論文 / 著書情報
Article / Book Information
題目(和文)

概日リズムに着目した肝毒性評価試験構築に関する研究

Title(English)
著者(和文)

守矢恒司

Author(English)

Koji Moriya

出典(和文)

学位:博士(工学),学位授与機関:東京工業大学,
報告番号:甲第11545号,
授与年月日:2020年5月31日,
学位の種別:課程博士,
審査員:田川 陽一,生駒 俊之,粂 昭苑,秦 猛志,藤枝 俊宣

Citation(English)

Degree:Doctor (Engineering),
Conferring organization: Tokyo Institute of Technology,
Report number:甲第11545号,
Conferred date:2020/5/31,
Degree Type:Course doctor,
Examiner:,,,,

学位種別(和文)

博士論文

Category(English)

Doctoral Thesis

種別(和文)

論文要旨

Type(English)

Summary

Powered by T2R2 (Science Tokyo Research Repository)ᨵゞ
5HYLVHG$XJXVW㸦 ㄽᩥ༤ኈ㸧(Dissertation Doctorate)          ㄽ ᩥ

 せ ᪨㸦࿴ᩥ2000Ꮠ⛬ᗘ㸧Dissertation Summary (approx. 2000 characters in Japanese)ሗ࿌␒ྕஎ ➨      ྕẶྡNameFor   Ᏺ▮ ᜏྖadministrativeuse only࠙ㅖゝࠚᆅ⌫ୖ࡟Ꮡᅾࡍࡿ⏕≀ࡣᆅ⌫ࡢ⮬㌿࡟ࡼࡿ᫨ኪࡢ⎔ቃኚ໬࡟ᛂࡌ࡚ࠊ⏕⌮ᶵ⬟ࡀ⣙㸰㸲᫬㛫࿘ᮇࠊᴫ᪥ࣜ ࢬ࣒ࢆ♧ࡍࠋ⏕యෆ࡛ࡣኴ㝧ගࢆ⥙⭷ࡀගࢩࢢࢼࣝ࡜ࡋ࡚ཷᐜࡋࠊどᗋୗ㒊࡟Ꮡᅾࡍࡿど஺ཫୖ᰾࡬࡜ఏ㐩ࡉࢀࡿࠋど஺ཫୖ᰾ࡣࡑࢀ࡟ᛂࡌ࡚ࠊᾮᛶᅉᏊࡸ⚄⤒ఏ㐩ࢆ௓ࡋ࡚඲㌟ࡢᮎᲈ⤌⧊ࡢᴫ᪥ࣜࢬ࣒ࢆ ྠㄪࡉࡏࡿࠋ⫢⮚ࡶᴫ᪥ࣜࢬ࣒ࡢไᚚࢆཷࡅࡿࡓࡵࠊ⫢ᶵ⬟ࡶኚືࡋ࡚࠸ࡿࡇ࡜࠿ࡽ⸆≀ᛂ⟅ᛶࡣᢞ୚᫬้࡛␗࡞ࡿࠋࡇࡢࡼ࠺࡞ಶయࡢᛂ⟅ᛶࢆ཯ᫎࡍࡿ⣽⬊ᇵ㣴⣔ࢆ⏝࠸ࡓẘᛶヨ㦂ࡢ☜❧࡟ࠊᴫ᪥ࣜࢬ࣒ ࢆ෌⌧ࡍࡿࡇ࡜ࡣẘᛶࢆ⢭☜࡟ホ౯ࡍࡿ࠺࠼࡛ᚲ㡲࡛࠶ࡿ࡜⪃࠼ࡿࠋ࠙ㄽᩥࡢᵓᡂ࡜ྛ❶ࡢせ᪨ࠚ➨1❶ ᗎㄽᮏㄽᩥࡢ⫼ᬒ࠾ࡼࡧ┠ⓗࢆ㏙࡭ࡓࠋᴫ᪥ࣜࢬ࣒ࡢ⏕⌮ⓗព⩏࠾ࡼࡧ࡟ศᏊ࣓࢝ࢽࢬ࣒࡟ࡘ࠸࡚ᴫㄝࡋࠊ⫢ ⮚ࡢᶵ⬟࠾ࡼࡧ⸆≀௦ㅰ㐣⛬࡟࠾ࡅࡿᴫ᪥ࣜࢬ࣒ࡢ㔜せᛶࢆ㏙࡭ࡓࠋ᪂⸆ࡸ໬ྜ≀ࡢ୍⯡ⓗ࡞⫢ẘᛶヨ㦂࡛ࡣᴫ᪥ࣜࢬ࣒࡟ࡼࡿ⫢ᶵ⬟ኚືࡣ⪃៖ࡉࢀ࡚࠸࡞࠸⌧≧ࢆ㏙࡭ࡓࠋᮏㄽᩥ࡛ࡣ⫢ẘᛶㄏⓎ≀㉁ ࡜ࡋ࡚㐣๫ᦤྲྀ࡟ࡼࡾ⫢㞀ᐖࢆㄏⓎࡍࡿゎ⇕㙠③๣࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥࢆ⏝࠸ࠊ࣐࢘ࢫಶయࡢ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥ⫢ẘᛶ࡟ᑐࡍࡿᛂ⟅ᛶࢆᣦᶆ࡟ࠊ⣽⬊ᇵ㣴⣔࡟࠾࠸࡚ࡶಶయྠᵝ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥ⫢ ẘᛶ࡟ᑐࡍࡿᛂ⟅ᛶࡀᴫ᪥ࣜࢬ࣒ࡢไᚚࢆཷࡅࡿ࠿࡝࠺࠿ࢆ᳨ドࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋࡓࠋ➨2❶ ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥࡢゎ⇕స⏝࡜⫢㞀ᐖㄏⓎ࡟࠾ࡅࡿᴫ᪥ࣜࢬ࣒ࡢᙳ㡪ᴫ᪥ࣜࢬ࣒ࢆᣢࡕࠊ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥࡢゎ⇕ຠᯝࡢᙳ㡪ࢆཷࡅࡿయࢆ ⤒᫬ⓗ࡟ᐃࡋࠊ᭱ప

