上原記念生命科学財団研究報告集, 34 (2020)ୖཎグᛕ⏕࿨⛉Ꮫ㈈ᅋ◊✲ሗ࿌㞟, 34㸦2020㸧

161. ࢜ࣝ࢞ࣀ࢖ࢻࢆ⏝࠸ࡓ⬡⫫⫢⅖ࡢከ⣽⬊㐃ᦠᇶ┙ࡢゎ᫂⡿ᒣ 㮚௓ᮾி་⛉ṑ⛉኱Ꮫ ⤫ྜ◊✲ᶵᵓ ඛ➃་ṑᕤᏛ๰ᡂ◊✲㒊㛛 ๰⏕་Ꮫࢥࣥࢯ࣮ࢩ࢔࣒
Key words㸸࢜ࣝ࢞ࣀ࢖ࢻ㸪㠀࢔ࣝࢥ࣮ࣝᛶ⬡⫫ᛶ⫢⅖㸪⥺⥔໬㸪iPS ⣽⬊㸪⣽⬊㛫┦஫స⏝⥴ ゝ㏆ᖺࠊ⏕ά⩦័⑓ࡸ⢾ᒀ⑓ࢆ⫼ᬒ࡜ࡋ࡚ࠊከ㔞㣧㓇Ṕࡀ↓࠸࡟ࡶ㛵ࢃࡽࡎ⫢⮚࡟⬡⫫ࡀ⵳✚ࡋ࡚ࡋࡲ࠺㠀࢔ࣝࢥ࣮ࣝᛶࡢ⬡⫫ᛶ⫢⑌ᝈࡀᛴቑࡋ࡚࠸ࡿࠋ⬡⫫ᛶ⫢⅖ࡢ୰࡛ࡶࠊ㠀࢔ࣝࢥ࣮ࣝᛶ⬡⫫ᛶ⫢⅖㸦non-alcoholic steatohepatitis㸸
NASH㸧ࡣࠊ༢⣧࡞⬡⫫⫢࠿ࡽⓎᒎࡋ࡚ࠊ⫢⮚ࡢ⅖⑕ࡸ⥺⥔໬ࢆక࠺㐍⾜ᛶࡢ៏ᛶ⑌ᝈ࡛࠶ࡿࠋNASH ࡣ㧗㢖ᗘ࡛  ⫢◳ኚࡸ⫢ࡀࢇࢆⓎ⑕ࡍࡿࡓࡵࠊ἞⒪௓ධࡀᚲ㡲࡛࠶ࡿࡶࡢࡢࠊ⌧ᅾࡲ࡛࡟᭷ຠᛶࡢ㧗࠸἞⒪ἲࡀᏑᅾࡋ࡞࠸ࡇ࡜ࡀၥ㢟࡜࡞ࡗ࡚࠸ࡿ㹙1㹛
ࠋNASH ࡟㛵ࡍࡿᚑ᮶ࡢ◊✲ࡣࠊ୺࡟ࣔࢹࣝື≀ࢆ⏝࠸࡚⾜ࢃࢀ࡚ࡁࡓࡀࠊࣄࢺࡢ⫢⅖࣭⥺⥔໬⑓ែࢆ༑ศ࡟෌⌧࡛ࡁ࡞࠸ࡓࡵ࡟ࠊࣄࢺ NASH ⑓ែࡢ⌮ゎࡸ⸆๣㛤Ⓨ࡟ࡣ⮳ࡗ࡚࠸࡞࠸ࠋࡲࡓࠊ⬡⫫⫢࠿ࡽ NASH ࡬⛣⾜ࡍࡿ㐍ᒎᗘྜ࠸࡟ࡣ኱ࡁ࡞ಶேᕪࡀ࠶ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠾ࡾ㹙2㹛ࠊಶࠎࡢᝈ⪅࡛㉳ࡇࡿ⅖⑕ࡸ⥺⥔໬࡞࡝ࡢ⑓ែࢆ෌⌧࡛ࡁࡿࣄࢺ⫢⮚ࣔࢹࣝࡢ㛤Ⓨࡣႚ⥭ࡢㄢ㢟࡜࡞ࡗ࡚࠸ࡿࠋ
NASH ࡢ⑓ែࡣࠊ⫢⣽⬊ࡢ⬡⫫⵳✚ࠊ⫢⮚ᖖᅾᛶ࣐ࢡࣟࣇ࢓࣮ࢪ ࡛࠶ࡿࢡࢵࣃ࣮⣽⬊ࡀ㉳Ⅼ࡜࡞ࡗ࡚ច㉳ࡉࢀࡿ
⅖⑕཯ᛂࠊ⫢ᫍ⣽⬊࡟ࡼࡿ⣽⬊እᇶ㉁ࡢ⏘⏕ࠊ࡜࠸ࡗࡓ」ᩘ✀㢮ࡢ⣽⬊ࡢᛂ⟅ࡀ㐃㙐ࡋ࡚ᙧᡂࡉࢀࡿࠋࡋࡓࡀࡗ࡚ࠊ ࡇࡢࡼ࠺࡞࣊ࢸࣟ࡞⣽⬊㛫┦஫స⏝ࡢほⅬ࠿ࡽࠊNASH Ⓨ⑕ᶵᵓࢆࣄࢺ࡛ࣞ࣋ࣝ⌮ゎࡍࡿࡇ࡜ࡣࠊ἞⒪㛤Ⓨࢆ   ๓㐍ࡉࡏࡿୖ࡛ᴟࡵ࡚㔜せ࡜⪃࠼ࡽࢀࡿࠋᮏ◊✲࡛ࡣࠊయෆ⎔ቃࢆᶍࡋࡓ୕ḟඖᇵ㣴⣔࡟࠾࠸࡚ࠊiPS ⣽⬊࡞࡝ࡢࣄࢺ
