研究者データベース
臼井 達哉USUI Tatsuyaウスイ タツヤ
| ■所属部署名 | 農学研究院 動物生命科学部門 | |||
| ■職名 | 准教授 | |||
Last Updated :2023/04/05
業績情報
氏名・連絡先
氏名
ウスイ タツヤ, 臼井 達哉, USUI Tatsuya生年
1985eメールアドレス
fu7085
go.tuat.ac.jp
主たる所属・職名
農学研究院 動物生命科学部門, 准教授
その他の所属
経歴
- 日本学術振興会
特別研究員(DC1・農学)
自 2011年04月01日, 至 2013年03月31日 - 日本学術振興会
特別研究員(PD・農学)
自 2013年04月01日, 至 2013年10月31日 - 山口大学
共同獣医学部・獣医毒性学
助教
自 2013年11月01日, 至 2017年08月31日 - スタンフォード大学
医学部
客員教員
自 2015年04月01日, 至 2015年09月30日
学歴
学位
免許・資格
教育・研究活動状況
- 近年、三次元の上皮組織構造を培養ディッシュ上で再現する方法としてマトリゲル三次元培養法(オルガノイド培養法)が開発されました。オルガノイド培養法によって、様々な組織の上皮細胞を三次元的に長期培養出来ることが示され、がん研究への応用が始まりました。 当研究室では、前立腺がん罹患犬の尿サンプルを用いて前立腺がんオルガノイドを作製する方法を見出し(Usui et al., Cancer Science, 2017)、このオルガノイドが、生体内のがん微小環境を高いレベルで再現できること、免疫不全マウスの体内で腫瘍を再形成すること、抗がん剤および放射線に対する感受性試験への応用が可能であることを明らかにしました。 また、ヒト、がんモデルマウス、犬由来の様々な組織(肺、大腸、前立腺、膀胱、海馬、肝臓)からオルガノイドを作製し、病態解析を行うことで新たながんを含めた様々な病態メカニズム解明や診断マーカーの開発を目指しています。イヌ膀胱がんオルガノイド培養法の確立
研究分野
研究キーワード
研究テーマ
担当授業科目
科学研究費助成事業
- 基盤研究(C)
中皮腫克服に向けた戦略：新規早期診断マーカーおよび治療ターゲット分子の探索研究
自 2021年, 至 2021年 - 基盤研究(B)
非アルコール性脂肪性肝疾患肝発がんラットモデルにおける選択的オートファジーの解明
自 2021年, 至 2021年 - 基盤研究(B)
尿サンプル由来オルガノイドを用いた多角的な研究:イヌ膀胱がん克服にむけて
自 2020年, 至 2023年 - 基盤研究(C)
中皮腫克服に向けた戦略：新規早期診断マーカーおよび治療ターゲット分子の探索研究
自 2020年, 至 2020年 - 基盤研究(B)
非アルコール性脂肪性肝疾患肝発がんラットモデルにおける選択的オートファジーの解明
自 2020年, 至 2020年 - 基盤研究(C)
音響キャビテーション誘導による空間選択的抗癌剤取り込み技術の研究
自 2018年, 至 2018年 - 若手研究(B)
三次元マウス肺がんモデルを用いたがん・血管内皮連関機構の解明
自 2017年, 至 2018年
論文
- Autophagy regulates levels of tumor suppressor enzyme protein phosphatase 6
Fujiwara, Nobuyuki; Shibutani, Shusaku; Sakai, Yusuke; Watanabe, Toshio; Kitabayashi, Issay; Oshima, Hiroko; Oshima, Masanobu; Hoshida, Hisashi; Akada, Rinji; Usui, Tatsuya; Ohama, Takashi; Sato, Koichi
CANCER SCIENCE
WILEY
Protein phosphatase 6 (PP6) is an essential serine/threonine protein phosphatase that acts as an important tumor suppressor. However, increased protein levels of PP6 have been observed in some cancer types, and they correlate with poor prognosis in glioblastoma. This raises a question about how PP6 protein levels are regulated in normal and transformed cells. In this study, we show that PP6 protein levels increase in response to pharmacologic and genetic inhibition of autophagy. PP6 associates with autophagic adaptor protein p62/SQSTM1 and is degraded in a p62-dependent manner. Accordingly, protein levels of PP6 and p62 fluctuate in concert under different physiological and pathophysiological conditions. Our data reveal that PP6 is regulated by p62-dependent autophagy and suggest that accumulation of PP6 protein in tumor tissues is caused at least partially by deficiency in autophagy.
2020年12月, 研究論文（学術雑誌）, 共同, 111, 12, 1347-9032, DOI(公開)(r-map), 4371, 4380 - Progenitor identification and SARS-CoV-2 infection in human distal lung organoids
Salahudeen, Ameen A.; Choi, Shannon S.; Rustagi, Arjun; Zhu, Junjie; van Unen, Vincent; De la O, Sean M.; Flynn, Ryan A.; Margalef-Catala, Mar; Santos, Antonio J. M.; Ju, Jihang; Batish, Arpit; Usui, Tatsuya; Zheng, Grace X. Y.; Edwards, Caitlin E.; Wagar, Lisa E.; Luca, Vincent; Anchang, Benedict; Nagendran, Monica; Nguyen, Khanh; Hart, Daniel J.; Terry, Jessica M.; Belgrader, Phillip; Ziraldo, Solongo B.; Mikkelsen, Tarjei S.; Harbury, Pehr B.; Glenn, Jeffrey S.; Garcia, K. Christopher; Davis, Mark M.; Baric, Ralph S.; Sabatti, Chiara; Amieva, Manuel R.; Blish, Catherine A.; Desai, Tushar J.; Kuo, Calvin J.