ࢆㄏᑟࡋࡓࠋ᭱పࡶࡋࡃࡣ᭱㧗࡟╔┠ࡋࡓࠋ࣐࢘ࢫࡢయ࿘ᮇ

௜㏆ࡢ᫬้࡟࠾࠸࡚ࠊ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥࢆᢞ୚ࡋࠊ⫢㞀ᐖ௜㏆࡛ᢞ୚ࡋࡓ࡜ࡇࢁࠊ㢧ⴭ࡞యపୗࢆ♧ࡋࠊ⏕໬Ꮫⓗゎᯒ࠾ࡼࡧ⫢⮚ࡢ⑓⌮ゎᯒ ࠿ࡽࡶ㢧ⴭ࡞⫢㞀ᐖࡢㄏᑟࡀࡳࡽࢀࡓࠋṇᖖࡢ࣐࢘ࢫ⫢⮚࡟࠾ࡅࡿ᫬ィ㑇ఏᏊ࠾ࡼࡧ࢔ࢭࢺ࢔࣑ࣀࣇ ࢙ࣥࡢ௦ㅰ㓝⣲ࡢⓎ⌧㔞ࢆ⤒᫬ⓗ࡟ᐃ㔞ゎᯒࡋࡓ࡜ࡇࢁࠊ᫬ィ㑇ఏᏊ࠾ࡼࡧ௦ㅰ㓝⣲ࡢኚືࡀ☜ㄆ࡛ࡁࠊ㢧ⴭ࡟ㄏᑟࡉࢀࡓ᫬้௜㏆࡛௦ㅰ㓝⣲ࡢⓎ⌧㔞ࡀ㧗࠸ࡇ࡜ࡀศ࠿ࡗࡓࠋ࣐࢘ࢫಶయ࡟࠾࠸࡚ࠊᢞ୚ ᫬้࡟ࡼࡾ⫢ẘᛶࡸ⏕⌮ᶵ⬟࡬ࡢᙳ㡪ࡀ␗࡞ࡿࡇ࡜ࢆ♧ࡋࠊ⫢ẘᛶᛂ⟅ᛶ࡟࠾ࡅࡿᴫ᪥ࣜࢬ࣒ࡢᚲ
せ ᛶ࡟ࡘ࠸࡚⪃ᐹࡋࡓࠋ➨3❶ ࣐࢘ࢫึ௦ᇵ㣴⫢⣽⬊ࡢㄪ〇᫬้࡟࠾ࡅࡿ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥㄏᑟ⫢ẘᛶ࡟ᑐࡍࡿឤཷᛶホ    ౯ᴫ᪥ࣜࢬ࣒ࢆไᚚࡍࡿ᪥↷࿘ᮇ࡟╔┠ࡋࠊᮅࡲࡓࡣኪ᫬้࡟⫢㞀ᐖࢆㄏⓎࡋࡓ࣐࢘ࢫ࡟࠾ࡅࡿ⫢ẘᛶホ ౯ࢆ⾜࡞࠸ࠊྠ᫬้࡟࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥࢆᭀ㟢࡛ࡁࡿࡼ࠺ࠊึ௦ᇵ㣴⫢⣽⬊ࢆㄪ〇ࡋࠊྠᵝ࡟⫢ẘᨵゞ
