https://orcid.org/0000-0003-2396-776X
Key Points
STAT5 is sufficient for IL-7 signaling in ETP-ALL. STAT5 supports growth, self-renewal, and chemoresistance of LSCs.
Pimozide enhances eradication of LSCs by chemotherapy.
Subjects:
Lymphoid Neoplasia
Abstract
Interleukin-7 (IL-7) supports the growth and chemoresistance of T-cell acute lymphoblastic leukemia (T-ALL), particularly the early T-cell precursor subtype (ETP-ALL), which frequently has activating mutations of IL-7 signaling. Signal transducer and activator of transcription (STAT5) is an attractive therapeutic target because it is almost universally activated in ETP-ALL, even in the absence of mutations of upstream activators such as the IL-7 receptor (IL-7R), Janus kinase, and Fms-like tyrosine kinase 3 (FLT3). To examine the role of activated STAT5 in ETP-ALL, we have used a Lmo2-transgenic (Lmo2Tg) mouse model in which we can monitor chemoresistant preleukemia stem cells (pre-LSCs) and leukemia stem cells (LSCs) that drive T-ALL development and relapse following chemotherapy. Using IL-7R-deficient Lmo2Tg mice, we show that IL-7 signaling was not required for the formation of pre-LSCs but essential for their expansion and clonal evolution into LSCs to generate T-ALL. Activated STAT5B was sufficient for the development of T-ALL in IL-7R-deficient Lmo2Tg mice, indicating that inhibition of STAT5 is required to block the supportive signals provided by IL-7. To further understand the role of activated STAT5 in LSCs of ETP-ALL, we developed a new transgenic mouse that enables T-cell specific and doxycycline-inducible expression of the constitutively activated STAT5B1∗6 mutant. Expression of STAT5B1∗6 in T cells had no effect alone but promoted expansion and chemoresistance of LSCs in Lmo2Tg mice. Pharmacologic inhibition of STAT5 with pimozide-induced differentiation and loss of LSCs, while enhancing response to chemotherapy. Furthermore, pimozide significantly reduced leukemia burden in vivo and overcame chemoresistance of patient-derived ETP-ALL xenografts. Overall, our results demonstrate that STAT5 is an attractive therapeutic target for eradicating LSCs in ETP-ALL.
Subjects:
Lymphoid Neoplasia
References
1.
Marks
DI
, Rowntree
C
. Management of adults with T-cell lymphoblastic leukemia
. Blood
. 2017
;129
(9
):1134
-1142
.2.
Waanders
E
, Gu
Z
, Dobson
SM
, et al. Mutational landscape and patterns of clonal evolution in relapsed pediatric acute lymphoblastic leukemia
. Blood Cancer Discov
. 2020
;1
(1
):96
-111
.3.
Silverman
LB
. Balancing cure and long-term risks in acute lymphoblastic leukemia
. Hematology Am Soc Hematol Educ Program
. 2014
;2014
(1
):190
-197
.4.
Coustan-Smith
E
, Mullighan
CG
, Onciu
M
, et al. Early T-cell precursor leukaemia: a subtype of very high-risk acute lymphoblastic leukaemia
. Lancet Oncol
. 2009
;10
(2
):147
-156
.5.
Liu
Y
, Easton
J
, Shao
Y
, et al. The genomic landscape of pediatric and young adult T-lineage acute lymphoblastic leukemia
. Nat Genet
. 2017
;49
(8
):1211
-1218
.6.
Zhang
J
, Ding
L
, Holmfeldt
L
, et al. The genetic basis of early T-cell precursor acute lymphoblastic leukaemia
. Nature
. 2012
;481
(7380
):157
-163
.7.
Wingelhofer
B
, Neubauer
HA
, Valent
P
, et al. Implications of STAT3 and STAT5 signaling on gene regulation and chromatin remodeling in hematopoietic cancer
. Leukemia
. 2018
;32
(8
):1713
-1726
.8.
Igelmann
S
, Neubauer
HA
, Ferbeyre
G
. STAT3 and STAT5 activation in solid cancers
. Cancers (Basel)
. 2019
;11
(10
):1428
.9.
