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152. Cyclization of Single-Chain Fv Antibodies Markedly Suppressed Their Characteristic Aggregation Mediated by Inter-Chain VH-VL Interactions

Author

Yamauchi, Soichiro, primary, Kobashigawa, Yoshihiro, additional, Fukuda, Natsuki, additional, Teramoto, Manaka, additional, Toyota, Yuya, additional, Liu, Chenjiang, additional, Ikeguchi, Yuka, additional, Sato, Takashi, additional, Sato, Yuko, additional, Kimura, Hiroshi, additional, Masuda, Takeshi, additional, Ohtsuki, Sumio, additional, Noi, Kentaro, additional, Ogura, Teru, additional, and Morioka, Hiroshi, additional

Published
2019
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153. Tandem Mass Spectrometry Imaging Reveals Distinct Accumulation Patterns of Steroid Structural Isomers in Human Adrenal Glands

Author

Takeo, Emi, primary, Sugiura, Yuki, additional, Uemura, Tatsuki, additional, Nishimoto, Koshiro, additional, Yasuda, Masanori, additional, Sugiyama, Eiji, additional, Ohtsuki, Sumio, additional, Higashi, Tatsuya, additional, Nishikawa, Tetsuo, additional, Suematsu, Makoto, additional, Fukusaki, Eiichiro, additional, and Shimma, Shuichi, additional

Published
2019
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154. Large-Scale Quantitative Comparison of Plasma Transmembrane Proteins between Two Human Blood–Brain Barrier Model Cell Lines, hCMEC/D3 and HBMEC/ciβ

Author

Masuda, Takeshi, primary, Hoshiyama, Toshiki, additional, Uemura, Tatsuki, additional, Hirayama-Kurogi, Mio, additional, Ogata, Seiryo, additional, Furukawa, Arisu, additional, Couraud, Pierre-Olivier, additional, Furihata, Tomomi, additional, Ito, Shingo, additional, and Ohtsuki, Sumio, additional

Published
2019
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162. Attenuation of prostaglandin E2 elimination across the mouse blood-brain barrier in lipopolysaccharide-induced inflammation and additive inhibitory effect of cefmetazole

Author

Akanuma Shin-ichi, Uchida Yasuo, Ohtsuki Sumio, Tachikawa Masanori, Terasaki Tetsuya, and Hosoya Ken-ichi

Subjects
Blood-brain barrier, lipopolysaccharide, inflammation, multidrug resistance-associated protein, MRP4, Oat3, Oatp1a4, PGE2, prostaglandin, transporter, Neurology. Diseases of the nervous system, RC346-429
Abstract

Abstract Background Peripheral administration of lipopolysaccharide (LPS) induces inflammation and increases cerebral prostaglandin E2 (PGE2) concentration. PGE2 is eliminated from brain across the blood-brain barrier (BBB) in mice, and this process is inhibited by intracerebral or intravenous pre-administration of anti-inflammatory drugs and antibiotics such as cefmetazole and cefazolin that inhibit multidrug resistance-associated protein 4 (Mrp4/Abcc4)-mediated PGE2 transport. The purpose of this study was to examine the effect of LPS-induced inflammation on PGE2 elimination from brain, and whether antibiotics further inhibit PGE2 elimination in LPS-treated mice. Methods [3H]PGE2 elimination across the BBB of intraperitoneally LPS-treated mice was assessed by the brain efflux index (BEI) method. Transporter protein amounts in brain capillaries were quantified by liquid chromatography-tandem mass spectrometry. Results The apparent elimination rate of [3H]PGE2 from brain was lower by 87%, in LPS-treated mice compared with saline-treated mice. The Mrp4 protein amount was unchanged in brain capillaries of LPS-treated mice compared with saline-treated mice, while the protein amounts of organic anion transporter 3 (Oat3/Slc22a8) and organic anion transporting polypeptide 1a4 (Oatp1a4/Slco1a4) were decreased by 26% and 39%, respectively. Either intracerebral or intravenous pre-administration of cefmetazole further inhibited PGE2 elimination in LPS-treated mice. However, intracerebral or intravenous pre-administration of cefazolin had little effect on PGE2 elimination in LPS-treated mice, or in LPS-untreated mice given Oat3 and Oatp1a4 inhibitors. These results indicate that peripheral administration of cefmetazole inhibits PGE2 elimination across the BBB in LPS-treated mice. Conclusion PGE2 elimination across the BBB is attenuated in an LPS-induced mouse model of inflammation. Peripheral administration of cefmetazole further inhibits PGE2 elimination in LPS-treated mice.