ࡢᖿ⣽⬊࠿ࡽ࣑ࢽ⮚ჾࢆᵓ⠏ࡍࡿ࢜ࣝ࢞ࣀ࢖ࢻᢏ⾡ࢆά⏝ࡍࡿࡇ࡜࡛ࠊ1㸬ࣄࢺ iPS ⣽⬊࠿ࡽࠊNASH ⑓ែ࡟≉ᚩⓗ࡞⬡⫫⵳✚ࠊ⅖⑕ࠊ⥺⥔໬ࢆㄏᑟ࡛ࡁࡿከ⣽⬊⣔⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ๰ฟࡍࡿࡓࡵࡢᇵ㣴ࢩࢫࢸ࣒ࢆ㛤Ⓨࡍࡿ࡜࡜ࡶ࡟ࠊ
2㸬ࣄࢺ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ⏝࠸࡚ NASH ࡢ⑓ែ㐍ᒎࡢ㉳Ⅼ࡜࡞ࡿ⣽⬊㛫┦஫స⏝ࡢ◚⥢ࢆࢩࢢࢼࣝศᏊ࡛ࣞ࣋ࣝ ᫂ࡽ࠿࡟ࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋࡓࠋ᪉ ἲ1㸬 ࣄࢺ iPS ⣽⬊࠿ࡽ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡬ࡢศ໬ㄏᑟ
ࣄࢺ iPS ⣽⬊ࡢᇵ㣴ࡣࠊࣇ࢕࣮ࢲ࣮ࣇ࣮ࣜࡢ᮲௳࡟࡚⾜ࡗࡓࠋࣄࢺ iPS ⣽⬊࠿ࡽ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡬ࡢศ໬ㄏᑟࡣࠊ
௨ୗࡢࡼ࠺࡟⾜ࡗࡓࠋࡍ࡞ࢃࡕࠊࣄࢺ iPS ⣽⬊࡟ Activin AࠊBMP4ࠊ࢘ࢩ⫾ඣ⾑Ύࢆῧຍࡍࡿࡇ࡜࡟ࡼࡾࠊ⬇యෆ  ⬇ⴥ⣽⬊ࢆㄏᑟࡋࡓ㹙3㹛ࠋࡑࡢᚋࠊWnt ࢔ࢦࢽࢫࢺࠊFGF4ࠊBMP ⤒㊰㜼ᐖ๣ࢆῧຍࡍࡿࡇ࡜࡟ࡼࡾࠊᚋ᪉๓⭠    ࢫࣇ࢙ࣟ࢖ࢻࢆᚓࡓ㹙4㹛ࠋᚓࡽࢀࡓᚋ᪉๓⭠ࢫࣇ࢙ࣟ࢖ࢻࢆ Matrigel ࡬ໟᇙࡋࠊࣞࢳࣀ࢖ࣥ㓟ࢆྵࡴᇵᆅ࡛ᇵ㣴ࡋࡓࠋḟ࡟ࠊHGFࠊOncostatin MࠊDexamethason ࢆῧຍࡋࡓ⫢⣽⬊ᇵ㣴⏝ࡢᇵᆅ HCM㸦Lonza㸧࡛ᇵ㣴ࡍࡿࡇ࡜࡛ࠊ   ⫢⣽⬊ศ໬ࢆಁ㐍ࡋࠊ᭱⤊ⓗ࡟㠀ᐇ㉁⣽⬊ࢆྵࡴ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆᵓ⠏ࡋࡓࠋ2㸬 