Radiochemistry (Hari Lab.)

Staff

  • Prof. Dr. Yoshiyuki Hari
  • Dr. Yuta Ito
  • Dr. Takashi Osawa

 

Research project

The research in our group is in the area of organic and bioorganic chemistry, and currently focuses on the development of nucleic acid analogs capable of targeting single-stranded RNA or double-stranded DNA in a sequence-specific fashion. To achieve the objective, synthetic organic chemistry and analytical chemistry are conducted.

(Keywords: Nucleic acid chemistry, Organic synthesis, Bioorganic chemistry, Medicinal chemistry)

 

Selected recent publications

Original articles

  1. T. Osawa, Y. Hari*, M. Dohi, Y. Matsuda, S. Obika*, Synthesis and properties of the 5-methyluridine derivative of 3,4-dihydro-2H-pyran-bridged nucleic acid (DpNA), J. Org. Chem. 2015, 80, 10474-10481.
  2. Y. Hari*, S. Kashima, Y. Matsuda, A. Sakata, R. Takamine, S. Ijitsu, S. Obika*, Base pair recognition ability of 2-(methylamino)pyrimidin-4-yl nucleobase in parallel triplex DNA, Heterocycles 2015, 90, 432-441.
  3. M. Akabane-Nakata, S. Obika*, Y. Hari*, Synthesis of oligonucleotides containing N,N-disubstituted 3-deazacytosine nucleobases by post-elongation modification and their triplex-forming ability with double-stranded DNA, Org. Biomol. Chem. 2014, 12, 9011-9015.
  4. Y. Hari*, S. Ijitsu, M. Akabane-Nakata, T. Yoshida, S. Obika*, Kinetic study of the binding of triplex-forming oligonucleotides containing partial cationic modifications to double-stranded DNA, Bioorg. Med. Chem. Lett. 2014, 24, 3046-3049.
  5. Y. Hari*, M. Akabane, S. Obika*, 2′,4′-BNA bearing a chiral guanidinopyrrolidine-containing nucleobase with potent ability to recognize the CG base pair in parallel-motif DNA triplex, Chem. Commun. 2013, 49, 7421-7423.
  6. Y. Hari*, T. Morikawa, T. Osawa, S. Obika*, Synthesis and properties of 2′-O,4′-C-ethyleneoxy bridged 5-methyluridine, Org. Lett. 2013, 15, 3702-3705.
  7. Y. Hari*, M. Nakahara, S. Obika*, Triplex-forming ability of oligonucleotides containing 1-aryl-1,2,3-triazole nucleobases linked via a two atom-length spacer, Bioorg. Med. Chem. 2013, 21, 5583-5588.

Review articles / Books

  1. Y. Hari*, S. Obika*, Synthesis and properties of 2′,4′-bridged nucleic acids containing multiple heteroatoms in the bridges, J. Synth. Org. Chem., Jpn. 2016, 74, 141-153 (in Japanese).
  2. Y. Hari*, Development of artificial nucleic acid that recognizes a CG base pair in triplex DNA formation, Yakugaku Zasshi 2013, 133, 1201-1208 (in Japanese).
  3. Y. Hari*, S. Obika, T. Imanishi*, Towards the sequence-selective recognition of double-stranded DNA containing pyrimidine-purine interruptions by triplex-forming oligonucleotides, Eur. J. Org. Chem. 2012, 2875-2887.

Molecular and Cellular Physiology (Fukada Lab)

Staff
Prof. Dr. Toshiyuki Fukada

 

Research project

Work in my research group has demonstrated that zinc transporter-mediated zinc signals are indispensible for normal cellular functions, and impairment of zinc signaling causes diseases. “Zinc signaling” is an emerging research field and a novel platform in life sciences, so that I would like to explore the zinc signaling at molecular level, in particular about roles and mechanisms of zinc signaling involved in path-physiological conditions of epithelium, neurological diseases, and cancer. Development of regulators for zinc signaling is also one of my research goals.

 

Recent publications

Original article

1.Kamimura, D., K. Katsunuma, Y. Arima, T. Atsumi, J.J Jiang, H. Bando, J. Meng, L. Sabharwal, A. Stpfkova, N. Nishikawa, H. Suzuki, H. Ogura, N. Ueda, M. Harada, J. Kobayashi, T. Hasegawa, H. Yoshida, H. Koseki, I. Miura, S. Harada, K. Nishida, H. Kitamura, T. Fukada, T. Hirano, and M. Murakami. mKDEL receptor 1 regulates T-cell homeostasis via PP1 that is a key phosphatase for ISR.