Published
2011
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163. 1α,25-Dihydroxyvitamin D3 enhances cerebral clearance of human amyloid-β peptide(1-40) from mouse brain across the blood-brain barrier

Author

Ito Shingo, Ohtsuki Sumio, Nezu Yasuko, Koitabashi Yusuke, Murata Sho, and Terasaki Tetsuya

Subjects
Neurology. Diseases of the nervous system, RC346-429
Abstract

Abstract Background Cerebrovascular dysfunction has been considered to cause impairment of cerebral amyloid-β peptide (Aβ) clearance across the blood-brain barrier (BBB). Further, low levels of vitamin D are associated with increased risk of Alzheimer's disease, as well as vascular dysfunction. The purpose of the present study was to investigate the effect of 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3), an active form of vitamin D, on cerebral Aβ clearance from mouse brain. Methods The elimination of [125I]hAβ(1-40) from mouse brain was examined by using the Brain Efflux Index method to determine the remaining amount of [125I]hAβ(1-40) radioactivity after injection into the cerebral cortex. [125I]hAβ(1-40) internalization was analyzed using conditionally immortalized mouse brain capillary endothelial cells (TM-BBB4). Results Twenty-four hours after intraperitoneal injection of 1,25(OH)2D3 (1 μg/mouse), [125I]hAβ(1-40) elimination from mouse brain was increased 1.3-fold, and the level of endogenous Aβ(1-40) in mouse brain was reduced. These effects were observed at 24 h after i.p. injection of 1,25(OH)2D3, while no significant effect was observed at 48 or 72 h. Vitamin D receptor (VDR) mRNA was detected in mouse brain capillaries, suggesting that 1,25(OH)2D3 has a VDR-mediated genomic action. Furthermore, forskolin, which activates mitogen-activated protein kinase kinase (MEK), enhanced [125I]hAβ(1-40) elimination from mouse brain. Forskolin also enhanced [125I]hAβ(1-40) internalization in TM-BBB4 cells, and this enhancement was inhibited by a MEK inhibitor, suggesting involvement of non-genomic action. Conclusions The active form of vitamin D, 1,25(OH)2D3, appears to enhance brain-to-blood Aβ(1-40) efflux transport at the BBB through both genomic and non-genomic actions. Compounds activating these pathways may be candidate agents for modulating Aβ(1-40) elimination at the BBB.

Published
2011
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164. Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis.

Author

Ogata, Seiryo, Ito, Shingo, Masuda, Takeshi, and Ohtsuki, Sumio

Abstract

Isolated brain capillaries are essential for analyzing the changes of protein expressions at the blood–brain barrier (BBB) under pathological conditions. The standard brain capillary isolation methods require the use of at least five mouse brains in order to obtain a sufficient amount and purity of brain capillaries. The purpose of this study was to establish a brain capillary isolation method from a single mouse brain for protein expression analysis. We successfully isolated brain capillaries from a single frozen mouse brain by using a bead homogenizer in the brain homogenization step and combination of cell strainers and glass beads in the purification step. Western blot and proteomic analysis showed that proteins expressed at the BBB in mouse brain capillaries isolated by the developed method were more enriched than those isolated from a pool of five mouse brains by the standard method. By using the developed method, we further verified the changes in expression of BBB proteins in Glut1 -deficient mouse. The developed method is useful for the analysis of various mice models with low numbers and enables us to understand, in more detail, the physiology and pathology of BBB. [ABSTRACT FROM AUTHOR]

Published
2021
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170. SIRT7 has a critical role in bone formation by regulating lysine acylation of SP7/Osterix