ࣄࢺ iPS ⣽⬊⏤᮶⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢච␿ᰁⰍ࡜ࠊ㏱㐣ᆺ㟁Ꮚ㢧ᚤ㙾࡟ࡼࡿゎᯒ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆࣃ࣒ࣛ࣍ࣝ࢔ࣝࢹࣄࢻ࡛ᅛᐃࡋࡓᚋࠊࣃࣛࣇ࢕ࣥษ∦ࢆస〇ࡋࠊྛ✀࣐࣮࣮࢝ᢠయࢆ⏝࠸ࡓච␿ᰁⰍࢆ⾜ࡗࡓࠋࡲࡓࠊ࢜ࣝ࢞ࣀ࢖ࢻࢆᵓᡂࡍࡿ⣽⬊ࡢᙧែⓗ≉ᚩࢆゎᯒࡍࡿࡓࡵ࡟ࠊᅛᐃࡋࡓ࢜ࣝ࢞ࣀ࢖ࢻࡢ㉸ⷧษ∦ࢆస〇ࡋࠊ㏱㐣㟁Ꮚ㢧ᚤ㙾㸦TEM㸧࡟ࡼࡗ࡚ほᐹࡋࡓࠋ3㸬 ࣄࢺ iPS ⣽⬊⏤᮶⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟࠾ࡅࡿ㑇ఏᏊⓎ⌧ゎᯒ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࠿ࡽࢺ࣮ࢱࣝ RNA ࢆᢳฟࡋࡓᚋࠊ㏫㌿෗㓝⣲ࢆ⏝࠸࡚ cDNA ࢆྜᡂࡋࡓࠋྛᶆⓗ㑇ఏᏊ࡟ᑐࡍࡿ
ࣉࣛ࢖࣐࣮ࢆ⏝࠸࡚ࠊSYBR Green ࢖ࣥࢱ࣮࣮࢝ࣞࢱ࣮ἲ࡟ࡼࡿᐃ㔞 PCR ࢆ⾜ࡗࡓࠋ1ୖཎグᛕ⏕࿨⛉Ꮫ㈈ᅋ◊✲ሗ࿌㞟, 34㸦2020㸧㻌

4㸬 ࣄࢺ iPS ⣽⬊⏤᮶⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢࢩࣥࢢࣝࢭࣝࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒ゎᯒ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ㓝⣲ฎ⌮࡟ࡼࡗ࡚༢୍⣽⬊໬ࡋࡓᚋࠊྜィ⣙ 20,000 ⣽⬊࠿ࡽ 10X Genomics Chromium  ࣉࣛࢵࢺࣇ࢛࣮࣒ࢆ⏝࠸࡚ࠊࢩࣥࢢࣝࢭࣝ RNA-seq ࣛ࢖ࣈࣛࣜࢆస〇ࡋࡓࠋIllumina HiSeq X ࢆ⏝࠸࡚ࣛ࢖ࣈࣛࣜࡢࢩ࣮ࢣࣥࢫࢹ࣮ࢱࢆྲྀᚓࡋࡓᚋࠊCell Ranger