研究者詳細 - 井藤　彰

論文 - 井藤　彰

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Intramyocardial Transplantation ofHuman iPS Cell–Derived CardiacSpheroids Improves Cardiac Function inHeart Failure Animals. 査読有り

JACC: Basic to Translational Science 36 巻 ( 4 ) 頁： 101 - 108 2021年

Heat dose based large tumor treatment with multiple site injections of heat-generating nanoparticles dispersible within tumor tissue 査読有り

2. Tomio Morino, Akira Ito, Toshiki Etani, Taku Naiki, Noriyasu Kawai, Takahiro Yasui

Thermal Medicine 36 巻 ( 4 ) 頁： 104 - 108 2020年12月

腫瘍内分散性と発熱性を有するマグネタイト正電荷脂質複合粒子の腫瘍内マルチサイト投与法を用いた大型腫瘍の治療条件とその完全退縮効果

森野富夫, 井藤彰, 惠谷俊紀, 内木拓, 河合憲康, 安井孝周

36 巻 ( 4 ) 頁： 101 - 108 2020年12月

記述言語：英語出版者・発行元：日本ハイパーサーミア学会

腫瘍内に局所投与したマグネタイト正電荷脂質複合粒子（MCL粒子）に交番磁場を照射し，その発熱誘導により癌を治療する方法が，本邦で開発された．筆者らは，前報において，腫瘍体積当たりの発熱量（J/cm3 tumor volume）がインビボの抗腫瘍効果に相関することを示し，その施術管理指標（ヒートドーズ，J/cm3 tumor volume）としての有用性を報告した．更に，体積が1.36 cm3の腫瘍の単一サイトに45 mgのMCL粒子を投与した条件において，完全退縮に必要なヒートドーズの基準値（700～850 J/cm3）を示した．本研究の目的は，より大型の臨床腫瘍の治療を想定して，MCL粒子を腫瘍の複数サイトへ投与する際の治療条件をデザインし，その抗腫瘍効果を検証することにある．動物腫瘍モデルには，化学物質で誘発したラット乳腺腫瘍（2.19～3.81 cm3）を用いた．ラット乳腺腫瘍の治療条件は，MCL粒子を投与した複数のサイトで，前報の単一サイト投与の治療条件を再現することを基本コンセプトとして，デザインした．投与サイト数は，腫瘍体積（cm3）を除した値が1.36 cm3/siteに近似するように設定し，MCL粒子は腫瘍内の投与サイトが均等間隔となるように注射針を穿刺し，緩徐に同時投与した．ヒートドーズ（J/cm3）は，前報の基準値に近似するように照射条件を調節し，同様に30分3連日の磁場照射を行った．この結果，治療開始21日目に全例で完全退縮が確認され，大型腫瘍に対する複数サイト投与と照射条件のデザイン法の妥当性が示された．更に，技術的，実務的に最良と考えられる定型的な治療条件（投与量，ヒートドーズ，照射プロトコール）を規定して，当該条件で誘導される壊死領域（cm3 necrosed tumor volume/site）の推定法を示し，その臨床活用法を論じた．

DOI： 10.3191/thermalmed.36.101

CiNii Research

ハイパーサーミアの新展開 -基礎医学・臨床応用に関する最近の進歩- 査読有り

近藤隆、鈴木文男、水上達治、斎藤淳一、河合憲康、井藤　彰

放射線生物研究 55 巻 ( 3 ) 頁： 210 - 228 2020年9月

Magnetic heating of nanoparticles as a scalable cryopreservation technology for human induced pluripotent stem cells

Ito Akira, Yoshioka Kantaro, Masumoto Shinya, Sato Keiichiro, Hatae Yuki, Nakai Tomoki, Yamazaki Takashi, Takahashi Masazumi, Tanoue Shota, Horie Masanobu