NATURE
NATURE RESEARCH
The distal lung contains terminal bronchioles and alveoli that facilitate gas exchange. Three-dimensional in vitro human distal lung culture systems would strongly facilitate the investigation of pathologies such as interstitial lung disease, cancer and coronavirus disease 2019 (COVID-19) pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here we describe the development of a long-term feeder-free, chemically defined culture system for distal lung progenitors as organoids derived from single adult human alveolar epithelial type II (AT2) or KRT5(+) basal cells. AT2 organoids were able to differentiate into AT1 cells, and basal cell organoids developed lumens lined with differentiated club and ciliated cells. Single-cell analysis of KRT5(+) cells in basal organoids revealed a distinct population of ITGA6(+)ITGB4(+) mitotic cells, whose offspring further segregated into a TNFRSF12A(hi) subfraction that comprised about ten per cent of KRT5(+) basal cells. This subpopulation formed clusters within terminal bronchioles and exhibited enriched clonogenic organoid growth activity. We created distal lung organoids with apical-out polarity to present ACE2 on the exposed external surface, facilitating infection of AT2 and basal cultures with SARS-CoV-2 and identifying club cells as a target population. This long-term, feeder-free culture of human distal lung organoids, coupled with single-cell analysis, identifies functional heterogeneity among basal cells and establishes a facile in vitro organoid model of human distal lung infections, including COVID-19-associated pneumonia. A long-term culture method for organoids derived from single adult human lung cells is used to identify progenitor cells and study SARS-CoV-2 infection.
2020年12月24日, 研究論文（学術雑誌）, 共同, 588, 7839, 0028-0836, DOI(公開)(r-map), 670, + - Establishment of 2.5D organoid culture model using 3D bladder cancer organoid culture
Abugomaa, Amira; Elbadawy, Mohamed; Yamanaka, Megumi; Goto, Yuta; Hayashi, Kimika; Mori, Takashi; Uchide, Tsuyoshi; Azakami, Daigo; Fukushima, Ryuji; Yoshida, Toshinori; Shibutani, Makoto; Yamashita, Risako; Kobayashi, Mio; Yamawaki, Hideyuki; Shinohara, Yuta; Kaneda, Masahiro; Usui, Tatsuya; Sasaki, Kazuaki
SCIENTIFIC REPORTS
NATURE PUBLISHING GROUP
Three-dimensional (3D) organoid culture holds great promises in cancer precision medicine. However, Matrigel and stem cell-stimulating supplements are necessary for culturing 3D organoid cells. It costs a lot of money and consumes more time and effort compared with 2D cultured cells. Therefore, the establishment of cheaper and Matrigel-free organoid culture that can maintain the characteristics of a part of 3D organoids is demanded. In the previous study, we established a dog bladder cancer (BC) 3D organoid culture system by using their urine samples. Here, we successfully isolated cells named 2.5D organoid from multiple strains of dog BC 3D organoids using 2.5 organoid media. The cell proliferation speed of 2.5D organoids was faster than parental 3D organoid cells. The expression pattern of stem cell markers was close to 3D organoids. Injection of 2.5D organoid cells into immunodeficient mice formed tumors and showed the histopathological characteristics of urothelial carcinoma similar to the injection of dog BC 3D organoids. The 2.5D organoids had a similar sensitivity profile for anti-cancer drug treatment to their parental 3D organoids. These data suggest that our established 2.5D organoid culture method might become a reasonable and useful tool instead of 3D organoids in dog BC research and therapy.
2020年06月10日, 研究論文（学術雑誌）, 共同, 10, 1, 2045-2322, DOI(公開)(r-map) - Development of Prostate Cancer Organoid Culture Models in Basic Medicine and Translational Research
Elbadawy, Mohamed; Abugomaa, Amira; Yamawaki, Hideyuki; Usui, Tatsuya; Sasaki, Kazuaki
CANCERS
MDPI
Prostate cancer (PC) is the most prevalent cancer in men and the second main cause of cancer-related death in Western society. The lack of proper PC models that recapitulate the molecular and genomic landscape of clinical disease has hampered progress toward translational research to understand the disease initiation, progression, and therapeutic responses in each patient. Although several models have been developed, they hardly emulated the complicated PC microenvironment. Precision medicine is an emerging approach predicting appropriate therapies for individual cancer patients by means of various analyses of individual genomic profiling and targeting specific cancer pathways. In PC, precision medicine also has the potential to impose changes in clinical practices. Here, we describe the various PC models with special focus on PC organoids and their values in basic medicine, personalized therapy, and translational researches in vitro and in vivo, which could help to achieve the full transformative power of cancer precision medicine.
2020年04月, 研究論文（学術雑誌）, 共同, 12, 4, DOI(公開)(r-map) - Efficacy of primary liver organoid culture from different stages of non-alcoholic steatohepatitis (NASH) mouse model
Elbadawy, Mohamed; Yamanaka, Megumi; Goto, Yuta; Hayashi, Kimika; Tsunedomi, Ryouichi; Hazama, Shoichi; Nagano, Hiroaki; Yoshida, Toshinori; Shibutani, Makoto; Ichikawa, Ryo; Nakahara, Junta; Omatsu, Tsutomu; Mizutani, Tetsuya; Katayama, Yukie; Shinohara, Yuta; Abugomaa, Amira; Kaneda, Masahiro; Yamawaki, Hideyuki; Usui, Tatsuya; Sasaki, Kazuaki
BIOMATERIALS
ELSEVIER SCI LTD
Non-alcoholic steatohepatitis (NASH) is associated with liver fibrosis and cirrhosis, which eventually leads to hepatocellular carcinoma. Although several animal models were developed to understand the mechanisms of NASH pathogenesis and progression, it remains obscure. A 3D organoid culture system can recapitulate organ structures and maintain gene expression profiles of original tissues. We therefore tried to generate liver organoids from different degrees [defined as mild (NASH A), moderate (NASH B) and severe (NASH C)] of methionine- and choline-deficient diet-induced NASH model mice and analyzed the difference of their architecture, cell components, organoid-forming efficacy, and gene expression profiles. Organoids from each stage of NASH model mice were successfully generated. Interestingly, epithelial-mesenchymal transition was observed in NASH C organoids. Expression of Collagen I and an activated hepatic stellite cell marker, alpha-sma was upregulated in the liver organoids from NASH B and C mice. The analysis of RNA sequencing revealed that several novel genes were upregulated in all NASH liver organoids. These results suggest that our generated liver organoids from different stages of NASH diseased mice might become a useful tool for in vitro studies of the molecular mechanism of NASH development and also for identifying novel biomarkers for early diagnosis of NASH disease.
