Key Points

  • Utx deficiency genetically compromises various metabolic and signaling pathways and phenotypically induces hematopoietic aging.

  • UTX maintains hematopoietic stem cell function via both demethylase-dependent and -independent epigenetic programming.

Abstract

Epigenetic regulation is essential for the maintenance of the hematopoietic system, and its deregulation is implicated in hematopoietic disorders. In this study, UTX, a demethylase for lysine 27 on histone H3 (H3K27) and a component of COMPASS-like and SWI/SNF complexes, played an essential role in the hematopoietic system by globally regulating aging-associated genes. Utx-deficient (UtxΔ/Δ) mice exhibited myeloid skewing with dysplasia, extramedullary hematopoiesis, impaired hematopoietic reconstituting ability, and increased susceptibility to leukemia, which are the hallmarks of hematopoietic aging. RNA-sequencing (RNA-seq) analysis revealed that Utx deficiency converted the gene expression profiles of young hematopoietic stem-progenitor cells (HSPCs) to those of aged HSPCs. Utx expression in hematopoietic stem cells declined with age, and UtxΔ/Δ HSPCs exhibited increased expression of an aging-associated marker, accumulation of reactive oxygen species, and impaired repair of DNA double-strand breaks. Pathway and chromatin immunoprecipitation analyses coupled with RNA-seq data indicated that UTX contributed to hematopoietic homeostasis mainly by maintaining the expression of genes downregulated with aging via demethylase-dependent and -independent epigenetic programming. Of note, comparison of pathway changes in UtxΔ/Δ HSPCs, aged muscle stem cells, aged fibroblasts, and aged induced neurons showed substantial overlap, strongly suggesting common aging mechanisms among different tissue stem cells.