Kucuk
C
, Jiang
B
, Hu
X
, et al. Activating mutations of STAT5B and STAT3 in lymphomas derived from gammadelta-T or NK cells
. Nat Commun
. 2015
;6
:6025
.10.
Bandapalli
OR
, Schuessele
S
, Kunz
JB
, et al. The activating STAT5B N642H mutation is a common abnormality in pediatric T-cell acute lymphoblastic leukemia and confers a higher risk of relapse
. Haematologica
. 2014
;99
(10
):e188
-192
.11.
Johnson
SE
, Shah
N
, Bajer
AA
, LeBien
TW
. IL-7 activates the phosphatidylinositol 3-kinase/AKT pathway in normal human thymocytes but not normal human B cell precursors
. J Immunol
. 2008
;180
(12
):8109
-8117
.12.
Barata
JT
, Silva
A
, Brandao
JG
, Nadler
LM
, Cardoso
AA
, Boussiotis
VA
. Activation of PI3K is indispensable for interleukin 7-mediated viability, proliferation, glucose use, and growth of T cell acute lymphoblastic leukemia cells
. J Exp Med
. 2004
;200
(5
):659
-669
.13.
Silva
AP
, Almeida
ARM
, Cachucho
A
, et al. Overexpression of wild type IL-7Ralpha promotes T-cell acute lymphoblastic leukemia/lymphoma
. Blood
. 2021
;138
(12
):1040
-1052
.14.
Ribeiro
D
, Melao
A
, van Boxtel
R
, et al. STAT5 is essential for IL-7-mediated viability, growth, and proliferation of T-cell acute lymphoblastic leukemia cells
. Blood Adv
. 2018
;2
(17
):2199
-2213
.15.
Ribeiro
D
, Melao
A
, Barata
JT
. IL-7R-mediated signaling in T-cell acute lymphoblastic leukemia
. Adv Biol Regul
. 2013
;53
(2
):211
-222
.16.
Tremblay
CS
, Brown
FC
, Collett
M
, et al. Loss-of-function mutations of Dynamin 2 promote T-ALL by enhancing IL-7 signalling
. Leukemia
. 2016
;30
(10
):1993
-2001
.17.
Bailey
E
, Li
L
, Duffield
AS
, Ma
HS
, Huso
DL
, Small
D
. FLT3/D835Y mutation knock-in mice display less aggressive disease compared with FLT3/internal tandem duplication (ITD) mice
. Proc Natl Acad Sci U S A
. 2013
;110
(52
):21113
-21118
.18.
Choudhary
C
, Brandts
C
, Schwable
J
, et al. Activation mechanisms of STAT5 by oncogenic Flt3-ITD
. Blood
. 2007
;110
(1
):370
-374
.19.
Kato
Y
, Iwama
A
, Tadokoro
Y
, et al. Selective activation of STAT5 unveils its role in stem cell self-renewal in normal and leukemic hematopoiesis
. J Exp Med
. 2005
;202
(1
):169
-179
.20.
Haetscher
N
, Feuermann
Y
, Wingert
S
, et al. STAT5-regulated microRNA-193b controls haematopoietic stem and progenitor cell expansion by modulating cytokine receptor signalling
. Nat Commun
. 2015
;6
:8928
.21.
Fatrai
S
, Wierenga
ATJ
, Daenen
SMGJ
, Vellenga
E
, Schuringa
JJ
. Identification of HIF2alpha as an important STAT5 target gene in human hematopoietic stem cells
. Blood
. 2011
;117
(12
):3320
-3330
.22.
Kollmann
S
, Grundschober
E
, Maurer
B
, et al. Twins with different personalities: STAT5B-but not STAT5A-has a key role in BCR/ABL-induced leukemia
. Leukemia
. 2019
;33
(7
):1583
-1597
.23.
Harir
N
, Pecquet
C
, Kerenyi
M
, et al. Constitutive activation of Stat5 promotes its cytoplasmic localization and association with PI3-kinase in myeloid leukemias
. Blood
. 2007
;109
(4
):1678
-1686
.24.
Schepers
H
, Wierenga
ATJ
, Vellenga
E
, Schuringa
JJ
. STAT5-mediated self-renewal of normal hematopoietic and leukemic stem cells
. JAKSTAT
. 2012
;1
(1
):13
-22
.25.