2020年04月, 研究論文（学術雑誌）, 共同, 237, 0142-9612, DOI(公開)(r-map) - Emerging Roles of Cancer Stem Cells in Bladder Cancer Progression, Tumorigenesis, and Resistance to Chemotherapy: A Potential Therapeutic Target for Bladder Cancer
Abugomaa, Amira; Elbadawy, Mohamed; Yamawaki, Hideyuki; Usui, Tatsuya; Sasaki, Kazuaki
CELLS
MDPI
Bladder cancer (BC) is a complex and highly heterogeneous stem cell disease associated with high morbidity and mortality rates if it is not treated properly. Early diagnosis with personalized therapy and regular follow-up are the keys to a successful outcome. Cancer stem cells (CSCs) are the leading power behind tumor growth, with the ability of self-renewal, metastasis, and resistance to conventional chemotherapy. The fast-developing CSC field with robust genome-wide screening methods has found a platform for establishing more reliable therapies to target tumor-initiating cell populations. However, the high heterogeneity of the CSCs in BC disease remains a large issue. Therefore, in the present review, we discuss the various types of bladder CSC heterogeneity, important regulatory pathways, roles in tumor progression and tumorigenesis, and the experimental culture models. Finally, we describe the current stem cell-based therapies for BC disease.
2020年01月, 研究論文（学術雑誌）, 共同, 9, 1, DOI(公開)(r-map) - Efficacy of Juzen-taiho-to against vincristine-induced toxicity in dogs
Shinohara, Yuta; Nishino, Yoko; Yamanaka, Megumi; Ohmori, Keitaro; Elbadawy, Mohamed; Usui, Tatsuya; Sasaki, Kazuaki
JOURNAL OF VETERINARY MEDICAL SCIENCE
JAPAN SOC VET SCI
Vincristine, one of the anti-cancer drugs used in veterinary practice, has adverse hematological and gastrointestinal effects in dogs. Juzen-taiho-to is a traditional Chinese medicine used for patients with anorexia in human medicine. However, the protective effects of Juzen-taiho-to against anti-cancer drug-induced toxicity in dogs have not been investigated. We therefore examined whether the administration of Juzen-taiho-to to dogs affects gastric motility, and vincristine-induced gastrointestinal and hematological toxicity. The study was composed of three trials. In the first trial, Juzen-taiho-to (450 mg/kg/day) was orally administered to five dogs. In the second and third trials, vincristine (0.75 mg/m(2)) was intravenously administered to each dog in the absence or presence of Juzen-taiho-to (450 mg/kg/day). During these trials, gastric motility and blood parameters were assessed. Juzen-taiho-to increased gastric motility and improved vincristine-induced gastrointestinal, but not hematological, adverse effects in dogs. This study suggested that Juzen-taiho-to may be applicable for gastrointestinal care in dogs receiving chemotherapy.
2019年12月, 研究論文（学術雑誌）, 共同, 81, 12, 0916-7250, DOI(公開)(r-map), 1810, 1816 - Possible anti-oxidative effects of long-term administration of Juzen-taiho-to in dogs
Shinohara, Yuta; Oyama, Ayaka; Usui, Tatsuya; Sasaki, Kazuaki
JOURNAL OF VETERINARY MEDICAL SCIENCE
JAPAN SOC VET SCI
It is known that oxidative stress is related to disease in humans and dogs. Many traditional Chinese medicines have been reported to have anti-oxidative effects, but there are no reports that they have anti-oxidative effects in dogs. In this study, we examined the anti-oxidative effects of Juzen-taiho-to, a traditional Chinese medicine, in dogs. Five healthy female beagle dogs (38-41 months of age weighing 8.6-10.7 kg) were orally administered Juzen-taiho-to at 450 mg/kg with food for 28 days. Blood samples were taken from all five dogs on days 0, 7, 14, 21, and 28. Using the blood samples, improvement of the antioxidant level as assessed by the biological antioxidant potential (BAP), reduced oxidative stress level as assessed by derivatives of reactive oxygen metabolites (d-ROMs), and improvement of blood fluidity were examined. Regarding the antioxidant level and blood fluidity, no significant difference was observed, but the oxidative stress level on days 14, 21, and 28 was significantly lower than that on day 0. Thus, Juzen-taiho-to may have anti-oxidative effects in dogs by reducing oxidative stress and be useful for oxidative stress-related diseases in dogs.