5HYLVHG$XJXVWᛶ ヨ㦂ࢆ⾜࡞ࡗࡓࠋ࣐࢘ࢫಶయ࡛ࠊኪ᫬้࡟ᢞ୚ࡋࡓ⩌࡛㢧ⴭ࡟⫢㞀ᐖࢆㄏᑟࡉࢀࡓࠋึ௦ᇵ㣴⫢⣽⬊࡟ ࠾࠸࡚ࡶࠊಶయྠᵝࠊኪ᫬้࡟ㄪ〇ࡉࢀࡓ⩌࡛㢧ⴭ࡟⫢⣽⬊Ṛࡀㄏᑟࡉࢀࡓࠋ᫬ィ㑇ఏᏊࡢⓎ⌧㔞ࢆẚ ㍑ࡋࡓ࡜ࡇࢁࠊ࣐࢘ࢫ⫢⮚࡛ぢࡽࢀࡓⓎ⌧㔞ࡢ㐪࠸ࡀึ௦ᇵ㣴⫢⣽⬊࡛ࡶྠᵝ࡟ࡳࡽࢀࡓࠋࡲࡓ⸆≀௦ ㅰ㓝⣲㑇ఏᏊⓎ⌧㔞࡟࠾࠸࡚ࡶࠊኪ᫬้ࡢ࣐࢘ࢫ⫢⮚࠾ࡼࡧࡧ ࠊึ௦ᇵ㣴⫢⣽⬊࡛ࠊ➨I┦཯ᛂ࡟㛵୚ࡍ ࡿ௦ㅰ㓝⣲ࡀ㧗࠸Ⓨ⌧ࢆ♧ࡋࡓࠋࡲࡓゎẘ⬟ຊࡢᣦᶆ࡛࠶ࡿ⣽⬊ෆࢢࣝࢱࢳ࢜ࣥ⃰ᗘࡣᮅ᫬้ࡢ⩌࡛㧗࠸ࡇ࡜ࡀศ࠿ࡗࡓࠋㄪ〇ࡉࢀࡓึ௦ᇵ㣴⫢⣽⬊ࡣࠊ༢㞳┤๓ࡢᴫ᪥ࣜࢬ࣒ࢆಖᣢࡋࠊಶయྠᵝࡢ⫢ẘ ᛶࡀㄏⓎࡋࡓࡶࡢ࡜⪃࠼ࡿࠋࡘࡲࡾࠊ⣽⬊ᇵ㣴⣔࡟࠾࠸࡚ࡶᴫ᪥ࣜࢬ࣒ࡢᙳ㡪ࢆཷࡅࡿྍ⬟ᛶࢆ♧ࡋࡓࠋ➨4❶ ᴫ᪥ࣜࢬ࣒ࢆྠㄪࡉࡏࡓ⣽⬊ᇵ㣴⣔࡟࠾ࡅࡿ⫢ẘᛶホ౯ㄪ〇ࡉࢀࡓึ௦ᇵ㣴⫢⣽⬊ࡣࠊ᪥↷࿘ᮇ࡟ࡼࡾᴫ᪥ࣜࢬ࣒ࡀྠㄪࡋࡓ⫢⮚ࡀᇶ‽࡜࡞ࡗ࡚࠸ࡿࡓࡵࠊ⣽⬊ 㞟ᅋ࡜ࡋ࡚ᴫ᪥ࣜࢬ࣒ࡢ࿘ᮇᛶࡀ࠶ࡿ୍ᐃ᪉ྥ࡟ᐃࡲࡗ࡚࠸ࡿ࡜⪃࠼ࡿࠋࡋ࠿ࡋ㏻ᖖࡢ⣽⬊ᰴ࡛ࡣࠊྠ ㄪᅉᏊࡀ࡞ࡃࠊಶࠎࡢ⣽⬊ࡣ஘ࢀࡓ࿘ᮇᛶࢆᣢࡕࠊ⣔඲య࡜ࡋ࡚ぢ࠿ࡅୖࠊᴫ᪥ࣜࢬ࣒ࡀᾘኻࡋ࡚࠸
ࡿࠋࡑࡇ࡛ࠊ࣐࢘ࢫ⫢⒴⣽⬊ᰴHepa8F5ࢆ⏝࠸ࠊForskolin࡟ࡼࡿᴫ᪥ࣜࢬ࣒ࡢྠㄪࢆ⾜࡞ࡗࡓࠋ᫬ィ㑇ఏ Ꮚ࠾ࡼࡧ⸆≀௦ㅰ㓝⣲㑇ఏᏊࡢⓎ⌧㔞ゎᯒ࠿ࡽࠊ⣙24᫬㛫ࡢ࿘ᮇᛶࢆ⏕ࡳฟࡍࡇ࡜ࡀ࡛ࡁࡓࠋࡲࡓ࢔ࢭ ࢺ࢔࣑ࣀࣇ࢙ࣥ⫢ẘᛶࡣ⸆≀௦ㅰ㓝⣲ࡢⓎ⌧㔞࡟ኚື࡜୍⮴ࡋࡓࡇ࡜࠿ࡽࠊ⣽⬊ᇵ㣴⣔࡛ᴫ᪥ࣜࢬ࣒ࢆ෌⌧ࡍࡿࡇ࡜ࡢ㔜せᛶࢆ♧ࡋࡓࠋ