Bunting
KD
, Bradley
HL
, Hawley
TS
, Moriggl
R
, Sorrentino
BP
, Ihle
JN
. Reduced lymphomyeloid repopulating activity from adult bone marrow and fetal liver of mice lacking expression of STAT5
. Blood
. 2002
;99
(2
):479
-487
.26.
Wang
Z
, Li
G
, Tse
W
, Bunting
KD
. Conditional deletion of STAT5 in adult mouse hematopoietic stem cells causes loss of quiescence and permits efficient nonablative stem cell replacement
. Blood
. 2009
;113
(20
):4856
-4865
.27.
Maude
SL
, Dolai
S
, Delgado-Martin
C
, et al. Efficacy of JAK/STAT pathway inhibition in murine xenograft models of early T-cell precursor (ETP) acute lymphoblastic leukemia
. Blood
. 2015
;125
(11
):1759
-1767
.28.
Delgado-Martin
C
, Meyer
LK
, Huang
BJ
, et al. JAK/STAT pathway inhibition overcomes IL7-induced glucocorticoid resistance in a subset of human T-cell acute lymphoblastic leukemias
. Leukemia
. 2017
;31
(12
):2568
-2576
.29.
Kontro
M
, Kuusanmaki
H
, Eldfors
S
, et al. Novel activating STAT5B mutations as putative drivers of T-cell acute lymphoblastic leukemia
. Leukemia
. 2014
;28
(8
):1738
-1742
.30.
Abdelrahman
RA
, Begna
KH
, Al-Kali
A
, Hogan
WJ
, Litzow
MR
, Tefferi
A
. Revised assessment of response and long-term discontinuation rates among 111 patients with myelofibrosis treated with momelotinib or ruxolitinib
. Leukemia
. 2015
;29
(2
):498
-500
.31.
Tefferi
A
, Litzow
MR
, Pardanani
A
. Long-term outcome of treatment with ruxolitinib in myelofibrosis
. N Engl J Med
. 2011
;365
(15
):1455
-1457
.32.
Larson
RC
, Fisch
P
, Larson
TA
, et al. T cell tumours of disparate phenotype in mice transgenic for Rbtn-2
. Oncogene
. 1994
;9
(12
):3675
-3681
.33.
McCormack
MP
, Young
LF
, Vasudevan
S
, et al. The Lmo2 oncogene initiates leukemia in mice by inducing thymocyte self-renewal
. Science
. 2010
;327
(5967
):879
-883
.34.
Abdulla
H
, Vo
A
, Shields
BJ
, et al. T-ALL can evolve to oncogene independence
. Leukemia
. 2021
;35
(8
):2205
-2219
.35.
Tremblay
CS
, Chiu
SK
, Saw
J
, et al. Small molecule inhibition of dynamin-dependent endocytosis targets multiple niche signals and impairs leukemia stem cells
. Nat Commun
. 2020
;11
(1
):6211
.36.
Tremblay
CS
, Saw
J
, Chiu
SK
, et al. Restricted cell cycle is essential for clonal evolution and therapeutic resistance of pre-leukemic stem cells
. Nat Commun
. 2018
;9
(1
):3535
.37.
Haenebalcke
L
, Goossens
S
, Naessens
M
, et al. Efficient ROSA26-based conditional and/or inducible transgenesis using RMCE-compatible F1 hybrid mouse embryonic stem cells
. Stem Cell Rev Rep
. 2013
;9
(6
):774
-785
.38.
Onishi
M
, Nosaka
T
, Misawa
K
, et al. Identification and characterization of a constitutively active STAT5 mutant that promotes cell proliferation
. Mol Cell Biol
. 1998
;18
(7
):3871
-3879
.39.
Peschon
JJ
, Morrissey
PJ
, Grabstein
KH
, et al. Early lymphocyte expansion is severely impaired in interleukin 7 receptor-deficient mice
. J Exp Med
. 1994
;180
(5
):1955
-1960
.40.
de Boer
J
, Williams
A
, Skavdis
G
, et al. Transgenic mice with hematopoietic and lymphoid specific expression of Cre
. Eur J Immunol
. 2003
;33
(2
):314
-325
.41.