Nature Communications 6, Article number: 7474, 2015

2.Jenkitkasemwong, S., C-Y Wang, R. Coffey, W. Zhang, A. Chan, T. Biel, J-S. Kim, S. Hojyo, T. Fukada, and M. Knutson. SLC39A14 Is Required for the Development of Hepatocellular Iron Overload in Murine Models of Hereditary Hemochromatosis.

Cell Metabolism 22: 138–150, 2015

3.Hojyo, S., T. Miyai, H. Fujishiro, M. Kawamura, T. Yasuda, A. Hijikata, BH. Bin, T. Irié, J. Tanaka, T. Atsumi, M. Murakami, M. Nakayama, O. Ohara, S. Himeno, H. Yoshida, H. Koseki, T. Ikawa, K. Mishima, and ※T. Fukada. Zinc transporter SLC39A10/ZIP10 controls humoral immunity by modulating B cell receptor signal strength.

PNAS. 111:11786-91, 2014 (corresponding author)

4.Bin BH., S. Hojyo, T. Hosaka, J. Bhin, H. Kano, T. Miyai, M. Ikeda, T. Kimura-Someya Mikako Shirouzu, EG Cho, K. Fukue, T. Kambe, W. Ohashi, KH Kim, J. Seo, DH. Choi, YJ Nam, D. Hwang, A. Fukunaka, Y. Fujitani, S. Yokoyama, A. Superti-Furga, S. Ikegawa, TR Lee, and ※T. Fukada. Molecular pathogenic basis of Spondylocheirodysplastic Ehlers-Danlos syndrome caused by mutant ZIP13 proteins.

EMBO Molecular Medicine 6: 1028-1042, 2014 (corresponding author)

5.Miyai, T., S. Hojyo, T. Ikawa, M. Kawamura, T. Irié, H. Ogura, A. Hijikata, BH. Bin, T. Yasuda, H. Kitamura, M. Nakayama, O. Ohara, H.o Yoshida, H. Koseki, K. Mishima, and ※T. Fukada. Zinc transporter SLC39A10/ZIP10 facilitates antiapoptotic signaling during early B-cell development.

PNAS. 111:11780-85, 2014 (corresponding author)

 

Review article

  1. Hojyo S, Miyai T, ※Fukada T.

B-cell receptor strength and zinc signaling: Unraveling the role of zinc transporter ZIP10 in humoral immunity.

Receptor and Clinical Investigation 2: e387; 2015 (corresponding author)

2.Bin, BH., S. Hojyo, and ※Fukada T.

Spondylocheirodysplastic Ehlers-Danlos syndrome (SCD-EDS) and the mutant zinc transporter ZIP13.

Rare Diseases 2: e974982; 2014 (corresponding author)

 

Book

1.Fukada, T

Genetic study of zinc transporters and zinc signaling, edited by Collins J

Molecular, Genetic, and Nutritional Aspects of Major and Trace Minerals, ELSEVIER (in press) (corresponding author)

2.Fukada, T, Hojyo, S., and Bin, B.H.

Zinc signal in growth control and bone diseases, edited by Fukada T, and T. Kambe

Zinc Signals in Cellular Functions and Disorders, Springer, 249-267, 2014 (corresponding author).

Department of Public Health (Suzuki Lab.)

Prolonged endoplasmic reticulum stress alters placental morphology and causes low birth weight

Kawakami T, Yoshimi M, Kadota Y, Inoue M, Sato M and Suzuki S.

Toxicol Appl Pharmacol. 2014 Mar 1;275(2):134-44.

Highlights

 •Maternal exposure to excessive ER stress induced preterm birth and IUGR.

•Prolonged excessive ER stress altered the formation of the placental labyrinth.

•ER stress decreased GLUT1 mRNA expression in the placenta, but increased GLUT3.

•ER stress-induced IUGR causes decreased glycogen and altered glucose transport.

