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7. Construction of in vitro patient‑derived tumor models to evaluate anticancer agents and cancer immunotherapy

Author

Takahashi, Nobuhiko, primary, Higa, Arisa, additional, Hiyama, Gen, additional, Tamura, Hirosumi, additional, Hoshi, Hirotaka, additional, Dobashi, Yuu, additional, Katahira, Kiyoaki, additional, Ishihara, Hiroya, additional, Takagi, Kosuke, additional, Goda, Kazuhito, additional, Okabe, Naoyuki, additional, Muto, Satoshi, additional, Suzuki, Hiroyuki, additional, Shimomura, Kenju, additional, Watanabe, Shinya, additional, and Takagi, Motoki, additional

Published
2021
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10. An In Vitro System for Evaluating Molecular Targeted Drugs Using Lung Patient-Derived Tumor Organoids

Author

Takahashi, Nobuhiko, primary, Hoshi, Hirotaka, additional, Higa, Arisa, additional, Hiyama, Gen, additional, Tamura, Hirosumi, additional, Ogawa, Mayu, additional, Takagi, Kosuke, additional, Goda, Kazuhito, additional, Okabe, Naoyuki, additional, Muto, Satoshi, additional, Suzuki, Hiroyuki, additional, Shimomura, Kenju, additional, Watanabe, Shinya, additional, and Takagi, Motoki, additional

Published
2019
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11. Evaluation of anticancer agents using patient-derived tumor organoids characteristically similar to source tissues

Author

Tamura, Hirosumi, primary, Higa, Arisa, additional, Hoshi, Hirotaka, additional, Hiyama, Gen, additional, Takahashi, Nobuhiko, additional, Ryufuku, Masae, additional, Morisawa, Gaku, additional, Yanagisawa, Yuka, additional, Ito, Emi, additional, Imai, Jun-Ichi, additional, Dobashi, Yuu, additional, Katahira, Kiyoaki, additional, Soeda, Shu, additional, Watanabe, Takafumi, additional, Fujimori, Keiya, additional, Watanabe, Shinya, additional, and Takagi, Motoki, additional

Published
2018
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12. Construction of a novel cell-based assay for the evaluation of anti-EGFR drug efficacy against EGFR mutation

Author

Hoshi, Hirotaka, primary, Hiyama, Gen, additional, Ishikawa, Kosuke, additional, Inageda, Kiyoshi, additional, Fujimoto, Jiro, additional, Wakamatsu, Ai, additional, Togashi, Takushi, additional, Kawamura, Yoshifumi, additional, Takahashi, Nobuhiko, additional, Higa, Arisa, additional, Goshima, Naoki, additional, Semba, Kentaro, additional, Watanabe, Shinya, additional, and Takagi, Motoki, additional

Published
2016
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16. A unique mechanism of successful fertilization in a domestic bird

Author

Sasanami Tomohiro, Izumi Shunsuke, Sakurai Naoki, Hirata Toshifumi, Mizushima Shusei, Matsuzaki Mei, Hiyama Gen, Yorinaga Eriko, Yoshimura Takashi, Ukena Kazuyoshi, Tsutsui Kazuyoshi, Sasanami Tomohiro, Izumi Shunsuke, Sakurai Naoki, Hirata Toshifumi, Mizushima Shusei, Matsuzaki Mei, Hiyama Gen, Yorinaga Eriko, Yoshimura Takashi, Ukena Kazuyoshi, and Tsutsui Kazuyoshi

Abstract

publisher

Published
2015

17. Lactic acid is a sperm motility inactivation factor in the sperm storage tubules

Author

Matsuzaki Mei, Mizushima Shusei, Hiyama Gen, Hirohashi Noritaka, Shiba Kogiku, Inaba Kazuo, Suzuki Tomohiro, Dohra Hideo, Ohnishi Toshiyuki, Sato Yoshikatsu, Kohsaka Tetsuya, Ichikawa Yoshinobu, Atsumi Yusuke, Yoshimura Takashi, Sasanami Tomohiro, Matsuzaki Mei, Mizushima Shusei, Hiyama Gen, Hirohashi Noritaka, Shiba Kogiku, Inaba Kazuo, Suzuki Tomohiro, Dohra Hideo, Ohnishi Toshiyuki, Sato Yoshikatsu, Kohsaka Tetsuya, Ichikawa Yoshinobu, Atsumi Yusuke, Yoshimura Takashi, and Sasanami Tomohiro