Srinivas
S
, Watanabe
T
, Lin
CS
, et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus
. BMC Dev Biol
. 2001
;1
:4
.42.
Tremblay
M
, Tremblay
CS
, Herblot
S
, et al. Modeling T-cell acute lymphoblastic leukemia induced by the SCL and LMO1 oncogenes
. Genes Dev
. 2010
;24
(11
):1093
-1105
.43.
Szymanska
B
, Wilczynska-Kalak
U
, Kang
MH
, et al. Pharmacokinetic modeling of an induction regimen for in vivo combined testing of novel drugs against pediatric acute lymphoblastic leukemia xenografts
. PLoS One
. 2012
;7
(3
):e33894
.44.
Nelson
EA
, Walker
SR
, Xiang
M
, et al. The STAT5 inhibitor pimozide displays efficacy in models of acute myelogenous leukemia driven by FLT3 mutations
. Genes Cancer
. 2012
;3
(7-8
):503
-511
.45.
Moriggl
R
, Sexl
V
, Kenner
L
, et al. Stat5 tetramer formation is associated with leukemogenesis
. Cancer Cell
. 2005
;7
(1
):87
-99
.46.
Klammt
J
, Neumann
D
, Gevers
EF
, et al. Dominant-negative STAT5B mutations cause growth hormone insensitivity with short stature and mild immune dysregulation
. Nat Commun
. 2018
;9
(1
):2105
.47.
Brachet-Botineau
M
, Polomski
M
, Neubauer
HA
, et al. Pharmacological inhibition of oncogenic STAT3 and STAT5 signaling in hematopoietic cancers
. Cancers (Basel)
. 2020
;12
(1
):240
.48.
Nelson
EA
, Walker
SR
, Weisberg
E
, et al. The STAT5 inhibitor pimozide decreases survival of chronic myelogenous leukemia cells resistant to kinase inhibitors
. Blood
. 2011
;117
(12
):3421
-3429
.49.
Bar-Natan
M
, Nelson
EA
, Walker
SR
, Kuang
Y
, Distel
RJ
, Frank
DA
. Dual inhibition of Jak2 and STAT5 enhances killing of myeloproliferative neoplasia cells
. Leukemia
. 2012
;26
(6
):1407
-1410
.50.
Schafranek
L
, Nievergall
E
, Powell
JA
, et al. Sustained inhibition of STAT5, but not JAK2, is essential for TKI-induced cell death in chronic myeloid leukemia
. Leukemia
. 2015
;29
(1
):76
-85
.51.
Subramaniam
D
, Angulo
P
, Ponnurangam
S
, et al. Suppressing STAT5 signaling affects osteosarcoma growth and stemness
. Cell Death Dis
. 2020
;11
(2
):149
.52.
Wingelhofer
B
, Maurer
B
, Heyes
EC
, et al. Pharmacologic inhibition of STAT5 in acute myeloid leukemia
. Leukemia
. 2018
;32
(5
):1135
-1146
.53.
Armstrong
F
, Brunet de la Grange
P
, Gerby
B
, et al. NOTCH is a key regulator of human T-cell acute leukemia initiating cell activity
. Blood
. 2009
;113
(8
):1730
-1740
.54.
Deftos
ML
, He
YW
, Ojala
EW
, Bevan
MJ
. Correlating notch signaling with thymocyte maturation
. Immunity
. 1998
;9
(6
):777
-786
.55.
Govaerts
I
, Jacobs
K
, Vandepoel
R
, Cools
J
. JAK/STAT pathway mutations in T-ALL, including the STAT5B N642H mutation, are sensitive to JAK1/JAK3 inhibitors
. Hemasphere
. 2019
;3
(6
):e313
.56.
Gerby
B
, Tremblay
CS
, Tremblay
M
, et al. SCL, LMO1 and Notch1 reprogram thymocytes into self-renewing cells
. PLoS Genet
. 2014
;10
(12
):e1004768
.57.
Rokita
JL
, Rathi
KS
, Cardenas
MF
, et al. Genomic profiling of childhood tumor patient-derived xenograft models to enable rational clinical trial design
. Cell Rep
. 2019
;29
(6
):1675
-1689.e9
.58.