Abstract

The role of endoplasmic reticulum (ER) stress in pregnancy remains largely unknown. Pregnant mice were subcutaneously administered tunicamycin (Tun), an ER stressor, as a single dose [0, 50, and 100 μg Tun/kg/body weight (BW)] on gestation days (GDs) 8.5, 12.5, and 15.5. A high incidence (75%) of preterm delivery was observed only in the group treated with Tun 100 μg/kg BW at GD 15.5, indicating that pregnant mice during late gestation are more susceptible to ER stress on preterm delivery. We further examined whether prolonged in utero exposure to ER stress affects fetal development. Pregnant mice were subcutaneously administered a dose of 0, 20, 40, and 60 μg Tun/kg from GD 12.5 to 16.5. Tun treatment decreased the placental and fetal weights in a dose-dependent manner. Histological evaluation showed the formation of a cluster of spongiotrophoblast cells in the labyrinth zone of the placenta of Tun-treated mice. The glycogen content of the fetal liver and placenta from Tun-treated mice was lower than that from control mice. Tun treatment decreased mRNA expression of Slc2a1/glucose transporter 1 (GLUT1), which is a major transporter for glucose, but increased placental mRNA levels of Slc2a3/GLUT3. Moreover, maternal exposure to Tun resulted in a decrease in vascular endothelial growth factor receptor-1 (VEGFR-1), VEGFR-2, and placental growth factor. These results suggest that excessive and exogenous ER stress may induce functional abnormalities in the placenta, at least in part, with altered GLUT and vascular-related gene expression, resulting in low infant birth weight.

Department of Biochemistry (Kuzuhara Lab.)

Suppression of increased blood glucose levels in mice by Awa-ban tea following oral administration of mono- and disaccharides

Miki Hiasa, Megumi Kurokawa, Hiroshi Akita, Masatomo Harada, Kengo Niki, Kana Ohta, Masaki Shoji, Noriko Echigo, Takashi Kuzuhara

Journal of Functional Foods8, 188–192 (2014).

Highlights
•Awa-ban tea suppressed blood glucose levels after administration of maltose in mice
•Awa-ban tea suppressed blood glucose levels after administration of sucrose in mice
•Awa-ban tea suppressed blood glucose levels after administration of glucose in mice
•Awa-ban tea suppressed the area under the curve (AUC) of blood glucose by 72–83%
•Awa-ban tea is an alternative functional beverage to prevent diabetes

 

Awa-ban tea is a uniquely flavored, pickled and anaerobically fermented tea found only in the Tokushima prefecture in the Shikoku region of Japan. Diabetes is a major health problem worldwide. Here, we report that Awa-ban tea suppressed the increase in blood glucose levels after oral administration of maltose, sucrose, or glucose in mice (n = 9–54). Awa-ban tea suppressed the area under the curve (AUC) of blood glucose by 72–83%. Thus, we propose that Awa-ban tea can be used as an alternative functional beverage for diabetes prevention.

 

Department of Molecular Nutrition and Toxicology (Himeno Lab.)

Interleukin-6 enhances manganese accumulation in SH-SY5Y cells: implications of the up-regulation of ZIP14 and the down-regulation of ZnT10.

Hitomi Fujishiro Mari Yoshida Yuka Nakano and Seiichiro Himeno

Metallomics, 6, 944-949 (2014).

Exposure to an excess amount of manganese causes neurological symptoms similar to Parkinson’s disease. Zinc transporters such as Zrt, Irt-related protein 8 (ZIP8), and ZIP14 have been shown to have affinities for Mn2+ as well as Zn2+, but their roles in Mn2+ uptake in neuronal cells remain unclear. Recent studies have shown that another zinc transporter ZnT10 may be involved in manganese excretion. Here we examined the roles of ZIP8, ZIP14, and ZnT10 in the transport of manganese in human SH-SY5Y neuroblastoma cells. The introduction of siRNA of ZIP14 decreased the uptake of Mn2+, suggesting a significant role of ZIP14 in Mn2+ uptake in SH-SY5Y cells. The pretreatment of SH-SY5Y cells with interleukin-6 (IL-6) markedly increased the accumulation of manganese to approx. 3-fold that of the control, which could be partly explained by the increased uptake of Mn2+ due to the up-regulation of ZIP14 by IL-6. The treatment of SH-SH5Y cells with IL-6 clearly decreased both the mRNA and protein levels of ZnT10 with a concomitant decrease in the manganese excretion efficiency. These results suggest that both the up-regulation of ZIP14 and the down-regulation of ZnT10 by IL-6 might have enhanced the accumulation of manganese in SH-SY5Y cells. Our results provide new insight into the roles of zinc transporters in the aberrant manganese accumulation in neuronal cells, particularly in the presence of inflammatory cytokines such as IL-6.

 

Department of Microbiology (Nagahama Lab.)

Intracellular trafficking of Clostridium botulinum C2 toxin.

 

Masahiro Nagahama, Chihiro Takahashi, Kouhei Aoyanagi, Ryo Tashiro, Keiko Kobayashi,
Yoshihiko Sakaguchi, Kazumi Ishidoh, Jun Sakurai

Toxicon, 82, 76-82 (2014).