ࣃ࢖ࣉࣛ࢖ࣥ࡟ࡼࡗ࡚࣮ࣜࢻࣉࣟࢭࢩࣥࢢࢆᐇ᪋ࡋࡓࠋ⣽⬊㸭㑇ఏᏊࡢࣇ࢕ࣝࢱࣜࣥࢢࠊࢹ࣮ࢱ⤫ྜࠊࢫࢣ࣮ࣜࣥࢢࠊࢡࣛࢫࢱࣜࣥࢢ࠾ࡼࡧࢹ࣮ࢱྍど໬ࡣࠊSeurat v3 ࢆ⏝࠸࡚ᐇ᪋ࡋࡓࠋ⤖ ᯝ1㸬ࣄࢺ iPS ⣽⬊࠿ࡽࡢከ⣽⬊⣔⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢ๰ฟ
⅖⑕ࡸ⥺⥔໬ࢆ෌⌧ྍ⬟࡞⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ๰ฟࡍࡿࡓࡵ࡟ࠊࣄࢺ iPS ⣽⬊࠿ࡽࠊୖ⓶⣔ࡢ⫢⣽⬊࡟ຍ࠼࡚ࠊ⅖⑕ࢆច㉳ࡍࡿࢡࢵࣃ࣮⣽⬊ࡸ⥺⥔໬ࢆㄏᑟࡍࡿ⫢ᫍ⣽⬊࡞࡝ࡢ」ᩘ✀㢮ࡢ⣽⬊ࢆྠ᫬࡟ศ໬ࡉࡏࡿᡭἲࢆ࡜ࡗࡓࠋ   ࡍ࡞ࢃࡕࠊ⫢⮚ࡢⓎ⏕㉳※࡛࠶ࡿ๓⭠ෆ⬇ⴥࡢᙧᡂࡸࠊ࣐ࢡࣟࣇ࢓࣮ࢪ࣭⫢ᫍ⣽⬊ࡢศ໬࡟㔜せ࡜ࡉࢀࡿࣞࢳࣀ࢖ࣥ㓟ࢩࢢࢼࣝ࡟╔┠ࡋࠊ࢜ࣝ࢞ࣀ࢖ࢻࡢᙧᡂึᮇ࡟ࣞࢳࣀ࢖ࣥ㓟ࢆ୍㐣ⓗ࡟ᑟධࡍࡿࡇ࡜࡛ࠊ」ᩘ⣔㆕ࡢ⣽⬊ࢆྠ᫬࡟๰ฟࡍࡿᡭἲࢆ☜❧ࡋࡓ㸦ᅗ 1a㸧㹙5㹛ࠋྛ⣽⬊✀࣐࣮࣮࢝ࡢⓎ⌧ゎᯒࡢ⤖ᯝࠊᵓ⠏ࡉࢀࡓ࢜ࣝ࢞ࣀ࢖ࢻ࡟ࡣࠊ⫢⣽⬊  㸦AlbuminࠊE-cadherin 㝧ᛶ㸧࡟ຍ࠼࡚ࠊࢡࢵࣃ࣮⣽⬊㸦CD68ࠊCD11b 㝧ᛶ㸧ࡸ⫢ᫍ⣽⬊㸦VimentinࠊDesmin    㝧ᛶ㸧࡟㢮ఝࡋࡓ⣽⬊⩌ࡀྵࡲࢀ࡚࠸ࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗࡓ㸦ᅗ 1b㸧ࠋࡉࡽ࡟ࠊࢩࣥࢢࣝࢭࣝࢺࣛࣥࢫࢡࣜࣉࢺ࣮࣒
ゎᯒࡢ⤖ᯝ࠿ࡽࡶࠊᮏ࢜ࣝ࢞ࣀ࢖ࢻ⣔࡟࠾ࡅࡿ⫢⮚≉␗ⓗ࡞ከ⣽⬊⣔㆕ࡢᏑᅾࡀ⿬௜ࡅࡽࢀࡓࠋḟ࡟ࠊ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢᶵ⬟ⓗᵓ㐀ࡀⓎ㐩ࡋ࡚࠸ࡿ࠿࡝࠺࠿ࢆ᫂ࡽ࠿࡟ࡍࡿࡓࡵ࡟ࠊTEM ࢆ⏝࠸ࡓᚤ⣽ᵓ㐀ࡢ ほᐹࢆ⾜ࡗࡓࠋࡑࡢ⤖ᯝࠊ⫢⣽⬊㛫࡟ࡣࠊศἪࡉࢀࡓ⫹Ồࢆ⫹⟶࡬㐠ᦙࡍࡿࡓࡵࡢẟ⣽⫹⟶ࡀᙧᡂࡉࢀ࡚࠸ࡿࡇ࡜ࠊ ࡲࡓࠊ⫢ᫍ⣽⬊࡟ࡣࠊࡑࡢ≉ᚩ࡛࠶ࡿࣅࢱ࣑ࣥ A ㈓ⶶ⬡⫫⁲ࢆෆໟࡋࠊࢥ࣮ࣛࢤࣥศἪ⬟ࢆ᭷ࡋ࡚࠸ࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗࡓ㸦ᅗ 1c㸧ࠋ௚ࡢ⤖ᯝࡶేࡏࠊᮏᡭἲ࡛๰ฟࡉࢀࡿࣄࢺ iPS ⣽⬊⏤᮶⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡣࠊච␿⣔⣽⬊ࢆྵࡴ   㠀ᐇ㉁⣽⬊ࡀྵࡲࢀࡓከ⣽⬊⣔ࡢࣄࢺ⫢⮚ࣔࢹ࡛ࣝ࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ

ᅗ 1㸬ࣄࢺ iPS ⣽⬊࠿ࡽࡢከ⣽⬊⣔⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢ๰ฟa㸧࢜ࣝ࢞ࣀ࢖ࢻࡢ๰ฟ᪉ἲࡢᴫ␎ᅗࠋb㸧๰ฟࡉࢀࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢ᫂ど㔝ീ࡜⺯ගච␿ᰁⰍീࠋc㸧TEM ࢆ⏝࠸ࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟࠾ࡅࡿᚤ⣽ᵓ㐀ࡢほᐹࠋ2ୖཎグᛕ⏕࿨⛉Ꮫ㈈ᅋ◊✲ሗ࿌㞟, 34㸦2020㸧
㻌2㸬ࣄࢺ iPS ⣽⬊⏤᮶⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡬ࡢ⅖⑕࣭⥺⥔໬ࡢㄏᑟḟ࡟ࠊࣄࢺ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟ NASH ࡢ≉ᚩ࡛࠶ࡿ⅖⑕ࡸ⥺⥔໬ࢆㄏᑟࡍࡿࡓࡵࡢᇵ㣴᮲௳ࢆࠊ㐟㞳⬡⫫㓟ࢆ୰ᚰ࡟᳨ウࡋࡓࠋࡑࡢ⤖ᯝࠊ࢜ࣞ࢖ࣥ㓟ࡢῧຍ࡟ࡼࡾࠊ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ୰࡟⬡⫫⁲ࡀ㢧ⴭ࡟⵳✚ࡋࠊࡑࡢᚋࠊTNF-Șࡸ
