Epigenomic analysis of human retinal arterial and venous vascular endothelial cells

2020 Fiscal Year Annual Research Report

• Project/Area Number: 18K09398
• Research Institution: Jichi Medical University
• Principal Investigator: 蕪城 俊克 自治医科大学, 医学部, 教授 (00280941)
• Co-Investigator(Kenkyū-buntansha): 和田 洋一郎 東京大学, アイソトープ総合センター, 教授 (10322033)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 網膜血管内皮細胞 / ヒトゲノム / エピゲノム解析 / 網羅的遺伝子解析 / サイトカイン

Outline of Annual Research Achievements

我々は内因性ぶどう膜炎と比べ感染性ぶどう膜炎では蛍光眼底造影で高率に動脈炎、網膜動脈血管閉塞が観察されることを見いだした（Kaburaki T, et al. Jpn J Ophthalmol. 2020）。本研究では、ヒト網膜動脈および静脈血管内皮細胞の免疫学的特性を解明する目的で、ヒト眼球から網膜動脈および静脈血管内皮細胞を分離培養し、遺伝子発現解析およびエピゲノム解析を試みた。
まず我々は眼窩腫瘍などで眼球摘出となった6例の患者眼球から、網膜動脈血管・静脈血管を別々に採取し、それぞれの血管内皮細胞の初代培養を試みた。しかし、得られる細胞数が少ないことや、細胞のviabilityが低いことから、RNA抽出が困難であった。
そこで我々はヒト眼球から採取した網膜血管内皮細胞を用いた解析から、ヒト網膜毛細血管内皮細胞株（HREC, ACBRI 181、Cell systems)を用いた研究に変更した。我々は感染性と内因性ぶどう膜炎患者から採取した硝子体液に含まれる炎症性サイトカイン33種類の濃度を測定、統計学的に比較し、内因性ぶどう膜炎ではTNFαとそれによって発現誘導されるケモカインなどの発現が上昇するのに対し、感染性ぶどう膜炎では、ＴＮＦα以外にもIL-1βやインターフェロン(ＩＦＮ)も高値となることを見いだした（Fukunaga H, et al. Sci Rep 2020).そこでHRMECにTNFα、IL-1β、IFNγの複合刺激を行い、刺激後の遺伝子発現の変化をRNA sequenceで解析した。その結果、3種類の複合刺激によって発現が著明に上昇する遺伝子群を見いだした。現在、この遺伝子群のHRMECにおける役割の解析を行っている。

