD) domain. We constructed a recombinant protein PTD-ODD-HaloTag (POH) that is under the same ODD regulation as HIF alpha and contains protein transduction domain (PTD) and an interchangeable labeling system. Administration of near-infrared fluorescently labeled POH (POH-N) to mouse models of cancers allowed successful monitoring of HIF-active regions. Immunohistochemical analysis for intratumoral localization of POH probe revealed its specificity to HIF-active cells. Furthermore, lack of the PTD domain or a point mutation in the ODD domain abrogated the specificity of POH-N to HIF-active cells. Overall results indicate that POH is a practical probe specific to HIF-active cell in cancers and suggest its large potential for imaging and targeting of HIF-related diseases.
  • Yusuke Aoi, Takahiro Kuchimaru, Daisuke Maki, Fuminobu Sato, Toshiji Ikeda, Yushi Kato, Takayoshi Yamamoto, Toshiyuki Iida
    Japanese Journal of Applied Physics 48 5 0560011 - 0560014 2009年05月 [査読有り][通常論文]
     
    X-ray microbeam transport in radiophotoluminescent (RPL) glass was examined by fluorescence microscopy. A silver-activated phosphate glass dosimeter plate was irradiated with an X-ray microbeam emitted from a 50 kV micro focus X-ray tube equipped with a glass capillary. A RPL region (wavelength ~650 nm) was observed with UV light excitation along the X-ray microbeam track in the glass dosimeter plate. The divergence of the RPL track was small owing to the X-ray energy dependence of the divergence of the microbeam, which was determined from the critical angle of the X-ray reflection on the inner wall of the glass capillary. The image of the RPL track was analyzed using photon transport and light trace simulation codes. It was confirmed by comparison between the RPL observation and simulation results that the RPL image in the silver-activated phosphate glass accurately projected the energy-deposited track along the X-ray microbeam. © 2009 The Japan Society of Applied Physics.
  • T. Kuchimaru, F. Sato, Y. Aoi, T. Fujita, T. Ikeda, K. Shimizu, Y. Kato, T. Iida
    Radiation and Environmental Biophysics 47 4 535 - 540 2008年11月 [査読有り][通常論文]
     
    To identify individual cells exposed to a X-ray microbeam in a cell population, we developed a biocompatible microchamber-array chip using UV lithography of photopolymer SU-8. The center-to-center distance between microchambers is 50 μm including a wall of 15 μm height. Using the microchamber-array chip, we performed tracking of individual exposed cells. Sample cells loaded in a microchamber array were selectively irradiated with the X-ray microbeam under microscopic observation. All the irradiated cells were indexed by the array arrangement of the microchambers. For about 24 h of post-irradiation incubation, the irradiated cells were identified successfully by time-lapse observation. In addition, the induction of radiation effects was observed in identified cells using immunofluorescence. © 2008 The Author(s).
  • T. Kuchimaru, F. Sato, K. Honda, Y. Kato, T. Iida
    Radiation Measurements 43 1 S125 - S127 2008年08月 [査読有り][通常論文]
     
    Three-dimensional (3D) image processing for analysis of track etching in a Solid State Nuclear Track Detector (SSNTD) was performed. A SSNTD in contact with boron nitride films was exposed to epithermal and thermal neutrons. In chemical etching process, sequential images of etch-pits on the surface of the SSNTD were obtained with a developed observation system. By use of a digital-image-processing technique, 3D images were reconstructed from data on sequential surface images of the track detector for particles released due to 10 B (n, α)7 Li reactions. © 2008 Elsevier Ltd. All rights reserved.
  • Fuminobu Sato, T. Kuchimaru, T. Ikeda, K. Shimizu, Y. Kato, T. Yamamoto, T. Iida
    Radiation Measurements 43 2-6 912 - 916 2008年02月 [査読有り][通常論文]
     
    A commercially available radiophotoluminescent (RPL) glass dosimeter was used for the profile measurement of an X-ray microbeam. The X-ray microbeam of 10 μ m diameter was produced by a microfocus X-ray tube and a glass capillary. An RPL glass plate of 1 mm thickness was irradiated with the X-ray microbeam. After the irradiation, the RPL image for the X-ray microbeam on the glass plate was observed with a conventional fluorescence microscope. In the RPL image analysis, the RPL photons around 600 nm wavelength were detected by exposure to UV photons in the wavelength range from 330 to 380 nm. The profile of the X-ray microbeam was well measured with the RPL glass dosimeter and the RPL intensity was proportional to the X-ray fluence. Normal human fibroblasts cells, AG01522B, were grown on the RPL glass plate for single cell irradiation. A pre-selected living cell was irradiated by the X-ray microbeam and the absorbed dose was estimated to be about 3 Gy. After the irradiation, the targeting cell was stained with anti-γ-H2AX antibody, Alexa Fluor® 488 and propidium iodide, to visualize the effects on DNA double strand breaks in the nucleus of the cell. In fluorescence microscopy, foci in the nucleus corresponding to the phosphorylation of serine 139 of H2AX and the X-ray beam spot were simultaneously observed at the position of the targeting cell. © 2007 Elsevier Ltd. All rights reserved.
  • Fuminobu Sato, Takahiro Kuchimaru, Yushi Kato, Toshiyuki Iida
    Japanese Journal of Applied Physics 47 1 269 - 272 2008年01月 [査読有り][通常論文]
     
