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ガス分子を介した新しい生体制御機構の学際的アプローチによる
解明と医学への応用
研究代表者 末松 誠（慶應義塾大学・医学部・教授）
＜背景・目的＞
ガス分子は細胞膜を他の物質に比べて容易に通過して生体を構成する高分子と結合して特異的な機能を付与する分子群です。水素、メタン、一酸化炭素、アンモニアなどは太古の宇宙を構成する基本分子でした。細菌はガス分子を生存保証のための警報装置をオン・オフするための情報分子として使用しており、その「遺産」は高等動物でも生存に関わるしくみとして巧妙に利用されているはずです。しかしこれまでガス分子の生理作用や生体内挙動のメカニズムを包括的に解明することは技術的に極めて困難でした。本プロジェクトでは学際的アプローチを通じて細胞内に存在する非蛋白性低分子代謝物の全体像（＝メタボローム）が特定のガス分子の消長に応じてどのように変動するかを最新鋭の質量分析技術で探索し、標的候補分子のガス感受性を構造解析技術を駆使して分析することにより代謝制御に関わるメカニズムを解くことにより生理学的意義の解明を目指し、さらに得られた基盤知見を統合することにより医学応用に繋がる研究を推進します。
＜内容・方法＞
1. 赤血球のヘモグロビンを介した酸素センシング機構解明
(a) レーザー技術による細胞膜上の特異的分子破壊実験による解析
(b) 赤血球メタボロームの低酸素応答の解析による血管機能制御における意義の解明
2. 部位特異的遺伝子改変動物による酸素センサー分子機能の解明
(a) ガス分子の生成・受容・分配に関わる分子を望みの細胞・臓器で発現・消失させること
による機能解析
(b) 造血幹細胞の代謝の酸素依存性と増殖・分化シグナルとの連関解明
(c) 酸素感受性転写因子HIF-1のknockout miceを用いた臓器障害・炎症モデル解析による
新しい疾患制御法の開発
3. CO, NOなどのガス分子の新しい受容・情報伝達系の探索
(a) 最新のコンピュータ技術を駆使し高速で効率よく体内の低分子代謝物の全体像
（メタボローム）を解析できる系を用いてガス分子が最初に結合する標的分子候補を探索
(b) 候補分子とガス分子の相互作用を物理化学的構造解析技術を用いてガス分子の結合に
伴う蛋白質の構造・機能の変化を解析
(c) (b)の生理学的意義を２に示した手法で解析することにより理解
4. 生体内におけるO2, CO2, CO, NO, H2Sによる統合的代謝制御システムの解明
(a) NO, H2Sと低分子代謝化合物の複合体の生体内検出
(b) 病原微生物のエネルギー代謝系解析
＜期待される成果＞
虚血性臓器障害の病態制御法の創出、インテリジェント人工酸素運搬体（必要なところで効率よく酸素供給や血流を増やすことのできる人工の分子集合体）の開発、ヒトと代謝系のパラダイムが異なる病原微生物を死滅させるための新しい標的分子の探索などに繋がる基盤的技術の創出が期待できる。
関連する業績（1994〜現在）
Suematsu, M., Kashiwagi, S., Sano, T., Goda, N., Shinoda, Y., Ishimura, Y.
Carbon monoxide as an endogenous modulator of hepatic vascular perfusion.
Biochem. Biophys. Res. Commun. 205, 1333-1337, 1994.
Suematsu, M., Goda, N., Sano. T., Kashiwagi, S., Egawa, T., Shinoda, Y.,
Ishimura, Y. Carbon monoxide : An endogenous modulator of sinusoidal tone in the
perfused rat liver. J. Clin. Invest. 96, 2431-2437, 1995.
Suematsu, M., Wakabayashi, Y., Ishimura, Y. Gaseous monoxides: A novel regulator
of microvascular function in the liver. Cardiovasc. Res. (invited review) 32,
679-686, 1996.
Yamaguchi, T., Wakabayashi, Y., Tanaka, M., Sano, T., Ishikawa, H., Nakajima,
H., Suematsu, M., Ishimura, Y. Taurocholate is necessary for directional
excretion bilirubin into bile in the perfused rat liver. Am. J. Physiol. (Gastrointest.