➨5❶ ⥲ᣓ࠾ࡼࡧ௒ᚋࡢᒎᮃ⥲ᣓ࡜ࡋ࡚࡚ ࠊᮏㄽᩥࡢព⩏ࢆࡲ࡜ࡵࡿ࡜࡜ࡶ࡟ࠊ௒ᚋࡢᒎᮃࢆグࡋࡓࠋ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥ⫢ẘᛶ࡟ᑐࡍࡿᛂ⟅ᛶホ౯ࢆ㏻ࡋࠊ⣽⬊ᇵ㣴⣔࡟࠾࠸࡚ࡶᴫ᪥ࣜࢬ࣒ࡢไᚚࢆཷࡅࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋࠊ⫢ẘᛶホ౯ ࡟࠾ࡅࡿᴫ᪥ࣜࢬ࣒ࡢ㔜せᛶࢆぢฟࡋࡓࠋᮏ◊✲࡟ࡼࡾᚓࡽࢀࡓ▱ぢࡣࠊࡼࡾ⢭☜࡞⫢ẘᛶヨ㦂⣔ᵓ⠏࡟㈉⊩࡛ࡁࡿࡶࡢ࡛࠶ࡾࠊ⢭ᗘࡢ㧗࠸໬ྜ≀ࡢᏳ඲ᛶヨ㦂ࡀᐇ᪋࡛ࡁࡿࡶࡢ࡜ᮇᚅࡉࢀࡿࠋ

࠙Ⓨ⾲ㄽᩥࠚ࣭༤ኈㄽᩥ࡟㛵ࡍࡿཎⴭㄽᩥMoriya K, Tamai M, Koga T, Tanaka T and Tagawa YI.
Acetaminophen-induced hepatotoxicity of cultured hepatocytes depends on timing of isolation from lightcycle controlled mice. Genes to Cells. in press. doi:10.1111/gtc.12755
࣭ࡑࡢ௚ࡢㄽᩥImada T, Moriya K, Uchiyama M, Inukai N, Hitotsuyanagi M, Masuda A1, Suzuki T, Ayukawa S, Tagawa
YI, Dohmae N, Kohara M, Yamamura M, Kiga D. A Highly Bioactive Lys-Deficient IFN Leads toa SiteSpecific Di-PEGylated IFN with Equivalent Bioactivity to That of Unmodified IFN-ĮEACS Synth Biol.
2018. 7. 2537-2546
ഛ⪃㸸ㄽᩥせ᪨ࡣࠊ࿴ᩥ2000Ꮠ࡜ⱥᩥ300ㄒࢆ1㒊ࡎࡘᥦฟࡍࡿ࠿ࠊࡶࡋࡃࡣⱥᩥ800ㄒࢆ1㒊ᥦฟࡋ࡚ࡃࡔࡉ࠸ࠋ
Note: Dissertation summaries must be written in either of the following formats: (A) both in Japanese (approx. 2000 characters) and in
English (approx. 300 words), or (B) in English (approx. 800 words).
ὀព㸸ㄽᩥせ᪨ࡣࠊᮾᕤ኱ࣜࢧ࣮ࢳ࣏ࣜࢪࢺࣜ㸦T2R2㸧࡟࡚࢖ࣥࢱ࣮ࢿࢵࢺබ⾲ࡉࢀࡲࡍࡢ࡛ࠊබ⾲ྍ⬟࡞⠊ᅖࡢෆᐜ࡛సᡂࡋ࡚ࡃ
ࡔࡉ࠸ࠋ
Important: Dissertation summaries will be published online on the Tokyo Tech Research Repository (T2R2). Do not include
information treated as confidential under certain circumstances.