    An in-situ observation system was developed to record time-lapse images of etch pits formed on the surface of a solid-state nuclear track detector (SSNTD), CR-39. The CR-39 track detector was irradiated with alpha and Li particles produced by a neutron capture reaction, 10B(n,α)7Li. The time-lapse images of the etched surface of the detector were carefully observed to elucidate the etch pit formation process. Pit-evolution images were constructed by digital image processing of the data of the time-lapse images. In addition, a finite element model of SSNTD was used to simulate the etch pit formation. Pit-evolution image analysis and computational simulation were performed to reveal the etch pit formation obtained via the incidence angle of energetic particles on a CR-39. © 2008 The Japan Society of Applied Physics.
  • T. Kuchimaru, F. Sato, Y. Higashino, K. Shimizu, Y. Kato, T. Iida
    IEEE Transactions on Nuclear Science 53 3 1363 - 1366 2006年06月 [査読有り][通常論文]
     
    A tabletop micro X-ray beam irradiation system has been developed to study radiation effects on living cells. The microbeam system is composed of a micro focus X-ray tube, a capillary for X-ray guide, an X-ray semiconductor detector for fluorescent X-ray analysis, and an inverted microscope. A beam profile measurement was performed and the size of the focused beam was 10 μm in diameter [full-width at half-maximum (FWHM)] with a divergence of 5.1 mrad. The data on the microbeam measurement were approximately in agreement with a simulation of X-ray optical traces for the capillary. In addition, the microdosimetric characterization for single cell irradiation was performed with the beam profile measurement and a photon-electron transport simulation. The maximum of the dose rate for the sample cells set in the system was estimated to be 0.05 Gy/s. In a preliminary experiment, single yeast cells were irradiated with the X-ray microbeams, and the data on the survival rate of the cell samples for the X-ray doses were obtained. Then, the microbeam irradiation system is useful to investigate the radiation effects on the living cells. © 2006 IEEE.
  • T. Kuchimaru, F. Sato, Y. Higashino, K. Shimizu, Y. Kato, T. Iida
    IEEE Nuclear Science Symposium Conference Record 5 3005 - 3009 2005年 [査読有り][通常論文]
     
    A. tabletop micro X-ray beam irradiation system has been developed to research radiation effects on living cells. The microbeam system is composed of a micro focus X-ray tube, a capillary for X-ray guide, an X-ray semiconductor detector for fluorescent X-ray analysis and an inverted microscope. A beam profile measurement was performed and the size of the focused beam was 10 μm in diameter (FWHM) with the divergence of 5.1 mrad. The data on the microbeam measurement was approximately agreement with a simulation of X-ray optical traces for the designed capillary. In addition, the microdosimetric characterization for the single cell irradiation was performed with the beam profile measurement and a photon-electron transport simulation. The maximum of the dose rate for the sample cells set in the system was estimated to be 0.05 Gy/s. In a preliminary experiment, single yeast cell was irradiated with the X-ray microbeam, and the data on the survival rate of the cell samples for the X-ray doses was obtained. Then, the microbeam irradiation system is useful to research the radiation effects to the living cells. © 2005 IEEE.

MISC

  • Takuya Tsubaki, Tetsuya Kadonosono, Tadashi Shiozawa, Takahiro Kuchimaru, Takashi Ushiki, Masayoshi Masuko, Shinae Kizaka-Kondoh BLOOD 128 (22) 2016年12月 [査読無し][通常論文]
  • 石田 忠, 島本 拓弥, 口丸 高弘, 近藤 科江, 小俣 透 ロボティクス・メカトロニクス講演会講演概要集 2016 (0) 2P1 -19a1 2016年 [査読無し][通常論文]
     
    <p>We propose a micro-channel device to collect cancer cells with high migrating capability, which could cause metastasis. In order to collect cells attached on extracellular matrix, focal adhesions between cells and extracellular matrix should be broken. The desmosomes can be mechanically broken by using extension of cell culture surface. For this purpose, the device locally has a balloon under a cell culture surface. The extension rate of the cell culture surface is 40% with 300 μL water. Using the device, we could detach the high-migration cell, which has net displacement of 150 μm and total path of 250 μm, and collect it by the flow of cultural medium.</p>
  • 島本 拓弥, 石田 忠, 口丸 高弘, 近藤 科江, 小俣 透 「センサ・マイクロマシンと応用システム」シンポジウム論文集 電気学会センサ・マイクロマシン部門 [編] 32 1 -5 2015年10月 [査読無し][通常論文]
  • 石田 忠, 尾崎 展也, 口丸 高弘, 近藤 科江, 小俣 透 ロボティクス・メカトロニクス講演会講演概要集 2015 (0) _1P1 -L09_1-_1P1-L09_3 2015年 [査読無し][通常論文]
     