Liver Physiol.) 270, G1028-G1032, 1996
Goda, N., Suematsu, M., Mukai, M., Kiyokawa, K., Notori, M., Nozawa, S.,
Ishimura, Y. Endogenous nitric oxide production modulates
mitochondrion-dependent oxidative stress in cultured human placental
trophoblastic cells. Am. J. Physiol. (Heart & Circ. Physiol.) 272 (40),
H1893-1899, 1996.
Kashiwagi, S., Suematsu, M., Wakabayashi, Y., Koizumi, A., Tachibana, M.,
Kawada, N., Inoue, M., Ishimura, Y., Kaneko, A. Electrophysiological
characterization of cultured hepatic stellate cells in rats. Am. J. Physiol. (Gastrointest.
Liver Physiol.) 272, G742-G750, 1997.
Shiomi, M., Sano, T., Shinoda, Y., Wakabayashi, Y., Nimura, Y., Ishimura, Y.,
Suematsu, M. Nitric oxide suppression reversibly attenuates mitochondrial
dysfunction and cholestasis in endotoxemic rat liver. Hepatology 27, 108-115,
1998.
Goda, N., Suzuki, K., Naito, M., Takeoka, S., Tsuchida, E., Ishimura, Y.,
Tamatani, T., Suematsu, M. Distribution of heme oxygenase isoforms in rat liver:
Topographic basis for carbon monoxide-mediated microvascular relaxation. J. Clin.
Invest. 101, 604-612, 1998.
Suzuki, H., DeLano, F.A., Parks, D.A., Jamshidi, N., Granger, D.N., Ishii, H.,
Suematsu, M., Zweifach, B.W., Schmid-Schoenbein, G.W. Xanthine oxidase activity
associated with arterial blood pressure in spontaneously hypertensive rats.
Proc. Natl. Acad. Sci. USA 95, 4754-4759, 1998.
Shinoda, Y., Suematsu, M., Wakabayashi, Y., Goda, N., Kashiwagi, S., Shimmura,
S., Saito, S., Yamaguchi, T., Ishimura, Y. Carbon monoxide as a modulator of
bile canalicular contractility in cultured rat hepatocytes. Hepatology 28,
286-295, 1998.
Hokari, R., Miura, S., Fujimori, H., Tsuzuki, Y., Shigematsu, T., Higuchi, H.,
Kimura, H., Serizawa, H., Suematsu, M., Yagita, H., Granger, D.N., Ishii, H.
Nitric oxide modulates T lymphocyte migration in Peyer's patches and villus
submucosa of rat small intestine. Gastroenterology 115, 618-627, 1998.
Johnston, B., Burns, A.R., Suematsu, M., Issekutz, T.B., Woodman, R.C., Kubes,
P. Chronic inflammation upregulates chemokine receptors and induces neutrophil
migration to monocyte chemoattractant protein-1. J. Clin. Invest. 103,
1269-1276, 1999
Mori, M., Suematsu, M., Kyokane, T., Sano, T., Suzuki, H., Yamaguchi, T.,
Ishimura, Y., Ishii, H. Carbon monoxide mediates alterations in paracellular
permeability and vesicular transport in acetaminophen-treated perfused rat
liver. Hepatology 30, 160-168, 1999.
Wakabayashi, Y., Takamiya, R., Mizuki, A., Kyokane, T., Goda, N., Yamaguchi, T.,
Takeoka, S., Tsuchida, E., Suematsu, M., Ishimura, Y. Carbon monoxide
overproduced by heme oxygenase-1 causes a reduction of microvascular tone in
perfused rat liver. Am. J. Physiol. (Gastrointest. Liver Physiol.) 277,
G1088-1096, 1999.
Kumamoto, Y., Suematsu, M., Shimazu, M., Makino, N., Kato, Y., Sano, T.,
Wakabayashi, G., Ishimura, Y., Naito, M., Kitajima, M. Kupffer cell-independent
hyperacute oxidative stress preceding impairment of bile acid-dependent bile
formation in the cold storage-reperfused rat liver. Hepatology 30, 1454-1463,
1999.