2019年11月, 研究論文（学術雑誌）, 共同, 81, 11, 0916-7250, DOI(公開)(r-map), 1616, 1620 - Establishment of a novel experimental model for muscle-invasive bladder cancer using a dog bladder cancer organoid culture
Elbadawy, Mohamed; Usui, Tatsuya; Mori, Takashi; Tsunedomi, Ryouichi; Hazama, Shoichi; Nabeta, Rina; Uchide, Tsuyoshi; Fukushima, Ryuji; Yoshida, Toshinori; Shibutani, Makoto; Tanaka, Takaharu; Masuda, Sosuke; Okada, Rena; Ichikawa, Ryo; Omatsu, Tsutomu; Mizutani, Tetsuya; Katayama, Yukie; Noguchi, Shunsuke; Iwai, Satomi; Nakagawa, Takayuki; Shinohara, Yuta; Kaneda, Masahiro; Yamawaki, Hideyuki; Sasaki, Kazuaki
CANCER SCIENCE
WILEY
In human and dogs, bladder cancer (BC) is the most common neoplasm affecting the urinary tract. Dog BC resembles human muscle-invasive BC in histopathological characteristics and gene expression profiles, and could be an important research model for this disease. Cancer patient-derived organoid culture can recapitulate organ structures and maintains the gene expression profiles of original tumor tissues. In a previous study, we generated dog prostate cancer organoids using urine samples, however dog BC organoids had never been produced. Therefore we aimed to generate dog BC organoids using urine samples and check their histopathological characteristics, drug sensitivity, and gene expression profiles. Organoids from individual BC dogs were successfully generated, expressed urothelial cell markers (CK7, CK20, and UPK3A) and exhibited tumorigenesis in vivo. In a cell viability assay, the response to combined treatment with a range of anticancer drugs (cisplatin, vinblastine, gemcitabine or piroxicam) was markedly different in each BC organoid. In RNA-sequencing analysis, expression levels of basal cell markers (CK5 and DSG3) and several novel genes (MMP28, CTSE, CNN3, TFPI2, COL17A1, and AGPAT4) were upregulated in BC organoids compared with normal bladder tissues or two-dimensional (2D) BC cell lines. These established dog BC organoids might be a useful tool, not only to determine suitable chemotherapy for BC diseased dogs but also to identify novel biomarkers in human muscle-invasive BC. In the present study, for the 1st time, dog BC organoids were generated and several specifically upregulated organoid genes were identified. Our data suggest that dog BC organoids might become a new tool to provide fresh insights into both dog BC therapy and diagnostic biomarkers.
2019年09月02日, 研究論文（学術雑誌）, 共同, 110, 9, 1347-9032, DOI(公開)(r-map), 2806, 2821 - Emerging Roles of C-Myc in Cancer Stem Cell-Related Signaling and Resistance to Cancer Chemotherapy: A Potential Therapeutic Target Against Colorectal Cancer
Elbadawy, Mohamed; Usui, Tatsuya; Yamawaki, Hideyuki; Sasaki, Kazuaki
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Myc is a nuclear transcription factor that mainly regulates cell growth, cell cycle, metabolism, and survival. Myc family proteins contain c-Myc, n-Myc, and l-Myc. Among them, c-Myc can become a promising therapeutic target molecule in cancer. Cancer stem cells (CSCs) are known to be responsible for the therapeutic resistance. In the previous study, we demonstrated that c-Myc mediates drug resistance of colorectal CSCs using a patient-derived primary three-dimensional (3D) organoid culture. In this review, we mainly focus on the roles of c-Myc-related signaling in the regulation of CSCs, chemotherapy resistance, and colorectal cancer organoids. Finally, we introduce the various types of c-Myc inhibitors and propose the possibility of c-Myc as a therapeutic target against colorectal cancer.
2019年05月01日, 研究論文（学術雑誌）, 共同, 20, 9, DOI(公開)(r-map) - Pericardial Mesothelioma in a Dog: The Feasibility of Ultrasonography in Monitoring Tumor Progression
Nabeta, Rina; Nakagawa, Yuki; Chiba, Shiori; Hou Xiantao; Usui, Tatsuya; Suzuki, Kazuhiko; Furuya, Tetsuya; Fukushima, Ryuji; Uchide, Tsuyoshi
FRONTIERS IN VETERINARY SCIENCE
FRONTIERS MEDIA SA
A 6-year-old neutered male Yorkshire Terrier presented with recurrent pericardial effusion. Although clinical examinations including computed tomography were inconclusive, an exploratory thoracotomy revealed multiple small nodules and plaques on the inner surface of the pericardial sac (Day 1). A subtotal pericardiectomy was performed to prevent cardiac tamponade due to the increasing pericardial effusion, and the resected section of the pericardium was histopathologically diagnosed with mesothelioma. After surgery, chemotherapy with intrathoracic carboplatin was commenced. During the course of the treatment, a detailed follow-up ultrasonographic scan was performed to detect early lesions disseminated on the pleura, originating from the primary pericardial mesothelioma. On Day 101, the minute pleural nodules, which were disseminated lesions as predicted, were successfully imaged by ultrasonography. As the clinical stage advanced, the nodules were observed to gradually increase in size and number, implying tumor progression. These observations highlight the feasibility of ultrasonography in detecting minute disseminated lesions at an early stage, monitoring tumor progression, and thereby, predicting the prognosis of canine pericardial mesothelioma.
2019年04月18日, 研究論文（学術雑誌）, 共同, 6, 2297-1769, DOI(公開)(r-map), 121 - Novel Functions of Death-Associated Protein Kinases through Mitogen-Activated Protein Kinase-Related Signals.
Elbadawy M, Usui T, Yamawaki H, Sasaki K.
Int J Mol Sci.
2018年10月04日, 研究論文（学術雑誌）, 共同, 19, 10, DOI(公開)(r-map), E3031 - A stable association with PME‐1 may be dispensable for PP2A demethylation – implications for the detection of PP2A methylation and immunoprecipitation
Yabe R, Tsuji S, Mochida S, Ikehara T, Usui T, Ohama T, Sato K
FEBS Open Bio.
2018年08月01日, 8, 9, DOI(公開)(r-map), 1486, 1496 - Development of an Experimental Model for Analyzing Drug Resistance in Colorectal Cancer
Elbadawy, Mohamed; Usui, Tatsuya; Yamawaki, Hideyuki; Sasaki, Kazuaki
CANCERS
MDPI
Colorectal cancer (CRC) is one of the most common cancers, for which combination treatment of chemotherapy is employed. However, most patients develop drug resistance during the course of treatment. To clarify the mechanisms of drug resistance, various research models have been developed. Recently, we established a human CRC patients-derived three-dimensional (3D) culture system using an air-liquid interface organoid method. It contained numerous cancer stem cells and showed resistance to 5-fluorouracil and Irinotecan. In this review, we introduce conventional and our established models for studying drug resistance in CRC.
