Key Points

  • Conditional expression of HGAL in mice leads to lymphoma that resembles human GC B-type DLBCL.

  • Irrespective of the differential stage at which HGAL is expressed, it leads to a block in B-cell differentiation at the GC reaction.

Abstract

Diffuse large B-cell lymphomas (DLBCLs) are clinically and genetically heterogeneous tumors. Deregulation of diverse biological processes specific to B cells, such as B-cell receptor (BCR) signaling and motility regulation, contribute to lymphomagenesis. Human germinal center associated lymphoma (HGAL) is a B-cell–specific adaptor protein controlling BCR signaling and B lymphocyte motility. In normal B cells, it is expressed in germinal center (GC) B lymphocytes and promptly downregulated upon further differentiation. The majority of DLBCL tumors, primarily GC B-cell types, but also activated types, express HGAL. To investigate the consequences of constitutive expression of HGAL in vivo, we generated mice that conditionally express human HGAL at different stages of hematopoietic development using 3 restricted Cre-mediated approaches to initiate expression of HGAL in hematopoietic stem cells, pro-B cells, or GC B cells. Following immune stimulation, we observed larger GCs in mice in which HGAL expression was initiated in GC B cells. All 3 mouse strains developed DLBCL at a frequency of 12% to 30% starting at age 13 months, leading to shorter survival. Immunohistochemical studies showed that all analyzed tumors were of the GC B-cell type. Exon sequencing revealed mutations reported in human DLBCL. Our data demonstrate that constitutive enforced expression of HGAL leads to DLBCL development.