Suematsu, M., Ishimura, Y. The heme oxygenase-carbon monoxide system: A
regulator of hepatobiliary functions. Hepatology 31, 3-6, 2000.
Makino, N., Suematsu, M., Sugiura, Y., Morikawa, H., Shiomi, H., Goda, N, Sano,
T., Nimura, Y., Sugimachi, K., Ishimura, Y. Altered expression of heme
oxygenase-1 in the livers of patients with portal hypertensive diseases.
Hepatology 33, 32-42, 2001.
Kitchin, K.T., Anderson, W.L., Suematsu, M. An ELISA assay for heme oxygenase-1.
J. Immunol. Meth. 247, 153-161, 2001.
Kyokane, T., Norimizu, S., Taniai, H., Naito, M., Takeoka, S., Tsuchida, E.,
Nimura, Y., Ishimura, Y., Suematsu, M. Carbon monoxide from heme catabolism
protects against hepatobiliary dysfunction in endotoxin-treated rat liver.
Gastroenterology 120, 1227-1240, 2001.
Taniai, H., Suematsu, M., Suzuki, T., Hori, R., Ishimura, Y., Nimura, Y.
Protective roles of endothelin B receptor-mediated nitric oxide generation in
hepatobiliary dysfunction in anoxia-reoxygenated perfused rat liver. Hepatology
33, 894-901, 2001.
Imai T, Morita T, Shindo T, Nagai R, Yazaki Y, Kurihara H, Suematsu M, Katayama
S. Vascular smooth muscle cell-directed overexpression of heme oxygenase-1
elevates blood pressure through attenuation of nitric oxide-induced vasodilation
in mice.
Circ Res. 89(1):55-62, 2001.
Hirano K, Kobayashi T, Watanabe T, Yamamoto T, Hasegawa G, Hatakeyama K,
Suematsu M, Naito M. Role of heme oxygenase-1 and Kupffer cells in the
production of bilirubin in the rat liver. Arch Histol Cytol. 64(2):169-78, 2001.
Morisaki, H., Katayama, T., Kotake, Y., Ito, M., Ishimura, Y., Takeda, J.,
Suematsu, M. Roles of carbon monoxide in leukocyte and platelet dynamics in rat
mesentery during sevoflurane anesthesia. Anesthesiology 95, 192-199, 2001.
Nakayama, M., Takahashi, K., Komaru T., Fukuchi, M., Shioiri, H., Sato, K.,
Kitamuro T., Shirato, K., Yamaguchi, T., Suematsu, M., Shibahara, S. Increased
expression of heme oxygenase 1 and bilirubin accumulation in foam cells of
rabbit atherosclerotic lesions. Arterioscler Thromb Vasc Biol 21, 1373-1377,
2001.
Suzuki, T., Makino, R., Suematsu, M. Organic phosphates as a new class of
soluble guanylate cyclase inhibitors. FEBS Lett. 507, 49-53, 2001
Hori, R., Kashiba, M., Goda, N., Kajimura, M., Yachie, A., Toma, K., Makino, N.,
Soejima, A., Nagasawa, T., Nakabayashi, K., Suematsu, M. Gene transfer of H25A
mutant heme oxygenase-1 protects cells against hydroperoxide-induced
cytotoxicity. J. Biol. Chem. 277, 10712-10718, 2002.
Ozawa, N., Goda, N., Makino, N., Yamaguchi, T., Yoshimura, Y., Suematsu, M.
Leydig cell-derived heme oxygenase-1 regulates apoptosis of premeiotic germ
cells in response to stress. J. Clin. Invest. 109, 457-467, 2002
Kajimura, M., Goda, N., Suematsu, M. Organ design for generation and reception
of CO: Lessons from the liver. Antioxid Redox Signal 4, 633-637, 2002.
Suematsu, M. COHb: A stress-induced marker reflecting surgical insults ? J
Gastroenterol Hepatol (Editorial) 17, 519-520, 2002.