    Cancer cells inside tumors are under low glucose and oxygen concentration, which increase the migration capability and are assumed to be an origin of cancer metastasis. However, the cellular level studies of cancer cells inside tumors are difficult due to the limitation of the conventional biological approaches, which cannot generate the conditions inside tumors. In order to overcome the difficulty, we design and fabricate a microfluidic device that can control the glucose and oxygen concentrations inside a micro chamber. We achieved the concentration gradients similar to the gradient inside a tumor.
  • 高木 哲史, 石田 忠, 口丸 高弘 「センサ・マイクロマシンと応用システム」シンポジウム論文集 電気学会センサ・マイクロマシン部門 [編] 30 1 -4 2013年11月 [査読無し][通常論文]
  • Chiemi Nishida, Kaori Sato-Kusubata, Yoshihiko Tashiro, Ismael Gritli, Aki Sato, Makiko Ohki-Koizumi, Yohei Morita, Makoto Nagano, Takeharu Sakamoto, Naohiko Koshikawa, Takahiro Kuchimaru, Shinae Kizaka-Kondo, Motoharu Seiki, Hiromitsu Nakauchi, Heissig Beate, Koichi Hattori BLOOD 120 (21) 2012年11月 [査読無し][通常論文]
  • MT1-MMPはHIF介在性サイトカイン遺伝子転写調節により造血において重要な役割を果たす(MT1MMP plays a critical role in hematopoiesis by regulating HIF-mediated cytokine gene transcription)
    Nishida Chiemi, Kusubata Kaori, Tashiro Yoshihiko, Ismael Gritli, Sato Aki, Koizumi Makiko, Morita Yohei, Nagano Makoto, Sakamoto Takeharu, Koshikawa Naohiko, Kuchimaru Takahiro, Kondoh Shinae, Seiki Motoharu, Nakauchi Hiromitsu, Beate Heissig, Hattori Koichi 臨床血液 53 (9) 1104 -1104 2012年09月 [査読無し][通常論文]
  • Shinae Kondoh, Takahiro Kuchimaru, Tetsuya Kadonosono JOURNAL OF PHARMACOLOGICAL SCIENCES 118 23P -23P 2012年 [査読無し][通常論文]
  • Youshi Fujita, Takahiro Kuchimaru, Tetsuya Kadonosono, Shotaro Tanaka, Yoshiki Hase, Hidekazu Tomimoto, Masahiro Hiraoka, Shinae Kizaka-Kondoh, Masafumi Ihara, Masafumi Ihara, Ryosuke Takahashi PLoS ONE 7 (10) e48051 2012年 [査読有り][通常論文]
     
    Within the ischemic penumbra, blood flow is sufficiently reduced that it results in hypoxia severe enough to arrest physiological function. Nevertheless, it has been shown that cells present within this region can be rescued and resuscitated by restoring perfusion and through other protective therapies. Thus, the early detection of the ischemic penumbra can be exploited to improve outcomes after focal ischemia. Hypoxia-inducible factor (HIF)-1 is a transcription factor induced by a reduction in molecular oxygen levels. Although the role of HIF-1 in the ischemic penumbra remains unknown, there is a strong correlation between areas with HIF-1 activity and the ischemic penumbra. We recently developed a near-infrared fluorescently labeled-fusion protein, POH-N, with an oxygen-dependent degradation property identical to the alpha subunit of HIF-1. Here, we conduct in vivo imaging of HIF-active regions using POH-N in ischemic brains after transient focal cerebral ischemia induced using the intraluminal middle cerebral artery occlusion technique in mice. The results demonstrate that POH-N enables the in vivo monitoring and ex vivo detection of HIF-1-active regions after ischemic brain injury and suggest its potential in imaging and drug delivery to HIF-1-active areas in ischemic brains. © 2012 Fujita et al.
  • 近赤外光プローブを用いたHIF活性細胞のIn vivoイメージング
    口丸 高弘, 門之園 哲哉, 牛木 隆志, 近藤 科江, 平岡 真寛 JSMI Report 3 (2) 152 -152 2010年05月 [査読無し][通常論文]
  • Shinae Kondoh, Takahiro Kuchimaru, Tetsuya Kadonosono, Masahiro Hiraoka JOURNAL OF PHARMACOLOGICAL SCIENCES 112 26P -26P 2010年 [査読無し][通常論文]

受賞

  • 2019年01月 コニカミノルタ画像科学奨励賞
     
    受賞者: 口丸 高弘
  • 2017年09月 日本癌学会 日本癌学会奨励賞
     
    受賞者: 口丸 高弘
  • 2017年02月 東京工業大学 手島精一記念若手研究賞(藤野・中村賞)
     
    受賞者: 口丸 高弘
  • 2014年05月 日本分子イメージング学会 第9回日本分子イメージング学会総会・学術集会 最優秀発表賞
     
    受賞者: 口丸 高弘


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