Author

Fukuda, Masatoshi, primary, Yoshizawa, Tatsuya, additional, Karim, Md. Fazlul, additional, Sobuz, Shihab U., additional, Korogi, Wataru, additional, Kobayasi, Daiki, additional, Okanishi, Hiroki, additional, Tasaki, Masayoshi, additional, Ono, Katsuhiko, additional, Sawa, Tomohiro, additional, Sato, Yoshifumi, additional, Chirifu, Mami, additional, Masuda, Takeshi, additional, Nakamura, Teruya, additional, Tanoue, Hironori, additional, Nakashima, Kazuhisa, additional, Kobashigawa, Yoshihiro, additional, Morioka, Hiroshi, additional, Bober, Eva, additional, Ohtsuki, Sumio, additional, Yamagata, Yuriko, additional, Ando, Yukio, additional, Oike, Yuichi, additional, Araki, Norie, additional, Takeda, Shu, additional, Mizuta, Hiroshi, additional, and Yamagata, Kazuya, additional

Published
2018
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171. Involvement of Claudin-11 in Disruption of Blood-Brain, -Spinal Cord, and -Arachnoid Barriers in Multiple Sclerosis

Author

Uchida, Yasuo, primary, Sumiya, Tomohito, additional, Tachikawa, Masanori, additional, Yamakawa, Tatsuya, additional, Murata, Sho, additional, Yagi, Yuta, additional, Sato, Kazuki, additional, Stephan, Alice, additional, Ito, Katsuaki, additional, Ohtsuki, Sumio, additional, Couraud, Pierre-Olivier, additional, Suzuki, Takashi, additional, and Terasaki, Tetsuya, additional

Published
2018
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172. Characterization of P-Glycoprotein Humanized Mice Generated by Chromosome Engineering Technology: Its Utility for Prediction of Drug Distribution to the Brain in Humans

Author

Yamasaki, Yuki, primary, Kobayashi, Kaoru, additional, Okuya, Fuka, additional, Kajitani, Naoyo, additional, Kazuki, Kanako, additional, Abe, Satoshi, additional, Takehara, Shoko, additional, Ito, Shingo, additional, Ogata, Seiryo, additional, Uemura, Tatsuki, additional, Ohtsuki, Sumio, additional, Minegishi, Genki, additional, Akita, Hidetaka, additional, Chiba, Kan, additional, Oshimura, Mitsuo, additional, and Kazuki, Yasuhiro, additional

Published
2018
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174. Identification of blood biomarkers in glioblastoma by SWATH mass spectrometry and quantitative targeted absolute proteomics

Author

Miyauchi, Eisuke, primary, Furuta, Takuya, additional, Ohtsuki, Sumio, additional, Tachikawa, Masanori, additional, Uchida, Yasuo, additional, Sabit, Hemragul, additional, Obuchi, Wataru, additional, Baba, Tomoko, additional, Watanabe, Michitoshi, additional, Terasaki, Tetsuya, additional, and Nakada, Mitsutoshi, additional

Published
2018
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181. Application of Quantitative Targeted Absolute Proteomics to Profile Protein Expression Changes of Hepatic Transporters and Metabolizing Enzymes During Cholic Acid-Promoted Liver Regeneration

Author

Miura, Takayuki, primary, Tachikawa, Masanori, additional, Ohtsuka, Hideo, additional, Fukase, Koji, additional, Nakayama, Shun, additional, Sakata, Naoaki, additional, Motoi, Fuyuhiko, additional, Naitoh, Takeshi, additional, Katayose, Yu, additional, Uchida, Yasuo, additional, Ohtsuki, Sumio, additional, Terasaki, Tetsuya, additional, and Unno, Michiaki, additional

Published
2017
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182. Multi-laboratory assessment of reproducibility, qualitative and quantitative performance of SWATH-mass spectrometry

Author

Collins, Ben C., primary, Hunter, Christie L., additional, Liu, Yansheng, additional, Schilling, Birgit, additional, Rosenberger, George, additional, Bader, Samuel L., additional, Chan, Daniel W., additional, Gibson, Bradford W., additional, Gingras, Anne-Claude, additional, Held, Jason M., additional, Hirayama-Kurogi, Mio, additional, Hou, Guixue, additional, Krisp, Christoph, additional, Larsen, Brett, additional, Lin, Liang, additional, Liu, Siqi, additional, Molloy, Mark P., additional, Moritz, Robert L., additional, Ohtsuki, Sumio, additional, Schlapbach, Ralph, additional, Selevsek, Nathalie, additional, Thomas, Stefani N., additional, Tzeng, Shin-Cheng, additional, Zhang, Hui, additional, and Aebersold, Ruedi, additional