Takamiya, R., Murakami, M., Goda, N., Makino, N., Kajimura, M., Takamiya, Y.,
Yamaguchi, T., Ishimura, Y., Hozumi, N., Suematsu, M. Stabilization of mast
cells by heme oxygenase-1: an anti-inflammatory role. Am. J. Physiol. Heart
Circ. Physiol. 283, H861-H870, 2002.
Suematsu, M., Suzuki, H., DeLano, F.A., Schmid-Schoenbein, G.W. The inflammatory
aspect of the microcirculation in hypertension. Microcirculation 9, 259-276,
2002.
Morisaki, H., Katayama, T., Kotake, Y., Handa, M., Ikeda, Y., Takeda, J., Ito,
M., Suematsu, M. Carbon monoxide modulates endotoxin-induced microvascular
leukocyte adhesion through platelet-dependent mechanisms. Anesthesiology. 97,
701-709, 2002.
Kajimura, M., Shimoyama, M., Tsuyama, S., Suzuki, T., Ihara, H., Kozaki, S.,
Tsubota, K., Oguchi, Y, Suematsu, M.. Visualization of gaseous monoxide
reception by soluble guanylate cyclase in rat retina. FASEB J fj.02-0359fje.
2002.
Kashiwagi, S., Kajimura, M., Yoshimura, Y., Suematsu, M. Nonendothelial source
of nitric oxide in arterioles but not in venules: Alternative source revealed in
vivo by diaminofluorescein microfluorography. Circ Res 91, e55-e64, 2002.
Murai, M., Yoneyama, H., Ezaki, T., Suematsu, M., Terashima, Y., Harada, A.,
Hamada, H., Asakura, H., Ishikawa, H., Matsushima, K. Subendothelial dome of gut
Peyer’s patches is the essential anatomical site in initiating acute
graft-versus-host disease. Nat. Immunol. 4, 154-160, 2003.
Mitani F, Mukai K, Miyamoto H, Suematsu M, Ishimura Y. The undifferentiated cell
zone is a stem cell zone in adult rat adrenal cortex. Biochim Biophys Acta.
1619, 317-324, 2003.
Mukai, K., Mitani, F., Nagasawa, H., Suzuki, R., Suzuki, T., Suematsu, M.,
Ishimura, Y. An inverse correlation between expression of a preprocathepsin
B-related protein with cysteine-rich sequences and steroid 11β-hydroxylase in
adrenocortical cells. J. Biol. Chem. 278, 17084-17092, 2003.
Ogishima T, Kinoshita JY, Mitani F, Suematsu M, Ito A. Identification of outer
mitochondrial membrane cytochrome b5 as a modulator for androgen synthesis in
Leydig cells. J. Biol. Chem. 278, 21204-21211, 2003.
Kato, Y., Shimazu, M., Kondo, M., Kumamoto, Y., Wakabayashi, G., Kitajima, M.,
Suematsu, M. Bilirubin rinse: A simple protectant against the rat liver graft
injury mimicking heme oxygenase-1 preconditioning. Hepatology 38, 364-373, 2003.
Miyazawa, M., Suzuki, H., Masaoka, T., Kai, A., Suematsu, M., Nagata, H., Miura,
S., Ishii, H. Suppressed apoptosis in the inflamed gastric mucosa of
Helicobacter pylori-colonized iNOS-knockout mice. Free Radic. Biol. Med. 34,
121-1630, 2003
Suematsu, M. Quartet signal transducers in gas biology. Antioxid. Redox Signal.
5, 435-437, 2003.
Suematsu, M., Suganuma, K., Kashiwagi, S. Mechanistic probing of gaseous signal
transduction in microcirculation. Antioxid. Redox Signal. 5, 485-492, 2003.
Norimizu, S., Kudo, A., Ishikawa, K., Kajimura, M., Yamaguchi, T., Fujii, K.,
Taniai, H., Nimura, Y., Suematsu, M. Carbon monoxide stimulates mrp2-dependent
excretion of bilirubin-IXα into bile in the perfused rat liver. Antioxid. Redox
Signal. 5, 449-456, 2003.