IL-8 ࡞࡝ࡢ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡢⓎ⌧ࡀୖ᪼ࡍࡿࡇ࡜ࢆぢฟࡋࡓ㸦ᅗ 2aࠊb㸧㹙5㹛ࠋࡉࡽ࡟ࠊ࢜ࣞ࢖ࣥ㓟ࢆῧຍࡋ࡚
1㹼2 㐌㛫ᚋ࡟ࡣࠊI ᆺࢥ࣮ࣛࢤࣥ㝧ᛶ⥺⥔ࡀ⣽⬊እ࡟⵳✚ࡍࡿᵝᏊࡀほᐹࡉࢀ㸦ᅗ 2c㸧
ࠊin vitro ࡟࠾࠸࡚ NASH ࡢ⑓ែ࡟㔜せ࡞⅖⑕࠾ࡼࡧ⥺⥔໬ࢆ⤒᫬ⓗ࡟ࣄࢺ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟ㄏᑟྍ⬟࡛࠶ࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋᅗ 2㸬ࣄࢺ iPS ⣽⬊⏤᮶⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡬ࡢ⅖⑕࣭⥺⥔໬ࡢㄏᑟa㸧 ࢜ࣞ࢖ࣥ㓟ࠊࡲࡓࡣࣃ࣑ࣝࢳࣥ㓟ࢆฎ⌮ࡋࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟࠾ࡅࡿࠊBODIPY ࢆ⏝࠸ࡓ⬡⫫⵳✚ࡢྍど໬ࠋb㸧 ࢜ࣞ࢖ࣥ㓟㸦OA㸧ࠊࡲࡓࡣࣃ࣑ࣝࢳࣥ㓟㸦PA㸧ࢆฎ⌮ࡋࡓ㝿ࡢ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡢ㑇ఏᏊⓎ⌧㔞㸦ANOVA & Tukey’s testࠊ*P㸺0.05㸧ࠋc㸧 ࢜ࣞ࢖ࣥ㓟ࢆฎ⌮ࡋࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟࠾ࡅࡿCOL1A1ᰁⰍ࡟ࡼࡿ⥺⥔໬ࡢྍど໬ࠋ3㸬NASH ࡢ⑓ែ㐍ᒎࡢ㉳Ⅼ࡜࡞ࡿ⣽⬊㛫┦஫స⏝␗ᖖࡢ᥈⣴࡜ྠᐃᮏ NASH ࡢ⑓ែ㐍ᒎ࡟㛵୚ࡍࡿከ⣽⬊㛫┦஫స⏝ࢆྠᐃࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋ࡚ࠊ࢜ࣝ࢞ࣀ࢖ࢻ୰࡟Ⓨ⌧ࡍࡿศἪ