Published
2017
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187. Gene therapy for a mouse model of glucose transporter-1 deficiency syndrome

Author

Nakamura, Sachie, primary, Osaka, Hitoshi, additional, Muramatsu, Shin-ichi, additional, Takino, Naomi, additional, Ito, Mika, additional, Aoki, Shiho, additional, Jimbo, Eriko F., additional, Shimazaki, Kuniko, additional, Onaka, Tatsushi, additional, Ohtsuki, Sumio, additional, Terasaki, Tetsuya, additional, and Yamagata, Takanori, additional

Published
2017
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188. Downregulation of GNA13-ERK network in prefrontal cortex of schizophrenia brain identified by combined focused and targeted quantitative proteomics

Author

Hirayama-Kurogi, Mio, primary, Takizawa, Yohei, additional, Kunii, Yasuto, additional, Matsumoto, Junya, additional, Wada, Akira, additional, Hino, Mizuki, additional, Akatsu, Hiroyasu, additional, Hashizume, Yoshio, additional, Yamamoto, Sakon, additional, Kondo, Takeshi, additional, Ito, Shingo, additional, Tachikawa, Masanori, additional, Niwa, Shin-Ichi, additional, Yabe, Hirooki, additional, Terasaki, Tetsuya, additional, Setou, Mitsutoshi, additional, and Ohtsuki, Sumio, additional

Published
2017
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190. Involvement of Claudin-11 in Disruption of Blood-Brain, -Spinal Cord, and -Arachnoid Barriers in Multiple Sclerosis.

Author

Uchida, Yasuo, Sumiya, Tomohito, Tachikawa, Masanori, Yamakawa, Tatsuya, Murata, Sho, Yagi, Yuta, Sato, Kazuki, Stephan, Alice, Ito, Katsuaki, Ohtsuki, Sumio, Couraud, Pierre-Olivier, Suzuki, Takashi, and Terasaki, Tetsuya

Abstract

It is important to understand the molecular mechanisms of barrier disruption in the central nervous system (CNS) of patients with multiple sclerosis (MS). The purpose of the present study was to clarify whether claudin-11 is involved in the disruption of two endothelial barriers (blood-brain barrier (BBB) and blood-spinal cord barrier (BSCB)) and two epithelial barriers (blood-arachnoid barrier (BAB) and blood-CSF barrier (BCSFB)) in the CNS in MS. Immunohistochemical analysis revealed that, in both normal human and mouse, claudin-11 is co-localized with claudin-5 in the brain and spinal cord capillaries. The absolute protein expression level of claudin-11 was nearly equal to that of claudin-5 in rat brain capillaries, but was 2.81-fold greater in human brain capillaries. The protein expressions of claudin-11 were significantly downregulated in the brain and spinal cord capillaries of an MS patient and experimental autoimmune encephalomyelitis (EAE) mice. Specific downregulation of claudin-11 with siRNA significantly increased the transfer of membrane-impermeable FITC-dextran across human brain capillary endothelial cell (hCMEC/D3) monolayer. As for the epithelial barrier, claudin-11 protein expression was not decreased in choroid plexus epithelial cells forming the BCSFB in EAE mice, whereas it was decreased in brain and spinal cord meninges that form the BAB. Specific downregulation of claudin-11 with siRNA in a rat choroid plexus epithelial cell (TR-CSFB) monolayer significantly increased the permeability of FITC-dextran. In conclusion, our present findings indicate that claudin-11 expression at the BBB, BSCB, and BAB, but not the BCSFB, is downregulated in multiple sclerosis, impairing the functional integrity of these barriers. [ABSTRACT FROM AUTHOR]

Published
2019
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191. Combining Genomics To Identify the Pathways of Post-Transcriptional Nongenotoxic Signaling and Energy Homeostasis in Livers of Rats Treated with the Pregnane X Receptor Agonist, Pregnenolone Carbonitrile

