The study of control of immortalization of EBV-infected cells and the development of the immortalization inhibitor

• Project/Area Number: 19K08336
• 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: National Center for Child Health and Development
• Principal Investigator: Imadome Ken-Ichi 国立研究開発法人国立成育医療研究センター, 高度感染症診断部, 部長 (70392488)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: CD40シグナル / EBウイルス / PTLD / 移植後リンパ増殖症 / 不死化 / EBV / B細胞不死化 / CD40 / CD40L / がん化阻害薬 / 感染免疫 / 腫瘍ウイルス制御 / EBV-LPD/PTLD / ヒト化マウス

Outline of Research at the Start

EBウイルス(EBV)は移植医療後のリンパ増殖症 (PTLD)を引き起こす代表的なウイルスである。PTLDは、小児の臓器移植後に発生する悪性腫瘍の90% 以上を占め、移植医療成績を左右する重要な合併症である。PTLD発症は成人よりも10倍以上小児が多く、その予防と治療法の確立は急務である。現在、EBVに対する抗ウイルス薬はなく対処療法が主な治療になり、骨髄移植を選択しなくてはいけない症例も少なくない。本研究ではヒト化マウスを利用し、PTLD発症の鍵となるEBV感染細胞の不死化（がん化）制御機構の解明と不死化阻害薬の開発を目指し、PTLD発症予防法の確立を目標とする

Outline of Final Research Achievements

We used humanized mice in this study and aimed at the elucidation of the control of immortalization (transformation) of EBV-infected cells, and at the development of the immortalization inhibitor. And we aimed at the establishment of the PTLD prophylaxis.
We converted human cells from insect cells to manufacture CD40Ig of the CD40-signal inhibitor. CD40 signal was inhibited by CD40Ig which reduced the efficiency of immortalization. We enabled increase of the inhibition efficiency and the weight loss of the dose. Also, we paid attention to CD40 ligand (CD40L), and manufacture of CD40LIg which the signal inhibition to CD40L.As a result, manufacture of secretor CD40Ig and secretor CD40LIg using human cells and a collection were successful.

Academic Significance and Societal Importance of the Research Achievements

Epstein-Barr virus (EBウイルス：EBV)は移植医療後のリンパ増殖症 (PTLD: Posttransplant lymphoprolifirative disorder)を引き起こす代表的なウイルスである。PTLDは、小児の臓器移植後に発生する悪性腫瘍の90% 以上を占め、移植医療成績を左右する重要な合併症である。PTLD発症は成人よりも10倍以上小児が多く、その予防と治療法の確立は急務である。本研究の成果の実用化により、EBV未感染移植治療実施小児に対し、CD40Ig,CD40LIgを用いることでPTLD発症予防が可能になる初めての治療薬となることが考えられる。

