The protein structure determination of IL-36 related molecules and their clinical application.

• Project/Area Number: 18K07840
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Gifu University
• Principal Investigator: Ohnishi Hidenori 岐阜大学, 大学院医学系研究科, 教授 (60381620)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 蛋白立体構造解析 / 自己炎症性疾患 / 乾癬 / サイトカイン / 遺伝子変異

Outline of Final Research Achievements

Interleukin-36, which is associated with the self-defense system in dermis, is one of a molecule of IL-1 family. Recently, it was reported that generalized pustular psoriasis was caused by loss-of-functional mutation of IL36RN gene coding IL-36-receptor antagonist (IL-36Ra). In this study, we aimed a determination of the complex protein structure of IL-36 or IL-36Ra and IL-36-receptor α chain (IL-36R). Additionally, we tried to use the structural information for the clinical application. As the results, we established a purification method of recombinant proteins (IL-36, IL-36Ra and IL-36R). An IL-36R protein produced by the silkworm expression system could bind an IL-36Ra. Furthermore, we established an in vitro assay system to evaluate the variants of IL36RN whether is pathogenic or not. The protein structure determination will be continued in the next new research project.

Academic Significance and Societal Importance of the Research Achievements

IL-36受容体を標的とした抗体製剤の開発が進んでおり、IL-36が膿疱性乾癬の治療標的分子として有望であることは明らかである。蛋白立体構造の解明は、構造基盤による創薬につながる情報となりうる。また、患者遺伝子解析から様々なIL36RN遺伝子変異が同定されることを経験するが、既知遺伝子変異の場合、疾患関連性の判定は容易であるが、ときに新規のバリアントに遭遇することがある。その場合、なんらかのin vitro実験を行う必要があるため、一般臨床医には診断に至るまでの過程のハードルが著しく高くなってしまう。そこで詳細な構造情報があれば、病因変異かどうかの判定に利用することができる点でも有用である。