ᨵゞ
5HYLVHG$XJXVW㸦 ㄽᩥ༤ኈ㸧(Dissertation Doctorate)         ㄽ ᩥ

 せ ᪨ 㸦 ⱥ ᩥ 㸧㸦300ㄒ⛬ᗘ㸧Dissertation Summary (approx. 300 words in English)ሗ࿌␒ྕஎ ➨      ྕForẶྡNameᏲ▮ ᜏྖadministrativeuse only

Most physiological changes follow a daily cycle in animals because their circadian rhythm isadjusted by and synchronized to sunlight. In particular, the circadian rhythm affects liverfunctions, including pharmacokinetics and metabolism. Circadian rhythm has not been
considered significant in hepatotoxicity assays used in drug discovery and development. In thisstudy, to demonstrate the circadian effect on the hepatotoxicity testing, the response towards
acetaminophen (N-acetyl-p-aminophenol: APAP)-induced hepatotoxicity was examined in animaland cell culture model.
First, mice were more significantly induced hypothermia and liver injury by APAP at night thanthose during the day. The clock of APAP administration was consistent of the expression levelsof APAP-metabolizing enzyme. These results suggested that APAP-induced liver injury depended
on timing of APAP administration clock.
Second, it was examined whether APAP-induced hepatotoxicity depends on the retention ofcircadian rhythm after primary cultured hepatocytes which are isolated from the liver of mouse
maintained under a 12 h light cycle. The circadian rhythm was retained and mainly affected theexpression levels of phase I APAP-metabolizing enzymes and the levels of intracellularglutathione in primary hepatocytes after their isolation from the liver. These results indicatedthat circadian rhythm affected the cell culture model.
Finally, it was assessed whether the circadian rhythm of Hepa8F5, murine hepatoma cell line,
was synchronized by forskolin stimulation. The expression levels of circadian gene and APAPmetabolizing gene showed the circadian rhythm after forskolin. After synchronization, APAP
hepatotoxicity was dependent on the circadian rhythm of APAP-metabolizing enzyme in
Hepa8F5. These results indicated that the reproduction of circadian rhythm in cell culture system
is important for the hepatic assay model of cell culture.
In summary, the circadian rhythm affected the hepatic metabolic function of cultured cells
through the response against APAP-induced hepatotoxicity. This study demonstrates that it isnecessary to consider the exact construction of the hepatic assay model in relation to circadianrhythm when performing drug screens.
ഛ⪃㸸ㄽᩥせ᪨ࡣࠊ࿴ᩥ2000Ꮠ࡜ⱥᩥ300ㄒࢆ1㒊ࡎࡘᥦฟࡍࡿ࠿ࠊࡶࡋࡃࡣⱥᩥ800ㄒࢆ1㒊ᥦฟࡋ࡚ࡃࡔࡉ࠸ࠋ
Note: Dissertation summaries must be written in either of the following formats: (A) both in Japanese (approx. 2000 characters) and in
English (approx. 300 words), or (B) in English (approx. 800 words).
ὀព㸸ㄽᩥせ᪨ࡣࠊᮾᕤ኱ࣜࢧ࣮ࢳ࣏ࣜࢪࢺࣜ㸦T2R2㸧࡟࡚࢖ࣥࢱ࣮ࢿࢵࢺබ⾲ࡉࢀࡲࡍࡢ࡛ࠊබ⾲ྍ⬟࡞⠊ᅖࡢෆᐜ࡛సᡂࡋ࡚ࡃ
ࡔࡉ࠸ࠋ
Important: Dissertation summaries will be published online on the Tokyo Tech Research Repository (T2R2). Do not include
information treated as confidential under certain circumstances.