Research Products

• [Journal Article] Retinal vascular inflammatory and occlusive changes in infectious and non-infectious uveitis. (2020)
  • Author(s): Kaburaki T, Fukunaga H, Tanaka R, Nakahara H, Kawashima H, Shirahama S, Izawa H, Komae K, Takamoto M, Soga H, Aihara M.
  • Journal Title: Jpn J Ophthalmol. Volume: 64 Pages: 150-159
  • DOI: 10.1007/s10384-020-00717-4
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Genetic association of a gain of function interferon gamma receptor 1 (IFNGR1) polymorphism and the intergenic region LNCAROD/DKK1 with Behcet's disease. (2020)
  • Author(s): Louthrenoo W, Kasitanon N, Pathanapitoon K, Wangkaew S, Kuwata S, Nishi A, Kaburaki T, Tanaka R, Takeuchi F
  • Journal Title: Int J Rheum Dis Volume: 23 Pages: in press
  • DOI: 10.1111/1756-185X.13785.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Contribution of HLA-B*51:01 and -A*26:01 to Behcet's disease and their clinical association in Thai patients. (2020)
  • Author(s): Zhou HP, Tanaka R, Kaburaki T.
  • Journal Title: Ocul Immunol Inflamm. Volume: 28 Pages: in press
  • DOI: 10.1080/09273948.2020.1815796
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Comparisons of Clinical Features in Japanese Patients with Behcet's Uveitis Treated in the 1990s and the 2000s. (2020)
  • Author(s): Nakahara H, Kaburaki T, Tanaka R, Yoshida A, Takamoto M, Kawata M, Fujino Y, Kawashima H, Aihara M.
  • Journal Title: Ocul Immunol Inflamm. Volume: 28 Pages: 262-269
  • DOI: 10.1080/09273948.2018.1559928.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Clinical Features and Visual Outcome of Uveitis in Japanese Patients Younger than 18 Years. (2020)
  • Author(s): Kitano M, Tanaka R, Kaburaki T, Nakahara H, Shirahama S, Suzuki T, Komae K, Aihara M.
  • Journal Title: Ocul Immunol Inflamm. Volume: 28 Pages: in press
  • DOI: 10.1080/09273948.2020.1726972.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Fluorescein Angiography Scoring System Using Ultra-Wide-Field Fluorescein Angiography Versus Standard Fluorescein Angiography in Patients with Sarcoid Uveitis. (2020)
  • Author(s): Tanaka R, Kaburaki T, Yoshida A, Takamoto M, Miyaji T, Yamaguchi T.
  • Journal Title: Ocul Immunol Inflamm. Volume: 28 Pages: in press
  • DOI: 10.1080/09273948.2020.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Analysis of inflammatory mediators in the vitreous humor of eyes with pan-uveitis according to aetiological classification. (2020)
  • Author(s): Fukunaga H, Kaburaki T, Shirahama S, Tanaka R, Murata H, Sato T, Takeuchi M, Tozawa H, Urade Y, Katsura M, Kobayashi M, Wada Y, Soga H, Kawashima H, Kohro T, Aihara M.
  • Journal Title: Sci Rep. Volume: 10(1) Pages: 2783
  • DOI: 10.1038/s41598-020-59666-0.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Viral Loads in Ocular Fluids of Acute Retinal Necrosis Eyes Infected by Varicella-Zoster Virus Treated with Intravenous Acyclovir Treatment. (2020)
  • Author(s): Sato T, Yamamoto W, Tanaka A, Shimazaki H, Sugita S, Kaburaki T, Takeuchi M.
  • Journal Title: J Clin Med. Volume: 9 Pages: 1204.
  • DOI: 10.3390/jcm9041204.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Long Non-coding RNAs Involved in Pathogenic Infection. (2020)
  • Author(s): Shirahama S, Miki A, Kaburaki T, Akimitsu N.
  • Journal Title: Front Genet. Volume: 11 Pages: 454
  • DOI: 10.3389/fgene.2020.00454.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Involvement of autotaxin in the pathophysiology of elevated intraocular pressure in Posner-Schlossman syndrome. (2020)
  • Author(s): Igarashi N, Honjo M, Yamagishi R, Kurano M, Yatomi Y, Igarashi K, Kaburaki T, Aihara M.
  • Journal Title: Sci Rep. Volume: 14(10) Pages: 6265.
  • DOI: 10.1038/s41598-020-63284-1.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genetic control of CCL24, POR, and IL23R contributes to the pathogenesis of sarcoidosis. (2020)
  • Author(s): Meguro A, Ishihara M, Petrek M, Yamamoto K, Takeuchi M, Mrazek F, Kolek V, Benicka A, Yamane T, Shibuya E, Yoshino A, Isomoto A, Ota M, Yatsu K, Shijubo N, Nagai S, Yamaguchi E, Yamaguchi T, Namba K, Kaburaki T, et al.
  • Journal Title: Commun Biol. Volume: 3(1) Pages: 465.
  • DOI: 10.1038/s42003-020-01185-9.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparison of visual field defect progression in secondary Glaucoma due to anterior uveitis caused by three types of herpes viruses. (2020)
  • Author(s): Shirahama S, Kaburaki T, Takada S, Nakahara H, Tanaka R, Komae K, Fujino Y, Kawashima H, Aihara M.
  • Journal Title: Graefes Arch Clin Exp Ophthalmol. Volume: 258(3) Pages: 639-645.
  • DOI: 10.1007/s00417-019-04559-w.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The Relationship between Fluorescein Angiography Leakage after Infliximab Therapy and Relapse of Ocular Inflammatory Attacks in Ocular Behcet's Disease Patients. (2020)
  • Author(s): Shirahama S, Kaburaki T, Matsuda J, Tanaka R, Nakahara H, Komae K, Kawashima H, Aihara M.
  • Journal Title: Ocul Immunol Inflamm. Volume: 28 Pages: 1166-1170.
  • DOI: 10.1080/09273948.2019.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Long-Term Safety and Efficacy of Adalimumab in Patients with Noninfectious Intermediate Uveitis, Posterior Uveitis, or Panuveitis. (2020)
  • Author(s): Suhler EB, Jaffe GJ, Fortin E, Lim LL, Merrill PT, Dick AD, Brezin AP, Nguyen QD, Thorne JE, Van Calster J, Cimino L, Adan A, Goto H, Kaburaki T, Kramer M, Vitale AT, Kron M, Song AP, Liu J, Pathai S, Douglas KM, Schlaen A, Muccioli C, Van Velthoven MEJ, Zierhut M, Rosenbaum JT.
  • Journal Title: Ophthalmology Volume: 127 Pages: in press
  • DOI: 10.1016/j.ophtha.2020.10.036.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Long noncoding RNA U90926 is crucial for herpes simplex virus type 1 proliferation in murine retinal photoreceptor cells. (2020)
  • Author(s): Shirahama S, Onoguchi-Mizutani R, Kawata K, Taniue K, Miki A, Kato A, Kawaguchi Y, Tanaka R, Kaburaki T, Kawashima H, Urade Y, Aihara M, Akimitsu N
  • Journal Title: Sci Rep. Volume: 10(1) Pages: 19406.
  • DOI: 10.1038/s41598-020-76450-2.
• [Presentation] 眼炎症疾患の最近の進歩 －全身治療の進歩－ (2020)
  • Author(s): 蕪城 俊克
  • Organizer: 第124回日本眼科学会総会
  • Invited
• [Presentation] 前房からHSV-1 DNAを検出したヘルペス性角膜輪部炎の1例. (2020)
  • Author(s): 高野博子、蕪城俊克、武島 聡史、梯 彰弘.
  • Organizer: 第74回日本臨床眼科学会
• [Presentation] 難治性緑内障に対するアーメド緑内障バルブの手術成績 (2020)
  • Author(s): 御任真言、武島聡史、高木理那、田中克明、榛村真智子、木下望、髙野博子、蕪城俊克、梯彰弘.
  • Organizer: 第74回日本臨床眼科学会
• [Presentation] 蛍光眼底造影検査にてIRVAN症候群の診断に至った２例. (2020)
  • Author(s): 武斯斌、田中理恵、蕪城俊克、南貴弘、伊沢英知、外山琢、白濱新多朗、小野久子、曽我拓嗣
  • Organizer: 第74回日本臨床眼科学会