SCIENTIFIC REPORTS 10 巻 ( 1 ) 2020年8月

マグネタイト正電荷脂質複合粒子と交番磁場照射装置を用いた固形癌治療における新規な腫瘍内発熱量指標と施術条件のデザイン法

森野富夫, 田上祥太, 宮田周一郎, 平山鋼太郎, 井藤彰, 惠谷俊紀, 内木拓, 河合憲康, 安井孝周

36 巻 ( 2 ) 頁： 47 - 58 2020年7月

記述言語：英語出版者・発行元：日本ハイパーサーミア学会

腫瘍内に局所投与したナノサイズのマグネタイト正電荷脂質複合粒子（magnetite cationic liposomes, MCL）に交番磁場を照射し，その発熱誘導により癌を治療する方法が開発され，本邦で臨床研究が実施された．しかしながら，腫瘍温度は等しく上昇したにも拘わらず，その有効性は完全退縮～ほぼ無効まで広範囲に亘った．筆者らは，MCLのインビトロの殺細胞活性が培養液温度に相関しないこと，臨床効果が腫瘍体積当たりの投与量（mg-MCL/cm3 tumor volume）に相関することを示し，腫瘍温度に代わる指標として，腫瘍体積当たりの発熱量（J/cm3 tumor volume）を提言した．本研究の目的は，動物腫瘍モデルを用いて，その有用性を検証することにある．MCLの腫瘍内発熱量の算出を可能にするために，治療用装置で想定される腫瘍位置（mm）と出力（kW）をパラメータにして，MCLの発熱活性（J/g-MCL・min）を測定した．本装置を用いて，小型腫瘍（7 mm径）の完全退縮条件を再現し，その照射条件におけるMCLの発熱活性（J/g-MCL・min）に，腫瘍体積当たりの投与量（g-MCL/cm3）と照射時間（min）を乗じて，腫瘍内発熱量（J/cm3）を算出した．この結果，完全退縮に十分な1回照射当たりの腫瘍内発熱量は，大凡700～850 J/cm3であることが判明した．既に，温度指標では，大型腫瘍（15 mm径）における効果の減弱傾向が示されていたため，算出された腫瘍内発熱量を指標に，ラット大型腫瘍（13～16 mm径）の治療条件を机上でデザインし，その単回クールでの完全退縮効果を実証した．更に，温度指標では大型腫瘍の投与量設定（g-MCL/cm3）が各段に低いことが判明したため，測温部の温度上昇は伝熱により比較的容易に達成されるものの，低い投与量設定（g-MCL/cm3）に連動して腫瘍内発熱量（J/cm3）が低値となることが，効果減弱の要因と考察した．腫瘍内発熱量を指標とした治療条件のデザイン手法を示し，その臨床フィジビリティと装置開発の今後の方向性を論じた．

DOI： 10.3191/thermalmed.36.47

CiNii Research

若手化学工学者としての私の履歴書招待有り査読有り

井藤　彰

学術の動向 25 巻 ( 7 ) 頁： 98 - 102 2020年7月

若手化学工学者としての私の履歴書

井藤彰

学術の動向 25 巻 ( 7 ) 頁： 7_98 - 7_102 2020年7月

Heat dose index in vivo for cancer therapy using heat-generating nanoparticles named magnetite cationic liposomes and alternating magnetic field irradiator 査読有り

Tomio Morino, Shota Tanoue, Shuichiro Miyata, Kotaro Hirayama, Akira Ito, Toshiki Etani, Taku Naiki, Noriyasu Kawai, Takahiro Yasui

36 巻 ( 2 ) 頁： 47 - 58 2020年7月

Novel neuromuscular junction model in 2D and 3D myotubes co-cultured with induced pluripotent stem cell-derived motor neurons 査読有り

Yoshioka, K; Ito, A; Kawabe, Y; Kamihira, M

JOURNAL OF BIOSCIENCE AND BIOENGINEERING 129 巻 ( 4 ) 頁： 486 - 493 2020年4月

Targeted knock-in into the OVA locus of chicken cells using CRISPR/Cas9 system with homology-independent targeted integration 査読有り

Shi, M; Kawabe, Y; Ito, A; Kamihira, M

JOURNAL OF BIOSCIENCE AND BIOENGINEERING 129 巻 ( 3 ) 頁： 363 - 370 2020年3月

Medium temperature independent cytotoxicity of cell-adhesive heat-generating nanoparticles named magnetite cationic liposomes and its therapeutic use 査読有り

