Somatic <i>IRF4</i> mutations and thymic tropism in primary mediastinal large B-cell lymphoma

S Shinya Rai (2British Columbia Cancer, Centre for Lymphoid Cancer, Vancouver, Canada) G Gerben Duns (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) F Fabian Frontzek (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) J Jasper C. H. Wong (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) Y Yifan Yin M Michael Yu Li (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) M Makoto Kishida (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) M Manabu Fujisawa (2Institute of Medicine, University of Tsukuba, Department of Hematology, Tsukuba, Japan) S Shannon Healy (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) E Elena Viganò (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) A Aixiang Jiang (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) B Bruce Woolcock (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) A Adele Telenius (2British Columbia Cancer, Centre for Lymphoid Cancer, Vancouver, Canada) S Susana Ben-Neriah (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) B Barbara Meissner (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) M Merrill Boyle (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) H Hisae Nakamura (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) L Luke O’Brien (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) C Claudia Cassidy (1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada) A Andrew Lytle (7BC Cancer, Vancouver, Canada) P Pedro Farinha (6BC Cancer, Vancouver, BC, Canada) G Graham Slack (7BC Cancer, Vancouver, Canada) L Laura K. Hilton (4Centre for Lymphoid Cancer, BC Cancer, Vancouver, BC, Canada) R Ryan D. Morin (4Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada) L Leandro Venturutti (2BC Cancer Research Institute, Vancouver, Canada) T Tomohiro Aoki N Nicole Wretham (7Department of Experimental Therapeutics, British Columbia Cancer Research Institute, Vancouver, BC, Canada) J Jonathan W. Bush (4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada) L Laura Evgin K Kerry J. Savage (6BC Cancer, Vancouver, BC, Canada) D David W. Scott (5Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, Canada) C Christian Steidl (5Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, Canada)

Abstract

Abstract Disease-defining signatures in lymphomas, driven by intricate molecular mechanisms, have advanced molecular taxonomies, refined classification, and may guide clinical management; however, the role of these signatures in driving disease hallmarks, including subtype-specific organotropism, remains largely unexplored. Primary mediastinal large B-cell lymphoma (PMBCL) is an exemplary lymphoma characterized by disease manifestations in the thymic niche, unique genetic alterations, and immune escape. Here, we identified interferon regulatory factor 4 (IRF4)–C99R mutations uniquely occurring in PMBCL through mutational meta-analysis of large-scale data sets. By integrating multiomics approaches with genome editing in PMBCL cells, we revealed that IRF4-C99R contributes to a differentiation block phenotype. Specifically, we showed that IRF4-C99R reduces its binding to the interferon-stimulated response element (ISRE) motif within PRDM1, encoding a key transcriptional regulator of B-cell differentiation, resulting in decreased PRDM1 expression. Additionally, IRF4-C99R suppresses Traf2 and Nck-interacting kinase, a key interferon gamma (IFN-γ) pathway regulator, by impairing ISRE motif binding, thereby reducing IFN-γ signaling and increasing thymus and activation-regulated chemokine (TARC) expression, which drives TARC-mediated chemotaxis of T regulatory cells. We also revealed that IRF4-C99R upregulates ephrin type-B receptor 1 (EPHB1) through noncanonical activating protein 1–IRF composite motif binding and showed that overexpression of EPHB1 in an immunocompetent syngeneic lymphoma model influenced organotropism to favor thymic localization, without affecting overall tumor burden. IRF4-C99R mutation–induced phenotypes were validated in primary PMBCL tissues using single-nuclei RNA sequencing, confirming that the molecular mechanisms observed in vitro align with the pathophysiology of PMBCL in patients. Together, these findings demonstrate how a single genetic mutation orchestrates the coordinated regulation of hallmark traits including thymus-specific tropism in PMBCL.

Article Details

Journal Blood
Volume / Issue Vol. 146, Issue 13
Published September 25, 2025
Pages 1586-1600
ISSN 0006-4971
Publisher Elsevier BV

Journal Info

Blood

Elsevier BV

ISSN: 0006-4971 Health Sciences

Authors (32)

S

Shinya Rai

2British Columbia Cancer, Centre for Lymphoid Cancer, Vancouver, Canada

G

Gerben Duns

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

F

Fabian Frontzek

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

J

Jasper C. H. Wong

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

Y

Yifan Yin

M

Michael Yu Li

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

M

Makoto Kishida

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

M

Manabu Fujisawa

2Institute of Medicine, University of Tsukuba, Department of Hematology, Tsukuba, Japan

S

Shannon Healy

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

E

Elena Viganò

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

A

Aixiang Jiang

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

B

Bruce Woolcock

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

A

Adele Telenius

2British Columbia Cancer, Centre for Lymphoid Cancer, Vancouver, Canada

S

Susana Ben-Neriah

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

B

Barbara Meissner

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

M

Merrill Boyle

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

H

Hisae Nakamura

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

L

Luke O’Brien

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

C

Claudia Cassidy

1Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, BC, Canada

A

Andrew Lytle

7BC Cancer, Vancouver, Canada

P

Pedro Farinha

6BC Cancer, Vancouver, BC, Canada

G

Graham Slack

7BC Cancer, Vancouver, Canada

L

Laura K. Hilton

4Centre for Lymphoid Cancer, BC Cancer, Vancouver, BC, Canada

R

Ryan D. Morin

4Centre for Lymphoid Cancer, BC Cancer Research Institute, Vancouver, BC, Canada

L

Leandro Venturutti

2BC Cancer Research Institute, Vancouver, Canada

T

Tomohiro Aoki

N

Nicole Wretham

7Department of Experimental Therapeutics, British Columbia Cancer Research Institute, Vancouver, BC, Canada

J

Jonathan W. Bush

4Department of Pathology and Laboratory Medicine, The University of British Columbia, Vancouver, BC, Canada

L

Laura Evgin

K

Kerry J. Savage

6BC Cancer, Vancouver, BC, Canada

D

David W. Scott

5Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, Canada

C

Christian Steidl

5Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, Canada