ᅉᏊࡢ࠺ࡕࠊ࢜ࣞ࢖ࣥ㓟ฎ⌮࡟ࡼࡗ࡚Ⓨ⌧ࡀኚືࡍࡿࡶࡢࢆࠊᐃ㔞 PCR ἲ࡟ࡼࡗ࡚ࢫࢡ࣮ࣜࢽࣥࢢࡋࡓࠋࡑࡢ⤖ᯝࠊᡂ㛗ᅉᏊ X ࡀ࢜ࣞ࢖ࣥ㓟ࡢฎ⌮࡟ࡼࡗ࡚㢧ⴭ࡟ῶᑡࡍࡿࡇ࡜ࢆぢฟࡋࡓ㸦ᅗ 3a㸧ࠋࡉࡽ࡟ࠊ࢜ࣞ࢖ࣥ㓟ฎ⌮࡜࡜ࡶ࡟
ࣜࢥࣥࣅࢼࣥࢺ X ࢆῧຍࡍࡿ࡜ࠊ࢜ࣞ࢖ࣥ㓟༢⊂ฎ⌮⩌࡜ẚ㍑ࡋ࡚ࠊ࢜ࣝ࢞ࣀ࢖ࢻ୰ࡢ⬡⫫⵳✚ࡀపῶࡍࡿࡇ࡜ࠊ  ࡲࡓࠊ⥺⥔໬ࡀ㢧ⴭ࡟ᢚไࡉࢀࡿࡇ࡜ࡀ♧ࡉࢀࡓ㸦ᅗ 3b㸧ࠋᡂ㛗ᅉᏊ X ࡢ⣽⬊࡛ࣞ࣋ࣝࡢస⏝ࢆヲ⣽࡟᫂ࡽ࠿࡟ࡍࡿࡓࡵ࡟ࠊ࢜ࣞ࢖ࣥ㓟ࢆฎ⌮ࡋࡓ࢜ࣝ࢞ࣀ࢖ࢻࠊ࢜ࣞ࢖ࣥ㓟࡜ X ࢆྠ᫬ฎ⌮ࡋࡓ࢜ࣝ࢞ࣀ࢖ࢻࠊ࠾ࡼࡧ㝜ᛶᑐ㇟࡜ࡋ࡚ᮍฎ⌮ࡢ࢜ࣝ࢞ࣀ࢖ࢻࢆࢧࣥࣉࣝ࡜ࡋ࡚ࠊࢩࣥࢢࣝࢭࣝ RNA-seq ゎᯒࢆᐇ᪋ࡋࡓࠋ⌧ᅾࠊྛࢧࣥࣉࣝࢹ࣮ࢱࡢ⤫ྜ࡜ ࢡࣛࢫࢱ࣮ࡢ⣽⬊✀ྠᐃࡲ࡛᏶஢ࡋ࡚࠾ࡾ㸦ᅗ 3c㸧ࠊ่⃭౫Ꮡⓗ࡞㑇ఏᏊⓎ⌧ࡢᕪࢆᢳฟࡍࡿࡇ࡜࡟ࡼࡗ࡚ࠊ    ᡂ㛗ᅉᏊ X ࡀ࡝ࡢ⣽⬊✀࡟࡝ࡢࡼ࠺࡞ᛂ⟅ࢆᘬࡁ㉳ࡇࡍ⤖ᯝࠊ⥺⥔໬ࡀᢚไࡉࢀ࡚࠸ࡿࡢ࠿ࠊศᏊ࡛ࣞ࣋ࣝࡢ    ࣓࢝ࢽࢬ࣒ゎᯒࢆ㐍ࡵ࡚࠸ࡿࠋ
3ୖཎグᛕ⏕࿨⛉Ꮫ㈈ᅋ◊✲ሗ࿌㞟, 34㸦2020㸧㻌ᅗ 3㸬ࣄࢺ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ⏝࠸ࡓ NASH ࡟࠾ࡅࡿከ⣽⬊㐃ᦠᇶ┙ࡢゎᯒa㸧 ࢜ࣞ࢖ࣥ㓟㸦OA㸧ࠊࡲࡓࡣࣃ࣑ࣝࢳࣥ㓟㸦PA㸧ࢆฎ⌮ࡋࡓ㝿ࡢ X 㑇ఏᏊⓎ⌧㔞㸦ANOVA & Tukey’s testࠊ*P㸺0.05㸧ࠋb㸧 ࢜ࣞ࢖ࣥ㓟ࡲࡓࡣ X ࢆฎ⌮ࡋࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ࡟࠾ࡅࡿ⥺⥔໬ࡢኚ໬ࠋc㸧 ࢜ࣞ࢖ࣥ㓟ࡲࡓࡣ X ࢆฎ⌮ࡋࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࡢࢩࣥࢢࣝࢭࣝ RNA-seq ゎᯒࠋฎ⌮ࡈ࡜㸦ᕥ㸧ࠊ࠶ࡿ࠸ࡣࢡࣛࢫࢱ࣮ࡈ࡜㸦ྑ㸧࡟Ⰽศࡅࡋࡓ UMAP ࢆ♧ࡋࡓࠋ⪃ ᐹࡇࢀࡲ࡛࡟ࠊࣄࢺ iPS ⣽⬊࠿ࡽ⫢ᫍ⣽⬊ࡸࢡࢵࣃ࣮⣽⬊ࢆ஧ḟඖᇵ㣴⣔࡛ศ໬ㄏᑟࡍࡿᡭἲ㹙6㸪7㹛ࡸࠊࣄࢺ⫢⣽⬊࡜㠀ᐇ㉁⣽⬊ࢆඹᇵ㣴ࡍࡿᡭἲ㹙8㹛ࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀࠊ࠸ࡎࢀࡶ⬡⫫⵳✚࣭⅖⑕࣭⥺⥔໬࡜࠸ࡗࡓ」ᩘࡢ⑓ែࡀ  㛵୚ࡍࡿ⬡⫫ᛶ⫢⅖ࡢ⾲⌧ᆺࢆ෌⌧ࡍࡿ࡟ࡣ⮳ࡗ࡚࠸࡞࠸ࠋࡑࢀ࡟ᑐࡋ࡚ࠊᮏ◊✲࡟࠾࠸࡚㛤Ⓨࡋࡓ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻ
ࢩࢫࢸ࣒࡛ࡣࠊ⫢ᫍ⣽⬊ࡸච␿⣔⣽⬊࡞࡝ࡢ㠀ᐇ㉁⣽⬊ࢆྵࡴከ⣽⬊⣔㆕ࢆྠ᫬࡟ศ໬ࡉࡏࡿ࢔ࣉ࣮ࣟࢳࢆ࡜ࡿࡇ࡜࡟ࡼࡾࠊᵝࠎ࡞ࣄࢺ iPS ⣽⬊ᰴ࠿ࡽ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ෌⌧ᛶࡼࡃᵓ⠏ࡍࡿࡇ࡜ࡀྍ⬟࡜࡞ࡗࡓࠋࡲࡓࠊNASH ࡟   ≉ᚩⓗ࡞⅖⑕ࡸ⥺⥔໬ࡢึᮇ⑓ែࢆ in vitro ࡛ㄏᑟ࡛ࡁࠊ⣽⬊㛫┦஫స⏝ࡢほⅬ࠿ࡽ NASH ࡢ⑓ែ㐍ᒎࡢヲ⣽ࢆ  ศᏊ⏕≀Ꮫ࣭⣽⬊⏕≀Ꮫⓗ࡟㏣㊧ྍ⬟࡞ࣄࢺ⫢⮚ࣔࢹࣝ࡜࡞ࡿࡇ࡜ࡀᮇᚅࡉࢀࡿࠋᮏᡭἲࡢ⫢⮚࢜ࣝ࢞ࣀ࢖ࢻࢆ⏝࠸ࡓศἪᅉᏊࢫࢡ࣮ࣜࢽࣥࢢࡢ⤖ᯝࠊᡂ㛗ᅉᏊ X ࡀ⫢⮚ࡢ⥺⥔໬ࢆᢚไࡍࡿ   ࣘࢽ࣮ࢡ࡞άᛶࢆ᭷ࡋ࡚࠸ࡿࡇ࡜ࡀぢฟࡉࢀࡓࠋ௒ᚋࠊX ࡢᢠ⥺⥔໬స⏝ࡢᇶ㍈࡜࡞ࡿ⣽⬊✀ࡸ㈐௵ࢩࢢࢼࣝศᏊࡀ
≉ᐃࡉࢀࢀࡤࠊᮏ◊✲ࡢᡂᯝࡣࣄࢺ NASH ἞⒪࡟ᑐࡋ࡚⮫ᗋ⑓ែ࡬ࡢእᤄᛶ࡟ඃࢀࡓ๰⸆ࢶ࣮ࣝࢆᥦ౪࡛ࡁࠊ἞⒪ἲࡢ㛤Ⓨ࡟௒ᚋ኱ࡁࡃ㈉⊩ࡍࡿ࡜⪃࠼ࡽࢀࡿࠋ4ୖཎグᛕ⏕࿨⛉Ꮫ㈈ᅋ◊✲ሗ࿌㞟, 34㸦2020㸧㻌ඹྠ◊✲⪅࣭ㅰ㎡ᮏ◊✲ࡢඹྠ◊✲⪅ࡣࠊᮾி་⛉ṑ⛉኱Ꮫࣜࢧ࣮ࢳࢥ࢔ࢭࣥࢱ࣮ࡢ㓇ᕳ᭷㔛ᏊẶࠊྠ኱Ꮫྡ㄃ᩍᤵࡢ࿴Ề೺஧㑻༤ኈࠊྠ኱Ꮫ⤫ྜ◊✲ᶵᵓᩍᤵࡢṊ㒊㈗๎༤ኈ࡛࠶ࡿࠋᮏ◊✲ࡢࡈᨭ᥼ࢆ㈷ࡾࡲࡋࡓୖཎグᛕ⏕࿨⛉Ꮫ㈈ᅋ࡟῝ࡃឤㅰ  ࠸ࡓࡋࡲࡍࠋᩥ ⊩
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