2018年06月, 研究論文（学術雑誌）, 共同, 10, 6, 2072-6694, DOI(公開)(r-map) - Hedgehog Signals Mediate Anti-Cancer Drug Resistance in Three-Dimensional Primary Colorectal Cancer Organoid Culture
Usui, Tatsuya; Sakurai, Masashi; Umata, Koji; Elbadawy, Mohamed; Ohama, Takashi; Yamawaki, Hideyuki; Hazama, Shoichi; Takenouchi, Hiroko; Nakajima, Masao; Tsunedomi, Ryouichi; Suzuki, Nobuaki; Nagano, Hiroaki; Sato, Koichi; Kaneda, Masahiro; Sasaki, Kazuaki
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Colorectal cancer is one of the most common causes of cancer death worldwide. In patients with metastatic colorectal cancer, combination treatment with several anti-cancer drugs is employed and improves overall survival in some patients. Nevertheless, most patients with metastatic disease are not cured owing to the drug resistance. Cancer stem cells are known to regulate resistance to chemotherapy. In the previous study, we established a novel three-dimensional organoid culture model from tumor colorectal tissues of human patients using an air-liquid interface (ALI) method, which contained numerous cancer stem cells and showed resistance to 5-fluorouracil (5-FU) and Irinotecan. Here, we investigate which inhibitor for stem cell-related signal improves the sensitivity for anti-cancer drug treatment in tumor ALI organoids. Treatment with Hedgehog signal inhibitors (AY9944, GANT61) decreases the cell viability of organoids compared with Notch (YO-01027, DAPT) and Wnt (WAV939, Wnt-C59) signal inhibitors. Combination treatment of AY9944 or GANT61 with 5-FU, Irinotecan or Oxaliplatin decreases the cell viability of tumor organoids compared with each anti-cancer drug alone treatment. Treatment with AY9944 or GANT61 inhibits expression of stem cell markers c-Myc, CD44 and Nanog, likely through the decrease of their transcription factor, GLI-1 expression. Combination treatment of AY9944 or GANT61 with 5-FU or Irinotecan also prevents colony formation of colorectal cancer cell lines HCT116 and SW480. These findings suggest that Hedgehog signals mediate anti-cancer drug resistance in colorectal tumor patient-derived ALI organoids and that the inhibitors are useful as a combinational therapeutic strategy against colorectal cancer.
2018年04月, 研究論文（学術雑誌）, 共同, 19, 4, 1422-0067, DOI(公開)(r-map) - Stemness Is Enhanced in Gastric Cancer by a SET/PP2A/E2F1 Axis
Enjoji, Shuhei; Yabe, Ryotaro; Tsuji, Shunya; Yoshimura, Kazuhiro; Kawasaki, Hideyoshi; Sakurai, Masashi; Sakai, Yusuke; Takenouchi, Hiroko; Yoshino, Shigefumi; Hazama, Shoichi; Nagano, Hiroaki; Oshima, Hiroko; Oshima, Masanobu; Vitek, Michael P.; Matsuura, Tetsuya; Hippo, Yoshitaka; Usui, Tatsuya; Ohama, Takashi; Sato, Koichi
MOLECULAR CANCER RESEARCH
AMER ASSOC CANCER RESEARCH
Gastric cancer is the fifth most common malignancy and the third leading cause of cancer-related deaths worldwide. Chemotherapies against gastric cancer often fail, with cancer recurrence due potentially to the persistence of cancer stem cells. This unique subpopulation of cells in tumors possesses the ability to self-renew and dedifferentiate. These cancer stem cells are critical for initiation, maintenance, metastasis, and relapse of cancers; however, the molecular mechanisms supporting cancer stemness remain largely unknown. Increased kinase and decreased phosphatase activity are hallmarks of oncogenic signaling. Protein phosphatase 2A (PP2A) functions as a tumor-suppressor enzyme, and elevated levels of SET/I2PP2A, an endogenous PP2A protein inhibitor, are correlated with poor prognosis of several human cancers. Here, it was determined that SET expression was elevated in tumor tissue in a gastric cancer mouse model system, and SET expression was positively correlated with poor survival of human gastric cancer patients. Mechanistically, SET knockdown decreased E2F1 levels and suppressed the stemness of cancer cell lines. Immunoprecipitations show SET associated with the PP2A-B56 complex, and the B56 subunit interacted with the E2F1 transcription factor. Treatment of gastric cancer cells with the SET-targeting drug OP449 increased PP2A activity, decreased E2F1 protein levels, and suppressed stemness of cancer cells. These data indicate that a SET/PP2A/E2F1 axis regulates cancer cell stemness and is a potential target for gastric cancer therapy.
2018年03月, 研究論文（学術雑誌）, 共同, 16, 3, 1541-7786, DOI(公開)(r-map), 554, 563 - Preparation of human primary colon tissue-derived organoid using air liquid interface culture
Usui T, Sakurai M, Umata K, Yamawaki H, Ohama T, Sato K.
