Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/adbi.202300159

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Abstract

Tumors contain various stromal cells, such as immune cells, endothelial cells, and fibroblasts, which contribute to the development of a tumor‐specific microenvironment characterized by hypoxia and inflammation, and are associated with malignant progression. In this study, we investigated the activity of intratumoral hypoxia‐inducible factor (HIF), which functions as a master regulator of the cellular response to hypoxia and inflammation. We constructed the HIF activity‐monitoring reporter gene hypoxia‐response element‐Venus‐Akaluc (HVA) that expresses the green fluorescent protein Venus and modified firefly luciferase Akaluc in a HIF activity‐dependent manner, and created transgenic mice harboring HVA transgene (HVA‐Tg). In HVA‐Tg, HIF‐active cells can be visualized using AkaBLI, an ultra‐sensitive in vivo bioluminescence imaging technology that produces an intense near‐infrared light upon reaction of Akaluc with the D‐luciferin analog AkaLumine‐HCl. By orthotopic transplantation of E0771, a mouse triple negative breast cancer cell line without a reporter gene, into HVA‐Tg, we succeeded in noninvasively monitoring bioluminescence signals from HIF‐active stromal cells as early as 8 days after transplantation. The HIF‐active stromal cells initially clustered locally and then spread throughout the tumors with growth. Immunohistochemistry and flow cytometry analyses revealed that CD11b⁺F4/80⁺ macrophages were the predominant HIF‐active stromal cells in E0771 tumors. These results indicate that HVA‐Tg is a useful tool for spatiotemporal analysis of HIF‐active tumor stromal cells, facilitating investigation of the roles of HIF‐active tumor stromal cells in tumor growth and malignant progression.

DOI： 10.1111/cas.15907

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

Triple-negative breast cancer (TNBC) is an aggressive and highly heterogenous disease with no well-defined therapeutic targets. Treatment options are thus limited and mortality is significantly higher compared with other breast cancer subtypes. Mammary gland tissue-resident macrophages (MGTRMs) are found to be the most abundant stromal cells in early TNBC before angiogenesis. We therefore aimed to explore novel therapeutic approaches for TNBC by focusing on MGTRMs. Local depletion of MGTRMs in mammary gland fat pads the day before TNBC cell transplantation significantly reduced tumor growth and tumor-associated macrophage (TAM) infiltration in mice. Furthermore, local depletion of MGTRMs at the site of TNBC resection markedly reduced recurrence and distant metastases, and improved chemotherapy outcomes. This study demonstrates that MGTRMs are a major TAM resource and play pivotal roles in the growth and malignant progression of TNBC. The results highlight a possible novel anti-cancer approach targeting tissue-resident macrophages.

DOI： 10.1038/s42003-023-04525-7

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Publishing type：Research paper (international conference proceedings)  

DOI： 10.1117/12.2649659

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Authorship：Last author,　Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

Abstract

Cancer recurrence due to tumor cell quiescence after therapy and long-term remission is associated with cancer-related death. Previous studies have used cell models that are unable to return to a proliferative state; thus, the transition between quiescent and proliferative states is not well understood. Here, we report monolayer cancer cell models wherein the human non-small cell lung carcinoma cell line H2228 and pancreatic cancer cell line AsPC-1 can be reversibly induced to a quiescent state under hypoxic and serum-starved (HSS) conditions. Transcriptome and metabolome dual-omics profiles of these cells were compared with those of the human lung adenocarcinoma cell line A549, which was unable to enter a quiescent state under HSS conditions. The quiescence-inducible cells had substantially lower intracellular pyruvate and ATP levels in the quiescent state than in the proliferative state, and their response to sudden demand for energy was dramatically reduced. Furthermore, in quiescence-inducible cells, the transition between quiescent and proliferative states of these cells was regulated by the balance between the proliferation-promoting Ras and Rap1 signaling and the suppressive AGE/RAGE signaling. These cell models elucidate the transition between quiescent and proliferative states, allowing the development of drug-screening systems for quiescent tumor cells.