Abstract

publisher

Published
2015

18. Lactic acid is a sperm motility inactivation factor in the sperm storage tubules

Author

Matsuzaki, Mei, primary, Mizushima, Shusei, additional, Hiyama, Gen, additional, Hirohashi, Noritaka, additional, Shiba, Kogiku, additional, Inaba, Kazuo, additional, Suzuki, Tomohiro, additional, Dohra, Hideo, additional, Ohnishi, Toshiyuki, additional, Sato, Yoshikatsu, additional, Kohsaka, Tetsuya, additional, Ichikawa, Yoshinobu, additional, Atsumi, Yusuke, additional, Yoshimura, Takashi, additional, and Sasanami, Tomohiro, additional

Published
2015
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19. A unique mechanism of successful fertilization in a domestic bird

Author

Sasanami, Tomohiro, primary, Izumi, Shunsuke, additional, Sakurai, Naoki, additional, Hirata, Toshifumi, additional, Mizushima, Shusei, additional, Matsuzaki, Mei, additional, Hiyama, Gen, additional, Yorinaga, Eriko, additional, Yoshimura, Takashi, additional, Ukena, Kazuyoshi, additional, and Tsutsui, Kazuyoshi, additional

Published
2015
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22. The birth of quail chicks after intracytoplasmic sperm injection

Author

Mizushima Shusei, Hiyama Gen, Shiba Kogiku, Inaba Kazuo, Dohra Hideo, Ono Tamao, Shimada Kiyoshi, Sasanami Tomohiro, Mizushima Shusei, Hiyama Gen, Shiba Kogiku, Inaba Kazuo, Dohra Hideo, Ono Tamao, Shimada Kiyoshi, and Sasanami Tomohiro

Published
2014

39. Construction of a novel cell-based assay for the evaluation of anti-EGFR drug efficacy against EGFR mutation.

Author

Hoshi H, Hiyama G, Ishikawa K, Inageda K, Fujimoto J, Wakamatsu A, Togashi T, Kawamura Y, Takahashi N, Higa A, Goshima N, Semba K, Watanabe S, and Takagi M

Subjects
Afatinib, Antineoplastic Agents therapeutic use, Cell Culture Techniques, Cells, Cultured, Drug Resistance, Neoplasm drug effects, Erlotinib Hydrochloride pharmacology, Gefitinib, Humans, Mutation, Protein Kinase Inhibitors therapeutic use, Quinazolines pharmacology, Transfection, Antineoplastic Agents isolation & purification, Drug Evaluation, Preclinical methods, ErbB Receptors antagonists & inhibitors, ErbB Receptors genetics, Protein Kinase Inhibitors isolation & purification
Abstract

Epidermal growth factor receptor (EGFR) overexpression and EGFR-mediated signaling pathway dysregulation have been observed in tumors from patients with various cancers, especially non-small cell lung cancer. Thus, several anti-EGFR drugs have been developed for cancer therapy. For patients with known EGFR activating mutations (EGFR exon 19 in-frame deletions and exon 21 L858R substitution), treatment with an EGFR tyrosine kinase inhibitor (EGFR TKI; gefitinib, erlotinib or afatinib) represents standard first-line therapy. However, the clinical efficacy of these TKIs is ultimately limited by the development of acquired drug resistance such as by mutation of the gatekeeper T790 residue (T790M). To overcome this acquired drug resistance and develop novel anti-EGFR drugs, a cell-based assay system for EGFR TKI resistance mutant-selective inhibitors is required. We constructed a novel cell-based assay for the evaluation of EGFR TKI efficacy against EGFR mutation. To this end, we established non-tumorigenic immortalized breast epithelial cells that proliferate dependent on EGF (MCF 10A cells), which stably overexpress mutant EGFR. We found that the cells expressing EGFR containing the T790M mutation showed higher resistance against gefitinib, erlotinib and afatinib compared with cells expressing wild-type EGFR. In contrast, L858R mutant-expressing cells exhibited higher TKI sensitivity. The effect of T790M-selective inhibitors (osimertinib and rociletinib) on T790M mutant-expressing cells was significantly higher than gefitinib and erlotinib. Finally, when compared with commercially available isogenic MCF 10A cell lines carrying introduced mutations in EGFR, our EGFR mutant-overexpressing cells exhibited obviously higher responsiveness to EGFR TKIs depending on the underlying mutations because of the higher levels of EGFR phosphorylation in the EGFR mutant-overexpressing cells than in the isogenic cell lines. In conclusion, we successfully developed a novel cell-based assay for evaluating the efficacy of anti-EGFR drugs against EGFR mutation.