Schaefer, S., Kajimura, M., Tsuyama, S., Uchida, K., Sato, E., Inoue, M.,
Suematsu, M., Watanabe, K. Aberrant utilization of nitric oxide and regulation
of soluble guanylate cyclase in rat diabetic retinopathy. Antioxid. Redox
Signal. 457-466, 2003.
Egawa, T., Yoshioka, S., Takahashi, S., Hori, H., Nagano, S., Shimada, H.,
Ishimori, K., Morishima, I., Suematsu, M., Ishimura, Y. Kinetic and
spectroscopic characterization of hydroperoxy compound in the reaction of native
myoglobin with hydrogen peroxide. J. Biol. Chem. 278, 41597-41606, 2003.
Nagano, S., Shimada, H., Tarumi, A., Hishiki, T., Kimata-Ariga, Y., Egawa, T.,
Suematsu, M., Park, S.Y., Adachi, S., Shiro, Y., Ishimura, Y. Infrared
spectroscopic and mutational studies on putidaredoxin-induced conformational
changes in ferrous CO-P450cam. Biochemistry 42, 14507-14514. 2003
Watanabe, T., Hasegawa, G., Yamamoto, T., Hatakeyama, K., Suematsu, M., Naito,
M. Expression of heme oxygenase-1 in rat ontogeny. Arch. Histol. Cytol. 66,
155-162, 2003.
Katagiri, H., Ito, Y., Ishii, K., Hayashi, I., Suematsu, M., Yamashita, S.,
Murata, T., Narumiya, S., Kakita, A., Majima, M. Involvement of thromboxane
derived from cyclooxygenase-1 and -2 in hepatic microcirculatory dysfunction
during endotoxemia in mice. Hepatology 39, 139-150, 2004.
Tomita, K., Azuma, T., Kitamura, N., Nishida, J., Inokuchi, S., Nishimura, T.,
Suematsu, M., Ishii, H. Pioglitazone prevents alcohol-induced fatty liver in
rats through upregulation of c-Met. Gastroenterology 126, 873-885, 2004.
Kudo, A., Kashiwagi, S., Yoshimura, Y., Uchida, K., Arii, S., Suematsu, M.
Kupffer cells alter organic anion transport via multidrug resistance protein 2
in the post-cold ischemic rat liver. Hepatology 39, 1099-1109, 2004
Sakai, H., Masada, Y., Horinouchi, H., Ikeda, E., Sou, K., Takeoka, S., Suematsu,
M., Takaori, M., Kobayashi, K., Tsuchida, E. Physiologic capacity of
reticuloendothelial system for degradation of hemoglobin-vesicles (Artificial
oxygen carriers) after massive intravenous doses by dialy repeated infusions for
14 days. J. Pharmacol. Exp. Ther. 311 (3): 874-884, 2004
Mitani, F., Ogishima, T., Mukai, K., Hoshino, R., Watanabe, K., Suematsu, M.
Possible participation of outer mitochondrial membrane cytochrome B5 in
steroidogenesis in zona glomerulosa of rat adrenal cortex. Endocr. Res. 30(4),
639-644, 2004.
Fujii, K., Sakuragawa, T., Kashiba, M., Sugiura, Y., Maruyama, K., Goda, N.,
Nimura, Y., Suematsu, M. Hydrogen sulfide as an endogenous modulator of biliary
bicarbonate excretion in the rat liver. Antioxid Redox Signal. 7(5-6), 788-794,
2005.
Sugiura, Y., Kashiba, M., Hoshikawa, K., Sasaki, R., Saito, K., Kimura, H.,
Maruyama, K., Goda, N., Suematsu, M. Cadmium exposure alters metabolomics of
sulfur-containing amino acids in rat testes. Antioxid. Redox Signaling 7(5-6),
781-787, 2005.
Tian, J., Bryk, R., Itoh, M., Suematsu, M., Nathan, C. Variant tricarboxylic
acid cycle in Mycobacterium tuberculosis: Identification of α-ketoglutarate
decarboxylase. Proc. Natl. Acad. Sci. USA 2005, in press.
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