Tomio Morino, Hiroshi Takase, Toshiki Etani, Taku Naiki, Noriyasu Kawai, Akira Ito, Takahiro Yasui

Thermal Medicine 36 巻 ( 1 ) 頁： 25 - 34 2020年3月

Cre-Mediated Transgene Integration in Chinese Hamster Ovary Cells Using Minicircle DNA Vectors. 査読有り国際誌

Wang X, Kawabe Y, Hada T, Ito A, Kamihira M

Biotechnology journal 13 巻 ( 7 ) 頁： e1800063 2018年7月

Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems 査読有り

Yoshinori Kawabe, Shinya Komatsu, Shodai Komatsu, Mai Murakami, Akira Ito, Tetsushi Sakuma, Takahiro Nakamura, Takashi Yamamoto, Masamichi Kamihira

Journal of Bioscience and Bioengineering 125 巻 ( 5 ) 頁： 599 - 605 2018年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier B.V.

Chinese hamster ovary (CHO) cells have been used as host cells for the production of pharmaceutical proteins. For the high and stable production of target proteins, the transgene should be integrated into a suitable genomic locus of host cells. Here, we generated knock-in CHO cells, in which transgene cassettes without a vector backbone sequence were integrated into the hprt locus of the CHO genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) systems. We investigated the efficiency of targeted knock-in of transgenes using these systems. As a practical example, we generated knock-in CHO cells producing an scFv-Fc antibody using the CRIS-PITCh system mediated by microhomology sequences for targeting. We found that the CRIS-PITCh system can facilitate targeted knock-in for CHO cell engineering.

DOI： 10.1016/j.jbiosc.2017.12.003

Scopus

PubMed

Characterization of genetically engineered mouse hepatoma cells with inducible liver functions by overexpression of liver-enriched transcription factors 査読有り

Hideaki Yamamoto, Jane Marie Tonello, Takanori Sambuichi, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira

Journal of Bioscience and Bioengineering 125 巻 ( 1 ) 頁： 131 - 139 2018年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier B.V.

New cell sources for the research and therapy of organ failure could significantly alleviate the shortage of donor livers that are available to patients who suffer from liver disease. Liver carcinoma derived cells, or hepatoma cells, are the ideal cells for developing bioartificial liver systems. Such cancerous liver cells are easy to prepare in large quantities and can be maintained over long periods under standard culture conditions, unlike primary hepatocytes. However, hepatoma cells possess only a fraction of the functions of primary hepatocytes. In a previous study, by transducing cells with liver-enriched transcription factors that could be inducibly overexpressed—hepatocyte nuclear factor (HNF)1α, HNF1β, HNF3β [FOXA2], HNF4α, HNF6, CCAAT/enhancer binding protein (C/EBP)α, C/EBPβ and C/EBPγ—we created mouse hepatoma cells with high liver-specific gene expression called the Hepa/8F5 cell line. In the present study, we performed functional and genetic analyses to characterize the Hepa/8F5 cell line. Further, in three-dimensional cultures, the function of these cells improved significantly compared to parental cells. Ultimately, these cells might become a new resource that can be used in basic and applied hepatic research.

DOI： 10.1016/j.jbiosc.2017.07.011

Scopus

PubMed

温熱誘導型遺伝子発現システムの開発と応用招待有り査読有り

井藤　彰

放射線生物研究 53 巻 ( 2 ) 頁： 104-114 2018年

ハイパーサーミアに伴う免疫賦活のメカニズム招待有り査読有り

井藤彰

放射線生物研究 53 巻 ( 2 ) 頁： 104-114 2018年

Three-dimensional culture of a genetically modified hepatoma cell line using macroporous gelatin beads 査読有り

Jane Marie Tonello, Saori Kawashima, Kazuki Sato, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira

CYTOTECHNOLOGY 69 巻 ( 6 ) 頁： 925 - 931 2017年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SPRINGER