Research Products

• [Journal Article] Immunization with Epstein-Barr Virus Core Fusion Machinery Envelope Proteins Elicit High Titers of Neutralizing Activities and Protect Humanized Mice from Lethal Dose EBV Challenge. (2021)
  • Author(s): Cui X., Cao Z., Ishikawa Y., Cui S., Imadome KI., Snapper C. M.
  • Journal Title: Vaccines (Basel) Volume: 9(3) Issue: 3 Pages: 285293-285293
  • DOI: 10.3390/vaccines9030285
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Donor-derived myelodysplastic syndrome after allogeneic stem cell transplantation in a family with germline GATA2 mutation (2020)
  • Author(s): Sakata Naoki、Okano Munehiro、Masako Ryujin、Tanaka Ai、Yamashita Yuhei、Karasuno Takahiro、Imadome Ken-Ichi、Okada Mitsuru、Sugimoto Keisuke
  • Journal Title: International Journal of Hematology Volume: 113 Pages: 290-296
  • Peer Reviewed / Open Access
• [Journal Article] Dasatinib exacerbates splenomegaly of mice inoculated with Epstein-Barr virus-infected lymphoblastoid cell lines. (2020)
  • Author(s): 2)Kotaki R, Kawashima M, Yamamoto Y, Higuchi H, Nagashima E, Kurosaki N, Takamatsu M, Kikuti YY, Imadome KI, Nakamura N, *Kotani A.
  • Journal Title: Sci Rep. Volume: 10(1): Issue: 1 Pages: 4355-4355
  • DOI: 10.1038/s41598-020-61300-y
  • Peer Reviewed / Open Access
• [Journal Article] Parvovirus B19-Infected Tubulointerstitial Nephritis in Hereditary Spherocytosis (2020)
  • Author(s): Nishiyama Kei、Watanabe Yuka、Ishimura Masataka、Tetsuhara Kenichi、Imai Takashi、Kanemasa Hikaru、Ueki Kenji、Motomura Yoshitomo、Kaku Noriyuki、Sakai Yasunari、Imadome Ken-Ichi、Ohga Shouichi
  • Journal Title: Open Forum Infectious Diseases Volume: 7 Pages: 7-14
  • Peer Reviewed / Open Access
• [Journal Article] A case of human herpesvirus 6 encephalitis following pediatric hematopoietic stem cell transplantation: early diagnosis and treatment matters (2020)
  • Author(s): Sakamoto Atsushi、Yamada Masaki、Tsujimoto Shin-ichi、Osumi Tomoo、Arai Katsuhiro、Tomizawa Daisuke、Ishiguro Akira、Matsumoto Kimikazu、Imadome Ken-Ichi、Kato Motohiro
  • Journal Title: International Journal of Hematology Volume: 112 Pages: 751-754
  • Peer Reviewed / Open Access
• [Journal Article] Recurrent Enteritis Associated with Epstein-Barr Virus-positive CD4⁺ T-cell Lymphoproliferative Disorder after Autologous Stem Cell Transplantation (2020)
  • Author(s): Kawaguchi Shinichiro、Sato Kazuya、Oshiro Hisashi、Imadome Ken-Ichi、Kanda Yoshinobu
  • Journal Title: Internal Medicine Volume: 59 Pages: 2565-2569
  • NAID: 130007926290
  • Peer Reviewed / Open Access
• [Journal Article] Increased Granulopoiesis in the Bone Marrow following Epstein-Barr Virus Infection. (2019)
  • Author(s): Katahira Y, Higuchi H, Matsushita H, Yahata T, Yamamoto Y, Koike R, Ando K, Sato K, Imadome K, Kotani A.
  • Journal Title: Sci Rep Volume: 9 Issue: 1 Pages: 13445-13445
  • DOI: 10.1038/s41598-019-49937-w
  • NAID: 120006960403
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Shift in Epstein?Barr virus (EBV)‐infected cells in chronic active EBV disease (2019)
  • Author(s): Sakaki Shinichiro、Imadome Ken‐Ichi、Kawano Fuyuko、Nakadate Hisaya、Ishiguro Akira
  • Journal Title: Pediatrics International Volume: 61 Issue: 8 Pages: 825-826
  • DOI: 10.1111/ped.13935
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Systemic Epstein-Barr Virus-Positive T/NK Lymphoproliferative Diseases With SH2D1A/XIAP Hypomorphic Gene Variants (2019)
  • Author(s): Ishimura M, Eguchi K, Shiraishi A, Sonoda M, Azuma Y, Yamamoto H, Imadome K, Ohga S
  • Journal Title: Frontiers in Pediatrics Volume: 7 Pages: 183-183
  • DOI: 10.3389/fped.2019.00183
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Epstein-Barr Virus-Associated γδ T-Cell Lymphoproliferative Disorder Associated With Hypomorphic IL2RG Mutation (2019)
  • Author(s): Tanita K, Hoshino A, Imadome K, Kamiya T, Inoue K, Okano T, Yeh TW, Yanagimachi M, Shiraishi A, Ishimura M, Schober T, Rohlfs M, Takagi M, Imai K, Takada H, Ohga S, Klein C, Morio T, Kanegane H.
  • Journal Title: Front Pediatr Volume: 7 Pages: 15-15
  • DOI: 10.3389/fped.2019.00015
  • Peer Reviewed / Open Access / Int'l Joint Research