Research Products

• [Journal Article] Successful treatment of a novel type I interferonopathy due to a de novo PSMB9 gene mutation with a Janus kinase inhibitor (2021)
  • Author(s): Kataoka S, Kawashima N, Okuno Y, Muramatsu H, Miwata S, Narita K, Hamada M, Murakami N, Taniguchi R, Ichikawa D, Kitazawa H, Suzuki K, Nishikawa E, Narita A, Nishio N, Yamamoto H, Fukasawa Y, Kato T, Yamamoto H, Natsume J, Kojima S, Nishio I, Taketani T, Ohnishi H, Tahakashi Y.
  • Journal Title: Journal of Allergy and Clinical Immunology Volume: - Issue: 2 Pages: 639-644
  • DOI: 10.1016/j.jaci.2021.03.010
  • NAID: 120007192018
  • Peer Reviewed
• [Journal Article] A Novel Homozygous Mutation Destabilizes IKKβ and Leads to Human Combined Immunodeficiency (2021)
  • Author(s): Qin Tao、Jia Yanjun、Liu Yuhang、Dai Rongxin、Zhou Lina、Okada Satoshi、Tsumura Miyuki、Ohnishi Hidenori、Kato Zenichiro、Kanegane Hirokazu、Sun Xiulian、Zhao Xiaodong
  • Journal Title: Frontiers in Immunology Volume: 11 Pages: 517544-517544
  • DOI: 10.3389/fimmu.2020.517544
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional analysis of novel A20 variants in patients with atypical inflammatory diseases (2021)
  • Author(s): Kadowaki S, Hashimoto K, Nishimura T, Kashimada K, Kadowaki T, Kawamoto N, Imai K, Okada S, Kanegane H, Ohnishi H.
  • Journal Title: Arthritis Res Ther Volume: 23 Issue: 1 Pages: 52-52
  • DOI: 10.1186/s13075-021-02434-w
  • Peer Reviewed / Open Access
• [Journal Article] Clinical and Immunological Heterogeneity in Japanese Patients with Gain-of-Function Variants in STAT3 (2021)
  • Author(s): Tanita Kay、Sakura Fumiaki、Nambu Ryusuke、Tsumura Miyuki、Imanaka Yusuke、Ohnishi Hidenori、Kato Zenichiro、Pan Jie、Hoshino Akihiro、Suzuki Koji、Yasutomi Motoko、Umetsu Shuichiro、Okada Chizuru、Takagi Masatoshi、Imai Kohsuke、Ohara Osamu、Muise Alexo M.、Okada Satoshi、Morio Tomohiro、Kanegane Hirokazu
  • Journal Title: Journal of Clinical Immunology Volume: - Issue: 4 Pages: 780-790
  • DOI: 10.1007/s10875-021-00975-y
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Case of Muckle-Wells syndrome with erythema dominantly infiltrated by lymphocytes. (2021)
  • Author(s): Kadowaki S, Kimura T, Shiraki M, Mizutani Y, Nakama M, Kobayashi K, Suzui N, Kawamoto N, Ohnishi H, Seishima M.
  • Journal Title: J Dermatol. Volume: 48 Issue: 2
  • DOI: 10.1111/1346-8138.15686
  • Peer Reviewed
• [Journal Article] IRAK4 Deficiency Presenting with Anti-NMDAR Encephalitis and HHV6 Reactivation (2020)
  • Author(s): Nishimura Shiho、Kobayashi Yoshiyuki、Ohnishi Hidenori、Moriya Kunihiko、Tsumura Miyuki、Sakata Sonoko、Mizoguchi Yoko、Takada Hidetoshi、Kato Zenichiro、Sancho-Shimizu Vanessa、Picard Capucine、Irani Sarosh R.、Ohara Osamu、Casanova Jean-Laurent、Puel Anne、Ishikawa Nobutsune、Okada Satoshi、Kobayashi Masao
  • Journal Title: Journal of Clinical Immunology Volume: 41 Issue: 1 Pages: 125-135
  • DOI: 10.1007/s10875-020-00885-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immunophenotyping of A20 haploinsufficiency by multicolor flow cytometry. (2020)
  • Author(s): Kadowaki T, Ohnishi H, Kawamoto N, Kadowaki S, Hori T, Nishimura K, Kobayashi C, Shigemura T, Ogata S, Inoue Y, Hiejima E, Izawa K, Matsubayashi T, Matsumoto K, Imai K, Nishikomori R, Ito S, Kanegane H, Fukao T.
  • Journal Title: Clin Immunol. Volume: 216 Pages: 108441-108441
  • Peer Reviewed