Highlights

Clostridium botulinum C2 toxin (C2I and C2IIa) is delivered to early endosomes.
•The delivery of C2I to the cytoplasm occurs in early endosomes.
•C2IIa was sent back to the plasma membranes through recycling endosomes.

Abstract

Clostridium botulinum C2 toxin is a binary toxin composed of an enzymatic component (C2I) and binding component (C2II). The activated binding component (C2IIa) forms heptamers and the oligomer with C2I is taken up by receptor-mediated endocytosis. We investigated the intracellular trafficking of C2 toxin. When MDCK cells were incubated with C2I and C2IIa at 37 °C, C2I colocalized with C2IIa in cytoplasmic vesicles at 5 min, and C2I then disappeared (15 min incubation and later), and C2IIa was observed in the vesicles. Internalized C2I and C2IIa were transported to early endosomes. Some of both components were returned to the plasma membrane through recycling endosomes, whereas the rest of C2IIa was transported to late endosomes and lysosomes for degradation. Bafilomycin A1, an endosomal acidification inhibitor, caused the accumulation of C2IIa in endosomes, and both nocodazole and colchicine, microtubule-disrupting agents, restricted C2IIa’s movement in the cytosol. These results indicated that an internalized C2I and C2IIa complex was delivered to early endosomes, and that subsequent delivery of C2I to the cytoplasm occurred in early endosomes. C2IIa was either sent back to the plasma membranes through recycling endosomes or transported to late endosomes and lysosomes for degradation.

Department of Medicinal Chemistry (Tsunoda Lab.)

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Eur. J. Org. Chem 2013, 36, 8208-8213 (2013).

A Method to Prepare Optically Active Acyclic α-Benzyl Ketones by Thermodynamically Controlled Deracemization

Kaku, H., Imai, T., Kondo, R., Mamba, S., Watanabe, Y., Inai, M., Nishii, T., Horikawa, M., Tsunoda, T.

Thermodynamically controlled deracemization of some acyclic ketones bearing a chiral center at the position α to the carbonyl group was satisfactorily achieved. Acyclic ketones with high optical purities could be isolated after treatment of the racemic ketones with base in aqueous MeOH in the presence of (–)-(2R,3R)-trans-2,3-bis(hydroxydiphenylmethyl)-1,4-dioxaspiro[5.4]decane (1a). The efficiency of the deracemization was appreciably influenced by the ratio of H2O/MeOH used as solvent.

Department of Physical Chemistry (Fukuyama Lab.)

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Chem. Pharm. Bull.62, 122-124 (2014).

A New Spiroindene Pigment from the Medicinal Fungus Phellinus ribis

Miwa KuboYuhong LiuMami IshidaKenichi HaradaYoshiyasu Fukuyama

A new spiroindene pigment, phelliribsin A, was isolated from the medicinal fungus Phellinus ribis, and its structure was determined by two dimensional (2D)-NMR methods. Phelliribsin A is an unprecedented spiroindene compound, and was found to have cytotoxic activity against PC12 cells at a concentration of 30 µM.

Department of Analytical Chemistry (Tanaka Lab.)

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Heterocycles89, 1369-1391 (2014).
Reductive Cyclization Reactions to Bicyclic Compounds Using Samarium Diiodide

Motoo Tori and Masakazu Sono

This review covers reports on the one-electron reductive cyclization reactions affording bicyclic compounds, such as hydrindans, perhydronaphthalenes, perhydroguaianes, and other systems including five-, six-, seven-, and eight-membered carbocycles using SmI2. The substrate is aldehyde, ketone, or ester. The effect of additives, such as H2O, MeOH, HMPA, and NiI2, was studied. The mechanistic aspects recently investigated are also introduced.

Department of Physical Chemistry (Fukuyama Lab.)

 

47

Tetrahedron, 69, 6959-6968 (2013).

Total Synthesis of Riccardin C and (±)-Cavicularin via Pd-Catalyzed Ar-Ar Corss Couplings.
K. Harada, K. Makino, N. Shima, H. Okuyama, T. Esumi, M. Kubo, H. Hioki, Y. Asakawa, Y. Fukuyama.

Riccardin C, a specific LXR α agonist, is a representative macrocyclic bisbibenzyl-type natural product. As part of our synthetic studies on macrocyclic bisbibenzyls, the synthesis of riccardin C and its analog cavicularin was examined. The total synthesis of riccardin C was accomplished via a Pd-catalyzed intramolecular Suzuki-Miyaura coupling as the key macrocyclization step. This synthetic strategy was also extended in the synthesis of (±)-cavicularin, which was then attained by constructing the dihydrophenanthrene moiety using a Pd-catalyzed Ar-Ar coupling reaction.