REFERENCES

REFERENCES
1.
Coiffier
B
,
Lepage
E
,
Briere
J
, et al
.
CHOP chemotherapy plus rituximab compared with CHOP alone in elderly patients with diffuse large-B-cell lymphoma
.
N Engl J Med
.
2002
;
346
(
4
):
235
-
242
.
2.
Alizadeh
AA
,
Eisen
MB
,
Davis
RE
, et al
.
Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling
.
Nature
.
2000
;
403
(
6769
):
503
-
511
.
3.
Lenz
G
,
Wright
G
,
Dave
SS
, et al;
Lymphoma/Leukemia Molecular Profiling Project
.
Stromal gene signatures in large-B-cell lymphomas
.
N Engl J Med
.
2008
;
359
(
22
):
2313
-
2323
.
4.
Morin
RD
,
Johnson
NA
,
Severson
TM
, et al
.
Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin
.
Nat Genet
.
2010
;
42
(
2
):
181
-
185
.
5.
Rosenwald
A
,
Wright
G
,
Chan
WC
, et al;
Lymphoma/Leukemia Molecular Profiling Project
.
The use of molecular profiling to predict survival after chemotherapy for diffuse large-B-cell lymphoma
.
N Engl J Med
.
2002
;
346
(
25
):
1937
-
1947
.
6.
Lenz
G
,
Wright
GW
,
Emre
NC
, et al
.
Molecular subtypes of diffuse large B-cell lymphoma arise by distinct genetic pathways
.
Proc Natl Acad Sci USA
.
2008
;
105
(
36
):
13520
-
13525
.
7.
Chapuy
B
,
Stewart
C
,
Dunford
AJ
, et al
.
Molecular subtypes of diffuse large B cell lymphoma are associated with distinct pathogenic mechanisms and outcomes [published correction appears in Nat Med. 2018;24(8):1290-1291]
.
Nat Med
.
2018
;
24
(
5
):
679
-
690
.
8.
Schmitz
R
,
Wright
GW
,
Huang
DW
, et al
.
Genetics and pathogenesis of diffuse large B-cell lymphoma
.
N Engl J Med
.
2018
;
378
(
15
):
1396
-
1407
.
9.
Lenz
G
,
Staudt
LM
.
Aggressive lymphomas
.
N Engl J Med
.
2010
;
362
(
15
):
1417
-
1429
.
10.
Miao
Y
,
Medeiros
LJ
,
Li
Y
,
Li
J
,
Young
KH
.
Genetic alterations and their clinical implications in DLBCL
.
Nat Rev Clin Oncol
.
2019
;
16
(
10
):
634
-
652
.
11.
Knittel
G
,
Liedgens
P
,
Korovkina
D
, et al;
German International Cancer Genome Consortium Molecular Mechanisms in Malignant Lymphoma by Sequencing Project Consortium
.
B-cell-specific conditional expression of Myd88p.L252P leads to the development of diffuse large B-cell lymphoma in mice
.
Blood
.
2016
;
127
(
22
):
2732
-
2741
.
12.
Knies
N
,
Alankus
B
,
Weilemann
A
, et al
.
Lymphomagenic CARD11/BCL10/MALT1 signaling drives malignant B-cell proliferation via cooperative NF-κB and JNK activation
.
Proc Natl Acad Sci USA
.
2015
;
112
(
52
):
E7230
-
E7238
.
13.
Healy
JA
,
Nugent
A
,
Rempel
RE
, et al
.
GNA13 loss in germinal center B cells leads to impaired apoptosis and promotes lymphoma in vivo
.
Blood
.
2016
;
127
(
22
):
2723
-
2731
.
14.
Muppidi
JR
,
Schmitz
R
,
Green
JA
, et al
.
Loss of signalling via Gα13 in germinal centre B-cell-derived lymphoma
.
Nature
.
2014
;
516
(
7530
):
254
-
258
.
15.
Lossos
IS
,
Alizadeh
AA
,
Rajapaksa
R
,
Tibshirani
R
,
Levy
R
.
HGAL is a novel interleukin-4-inducible gene that strongly predicts survival in diffuse large B-cell lymphoma
.
Blood
.
2003
;
101
(
2
):
433
-
440
.
16.
Lu
X
,
Chen
J
,
Malumbres
R
,
Cubedo Gil
E
,
Helfman
DM
,
Lossos
IS
.
HGAL, a lymphoma prognostic biomarker, interacts with the cytoskeleton and mediates the effects of IL-6 on cell migration
.
Blood
.
2007
;
110
(
13
):
4268
-
4277
.
17.
Schenten
D
,
Egert
A
,
Pasparakis
M
,
Rajewsky
K
.
M17, a gene specific for germinal center (GC) B cells and a prognostic marker for GC B-cell lymphomas, is dispensable for the GC reaction in mice
.
Blood
.
2006
;
107
(
12
):
4849
-
4856
.
18.
Lu
X
,
Kazmierczak
K
,
Jiang
X
, et al
.
Germinal center-specific protein human germinal center associated lymphoma directly interacts with both myosin and actin and increases the binding of myosin to actin
.
FEBS J
.
2011
;
278
(
11
):
1922
-
1931
.
19.
Jiang
X
,
Lu
X
,
McNamara
G
, et al
.
HGAL, a germinal center specific protein, decreases lymphoma cell motility by modulation of the RhoA signaling pathway
.
Blood
.
2010
;
116
(
24
):
5217
-
5227
.
20.
Jiang
X
,
Lu
X
,
Zhang
Y
, et al
.
Interplay between HGAL and Grb2 proteins regulates B-cell receptor signaling