• [Journal Article] The IL1RN Mutation Creating the Most-Upstream Premature Stop Codon Is Hypomorphic Because of a Reinitiation of Translation. (2020)
  • Author(s): Moriya K, Kadowaki S, Nakano T, Akarcan SE, Kutukculer N, Aksu G, Sasahara Y, Kure S, Ohnishi H, Casanova JL, Puel A, Fukao T.
  • Journal Title: J Clin Immunol. Volume: 40 Issue: 4 Pages: 643-645
  • DOI: 10.1007/s10875-020-00770-1
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Haploinsufficiency of A20 with a novel mutation of deletion of exons 2-3 of TNFAIP3. (2020)
  • Author(s): Shimizu Maho、Matsubayashi Tadashi、Ohnishi Hidenori、Nakama Mina、Izawa Kazushi、Honda Yoshitaka、Nishikomori Ryuta
  • Journal Title: Modern Rheumatology Volume: e-pub Issue: 2 Pages: 1-5
  • DOI: 10.1080/14397595.2020.1719595
  • Peer Reviewed
• [Journal Article] Impetigo herpetiformis with a CARD14 Thr79Ile variant successfully treated with granulocyte and monocyte adsorption apheresis. (2020)
  • Author(s): Fujii Kento、Takahashi Tomoko、Matsuyama Kanako、Fujii Asami、Mizutani Yoko、Ohnishi Hidenori、Seishima Mariko
  • Journal Title: The Journal of Dermatology Volume: 47 Issue: 3
  • DOI: 10.1111/1346-8138.15241
  • Peer Reviewed
• [Journal Article] The Long-Term Efficacy of Cochlear Implantation for Hearing Loss in Muckel-Wells Syndrome. (2019)
  • Author(s): Ogawa B, Aoki M, Ohnishi H, Ohashi T, Hayashi H, Kuze B, Ito Y.
  • Journal Title: J Int Adv Otol. Volume: 15 Issue: 3 Pages: 454-458
  • DOI: 10.5152/iao.2019.6871
  • Peer Reviewed / Open Access
• [Journal Article] Immunologic Effects of Sirolimus in Patients With Vascular Anomalies. (2019)
  • Author(s): Nozawa Akifumi、Ozeki Michio、Yasue Shiho、Endo Saori、Kawamoto Norio、Ohnishi Hidenori、Fumino Shigehisa、Furukawa Taizo、Tajiri Tatsuro、Maekawa Takanobu、Fujino Akihiro、Souzaki Ryota、Fukao Toshiyuki
  • Journal Title: Journal of Pediatric Hematology/Oncology Volume: e-pub Issue: 5 Pages: 1-1
  • DOI: 10.1097/mph.0000000000001650
  • Peer Reviewed / Open Access
• [Journal Article] Prominent Dermal Langerhans Cells in an Omenn Syndrome Patient With a Novel Mutation in the IL2RG Gene (2019)
  • Author(s): Ibusuki Atsuko、Nishikawa Takuro、Hiraki Tsubasa、Okano Tsubasa、Imai Kohsuke、Kanegane Hirokazu、Ohnishi Hidenori、Kato Zenichiro、Fujii Kazuyasu、Tanimoto Akihide、Kawano Yoshifumi、Kanekura Takuro
  • Journal Title: The Journal of Dermatology Volume: 46 Issue: 11 Pages: 1019-1023
  • DOI: 10.1111/1346-8138.15054
  • Peer Reviewed / Open Access
• [Journal Article] Autosomal dominant Hashimoto’s thyroiditis with a mutation in <i>TNFAIP3</i> (2019)
  • Author(s): Hori Tomohiro、Ohnishi Hidenori、Kadowaki Tomonori、Kawamoto Norio、Matsumoto Hideki、Ohara Osamu、Fukao Toshiyuki
  • Journal Title: Clinical Pediatric Endocrinology Volume: 28 Issue: 3 Pages: 91-96
  • DOI: 10.1297/cpe.28.91
  • NAID: 130007681720
  • ISSN: 0918-5739, 1347-7358
  • Peer Reviewed / Open Access
• [Journal Article] An innate interaction between IL-18 and the propeptide that inactivates its precursor form (2019)
  • Author(s): Naotaka Tsutsumi, Ayumi Yokota, Takeshi Kimura, Zenichiro Kato, Toshiyuki Fukao, Masahiro Shirakawa, Hidenori Ohnishi & Hidehito Tochio
  • Journal Title: Scientific Reportsvolume Volume: 9 Issue: 1 Pages: 6160-6160
  • DOI: 10.1038/s41598-019-42661-5
  • NAID: 120007006235
  • Peer Reviewed / Open Access