Current Protocols in Toxicology
2018年02月21日, 75, DOI(公開)(r-map), 22.6.1, 22.6.7 - Establishment of a dog primary prostate cancer organoid using the urine cancer stem cells
Usui, Tatsuya; Sakurai, Masashi; Nishikawa, Shimpei; Umata, Koji; Nemoto, Yuki; Haraguchi, Tomoya; Itamoto, Kazuhito; Mizuno, Takuya; Noguchi, Shunsuke; Mori, Takashi; Iwai, Satomi; Nakagawa, Takayuki; Yamawaki, Hideyuki; Ohama, Takashi; Sato, Koichi
CANCER SCIENCE
WILEY
Dog spontaneously develop prostate cancer (PC) like humans. Because most dogs with PC have a poor prognosis, they could be used as a translational model for advanced PC in humans. Stem cell-derived 3-D organoid culture could recapitulate organ structures and physiology. Using patient tissues, a human PC organoid culture system was established. Recent study has shown that urine cells also possess the characteristic of stem cells. However, urine cell-derived PC organoids have never been produced. Therefore, we generated PC organoids using the dog urine samples. Urine organoids were successfully generated from each dog with PC. Each organoid showed cystic structures and resembled the epithelial structures of original tissues. Expression of an epithelial cell marker, E-cadherin, and a myofibloblast marker, -SMA, was observed in the urine organoids. The organoids also expressed a basal cell marker, CK5, and a luminal cell marker, CK8. CD49f-sorted basal cell organoids rapidly grew compared with CD24-sorted luminal cell organoids. The population of CD44-positive cells was the highest in both organoids and the original urine cells. Tumors were successfully formed with the injection of the organoids into immunodeficient mice. Treatment with a microtubule inhibitor, docetaxel, but not a cyclooxygenase inhibitor, piroxicam, and an mTOR inhibitor, rapamycin, decreased the cell viability of organoids. Treatment with a Hedgehog signal inhibitor, GANT61, increased the radiosensitivity in the organoids. These findings revealed that PC organoids using urine might become a useful tool for investigating the mechanisms of the pathogenesis and treatment of PC in dogs.
2017年12月, 研究論文（学術雑誌）, 共同, 108, 12, 1349-7006, DOI(公開)(r-map), 2383, 2392 - The role of SET/I2PP2A in canine mammary tumors.
Kake S, Tsuji S, Enjoji S, Hanasaki S, Hayase H, Yabe R, Tanaka Y, Nakagawa T, Liu HP, Chang SC, Usui T, Ohama T, Sato K.
Scientific Reports
2017年06月, 7, 1, DOI(公開)(r-map), 4279 - Establishment of a novel three-dimensional primary culture model for hippocampal neurogenesis
Usui T, Sakurai M, Kawasaki H, Ohama T, Yamawaki H, Sato K
Physiological Reports
2017年06月05日, 5, 12, DOI(公開)(r-map), e13318 - Regulation of intestinal myofibroblasts by KRas-mutated colorectal cancer cells through heparin-binding epidermal growth factor-like growth factor
Kawasaki, Hideyoshi; Saotome, Takuya; Usui, Tatsuya; Ohama, Takashi; Sato, Koichi
ONCOLOGY REPORTS
SPANDIDOS PUBL LTD
In colorectal cancer, gain-of-function mutations in KRas play a critical role in malignant transformation. Tumor growth in colorectal cancer is known to be promoted by the intestinal myofibroblasts (IMFs) that localize adjacent to the cancer cells, but the mechanisms of interaction between KRasmutated cancer cells and the myofibroblasts remain unclear. Here, we investigated the effects of KRas-mutated cells on the behavior of myofibroblasts by using mouse primary IMFs and cells of an IMF cell line (LmcMF) and a mouse colon epithelial cell line (aMoCl). Conditioned medium (CM) was collected from aMoC1 cells overexpressing a control vector or KRasV12 vector (KRasV12-CM), and the effects of KRasV12-CM on IMFs were analyzed by performing proliferation assays, wound-healing assays, Boyden chamber assays, and western blotting. Whereas KRasV12-CM exerted little effect on the differentiation and proliferation of primary IMFs, the CM promoted migration of both primary IMFs and LmcMF cells. In KRasV12-overexpressing aMoCl cells, mRNA expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) was higher than in mock-transfected aMoC1 cells, and HB-EGF promoted the migration of primary IMFs and LmcMF cells. Moreover, KRasV12-CM-induced IMF migration was suppressed by dacomitinib, an inhibitor of HB-EGF receptors. Notably, in LmcMF cells, both KRasV12-CM and HB-EGF activated extracellular signal-regulated kinase (ERK) and c-jun N-terminal kinase (JNK), whereas KRasV12CM-induced migration of IMFs was suppressed following treatment with either an ERK inhibitor (FR180204) or a JNK inhibitor (SP600125). These results suggest that HB-EGF secreted from KRas-mutated colorectal cancer cells promotes IMF migration through ERK and JNK activation, which, in turn, could support cancer progression.
2017年05月, 研究論文（学術雑誌）, 共同, 37, 5, 1021-335X, DOI(公開)(r-map), 3128, 3136 - Regulation of intestinal myofibroblasts by KRas-mutated colorectal cancer cells through heparin-binding epidermal growth factor-like growth factor.
Kawasaki H, Saotome T, Usui T, Ohama T, Sato K.
ONCOLOGY REPORTS
2017年03月, 37, 5, DOI(公開)(r-map), 3128, 3136 - Death-associated protein kinase 3 controls the tumor progression of A549 cells through ERK MAPK/c-Myc signaling
Kake, Satoru; Usui, Tatsuya; Ohama, Takashi; Yamawaki, Hideyuki; Satoi, Koichi
ONCOLOGY REPORTS
SPANDIDOS PUBL LTD
Death-associated protein kinases (DAPKs) are members of the serine/threonine protein kinase family, which regulate cell death. Although DAPK3 has been implicated as a tumor suppressor, a recent study revealed an oncogenic role of DAPK3. However, the role of DAPK3 in non-small cell lung cancer (NSCLC) remains unclear. Therefore, we examined whether DAPK3 controls the progression of NSCLC using the NSCLC cell line, A549. We generated A549 cells stably expressing small hairpin RNA (shRNA) targeting DAPK3. In the A549 cells, the protein level of DAPK3 was decreased and the cell proliferation was inhibited. DAPK3 knockdown caused WC-, cell cycle arrest as assessed by flow cytometric assay and reduced cyclin D1 expression in A549 cells. Phosphorylation of ERK and c-Myc, but not Akt and JNK, was inhibited by DAPK3 knockdown. Cell migration and invasion were also inhibited by DAPK3 knockdown as determined by a Boyden chamber assay and an invasion assay, respectively. Moreover, DAPK3 knockdown inhibited anchorage-independent cell growth as determined by soft-agar colony formation assay. In a mouse xenograft model, tumors derived from DAPK3-knockdown cells exhibited reduced tumor growth. The present results demonstrated for the first time that DAPK3 controls proliferation, migration, invasion, soft-agar colony formation and tumor growth through activation of ERK/c-Myc signaling in A549 cells. These findings indicate that DAPK3 may be a novel target for the treatment of NSCLC.