DOI： 10.1038/s41598-022-14272-0

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Other Link： https://www.nature.com/articles/s41598-022-14272-0

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/978-1-0716-2473-9_22.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SAGE Publications  

Photoacoustic (PA) technology can be used for non-invasive imaging of blood vessels. In this paper, we report on our prototype PA imaging system with a newly designed ultrasound sensor and its visualization performance of microvascular in animal. We fabricated an experimental system for animals using a high-frequency sensor. The system has two modes: still image mode by wide scanning and moving image mode by small rotation of sensor array. Optical test target, euthanized mice and rats, and live mice were used as objects. The results of optical test target showed that the spatial resolution was about two times higher than that of our conventional prototype. The image performance in vivo was evaluated in euthanized healthy mice and rats, allowing visualization of detailed blood vessels in the liver and kidneys. In tumor-bearing mice, different results of vascular induction were shown depending on the type of tumor and the method of transplantation. By utilizing the video imaging function, we were able to observe the movement of blood vessels around the tumor. We have demonstrated the feasibility of the system as a less invasive animal experimental device, as it can acquire vascular images in animals in a non-contrast and non-invasive manner.

DOI： 10.1177/01617346221099201

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Other Link： http://journals.sagepub.com/doi/full-xml/10.1177/01617346221099201

Language：English   Publishing type：Research paper (scientific journal)  

In this study, we proposed a droplet-based valveless microfluidic system that has the necessary functions to perform the binding, washing, eluting, and collecting processes of phage-display screening against spheroids, which can be expected to present a similar repertoire and number of membrane proteins as in vivo. Although spheroids have much larger sizes than single cells, spheroids are difficult to manipulate through manual operation. The proposed microfluidic system actively controls the position and velocity of droplets using a camera, three air pumps, and three liquid pumps to perform the processes for phage-display screening. The cross section of the microchannel is large in width and height for the passage of spheroids. Valves that can close such a large cross-sectional microchannel are not readily available. Thus, we proposed valveless flow control using liquid pumps. In addition, the proposed microfluidic system involves complex flow channels with airflow subchannels to perform phage-display screening. For washing, nonspecific-binding phages remaining in the flow channels must be minimized. The proposed microfluidic system can perform selective blocking and flush washing. Selective blocking can prevent the airflow channels from becoming hydrophilic with blocking liquid, and flush washing can flush phages remaining in the flow channel. We experimentally verified the functions of the developed microfluidic device based on the proposed system.

DOI： 10.1063/5.0085459

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publishing type：Research paper (scientific journal)  

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Authorship：Last author   Language：English   Publishing type：Research paper (scientific journal)  

Small antibody mimetics that contain high-affinity target-binding peptides can be lower cost alternatives to monoclonal antibodies (mAbs). We have recently developed a method to create small antibody mimetics called FLuctuation-regulated Affinity Proteins (FLAPs), which consist of a small protein scaffold with a structurally immobilized target-binding peptide. In this study, to further develop this method, we established a novel screening system for FLAPs called monoclonal antibody-guided peptide identification and engineering (MAGPIE), in which a mAb guides selection in two manners. First, antibody-guided design allows construction of a peptide library that is relatively small in size, but sufficient to identify high-affinity binders in a single selection round. Second, in antibody-guided screening, the fluorescently labeled mAb is used to select mammalian cells that display FLAP candidates with high affinity for the target using fluorescence-activated cell sorting. We demonstrate the reliability and efficacy of MAGPIE using daclizumab, a mAb against human interleukin-2 receptor alpha chain (CD25). Three FLAPs identified by MAGPIE bound CD25 with dissociation constants of approximately 30 nM as measured by biolayer interferometry without undergoing affinity maturation. MAGPIE can be broadly adapted to any mAb to develop small antibody mimetics.

DOI： 10.1038/s41598-021-01603-w

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Publishing type：Research paper (scientific journal)   Publisher：OAE Publishing Inc.  