Hepatoma cells are a candidate cell source for bio-artificial livers. However, they exhibit reduced liver functions compared with primary hepatocytes. In our previous study, genetically engineered mouse hepatoma cells were created by transduction with vectors mediating inducible overexpression of eight liver-enriched transcription factors. Upon the induction of the liver-enriched transcription factors transduced, the cells expressed both phenotypic and genotypic liver functions at high levels. In the present study, we performed three-dimensional culture of these cells using macroporous gelatin beads. When immobilized on the macroporous gelatin beads, these cells exhibited further enhancement in liver functionality, including increased albumin secretion, ammonia removal and cytochrome P450 activity. The levels of these functions were significantly enhanced compared to monolayer culture. The method is simple and scalable, and provides highly functional cells that can be used in basic and applied fields of hepatic research.

DOI： 10.1007/s10616-017-0117-0

Web of Science

PubMed

Accumulative scFv-Fc antibody gene integration into the &IThprt&IT chromosomal locus of Chinese hamster ovary cells 査読有り

Xue Wang, Yoshinori Kawabe, Risa Kato, Takeshi Hada, Akira Ito, Yoshimasa Yamana, Masako Kondo, Masamichi Kamihira

JOURNAL OF BIOSCIENCE AND BIOENGINEERING 124 巻 ( 5 ) 頁： 583 - 590 2017年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SOC BIOSCIENCE BIOENGINEERING JAPAN

We have previously developed an accumulative site-specific gene integration system (AGIS) using Cre-recombinase and mutated loxP sites. AGIS enables repeated transgene integration into a predetermined chromosomal site in mammalian cells. However, the process of establishing cells with multiple integrated copies of the transgene is still time-consuming. In the present study, we describe an improved version of AGIS that facilitates and accelerates the establishment of high-producer Chinese hamster ovary (CHO) cells. Two donor vectors were simultaneously introduced into the cells in a single transfection. Cells with successfully targeted transgene integration were screened based on a change in the color of the reporter fluorescent protein that they express. Repeated rounds of integration allowed the transgene copy number to be increased. As a model, an scFv-Fc antibody gene was integrated into the hprt locus of the CHO cell genome. After three rounds of integration, a high-producer CHO cell clone with six copies of the scFv-Fc gene was successfully established. scFv-Fc productivity was approximately four-fold greater than a control cell line harboring a single copy of the transgene. This newly designed AGIS procedure should facilitate the development of producer cells suitable for biopharmaceutical protein production. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2017.05.017

Web of Science

PubMed

Hypoxia-responsive transgene expression system using RTP801 promoter and synthetic transactivator fused with oxygen-dependent degradation domain 査読有り

Akihiko Ono, Akira Ito, Tomonaga Sato, Masaki Yamaguchi, Taiga Suzuki, Yoshinori Kawabe, Masamichi Kamihira

JOURNAL OF BIOSCIENCE AND BIOENGINEERING 124 巻 ( 1 ) 頁： 115 - 124 2017年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SOC BIOSCIENCE BIOENGINEERING JAPAN

Precise control of gene expression using an artificial gene circuit is a major challenge in the application of synthetic biology. Here, we designed a hypoxia-responsive transgene expression system by combining a hypoxia-inducible RTP801 promoter and a tetracycline-responsive transactivator fused with an oxygen-dependent degradation domain (TA-ODD). The reporter gene expression was highly induced by hypoxia when a transactivator-expression plasmid, pRTP801/TA-ODD, harboring a TA-ODD gene driven by the RTP801 promoter, was cotransfected with a reporter plasmid, pTRE/EGFP, harboring an EGFP gene controlled under the transactivator-responsive promoter. A stable cell line into which the expression units RTP801/TA-ODD and THE/EGFP had been introduced responded to hypoxia and expressed the reporter gene in an oxygen-concentration-dependent manner. Moreover, the cells demonstrated potential as sensors to detect hypoxic conditions in a three-dimensional tissue culture in vitro. These results indicate that the hypoxia-responsive transgene expression system is useful for constructing cell-based hypoxia detection systems. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2017.02.012

Web of Science

PubMed

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