2017年02月, 研究論文（学術雑誌）, 共同, 37, 2, 1021-335X, DOI(公開)(r-map), 1100, 1106 - Establishment of a Novel Model for Anticancer Drug Resistance in Three-Dimensional Primary Culture of Tumor Microenvironment
Usui T, Sakurai M, Enjoji S, Kawasaki H, Umata K, Ohama T, Fujiwara N, Yabe R, Tsuji S, Yamawaki H, Hazama S, Takenouchi H, Nakajima M, Tsunedomi R, Suzuki N, Nagano H, Sato.
Stem Cells International
2016年, DOI(公開)(r-map), 7053872 - Anti-tumor effects of perphenazine on canine lymphoma.
Tsuji S, Yabe R, Usui T, Mizuno T, Ohama T, Sato K.
The Journal of Veterinary Medical Science
2016年09月01日, 78, 8, DOI(公開)(r-map), 1293, 1298 - Regulation of Beclin 1 Protein Phosphorylation and Autophagy by Protein Phosphatase 2A (PP2A) and Death-associated Protein Kinase 3 (DAPK3).
Fujiwara N, Usui T, Ohama T, Sato K.
The Journal of Biological Chemistry
2016年05月13日, 291, 20, DOI(公開)(r-map), 10858, 10866 - Protein Phosphatase Methyl-Esterase PME-1 Protects Protein Phosphatase 2A from Ubiquitin/Proteasome Degradation.
Yabe R, Miura A, Usui T, Mudrak I, Ogris E, Ohama T, Sato K.
PLoS One
2015年12月, 10, 12, DOI(公開)(r-map), e0145226 - Eukaryotic elongation factor 2 kinase controls proliferation and migration of vascular smooth muscle cells.
Usui T, Nijima R, Sakatsume T, Otani K, Kameshima S, Okada M, Yamawaki H.
Acta Physiologica (Oxf).
2015年02月, 213, 2, DOI(公開)(r-map), 472, 480 - Death-associated protein kinase 3 mediates vascular structural remodelling via stimulating smooth muscle cell proliferation and migration.
Usui T, Sakatsume T, Nijima R, Otani K, Kazama K, Morita T, Kameshima S, Okada M, Yamawaki H.
Clinical Science(Lond)
2014年10月, 127, 8, DOI(公開)(r-map), 539, 548 - Brain-derived neurotrophic factor promotes angiogenic tube formation through generation of oxidative stress in human vascular endothelial cells.
Usui T, Naruo A, Okada M, Hayabe Y, Yamawaki H.
Acta Physiologica (Oxf).
2014年06月, 211, 2, DOI(公開)(r-map), 385, 394 - Zipper interacting protein kinase (ZIPK): function and signaling.
Usui T, Okada M, Yamawaki H.
Apoptosis.
2014年02月, 19, 2, DOI(公開)(r-map), 387, 391 - Histone deacetylase 4 controls neointimal hyperplasia via stimulating proliferation and migration of vascular smooth muscle cells.
Usui T, Morita T, Okada M, Yamawaki H.
Hypertension
2014年02月, 63, 2, DOI(公開)(r-map), 397, 403 - Eukaryotic elongation factor 2 kinase regulates the development of hypertension through oxidative stress-dependent vascular inflammation.
Usui T, Okada M, Hara Y, Yamawaki H.
American Journal of Physiology-Heart and Circulatory Physiology
2013年09月01日, 305, 5, DOI(公開)(r-map), H756, H768 - Death-associated protein kinase 3 mediates vascular inflammation and development of hypertension in spontaneously hypertensive rats.
Usui T, Okada M, Hara Y, Yamawaki H.