DOI： 10.20517/2394-47.2020.96

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Authorship：Last author   Language：English   Publishing type：Research paper (scientific journal)  

The current standard murine model of bone metastasis by using intracardiac injection (IC) has some limitations despite the great utility of this model. This fact emphasizes the need for a new murine model to accelerate basic research of bone metastasis. The present protocol provides instructions on caudal artery (CA) injection that is an easy-to-use method to reliably construct a murine bone metastasis model with a variety type of cancer cell lines. Bioluminescence imaging visualized that cancer cells injected via the caudal artery in the tail were efficiently delivered to a hind limb bone, where it is a common site affected with bone metastasis in mice. CA injection rarely causes stress-induced acute death in mice and enables us to inject a large number of cancer cells, thereby greatly increasing the frequency of bone metastasis in hind limb bones. Importantly, CA injection is technically as easy as tail vein injection and causes no lethal stress, indicating that it is a model that also contributes to animal welfare. CA injection model, therefore, could represent a powerful tool for many researchers to study molecular mechanisms of bone metastasis in mice.

DOI： 10.1007/978-1-0716-1258-3_4

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：OAE Publishing Inc.  

DOI： 10.20517/2394-4722.2020.104

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

Target-binding small proteins are promising alternatives to conventional monoclonal antibodies (mAbs), offering advantages in terms of tissue penetration and manufacturing costs. Recently, a design strategy to create small proteins called fluctuation-regulated affinity proteins (FLAPs) consisting of a structurally immobilized peptide from the complementarity-determining region (CDR) loops of mAbs (CDR-derived peptide) and a protein scaffold was developed. Because mAb paratopes are usually composed of multiple CDRs, FLAPs with multiple binding peptides may have an enhanced target-binding capability. Here, a strategy to create FLAPs bearing dual CDR-derived peptides (D-FLAPs) using the anti-human epithelial growth factor receptor type 2 (HER2) mAb trastuzumab as a basis is developed. Computationally selected CDR-derived peptides are first grafted onto two adjacent loops of the fibronectin type III domain (FN3) scaffold, yielding 80 D-FLAP candidates. After computational screening based on their similarity to the parental mAb with regard to the conformation of paratope residues, two candidates are selected. After further evaluation with ELISA, one D-FLAP with HYTTPP and GDGFYA peptides from CDR-L3 and CDR-H3 of the parental mAb, respectively, is found to bind HER2 with a dissociation constant of 58 nm. This method applies to various mAb drugs and allows the rational design of small protein alternatives.

DOI： 10.1002/biot.202000078

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Publishing type：Research paper (scientific journal)  

DOI： 10.3390/MI11050480

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DOI： 10.1039/d0ra00427h

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Authorship：Last author   Publishing type：Research paper (international conference proceedings)  

DOI： 10.1248/yakushi.19-00187-4

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Language：English  

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.

DOI： 10.1016/j.bmc.2019.115207

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3987/COM-19-S(F)47

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Authorship：Corresponding author   Language：English  

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.

DOI： 10.1038/s41598-020-57713-4

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Language：English  

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.

DOI： 10.1016/j.neuroscience.2019.06.025

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：American Chemical Society (ACS)  

DOI： 10.1021/acs.bioconjchem.9b00088

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：The Chemical Society of Japan  

電通大発のNIR生物発光基質アカルミネの構造に含窒素複素環構造を導入して、in vivoイメージングに有用な水溶性が高く、NIR発光性を維持した新規アナログの開発に成功した。

DOI： 10.1246/bcsj.20180350

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

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.

DOI： 10.3390/mi10020116

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/mi10010041

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Language：English   Publisher：日本癌学会  

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1111/cas.13702

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1038/s41467-018-05366-3

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Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.18632/oncotarget.24696

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Language：English   Publisher：PEPTIDE SCIENCE 2017 (I. Fujii (ed.))  

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Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1126/science.aaq1067

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.18632/oncotarget.24359

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Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese   Publisher：次世代がん治療 発症・転移メカニズムからがん免疫療法・ウィルス療法、診断法まで  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1002/pro.3089

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Language：Japanese   Publisher：バイオサイエンスとインダストリー  

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1038/srep34311

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Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese   Publisher：(一社)日本核医学会  

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Language：English  

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.