.
Blood Adv
.
2019
;
3
(
15
):
2286
-
2297
.
21.
Lu
X
,
Sicard
R
,
Jiang
X
, et al
.
HGAL localization to cell membrane regulates B-cell receptor signaling
.
Blood
.
2015
;
125
(
4
):
649
-
657
.
22.
Romero-Camarero
I
,
Jiang
X
,
Natkunam
Y
, et al
.
Germinal centre protein HGAL promotes lymphoid hyperplasia and amyloidosis via BCR-mediated Syk activation
.
Nat Commun
.
2013
;
4
(
1
):
1338
.
23.
Natkunam
Y
,
Hsi
ED
,
Aoun
P
, et al
.
Expression of the human germinal center-associated lymphoma (HGAL) protein identifies a subset of classic Hodgkin lymphoma of germinal center derivation and improved survival
.
Blood
.
2007
;
109
(
1
):
298
-
305
.
24.
Azambuja
D
,
Lossos
IS
,
Biasoli
I
, et al
.
Human germinal center-associated lymphoma protein expression is associated with improved failure-free survival in Brazilian patients with classical Hodgkin lymphoma
.
Leuk Lymphoma
.
2009
;
50
(
11
):
1830
-
1836
.
25.
Baecklund
E
,
Natkunam
Y
,
Backlin
C
, et al
.
Expression of the human germinal-centre-associated lymphoma protein in diffuse large B-cell lymphomas in patients with rheumatoid arthritis
.
Br J Haematol
.
2008
;
141
(
1
):
69
-
72
.
26.
George
SH
,
Gertsenstein
M
,
Vintersten
K
, et al
.
Developmental and adult phenotyping directly from mutant embryonic stem cells
.
Proc Natl Acad Sci USA
.
2007
;
104
(
11
):
4455
-
4460
.
27.
Mainardi
S
,
Mijimolle
N
,
Francoz
S
,
Vicente-Dueñas
C
,
Sánchez-García
I
,
Barbacid
M
.
Identification of cancer initiating cells in K-Ras driven lung adenocarcinoma
.
Proc Natl Acad Sci USA
.
2014
;
111
(
1
):
255
-
260
.
28.
Hobeika
E
,
Thiemann
S
,
Storch
B
, et al
.
Testing gene function early in the B cell lineage in mb1-cre mice
.
Proc Natl Acad Sci USA
.
2006
;
103
(
37
):
13789
-
13794
.
29.
Crouch
EE
,
Li
Z
,
Takizawa
M
, et al
.
Regulation of AID expression in the immune response
.
J Exp Med
.
2007
;
204
(
5
):
1145
-
1156
.
30.
Bolstad
BM
,
Irizarry
RA
,
Astrand
M
,
Speed
TP
.
A comparison of normalization methods for high density oligonucleotide array data based on variance and bias
.
Bioinformatics
.
2003
;
19
(
2
):
185
-
193
.
31.
Irizarry
RA
,
Bolstad
BM
,
Collin
F
,
Cope
LM
,
Hobbs
B
,
Speed
TP
.
Summaries of Affymetrix GeneChip probe level data
.
Nucleic Acids Res
.
2003
;
31
(
4
):
e15
.
32.
Irizarry
RA
,
Hobbs
B
,
Collin
F
, et al
.
Exploration, normalization, and summaries of high density oligonucleotide array probe level data
.
Biostatistics
.
2003
;
4
(
2
):
249
-
264
.
33.
Tusher
VG
,
Tibshirani
R
,
Chu
G
.
Significance analysis of microarrays applied to the ionizing radiation response
.
Proc Natl Acad Sci USA
.
2001
;
98
(
9
):
5116
-
5121
.
34.
Benjamini
Y
,
Drai
D
,
Elmer
G
,
Kafkafi
N
,
Golani
I
.
Controlling the false discovery rate in behavior genetics research
.
Behav Brain Res
.
2001
;
125
(
1-2
):
279
-
284
.
35.
Team
RDC
. A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria URL http://www.R-project.org/.
2010
.
36.
Gentleman
RC
,
Carey
VJ
,
Bates
DM
, et al
.
Bioconductor: open software development for computational biology and bioinformatics
.
Genome Biol
.
2004
;
5
(
10
):
R80
.
37.
Edgar
R
,
Domrachev
M
,
Lash
AE
.
Gene Expression Omnibus: NCBI gene expression and hybridization array data repository
.
Nucleic Acids Res
.
2002
;
30
(
1
):
207
-
210
.
38.
Mootha
VK
,
Lindgren
CM
,
Eriksson
KF
, et al
.
PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes
.
Nat Genet
.
2003
;
34
(
3
):
267
-
273
.
39.
Subramanian
A
,
Tamayo
P
,
Mootha
VK
, et al
.
Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles
.
Proc Natl Acad Sci USA
.
2005
;
102
(
43
):
15545
-
15550
.
40.
Liberzon
A
,
Birger
C
,
Thorvaldsdóttir
H
,
Ghandi
M
,
Mesirov
JP
,
Tamayo
P
.
The Molecular Signatures Database (MSigDB) hallmark gene set collection
.
Cell Syst
.
2015
;
1
(
6
):
417
-
425
.
41.
Shipp
MA
,
Ross
KN
,
Tamayo
P
, et al
.
Diffuse large B-cell lymphoma outcome prediction by gene-expression profiling and supervised machine learning
.
Nat Med
.
2002
;
8
(
1
):
68
-