• [Journal Article] Gain-of-function IKBKB mutation causes human combined immune deficiency (2018)
  • Author(s): Cardinez C, Miraghazadeh B, Tanita K, da Silva E, Hoshino A, Okada S, Chand R, Asano T, Tsumura M, Yoshida K, Ohnishi H, Kato Z, Yamazaki M, Okuno Y, Miyano S, Kojima S, Ogawa S, Andrews TD, Field MA, Burgio G,et al.
  • Journal Title: J Exp Med Volume: 215 Issue: 11 Pages: 2715-2724
  • DOI: 10.1084/jem.20180639
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Intramolecular interaction suggests an autosuppression mechanism for the innate immune adaptor protein MyD88 (2018)
  • Author(s): Uno Masatoshi、Watanabe-Nakayama Takahiro、Konno Hiroki、Akagi Ken-ichi、Tsutsumi Naotaka、Fukao Toshiyuki、Shirakawa Masahiro、Ohnishi Hidenori、Tochio Hidehito
  • Journal Title: Chemical Communications Volume: 54 Issue: 87 Pages: 12318-12321
  • DOI: 10.1039/c8cc06480f
  • Peer Reviewed
• [Journal Article] IKBA S32 Mutations Underlie Ectodermal Dysplasia with Immunodeficiency and Severe Noninfectious Systemic Inflammation (2018)
  • Author(s): Moriya K, Sasahara Y, Ohnishi H, Kawai T, Kanegane H.
  • Journal Title: J Clin Immunol Volume: 38 Issue: 5 Pages: 543-545
  • DOI: 10.1007/s10875-018-0522-y
  • Peer Reviewed
• [Journal Article] Hypoallergenic casein hydrolysate for peptide-based oral immunotherapy in cow's milk allergy (2018)
  • Author(s): Ueno HM, Kato T, Ohnishi H, Kawamoto N, Kato Z, Kaneko H, Kondo N, Nakano T.
  • Journal Title: J Allergy Clin Immunol Volume: 142 Issue: 1 Pages: 330-333
  • DOI: 10.1016/j.jaci.2018.04.005
  • Peer Reviewed
• [Presentation] The clinical and immunological profiles of haploinsufficiency of A20 in Japan. (2019)
  • Author(s): Ohnishi H., Kadowaki T., Kawamoto N., Hori T., Nishimura K., Kobayashi C., Shigemura T., Ogata S., Inoue Y., Kawai T., Hiejima E., Izawa K., Matsubayashi T., Takagi M., Imai K., Nishikomori R., Ito S., Heike T., Ohara O., Morio T, Kanegane H., Fukao T.
  • Organizer: The 10th Biannual Meeting of the International Society of Systemic Auto-Inflammatory Diseases
  • Int'l Joint Research
• [Presentation] A case of novel identified proteasome-related autoinflammation and immunodeficiency syndrome caused by PSMB9 mutation. (2019)
  • Author(s): Ohnishi H., Kataoka S., Muramatsu H., Kadoi E., Kanazawa N., Okada S., Honda Y., Izawa K., Nishikomori R., Taketani T., Hamazaki J., Murata S., Takahashi Y., Fukao T.
  • Organizer: The 10th Biannual Meeting of the International Society of Systemic Auto-Inflammatory Diseases
  • Int'l Joint Research
• [Presentation] What PID to suspect when a child present with difficult to treat mycobacterial infections? (2019)
  • Author(s): Ohnishi H.
  • Organizer: The 15th Congress of Asian Society for Pediatric Research (ASPR)
  • Invited
• [Presentation] 自己炎症性疾患の診断と治療およびイラリスの適正使用 (2018)
  • Author(s): 大西秀典
  • Organizer: 日本小児リウマチ学会総会・学術集会(第28回)
• [Presentation] Anti-NMDA receptor encephalitis associate with IRAK4 deficiency. (2018)
  • Author(s): Nishimura S, Kobayashi Y, Tsumura M, Ishikawa N, Ohnishi H, Takada H, Sancho-Shimizu V, Moriya K, Anne P, Capucine P, Irani S, Casanova JL, Okada S, Kobayashi M.
  • Organizer: ESID 2018 (European Society for Immunodeficiencies)
  • Int'l Joint Research
• [Book] 病態・治療論［14］ 小児疾患 (2019)
  • Author(s): 真部 淳、松藤 凡、小林 京子
  • Total Pages: 380
  • Publisher: 南江堂
  • ISBN: 9784524237555