Perry
JM
, Tao
F
, Roy
A
, et al. Overcoming Wnt-beta-catenin dependent anticancer therapy resistance in leukaemia stem cells
. Nat Cell Biol
. 2020
;22
(6
):689
-700
.59.
Riddell
J
, Gazit
R
, Garrison
BS
, et al. Reprogramming committed murine blood cells to induced hematopoietic stem cells with defined factors
. Cell
. 2014
;157
(3
):549
-564
.60.
van der Zwet
JCG
, Buijs-Gladdines
JGCAM
, Cordo
V
, et al. MAPK-ERK is a central pathway in T-cell acute lymphoblastic leukemia that drives steroid resistance
. Leukemia
. 2021
;35
(12
):3394
-3405
.61.
Maurer
B
, Nivarthi
H
, Wingelhofer
B
, et al. High activation of STAT5A drives peripheral T-cell lymphoma and leukemia
. Haematologica
. 2020
;105
(2
):435
-447
.62.
de Araujo
ED
, Erdogan
F
, Neubauer
HA
, et al. Structural and functional consequences of the STAT5B(N642H) driver mutation
. Nat Commun
. 2019
;10
(1
):2517
.63.
Pham
HTT
, Maurer
B
, Prchal-Murphy
M
, et al. STAT5BN642H is a driver mutation for T cell neoplasia
. J Clin Invest
. 2018
;128
(1
):387
-401
.64.
Vanden Bempt
M
, Demeyer
S
, Broux
M
, et al. Cooperative enhancer activation by TLX1 and STAT5 drives development of NUP214-ABL1/TLX1-positive T cell acute lymphoblastic leukemia
. Cancer Cell
. 2018
;34
(2
):271
-285.e7
.65.
de Bock
CE
, Demeyer
S
, Degryse
S
, et al. HOXA9 cooperates with activated JAK/STAT signaling to drive leukemia development
. Cancer Discov
. 2018
;8
(5
):616
-631
.66.
De Smedt
R
, Morscio
J
, Reunes
L
, et al. Targeting cytokine- and therapy-induced PIM1 activation in preclinical models of T-cell acute lymphoblastic leukemia and lymphoma
. Blood
. 2020
;135
(19
):1685
-1695
.67.
Padi
SKR
, Luevano
LA
, An
N
, et al. Targeting the PIM protein kinases for the treatment of a T-cell acute lymphoblastic leukemia subset
. Oncotarget
. 2017
;8
(18
):30199
-30216
.68.
Bhojwani
D
, Pui
CH
. Relapsed childhood acute lymphoblastic leukaemia
. Lancet Oncol
. 2013
;14
(6
):e205
-217
.69.
Li
Y
, Buijs-Gladdines
JGCAM
, Cante-Barrett
K
, et al. IL-7 receptor mutations and steroid resistance in pediatric T cell acute lymphoblastic leukemia: a genome sequencing study
. PLoS Med
. 2016
;13
(12
):e1002200
.70.
Meyer
LK
, Huang
BJ
, Delgado-Martin
C
, et al. Glucocorticoids paradoxically facilitate steroid resistance in T cell acute lymphoblastic leukemias and thymocytes
. J Clin Invest
. 2020
;130
(2
):863
-876
.71.
Britschgi
A
, Andraos
R
, Brinkhaus
H
, et al. JAK2/STAT5 inhibition circumvents resistance to PI3K/mTOR blockade: a rationale for cotargeting these pathways in metastatic breast cancer
. Cancer Cell
. 2012
;22
(6
):796
-811
.72.
Warsch
W
, Kollmann
K
, Eckelhart
E
, et al. High STAT5 levels mediate imatinib resistance and indicate disease progression in chronic myeloid leukemia
. Blood
. 2011
;117
(12
):3409
-3420
.73.
Wang
Z
, Mi
T
, Bradley
HL
, et al. Pimozide and imipramine blue exploit mitochondrial vulnerabilities and reactive oxygen species to cooperatively target high risk acute myeloid leukemia
. Antioxidants (Basel)
. 2021
;10
(6
):956
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