74
.
42.
Green
MR
,
Monti
S
,
Dalla-Favera
R
, et al
.
Signatures of murine B-cell development implicate Yy1 as a regulator of the germinal center-specific program
.
Proc Natl Acad Sci USA
.
2011
;
108
(
7
):
2873
-
2878
.
43.
Sander
S
,
Calado
DP
,
Srinivasan
L
, et al
.
Synergy between PI3K signaling and MYC in Burkitt lymphomagenesis
.
Cancer Cell
.
2012
;
22
(
2
):
167
-
179
.
44.
S eita
J
,
Sahoo
D
,
Rossi
DJ
, et al
.
Gene Expression Commons: an open platform for absolute gene expression profiling
.
PLoS One
.
2012
;
7
(
7
):
e40321
.
45.
Natkunam
Y
,
Lossos
IS
,
Taidi
B
, et al
.
Expression of the human germinal center-associated lymphoma (HGAL) protein, a new marker of germinal center B-cell derivation
.
Blood
.
2005
;
105
(
10
):
3979
-
3986
.
46.
Niu
H
,
Ye
BH
,
Dalla-Favera
R
.
Antigen receptor signaling induces MAP kinase-mediated phosphorylation and degradation of the BCL-6 transcription factor
.
Genes Dev
.
1;12
(
13
):
1953
-
1961
.
47.
Hong
JX
,
Wilson
GL
,
Fox
CH
,
Kehrl
JH
.
Isolation and characterization of a novel B cell activation gene
.
J Immunol
.
1993
;
150
(
9
):
3895
-
3904
.
48.
Bowman
EP
,
Campbell
JJ
,
Druey
KM
,
Scheschonka
A
,
Kehrl
JH
,
Butcher
EC
.
Regulation of chemotactic and proadhesive responses to chemoattractant receptors by RGS (regulator of G-protein signaling) family members
.
J Biol Chem
.
1998
;
273
(
43
):
28040
-
28048
.
49.
Han
JI
,
Huang
NN
,
Kim
DU
,
Kehrl
JH
.
RGS1 and RGS13 mRNA silencing in a human B lymphoma line enhances responsiveness to chemoattractants and impairs desensitization
.
J Leukoc Biol
.
2006
;
79
(
6
):
1357
-
1368
.
50.
Carreras
J
,
Kikuti
YY
,
Beà
S
, et al
.
Clinicopathological characteristics and genomic profile of primary sinonasal tract diffuse large B cell lymphoma (DLBCL) reveals gain at 1q31 and RGS1 encoding protein; high RGS1 immunohistochemical expression associates with poor overall survival in DLBCL not otherwise specified (NOS)
.
Histopathology
.
2017
;
70
(
4
):
595
-
621
.
51.
Chua
CEL
,
Tang
BL
.
Rab 10-a traffic controller in multiple cellular pathways and locations
.
J Cell Physiol
.
2018
;
233
(
9
):
6483
-
6494
.
52.
Gao
Y
,
Sun
B
,
Hu
J
, et al
.
Identification of gene modules associated with survival of diffuse large B-cell lymphoma treated with CHOP-based chemotherapy
.
Pharmacogenomics J
.
2020
;
20
(
5
):
705
-
716
.
53.
Basso
K
,
Klein
U
,
Niu
H
, et al
.
Tracking CD40 signaling during germinal center development
.
Blood
.
2004
;
104
(
13
):
4088
-
4096
.
54.
Guo
F
,
Luo
Y
,
Jiang
X
, et al
.
Recent BCR stimulation induces a negative autoregulatory loop via FBXO10 mediated degradation of HGAL
.
Leukemia
.
2020
;
34
(
2
):
553
-
566
.
55.
Shaffer
AL
,
Lin
KI
,
Kuo
TC
, et al
.
Blimp-1 orchestrates plasma cell differentiation by extinguishing the mature B cell gene expression program
.
Immunity
.
2002
;
17
(
1
):
51
-
62
.
56.
Dagan
LN
,
Jiang
X
,
Bhatt
S
,
Cubedo
E
,
Rajewsky
K
,
Lossos
IS
.
miR-155 regulates HGAL expression and increases lymphoma cell motility
.
Blood
.
2012
;
119
(
2
):
513
-
520
.
57.
Basso
K
,
Dalla-Favera
R
.
Roles of BCL6 in normal and transformed germinal center B cells
.
Immunol Rev
.
2012
;
247
(
1
):
172
-
183
.
58.
Cattoretti
G
,
Pasqualucci
L
,
Ballon
G
, et al
.
Deregulated BCL6 expression recapitulates the pathogenesis of human diffuse large B cell lymphomas in mice
.
Cancer Cell
.
2005
;
7
(
5
):
445
-
455
.
59.
Mandelbaum
J
,
Bhagat
G
,
Tang
H
, et al
.
BLIMP1 is a tumor suppressor gene frequently disrupted in activated B cell-like diffuse large B cell lymphoma
.
Cancer Cell
.
2010
;
18
(
6
):
568
-
579
.
60.
Pasqualucci
L
,
Compagno
M
,
Houldsworth
J
, et al
.
Inactivation of the PRDM1/BLIMP1 gene in diffuse large B cell lymphoma
.
J Exp Med
.
2006
;
203
(
2
):
311
-
317
.
61.
Reddy
A
,
Zhang
J
,
Davis
NS
, et al
.
Genetic and functional drivers of diffuse large B cell lymphoma
.
Cell
.
2017
;
171
(
2
):
481
-
494 e415
.
You do not currently have access to this content.

Sign in via your Institution

Sign In