Evolution of tumor subclones and T-cell dynamics underlie variable ibrutinib responses in Waldenström macroglobulinemia

H Hao Sun R Romanos Sklavenitis-Pistofidis S Shirong Liu X Xia Liu N Nicholas Tsakmaklis J John M. Hatcher (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) M Maria Luisa Guerrera (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) A Amanda Kofides (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) A Andres Ramirez-Gamero (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) A Abigail L. Peachey (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) S Shuqiang Li D Derin B. Keskin V Vipheaviny Chea N Nawoo Kim (2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA) H Haoxiang Lyu (2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA) W Wesley Lu (2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA) K Kenneth J. Livak K Kirsten Meid (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) A Alberto Guijosa (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) C Catherine A. Flynn (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) D Dominic Pizzarella (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) C Christopher J. Patterson (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) M Mu Hao (4State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China) S Shuhua Yi (4State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China) W Weiping Yuan (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences 1 , Beijing 100190,) A Andrew R. Branagan (Department of Medicine, Massachusetts General Hospital, Boston) C Catherine J. Wu I Irene M. Ghobrial L Lugui Qiu S Shayna R. Sarosiek (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) J Jorge J. Castillo (1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA) Z Zachary R. Hunter S Steven P. Treon

Abstract

Abstract To elucidate the molecular basis underlying differential responses and resistance to ibrutinib in Waldenström macroglobulinemia (WM), we conducted a prospective phase 2 trial of ibrutinib monotherapy in treatment-naïve patients. A total of 74 sequential bone marrow (BM) aspirates from 17 patients, collected from baseline through 48 treatment cycles, were profiled using single-cell multiomics. BM cells were segregated primarily into B-cell/plasma cell and T-cell compartments. Longitudinal clonal tracking of malignant B cells/plasma cells identified 3 distinct evolutionary patterns: evolution (early clone contraction with late clone expansion and increasing genomic complexity), devolution (early clone expansion with late clone contraction and genomic simplification), and no evolution (stable clonal architecture). The evolution pattern was strongly associated with disease progression, whereas devolution correlated with durable clinical response. Transcriptomic profiling of resistant clones enabled development and validation of the Waldenström ibrutinib prediction (WIP) score, which predicted treatment response at baseline. Within the WIP signature, LYN emerged as a key regulator; LYN knockdown or inhibition significantly increased WM cell sensitivity to ibrutinib, suggesting a rational combination strategy. In parallel, GZMB+ CD8+ effector-memory T cells expanded after treatment in patients with progressive disease and coexisted with tumor evolution. These cells exhibited persistently impaired cytotoxic programs (eg, GNLY), a dedifferentiated memory-like state, elevated PDCD1 expression, and reduced T-cell receptor diversity. Together, this study provides, to our knowledge, the first single-cell framework of tumor clonal evolution and T-cell dysfunction under ibrutinib in WM, introduces the WIP score as a predictive biomarker for treatment response, and identifies actionable tumor-intrinsic and immune mechanisms driving resistance. This trial was registered at www.ClinicalTrials.gov as NCT02604511.

Article Details

Journal Blood
Volume / Issue Vol. 147, Issue 20
Published May 14, 2026
Pages 2298-2314
ISSN 0006-4971
Publisher Elsevier BV

Journal Info

Blood

Elsevier BV

ISSN: 0006-4971 Health Sciences

Authors (33)

H

Hao Sun

R

Romanos Sklavenitis-Pistofidis

S

Shirong Liu

X

Xia Liu

N

Nicholas Tsakmaklis

J

John M. Hatcher

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

M

Maria Luisa Guerrera

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

A

Amanda Kofides

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

A

Andres Ramirez-Gamero

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

A

Abigail L. Peachey

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

S

Shuqiang Li

D

Derin B. Keskin

V

Vipheaviny Chea

N

Nawoo Kim

2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA

H

Haoxiang Lyu

2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA

W

Wesley Lu

2Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA

K

Kenneth J. Livak

K

Kirsten Meid

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

A

Alberto Guijosa

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

C

Catherine A. Flynn

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

D

Dominic Pizzarella

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

C

Christopher J. Patterson

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

M

Mu Hao

4State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China

S

Shuhua Yi

4State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China

W

Weiping Yuan

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences 1 , Beijing 100190,

A

Andrew R. Branagan

Department of Medicine, Massachusetts General Hospital, Boston

C

Catherine J. Wu

I

Irene M. Ghobrial

L

Lugui Qiu

S

Shayna R. Sarosiek

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

J

Jorge J. Castillo

1Bing Center for Waldenström’s Macroglobulinemia, Dana-Farber Cancer Institute, Boston, MA

Z

Zachary R. Hunter

S

Steven P. Treon