Author

Nagahori, Hirohisa, Nakamura, Kenji, Sumida, Kayo, Ito, Shingo, and Ohtsuki, Sumio

Abstract

Transcriptomic, proteomic, phosphoproteomic, and metabolomic analyses were combined to determine the role of pregnane X receptor (PXR) in nongenotoxic signaling and energy homeostasis in liver after rats were repeatedly orally dosed with the PXR agonist pregnenolone carbonitrile (PCN) for 7 days. Analyses of mRNAs and proteins in the supernatant, membrane, and cytosolic fractions of enlarged liver homogenates showed diverse expression profiles. Gene set enrichment analysis showed that the synchronous increase in mRNAs and proteins involved in chemical carcinogenesis and the response to drug was possibly mediated by the PXR pathway and proteasome core complex assembly was possibly mediated by the Nrf2 pathway. In addition, levels of proteins in the endoplasmic reticulum lumen and involved in the acute-phase response showed specific increase with no change in mRNA level, and those composed of the mitochondrial inner membrane showed specific decrease. The analysis of phosphorylated peptides of poly(A) RNA binding proteins showed a decrease in phosphorylation, possibly by casein kinase 2, which may be related to the regulation of protein expression. Proteins involved in insulin signaling pathways showed an increase in phosphorylation, possibly by protein kinase A, and those involved in apoptosis showed a decrease. Metabolomic analysis suggested the activation of the pentose phosphate and anaerobic glycolysis pathways and the increase of amino acid and fatty acid levels, as occurs in the Warburg effect. In conclusion, the results of combined analyses suggest that PXR’s effects are due to transcriptional and post-transcriptional regulation with alteration of nongenotoxic signaling pathways and energy homeostasis.

Published
2024
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192. High Expression of UGT1A1/1A6 in Monkey Small Intestine: Comparison of Protein Expression Levels of Cytochromes P450, UDP-Glucuronosyltransferases, and Transporters in Small Intestine of Cynomolgus Monkey and Human

Author

Akazawa, Takanori, Uchida, Yasuo, Miyauchi, Eisuke, Tachikawa, Masanori, Ohtsuki, Sumio, and Terasaki, Tetsuya

Abstract

Cynomolgus monkeys have been widely used for the prediction of drug absorption in humans. The purpose of this study was to clarify the regional protein expression levels of cytochromes P450 (CYPs), UDP-glucuronosyltransferases (UGTs), and transporters in small intestine of cynomolgus monkey using liquid chromatography–tandem mass spectrometry, and to compare them with the corresponding levels in human. UGT1A1 in jejunum and ileum were >4.57- and >3.11-fold and UGT1A6 in jejunum and ileum were >16.1- and >8.57-fold, respectively, more highly expressed in monkey than in human. Also, jejunal expression of monkey CYP3A8 (homologue of human CYP3A4) was >3.34-fold higher than that of human CYP3A4. Among apical drug efflux transporters, BCRP showed the most abundant expression in monkey and human, and the expression levels of BCRP in monkey and human were >1.74- and >1.25-fold greater than those of P-gp and >2.76- and >4.50-fold greater than those of MRP2, respectively. These findings should be helpful to understand species differences of the functions of CYPs, UGTs, and transporters between monkey and human. The UGT1A1/1A6 data would be especially important because it is difficult to identify isoforms responsible for species differences of intestinal glucuronidation by means of functional studies due to overlapping substrate specificity.

Published
2024
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193. Improving Proteomic Identification Using Narrow Isolation Windows with Zeno SWATH Data-Independent Acquisition

Author

Gu, Kongxin, Kumabe, Haruka, Yamamoto, Takumi, Tashiro, Naoto, Masuda, Takeshi, Ito, Shingo, and Ohtsuki, Sumio