DOI： 10.3390/mi7090155

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Language：Japanese   Publishing type：Research paper (scientific journal)  

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Authorship：Last author,　Corresponding author   Language：English  

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.

DOI： 10.1111/cas.12982

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3769/radioisotopes.65.247

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1038/ncomms11856

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/cgt.2016.13.

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Language：English   Publishing type：Research paper (scientific journal)  

PURPOSE: Hypoxia-inducible factor-1 (HIF-1) plays an important role in malignant tumour progression. For the imaging of HIF-1-active tumours, we previously developed a protein, POS, which is effectively delivered to and selectively stabilized in HIF-1-active cells, and a radioiodinated biotin derivative, (3-(123)I-iodobenzoyl)norbiotinamide ((123)I-IBB), which can bind to the streptavidin moiety of POS. In this study, we aimed to investigate the feasibility of the pretargeting method using POS and (123)I-IBB for rapid imaging of HIF-1-active tumours. METHODS: Tumour-implanted mice were pretargeted with POS. After 24 h, (125)I-IBB was administered and subsequently, the biodistribution of radioactivity was investigated at several time points. In vivo planar imaging, comparison between (125)I-IBB accumulation and HIF-1 transcriptional activity, and autoradiography were performed at 6 h after the administration of (125)I-IBB. The same sections that were used in autoradiographic analysis were subjected to HIF-1alpha immunohistochemistry. RESULTS: (125)I-IBB accumulation was observed in tumours of mice pretargeted with POS (1.6%ID/g at 6 h). This result is comparable to the data derived from (125)I-IBB-conjugated POS-treated mice (1.4%ID/g at 24 h). In vivo planar imaging provided clear tumour images. The tumoral accumulation of (125)I-IBB significantly correlated with HIF-1-dependent luciferase bioluminescence (R=0.84, p<0.01). The intratumoral distribution of (125)I-IBB was heterogeneous and was significantly correlated with HIF-1alpha-positive regions (R=0.58, p<0.0001). CONCLUSION: POS pretargeting with (123)I-IBB is a useful technique in the rapid imaging and detection of HIF-1-active regions in tumours.

DOI： 10.1007/s00259-010-1467-4

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<Purpose> : We aimed to evaluate the feasibility of using streptavidin–biotin-based pretargeting for positron emission tomography (PET) imaging of hypoxia-inducible factor (HIF)-1-active tumors. <Procedures> : We used POS, a genetically engineered form of streptavidin that selectively stabilizes in HIF-1-active cells, and (4-[18]F-fluorobenzoyl)norbiotinamide ([18]F-FBB), a radiolabeled biotin derivative, for performing a biodistribution study and for PET imaging. The tumoral [18]F-FBB accumulation was compared to the HIF-1-dependent luciferase bioluminescence and HIF-1α immunohistochemical signal. <Results> : [18]F-FBB accumulation was observed in POS-pretargeted tumors in mice (2.85 ± 0.55% injected dose per gram at 3 h), and clear PET images were obtained at the same time point. The tumoral [18]F-FBB accumulation positively correlated with luciferase bioluminescence (R = 0.72, P < 0.05), and most of the area showing [18]F-FBB accumulation corresponded to HIF-1α-positive areas. <Conclusion> : Pretargeting with POS and [18]F-FBB is an effective approach for PET imaging of HIF-1-active areas in tumors.

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Language：Japanese   Publisher：(公社)日本生化学会  

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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.

DOI： 10.1039/c5mb00327j

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DOI： 10.1242/bio.010918

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DOI： 10.1038/srep10657

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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.

DOI： 10.1038/srep09898

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

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.

DOI： 10.1016/j.jconrel.2015.01.011

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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-κB) in living cells relative to wild-type polyhedra with Ru carbonyls.

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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.

DOI： 10.1021/ic502159x

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DOI： 10.3390/mi6040409

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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.

DOI： 10.1021/ja508938f

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Language：Japanese   Publisher：羊土社  

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Language：Japanese   Publisher：羊土社  

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Language：English   Publisher：(一社)日本癌学会  

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DOI： 10.1007/s10147-013-0568-z

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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.

