Multiomics analysis reveals the genetic and epigenetic features of high-risk NK cell–type chronic active EBV infection

R Ryo Akazawa (1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan) T Takashi Mikami M Masaki Yamada I Itaru Kato H Hirohito Kubota S Satoshi Saida (1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan) Y Yoshinori Uchihara Y Yuriko Ishikawa (3Department of Advanced Medicine for Virus Infections, National Center for Child Health and Development, Tokyo, Japan) T Tatsuya Kamitori K Keiji Tasaka K Kiyotaka Isobe (1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan) T Tomoya Isobe K Kazushi Izawa K Katsutsugu Umeda (1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan) H Hidefumi Hiramatsu (1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan) K Keita Jinnouchi (9Department of Diagnostic Pathology, Kyoto University Hospital, Kyoto, Japan) M Masahiro Hirata (9Department of Diagnostic Pathology, Kyoto University Hospital, Kyoto, Japan) M Masakazu Fujimoto T Tomoo Daifu (10Department of Pediatrics, Japanese Red Cross Otsu Hospital, Otsu, Japan) H Hiroo Ueno (16Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan) S Seishiro Nodomi (5Department of Pediatrics, Kurashiki Central Hospital, Okayama, Japan) M Machiko Sawada H Hisanori Fujino (12Department of Pediatrics, Osaka Red Cross Hospital, Osaka, Japan) K Katsuyoshi Koh (23Japan Children’s Cancer Group ALL Committee, Nagoya, Japan) M Mitsuteru Hiwatari (14Department of Pediatrics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan) M Motohiro Kato (6Department of Pediatrics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan) H Hiroaki Goto I Ikumi Katano (17Laboratory Animal Research Department, Central Institute for Experimental Medicine and Life Science, Kawasaki, Japan) R Ryoji Ito (17Laboratory Animal Research Department, Central Institute for Experimental Medicine and Life Science, Kawasaki, Japan) M Mamoru Ito N Nobuyuki Kakiuchi M Masahiro M. Nakagawa Y Yuichi Shiraishi Y Yoshitaka Honda H Hiroyuki Yoshitomi (21Institute for the Advanced Study of Human Biology, Kyoto University, Kyoto, Japan) H Hideki Ueno (Department of Immunology, Graduate School of Medicine, Kyoto University) M Maho Sato (23Department of Hematology/Oncology, Osaka Women’s and Children’s Hospital, Osaka, Japan) S Satoru Miyano H Hironori Haga (9Department of Diagnostic Pathology, Kyoto University Hospital, Kyoto, Japan) A Akihisa Sawada (23Department of Hematology/Oncology, Osaka Women’s and Children’s Hospital, Osaka, Japan) K Ken-Ichi Imadome S Seishi Ogawa J Junko Takita

Abstract

Abstract Chronic active Epstein-Barr virus (EBV) infection (CAEBV) is an orphan disease characterized by the proliferation and infiltration of EBV-infected T/natural killer (NK) cells into multiple organs. Although CAEBV is a heterogeneous disease with diverse clinical courses, its pathogenesis remains poorly understood. In this study, we explored the molecular mechanisms underlying CAEBV by performing a comprehensive multiomics analysis, including genome, transcriptome, epigenome, and single-cell transcriptome and surface proteome analyses, of 65 patients with CAEBV. Methylation analysis identified 2 distinct subtypes of NK cell–type CAEBV based on the CpG island methylator phenotype (CIMP). In CIMP-positive CAEBV, regions associated with enhancer of zeste homolog 2 binding sites and histone H3 lysine 27 trimethylation exhibited increased DNA hypermethylation, resulting in downregulation of tumor suppressor and antiherpesvirus genes. CIMP-positive CAEBV had a particularly poor prognosis and displayed a “neoplastic” phenotype with a DNA methylation pattern similar to that of extranodal NK/T-cell lymphoma, a higher tumor mutation burden, and frequent copy number alterations. In addition, both in vitro and in vivo functional assays demonstrated that 5-azacytidine, a hypomethylating agent, was a potentially effective agent for high-risk CIMP-positive CAEBV. Finally, we established a method to effectively detect EBV-infected cells in single-cell analysis, suggesting that EBV-infected NK cells have tissue-resident properties and that innate and adaptive immunity to EBV is compromised in patients with CAEBV. The present findings provide insight into the complex molecular features of CAEBV and suggest potential molecular therapies.

Article Details

Journal Blood
Volume / Issue Vol. 146, Issue 19
Published November 06, 2025
Pages 2336-2349
ISSN 0006-4971
Publisher Elsevier BV

Journal Info

Blood

Elsevier BV

ISSN: 0006-4971 Health Sciences

Authors (43)

R

Ryo Akazawa

1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan

T

Takashi Mikami

M

Masaki Yamada

I

Itaru Kato

H

Hirohito Kubota

S

Satoshi Saida

1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan

Y

Yoshinori Uchihara

Y

Yuriko Ishikawa

3Department of Advanced Medicine for Virus Infections, National Center for Child Health and Development, Tokyo, Japan

T

Tatsuya Kamitori

K

Keiji Tasaka

K

Kiyotaka Isobe

1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan

T

Tomoya Isobe

K

Kazushi Izawa

K

Katsutsugu Umeda

1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan

H

Hidefumi Hiramatsu

1Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan

K

Keita Jinnouchi

9Department of Diagnostic Pathology, Kyoto University Hospital, Kyoto, Japan

M

Masahiro Hirata

9Department of Diagnostic Pathology, Kyoto University Hospital, Kyoto, Japan

M

Masakazu Fujimoto

T

Tomoo Daifu

10Department of Pediatrics, Japanese Red Cross Otsu Hospital, Otsu, Japan

H

Hiroo Ueno

16Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan

S

Seishiro Nodomi

5Department of Pediatrics, Kurashiki Central Hospital, Okayama, Japan

M

Machiko Sawada

H

Hisanori Fujino

12Department of Pediatrics, Osaka Red Cross Hospital, Osaka, Japan

K

Katsuyoshi Koh

23Japan Children’s Cancer Group ALL Committee, Nagoya, Japan

M

Mitsuteru Hiwatari

14Department of Pediatrics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

M

Motohiro Kato

6Department of Pediatrics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

H

Hiroaki Goto

I

Ikumi Katano

17Laboratory Animal Research Department, Central Institute for Experimental Medicine and Life Science, Kawasaki, Japan

R

Ryoji Ito

17Laboratory Animal Research Department, Central Institute for Experimental Medicine and Life Science, Kawasaki, Japan

M

Mamoru Ito

N

Nobuyuki Kakiuchi

M

Masahiro M. Nakagawa

Y

Yuichi Shiraishi

Y

Yoshitaka Honda

H

Hiroyuki Yoshitomi

21Institute for the Advanced Study of Human Biology, Kyoto University, Kyoto, Japan

H

Hideki Ueno

Department of Immunology, Graduate School of Medicine, Kyoto University

M

Maho Sato

23Department of Hematology/Oncology, Osaka Women’s and Children’s Hospital, Osaka, Japan

S

Satoru Miyano

H

Hironori Haga

9Department of Diagnostic Pathology, Kyoto University Hospital, Kyoto, Japan

A

Akihisa Sawada

23Department of Hematology/Oncology, Osaka Women’s and Children’s Hospital, Osaka, Japan

K

Ken-Ichi Imadome

S

Seishi Ogawa

J

Junko Takita