Hypertension
2012年10月, 60, 4, DOI(公開)(r-map), 1031, 1039
研究発表、招待講演等
- NASHモデルマウス由来肝臓オルガノイドの開発と創薬への応⽤
日本薬理学会年会
2021年03月08日, 口頭発表（招待・特別） - 三次元オルガノイド培養法を用いたオーダーメイド獣医療の開発と創薬への応用
第 4 回医薬品毒性機序研究会
2021年12月17日, 口頭発表（招待・特別） - イヌ膀胱がんオルガノイド培養法を用いた新規進行性膀胱がん実験モデルの確立
日本癌学会学術集会
2019年09月27日, ポスター発表 - Establishment of a culture method of dog bladder cancer organoids
Cell symposia organoid engineering
2019年08月27日, ポスター発表 - NASHモデルマウス由来肝臓オルガノイドの作製
第46回日本毒性学会学術年会
2019年06月26日, 口頭発表（一般） - ヒト大腸エアリキッドオルガノイドモデルを用いた大腸がん研究
第92回日本薬理学会年会
2019年03月16日, シンポジウム・ワークショップ パネル（公募） - イヌ膀胱がんオルガノイド培養法の樹⽴
第92回日本薬理学会年会
2019年03月15日, 口頭発表（一般） - テーラーメイド獣医療にむけた尿由来イヌ前立腺がんオルガノイド培養法の開発
第43回日本分子生物学会年会
2018年11月29日, ポスター発表 - Establishment of a dog primary prostate cancer organoid using the urine cancer stem cells
CMCB-2018 - BIT Congress Inc
2018年10月16日, 口頭発表（招待・特別） - 尿サンプル由来イヌ前立腺がんオルガノイド培養法の樹立
第161回日本獣医学会学術集会
2018年09月10日, 口頭発表（一般） - 三次元マウス肺がんと血管内皮細胞の共培養システムの樹立
第45回日本毒性学会学術年会
2018年07月18日, 口頭発表（一般） - Establishment of a prostate cancer organoid using the dog urine stem cells
第91回日本薬理学会年会は第18回国際薬理学・臨床薬理学会議（WCP2018）
2018年07月02日, ポスター発表 - Urine sample-derived cancer organoids enable tailor-made medicine of dog prostate cancer
EAVPT 2018: 14th International Congress of the European Association for Veterinary Pharmacology and Toxicology
2018年06月25日, 口頭発表（一般）
メディア報道
- コウモリが新型コロナの感染伝播に関与していない可能性、農工大がミニ臓器で確認
東京農工大学の大松勉准教授らが、ルーセットオオコウモリの腸を再現したミニ臓器(ミニ腸)の作製に成功し、それを使用した研究の結果、ルーセットオオコウモリが新型コロナウイルス感染症(COVID-19)の感染伝播に関与していないことが示唆されたと発表したことが紹介される。
マイナビニュース／gooニュース／ニコニコニュース／BIGLOBE／exciteニュース／マピオンニュース
自 2021年11月18日, 至 2021年11月18日 - 犬や猫のがん治療を支援 東京農工大発新興が始動
東京農工大学の山本晴大学院生らが、犬の膀胱癌や猫の乳がんの治療を支援するスタートアップを立ち上げたことが紹介される。
日経産業新聞
自 2021年03月31日, 至 2021年03月31日
The scientists, led by Tatsuya Usui, PhD, DVM, senior assistant professor from the Laboratory of Veterinary Pharmacology at the Tokyo University of Agriculture and Technology, published their findings “Establishment of 2.5D culture model using 3D bladder cancer organoid culture” in Scientific Reports.
自 2020年06月29日, 至 2020年06月29日- 高速培養可能で3Dオルガノイドの特徴をもつ「2.5次元培養細胞」の作出に成功－東京農工大
東京農工大学大学院共同獣医学専攻のアミラ・アブゴマ大学院生（博士課程1年）、同大農学研究院の佐々木一昭准教授、臼井達哉特任講師らの研究チームが、膀胱がんの3次元（3D）オルガノイド培養組織から細胞を分離し、新たな培養液成分を用いてゲルフリーの2次元環境下（平面上）で高速培養が可能かつ、3Dオルガノイド培養組織の特徴を維持する新たな2.5次元培養細胞（2.5Dオルガノイド）を作出することに成功したことが紹介される。
QLifePro
自 2020年06月12日, 至 2020年06月12日
The research team led by Senior Assistant Professor Tatsuya Usui from the Laboratory of Veterinary Pharmacology at the Tokyo University of Agriculture and Technology published their findings in the journal Scientific Reports on June 10th, 2020.
自 2020年06月10日, 至 2020年06月10日
Dr Tatsuya Usui, corresponding author of the paper and Senior Assistant Professor at the Laboratory of Veterinary Pharmacology, Tokyo University of Agriculture and Technology (TUAT), Japan, concluded:
自 2020年04月03日, 至 2020年04月03日- 「NASH」病態再現 創薬研究に寄与
臼井達哉特任講師らは山口大学などと共同で、脂肪肝の一種である「非アルコール性脂肪肝炎（NASH）」の病態を再現した小さな肝臓組織を作製したことが紹介される。
日本経済新聞／WEB
自 2020年02月24日, 至 2020年02月24日 - 非アルコール性脂肪肝炎の病態進行を再現するオルガノイド作出に成功－東京農工大ほか
東京農工大学が、病態ステージの異なる「非アルコール性脂肪肝炎」(以下、NASH)モデルマウスの肝臓組織から、NASH病態の特徴である肝線維化を再現した三次元培養組織を作出することに成功したと発表したことが紹介される。
Q Life
自 2020年02月14日, 至 2020年02月14日 - 犬の抗がん剤、尿で効果確認
東京農工大学の臼井達哉特任講師らの研究グループが、ぼうこうがんの犬の尿から、がんの細胞を短期間で増やす手法を確立したことが紹介される。
日経産業新聞
自 2019年08月01日, 至 2019年08月01日 - 膀胱がんの犬の尿から膀胱がん組織を再現、東京農工大学が成功
東京農工大学の臼井達哉特任講師らが、膀胱がん罹患犬の尿に含まれる微量ながん幹細胞から生体内の膀胱がん組織を培養ディッシュ上で再現する新たな実験モデルを確立したことが紹介される。
大学ジャーナル
自 2019年07月28日, 至 2019年07月28日 - 東京農工大、膀胱がん罹患犬の尿から膀胱がん組織の再現に成功
日経新聞電子版
自 2019年07月24日, 至 2019年07月24日
所属学協会
受賞
- 第8回 日本毒性学会技術賞
新規三次元培養法，エアリキッドインターフェイス(ALI)オルガノイド培養法の確立と毒性試験への応用
2018年06月26日 - 第31回 二宮善基記念賞
Histone deacetylase 4 controls neointimal hyperplasia via stimulating proliferation and migration of vascular smooth muscle cells
2016年06月 - 平成24年度 獣医学奨励賞
血管炎症性障害を介して高血圧症進展を制御する新規カルモジュリン関連蛋白質の探索と機能解析
2012年03月 - 第153回 日本獣医学会学術集会 大会長賞
血管炎症性障害を介して高血圧症進展を制御する新規カルモジュリン関連蛋白質の探索と機能解析
2012年03月 - 第2回 日本比較薬理学・毒性学会 奨励賞
ジヒドロピリジン系カルシウム拮抗薬CV-159の血管平滑筋細胞における抗炎症作用
2009年09月