DOI： 10.1371/journal.pone.0103397

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1111/cas.12391

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：がんの分子イメージング  

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DOI： 10.1007/s11307-013-0647-6

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DOI： 10.1089/ars.2011.4430

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DOI： 10.1016/j.nucmedbio.2012.12.008

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DOI： 10.1016/j.bbrc.2013.02.044

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DOI： 10.1039/c2md20134h

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DOI： 10.1182/blood-2011-11-390849

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DOI： 10.1371/journal.pone.0048051

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Language：Japanese   Publisher：(一社)日本核医学会  

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DOI： 10.1182/blood-2011-11-390849

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DOI： 10.1248/cpb.60.402

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DOI： 10.1021/bc2004704

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DOI： 10.1155/2012/262741

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DOI： 10.1111/j.1349-7006.2011.02057.x

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DOI： 10.1371/journal.pone.0026640

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DOI： 10.1007/s11307-010-0418-6

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DOI： 10.1254/jphs.10R20FM

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DOI： 10.1161/STROKEAHA.110.596379

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Language：Japanese   Publisher：(一社)日本神経学会  

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DOI： 10.1371/journal.pone.0015736

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DOI： 10.1007/s00259-010-1467-4

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DOI： 10.1073/pnas.1002372107

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DOI： 10.1371/journal.pone.0011114

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DOI： 10.1016/j.jconrel.2010.01.023

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DOI： 10.1152/ajpregu.00732.2009

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DOI： 10.1242/jcs.052498

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DOI： 10.1091/mbc.E09-05-0439

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DOI： 10.1016/j.biomaterials.2009.05.046

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DOI： 10.1038/mt.2009.119

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DOI： 10.1111/j.1349-7006.2009.01195.x

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DOI： 10.1016/j.addr.2009.01.006

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DOI： 10.1007/s00125-009-1331-x

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DOI： 10.2967/jnumed.108.061119

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DOI： 10.1158/1078-0432.CCR-08-2267

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DOI： 10.1074/jbc.M806653200

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DOI： 10.1117/12.808380

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DOI： 10.1293/tox.22.93

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Japanese Journal of Cancer and Chemotherapy Publishers Inc.  

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Youshi Fujita, Takashi Ushiki, Masafumi Ihara, Hidefumi Ito, Shinae Kizaka-Kondoh, Ryosuke Takahashi, 2009, &#039;Intravenously administered bone marrow cells alleviates white matter lesions in a model of chronic cerebral hypoperfusion&#039;, &lt;i&gt;Neuroscience Research&lt;/i&gt;, vol. 65, p. S123

DOI： 10.1016/j.neures.2009.09.586

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DOI： 10.1111/j.1349-7006.2008.00943.x

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Language：Japanese   Publisher：(一社)日本放射線影響学会  

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bbrc.2008.02.041

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DOI： 10.1042/BJ20070824

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DOI： 10.1021/bc7001665

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Language：English   Publishing type：Research paper (international conference proceedings)  

DOI： 10.1002/pat.1218

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Language：Japanese   Publisher：(一社)日本放射線影響学会  

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Authorship：Last author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/sj.onc.1210556

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DOI： 10.1016/j.bbrc.2007.06.149

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DOI： 10.1111/j.1349-7006.2007.00537.x

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DOI： 10.1158/0008-5472.CAN-06-2355

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Language：Japanese   Publisher：Japan Society for Bioscience, Biotechnology, and Agrochemistry  

DOI： 10.1271/kagakutoseibutsu1962.45.779

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J-GLOBAL

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J-GLOBAL

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.febslet.2006.09.025

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DOI： 10.1080/02841860500486648