Abstract

Data-independent acquisition (DIA) techniques such as sequential window acquisition of all theoretical mass spectra (SWATH) acquisition have emerged as the preferred strategies for proteomic analyses. Our study optimized the SWATH-DIA method using a narrow isolation window placement approach, improving its proteomic performance. We optimized the acquisition parameter combinations of narrow isolation windows with different widths (1.9 and 2.9 Da) on a ZenoTOF 7600 (Sciex); the acquired data were analyzed using DIA-NN (version 1.8.1). Narrow SWATH (nSWATH) identified 5916 and 7719 protein groups on the digested peptides, corresponding to 400 ng of protein from mouse liver and HEK293T cells, respectively, improving identification by 7.52 and 4.99%, respectively, compared to conventional SWATH. The median coefficient of variation of the quantified values was less than 6%. We further analyzed 200 ng of benchmark samples comprising peptides from known ratios ofEscherichia coli, yeast, and human peptides using nSWATH. Consequently, it achieved accuracy and precision comparable to those of conventional SWATH, identifying an average of 95,456 precursors and 9342 protein groups across three benchmark samples, representing 12.6 and 9.63% improved identification compared to conventional SWATH. The nSWATH method improved identification at various loading amounts of benchmark samples, identifying 40.7% more protein groups at 25 ng. These results demonstrate the improved performance of nSWATH, contributing to the acquisition of deeper proteomic data from complex biological samples.

Published
2024
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194. Multi-laboratory assessment of reproducibility, qualitative and quantitative performance of SWATH-mass spectrometry

Author

Collins, Ben C., primary, Hunter, Christie L., additional, Liu, Yansheng, additional, Schilling, Birgit, additional, Rosenberger, George, additional, Bader, Samuel L., additional, Chan, Daniel W., additional, Gibson, Bradford W., additional, Gingras, Anne-Claude, additional, Held, Jason M., additional, Hirayama-Kurogi, Mio, additional, Hou, Guixue, additional, Krisp, Christoph, additional, Larsen, Brett, additional, Lin, Liang, additional, Liu, Siqi, additional, Molloy, Mark P., additional, Moritz, Robert L., additional, Ohtsuki, Sumio, additional, Schlapbach, Ralph, additional, Selevsek, Nathalie, additional, Thomas, Stefani N., additional, Tzeng, Shin-Cheng, additional, Zhang, Hui, additional, and Aebersold, Ruedi, additional

Published
2016
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195. Identification of IGFBP2 and IGFBP3 As Compensatory Biomarkers for CA19-9 in Early-Stage Pancreatic Cancer Using a Combination of Antibody-Based and LC-MS/MS-Based Proteomics

Author

Yoneyama, Toshihiro, primary, Ohtsuki, Sumio, additional, Honda, Kazufumi, additional, Kobayashi, Makoto, additional, Iwasaki, Motoki, additional, Uchida, Yasuo, additional, Okusaka, Takuji, additional, Nakamori, Shoji, additional, Shimahara, Masashi, additional, Ueno, Takaaki, additional, Tsuchida, Akihiko, additional, Sata, Naohiro, additional, Ioka, Tatsuya, additional, Yasunami, Yohichi, additional, Kosuge, Tomoo, additional, Kaneda, Takashi, additional, Kato, Takao, additional, Yagihara, Kazuhiro, additional, Fujita, Shigeyuki, additional, Huang, Wilber, additional, Yamada, Tesshi, additional, Tachikawa, Masanori, additional, and Terasaki, Tetsuya, additional

Published
2016
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196. Front cover: Large-scale multiplex absolute protein quantification of drug-metabolizing enzymes and transporters in human intestine, liver, and kidney microsomes by SWATH-MS: Comparison with MRM/SRM and HR-MRM/PRM

Author

Nakamura, Kenji, primary, Hirayama-Kurogi, Mio, additional, Ito, Shingo, additional, Kuno, Takuya, additional, Yoneyama, Toshihiro, additional, Obuchi, Wataru, additional, Terasaki, Tetsuya, additional, and Ohtsuki, Sumio, additional

Published
2016
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200. Design and synthesis of a novel pre-column derivatization reagent with a 6-methoxy-4-quinolone moiety for fluorescence and tandem mass spectrometric detection and its application to chiral amino acid analysis

Author

Oyama, Tsubasa, primary, Negishi, Eiichi, additional, Onigahara, Hirohisa, additional, Kusano, Nao, additional, Miyoshi, Yurika, additional, Mita, Masashi, additional, Nakazono, Manabu, additional, Ohtsuki, Sumio, additional, Ojida, Akio, additional, Lindner, Wolfgang, additional, and Hamase, Kenji, additional

Published
2015
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