Published
2017
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40. Characterization and Expression of Turkey Prolactin Regulatory Element Binding in the Anterior Pituitary Gland and Pancreas During Embryogenesis.

Author

Hiyama G, Kansaku N, Wakui S, McQuaid R, and Zadworny D

Abstract

The PRL regulatory element-binding (PREB) protein is a transcription factor that was originally cloned from the rat anterior pituitary gland and characterized as a regulator of the PRL promoter. It is also strongly expressed in several extrapituitary tissues; however, its functional role is not well understood to date. In this study, we aimed to clone and characterize the turkey PREB gene and investigate its mRNA expression in the anterior pituitary gland and pancreas during embryogenesis. Based on the conserved sequence of chicken and mammalian PREB cDNAs, a turkey PREB cDNA fragment was obtained, and after sequencing of the fragment, the 5'-and 3'-ends of mRNA were amplified and determined. To identify the PREB gene structure, polymerase chain reaction (PCR) amplification was performed. The turkey PREB gene consists of 9 exons and 8 introns, and it encodes a 411-amino-acid protein. The expression of PREB mRNA in the anterior pituitary gland was measured during embryogenesis. Levels of PREB mRNA significantly increased at embryonic day 22, with maximum levels being detected on day 25 of ontogeny, which correlated with similar changes in levels of PRL mRNA. The highest level of PREB mRNA was detected on day 19 in the pancreas. However, the highest level of insulin mRNA was detected at embryonic day 25. These results indicate that PREB may be involved in the expression of PRL mRNA in the anterior pituitary gland, whereas insulin mRNA may be expressed independently of the expression of PREB mRNA in the pancreas during embryogenesis., (2016, Japan Poultry Science Association.)

Published
2016
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41. Sperm activation by heat shock protein 70 supports the migration of sperm released from sperm storage tubules in Japanese quail (Coturnix japonica).

Author

Hiyama G, Matsuzaki M, Mizushima S, Dohra H, Ikegami K, Yoshimura T, Shiba K, Inaba K, and Sasanami T

Subjects
Animals, Antibodies administration & dosage, Female, Fertilization drug effects, Fertilization in Vitro drug effects, Gene Expression, HSP70 Heat-Shock Proteins genetics, HSP70 Heat-Shock Proteins immunology, Male, Oviducts chemistry, Oviposition, Ovulation, RNA, Messenger analysis, Sperm Motility drug effects, Spermatozoa chemistry, Uterus drug effects, Voltage-Dependent Anion Channel 2 physiology, Coturnix, HSP70 Heat-Shock Proteins pharmacology, Oviducts physiology, Sperm Transport physiology, Spermatozoa drug effects, Spermatozoa physiology
Abstract

Systems for maintaining the viability of ejaculated sperm in the female reproductive tract are widespread among vertebrates and invertebrates. In birds, this sperm storage function is performed by specialized simple tubular invaginations called sperm storage tubules (SSTs) in the uterovaginal junction (UVJ) of the oviduct. Although the incidence and physiological reasons for sperm storage in birds have been reported extensively, the mechanisms of sperm uptake by the SSTs, sperm maintenance within the SSTs, and control of sperm release from the SSTs are poorly understood. In this study, we demonstrated that the highly conserved heat shock protein 70 (HSP70) stimulates sperm motility in vitro and also that HSP70 expressed in the UVJ may facilitate the migration of sperm released from the SSTs. Quantitative RT-PCR analysis demonstrated that the expression of HSP70 mRNA in the UVJ increases before ovulation/oviposition. Gene-specific in situ hybridization and immunohistochemical analysis with a specific antibody to HSP70 demonstrated that HSP70 is localized in the surface epithelium of the UVJ. Furthermore, injection of anti-HSP70 antibody into the vagina significantly inhibited fertilization in vivo. In addition, we found that recombinant HSP70 activates flagellar movement in the sperm and that the binding of recombinant HSP70 to the sperm surface is mediated through an interaction with voltage-dependent anion channel protein 2 (VDAC2). Our results suggest that HSP70 binds to the sperm surface by interacting with VDAC2 and activating sperm motility. This binding appears to play an important role in sperm migration within the oviduct.

Published
2013
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