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The genomic information obtained through the human genome project has been accelerating the analysis of the functions of various disease relevant genes. The high molecular weight biomolecules, including oligonucleotides, antisense nucleotides, small interference RNA and peptides, as well as genes (cDNA) and proteins, are becoming increasingly important for the development of molecular therapies. However, the potential of such information-rich macromolecules for therapeutic use has been limited by the poor permeability across the lipid bilayer of the cellular plasma membrane. Over the past decade, a unique activity of oligopeptides, known as protein transduction domains (PTDs) or cell penetrating peptides (CPPs), has made it possible to transduce biologically active macromolecules into living cells in vitro by conjugating a PTD to the desired macromolecule. Furthermore, this activity has also enabled the systemic delivery of bioactive macromolecules to all tissues in living animals. However, we are now confronted with the next difficulty delivering the macromolecules specifically to the therapeutic targets in vivo. In this review, we focus on the application of PTD to develop antitumor macromolecules and introduce several representative strategies to discriminate between tumor and normal tissue. In addition, we discuss the unique characteristics of breast cancer, which are expected to facilitate the application of PTD to develop novel protein therapy for breast cancer.

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DOI： 10.1210/en.2005-0679

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DOI： 10.1128/MCB.20.9.3266-3273.2000

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Language：English   Publisher：Noncegmented Negative Stranded Viruses  

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Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)  

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Authorship：Last author   Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)  

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Language：Japanese   Publisher：(一社)日本女性科学者の会  

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Language：Japanese   Publisher：The Japan Society of Vacuum and Surface Science  

DOI： 10.1380/vss.65.375

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：一般社団法人 日本女性科学者の会  

日本女性科学者の会では2020年9月13日、第13回学術大会として、SJWS奨励賞・功労賞贈呈式、文部科学大臣賞伝達式、および受賞記念講演会を行った。東京工業大学での現地開催と、Zoom機能を使ってweb配信するハイブリッドミーティングとしたため、その準備状況とノウハウなどをまとめる。

DOI： 10.5939/sjws.21005

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Other Link： https://ndlsearch.ndl.go.jp/books/R000000004-I031514168

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Other Link： https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20H02862/

Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese   Publisher：[東京] : Institute of Electrical Engineers of Japan  

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Other Link： https://ndlsearch.ndl.go.jp/books/R000000004-I031480595

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近年、標的結合小型蛋白質のデザイン技術の高度化と汎用化が進められている。筆者らが開発してきた抗体構造中の抗原結合ペプチドを利用した標的結合小型蛋白質のデザインについて、以下の項目に分けて概説した。1)標的結合小型蛋白質のデザインコンセプト、2)fluctuation-regulated affinity protein(FLAP)の創出、3)FLAPの高度化と医療応用、として述べた。

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Other Link： https://search.jamas.or.jp/default/link?pub_year=2020&ichushi_jid=J01459&link_issn=&doc_id=20200204230007&doc_link_id=1390565134825135616&url=https%3A%2F%2Fcir.nii.ac.jp%2Fcrid%2F1390565134825135616&type=CiNii&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00003_3.gif

Language：Japanese   Publisher：東京 : 上原記念生命科学財団  

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Language：Japanese   Publisher：ペプチド創薬の最前線  

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Language：Japanese   Publisher：東京 : ニューサイエンス社  

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Other Link： https://ndlsearch.ndl.go.jp/books/R000000004-I029104734

Language：Japanese   Publisher：[東京] : Institute of Electrical Engineers of Japan  

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Other Link： https://ndlsearch.ndl.go.jp/books/R000000004-I029904379

Language：Japanese   Publisher：生命科学系学会合同年次大会運営事務局  

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Language：Japanese   Publisher：生命科学系学会合同年次大会運営事務局  

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Publisher：株式会社医学書院  

DOI： 10.11477/mf.2425200694

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：一般社団法人 日本機械学会  

&lt;p&gt;In order to find applicable target molecules for molecular target anti-cancer drugs, molecules on the surface of cancer cells in a peptide screening process should be close to those in a human body. A cancer spheroid is a cancer cell aggregate and shows properties similar to tissues. We previously proposed and achieved a peptide screening method using floating spheroids which stirs peptides and spheroids in a liquid slug using an internal convection to promote the peptide adhesion onto the spheroids. In this study, we investigate adhesion efficiency dependent on the speed of the liquid slug in the range from 7.2 to 31 mm/s. At a low speed (7.2 mm/s), antibodies as a simulation of a peptide do not adhere on the spheroids due to weak internal convection in the liquid slug. At a high speed (31 mm/s), the antibodies hardly adhered on the spheroids. We found the speed around 20 mm/s showed the efficient adhesion.&lt;/p&gt;

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Language：English   Publishing type：Research paper, summary (international conference)  

Web of Science

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Language：Japanese   Publisher：Institute of Electrical Engineers of Japan  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese  

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Language：Japanese   Publisher：次世代のがん治療薬・診断のための研究開発  

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J-GLOBAL

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

&lt;p&gt;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.&lt;/p&gt;

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

&lt;p&gt;Peptides that can bind to normal cells should be removed in the screening of the tumor specific binding peptide. We developed a microfluidic device for screening of tumor specific binding peptide that has serially-connected chambers for normal cells and for cancer cells. The normal cell is to remove the non-specific binding peptides, and the cancer cell is to capture tumor specific binding peptides. Cell introduction into the chambers without cross-contamination and screening simulation using a fluorescence-labeled antibody are examined. In cell introduction experiment, contaminations of different types of cells were prevented. In the screening simulation, the trapped amount of the non-specific antibodies can be reduced to 46 % in the downstream chamber.&lt;/p&gt;

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Language：Japanese   Publisher：(公社)日本生化学会  

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Language：Japanese   Publisher：(公社)日本生化学会  

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Language：Japanese   Publisher：Institute of Electrical Engineers of Japan  

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Language：English   Publisher：日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese   Publisher：(株)北隆館  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：(株)北隆館  

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Language：Japanese  

CiNii Books

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Other Link： http://search.jamas.or.jp/link/ui/2016093005

Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

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.

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Language：Japanese   Publisher：(株)北隆館  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：日本癌学会  

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Language：English   Publisher：日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese   Publisher：(株)メディカルレビュー社  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：(一社)日本病理学会  

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Language：Japanese   Publisher：The Japan Society of Mechanical Engineers  

Cancer is the top leading cause of death in Japan. Anti-cancer drugs are one of the most popular treatments whereas they often have heavy side effects. Anti-cancer drugs with light or no side effects are under development using tumor specific binding peptides as guides to cancer cells. However, the number of candidate peptides for the tumor specific binding peptides are extremely large. An efficient peptide screening method is necessary to find them. For this purpose, we are developing a microfluidic device to uniformly disperse cancer cells into its micro chamber and screen the peptide form the large number of peptides. In this paper, we could uniformly disperse N87 cells into the micro chamber using a micro pillar array, and selectively trap yeasts with tumor specific binding peptides by N87 cells in the micro chamber.

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CiNii Research

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Other Link： https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-25250012/

Language：Japanese  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese   Publisher：(公社)日本生化学会  

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Language：Japanese   Publisher：(公社)日本生化学会  

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Language：Japanese   Publisher：(公社)日本生化学会  

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Language：Japanese   Publisher：(一社)日本癌治療学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：(公社)日本薬学会  

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Language：Japanese   Publisher：(株)北隆館  

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CiNii Research

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Language：Japanese   Publisher：寺田国際事務所先端医療技術研究所  

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Other Link： http://search.jamas.or.jp/link/ui/2014079630

Language：Japanese   Publisher：(公財)日本眼科学会  

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Language：Japanese   Publisher：(株)ニュー・サイエンス社  

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Language：Japanese   Publisher：東京 : ニューサイエンス社  

CiNii Books

CiNii Research

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Other Link： https://ndlsearch.ndl.go.jp/books/R000000004-I024141652

Language：Japanese   Publisher：日本眼薬理学会  

J-GLOBAL

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Language：English   Publisher：(一社)日本血液学会-東京事務局  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：English   Publisher：(一社)日本癌学会  

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Language：Japanese  

CiNii Books

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Other Link： http://search.jamas.or.jp/link/ui/2012183209

Language：Japanese   Publisher：日本分子イメージング学会  

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Language：Japanese   Publisher：日本分子イメージング学会  

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