REFERENCES

REFERENCES
1.
Simon
JA
,
Kingston
RE
.
Mechanisms of polycomb gene silencing: knowns and unknowns
.
Nat Rev Mol Cell Biol
.
2009
;
10
(
10
):
697
-
708
.
2.
Conway
E
,
Healy
E
,
Bracken
AP
.
PRC2 mediated H3K27 methylations in cellular identity and cancer
.
Curr Opin Cell Biol
.
2015
;
37
:
42
-
48
.
3.
Agger
K
,
Cloos
PA
,
Christensen
J
, et al
.
UTX and JMJD3 are histone H3K27 demethylases involved in HOX gene regulation and development
.
Nature
.
2007
;
449
(
7163
):
731
-
734
.
4.
Hong
S
,
Cho
YW
,
Yu
LR
,
Yu
H
,
Veenstra
TD
,
Ge
K
.
Identification of JmjC domain-containing UTX and JMJD3 as histone H3 lysine 27 demethylases
.
Proc Natl Acad Sci USA
.
2007
;
104
(
47
):
18439
-
18444
.
5.
Ford
DJ
,
Dingwall
AK
.
The cancer COMPASS: navigating the functions of MLL complexes in cancer [published correction appears in Cancer Genet. 2019;233-234:102]
.
Cancer Genet
.
2015
;
208
(
5
):
178
-
191
.
6.
Shilatifard
A
.
The COMPASS family of histone H3K4 methylases: mechanisms of regulation in development and disease pathogenesis
.
Annu Rev Biochem
.
2012
;
81
(
1
):
65
-
95
.
7.
Van der Meulen
J
,
Speleman
F
,
Van Vlierberghe
P
.
The H3K27me3 demethylase UTX in normal development and disease
.
Epigenetics
.
2014
;
9
(
5
):
658
-
668
.
8.
Miller
SA
,
Mohn
SE
,
Weinmann
AS
.
Jmjd3 and UTX play a demethylase-independent role in chromatin remodeling to regulate T-box family member-dependent gene expression
.
Mol Cell
.
2010
;
40
(
4
):
594
-
605
.
9.
Walport
LJ
,
Hopkinson
RJ
,
Vollmar
M
, et al
.
Human UTY(KDM6C) is a male-specific NE-methyl lysyl demethylase
.
J Biol Chem
.
2014
;
289
(
26
):
18302
-
18313
.
10.
Wang
C
,
Lee
JE
,
Cho
YW
, et al
.
UTX regulates mesoderm differentiation of embryonic stem cells independent of H3K27 demethylase activity
.
Proc Natl Acad Sci USA
.
2012
;
109
(
38
):
15324
-
15329
.
11.
Welstead
GG
,
Creyghton
MP
,
Bilodeau
S
, et al
.
X-linked H3K27me3 demethylase Utx is required for embryonic development in a sex-specific manner
.
Proc Natl Acad Sci USA
.
2012
;
109
(
32
):
13004
-
13009
.
12.
Lee
MG
,
Villa
R
,
Trojer
P
, et al
.
Demethylation of H3K27 regulates polycomb recruitment and H2A ubiquitination
.
Science
.
2007
;
318
(
5849
):
447
-
450
.
13.
Seenundun
S
,
Rampalli
S
,
Liu
QC
, et al
.
UTX mediates demethylation of H3K27me3 at muscle-specific genes during myogenesis
.
EMBO J
.
2010
;
29
(
8
):
1401
-
1411
.
14.
Wang
JK
,
Tsai
MC
,
Poulin
G
, et al
.
The histone demethylase UTX enables RB-dependent cell fate control
.
Genes Dev
.
2010
;
24
(
4
):
327
-
332
.
15.
Lee
S
,
Lee
JW
,
Lee
SK
.
UTX, a histone H3-lysine 27 demethylase, acts as a critical switch to activate the cardiac developmental program
.
Dev Cell
.
2012
;
22
(
1
):
25
-
37
.
16.
Thieme
S
,
Gyárfás
T
,
Richter
C
, et al
.
The histone demethylase UTX regulates stem cell migration and hematopoiesis
.
Blood
.
2013
;
121
(
13
):
2462
-
2473
.
17.
Jiang
W
,
Wang
J
,
Zhang
Y
.
Histone H3K27me3 demethylases KDM6A and KDM6B modulate definitive endoderm differentiation from human ESCs by regulating WNT signaling pathway
.
Cell Res
.
2013
;
23
(
1
):
122
-
130
.
18.
Morales Torres
C
,
Laugesen
A
,
Helin
K
.
Utx is required for proper induction of ectoderm and mesoderm during differentiation of embryonic stem cells
.
PLoS One
.
2013
;
8
(
4
):
e60020
.
19.
Denton
D
,
Aung-Htut
MT
,
Lorensuhewa
N
, et al
.
UTX coordinates steroid hormone-mediated autophagy and cell death
.
Nat Commun
.
2013
;
4
(
1
):
2916
.
20.
Shpargel
KB
,
Sengoku
T
,
Yokoyama
S
,
Magnuson
T
.
UTX and UTY demonstrate histone demethylase-independent function in mouse embryonic development
.
PLoS Genet
.
2012
;
8
(
9
):
e1002964
.
21.
Vandamme
J
,
Lettier
G
,
Sidoli
S
,
Di Schiavi
E
,
Nørregaard Jensen
O
,
Salcini
AE
.
The C. elegans H3K27 demethylase UTX-1 is essential for normal development, independent of its enzymatic activity
.
PLoS Genet
.
2012
;
8
(
5
):
e1002647
.
22.
Shpargel
KB
,
Starmer
J
,
Yee
D
,
Pohlers
M
,
Magnuson
T
.
KDM6 demethylase independent loss of histone H3 lysine 27 trimethylation during early embryonic development
.
PLoS Genet
.
2014
;
10
(
8
):
e1004507
.
23.
van Haaften
G
,
Dalgliesh
GL
,
Davies
H
, et al
.
Somatic mutations of the histone H3K27 demethylase gene UTX in human cancer
.
Nat Genet
.
2009
;
41
(
5
):
521
-
523
.
24.
Varela
I
,
Tarpey
P
,
Raine
K
, et al
.
Exome sequencing identifies frequent mutation of the SWI/SNF complex gene PBRM1 in renal carcinoma [published correction appears in Nature. 2012;484(7392):130]
.
Nature
.
2011
;
469
(
7331
):
539
-
542
.
25.
Gui
Y
,
Guo
G
,
Huang
Y
, et al
.
Frequent mutations of chromatin remodeling genes in transitional cell carcinoma of the bladder
.
Nat Genet
.
2011
;
43
(
9
):
875
-
878
.
26.
Grasso
CS
,
Wu
YM
,
Robinson
DR
, et al
.
The mutational landscape of lethal castration-resistant prostate cancer
.
Nature
.
2012
;
487
(
7406
):
239
-
243
.
27.
Jones
DT
,
Jäger
N
,
Kool
M
, et al
.
Dissecting the genomic complexity underlying medulloblastoma
.
Nature
.
2012
;
488
(
7409
):
100
-
105
.
28.
Mar
BG
,
Bullinger
L
,
Basu
E
, et al
.
Sequencing histone-modifying enzymes identifies UTX mutations in acute lymphoblastic leukemia
.
Leukemia
.
2012
;
26
(
8
):
1881
-
1883
.
29.
Robinson
G
,
Parker
M
,
Kranenburg
TA
, et al
.
Novel mutations target distinct subgroups of medulloblastoma
.
Nature
.
2012
;
488
(
7409
):
43
-
48
.
30.
Wang
L
,
Shilatifard
A
.
UTX Mutations in Human Cancer
.
Cancer Cell
.
2019
;
35
(
2
):
168
-
176
.
31.
Ribera
J
,
Zamora
L
,
Morgades
M
, et al;
Spanish PETHEMA Group and the Spanish Society of Hematology
.
Copy number profiling of adult relapsed B-cell precursor acute lymphoblastic leukemia reveals potential leukemia progression mechanisms
.
Genes Chromosomes Cancer
.
2017
;
56
(
11
):
810
-
820
.
32.
Stief
SM
,
Hanneforth
AL
,
Weser
S
, et al
.
Loss of KDM6A confers drug resistance in acute myeloid leukemia
.
Leukemia
.
2020
;
34
(
1
):
50
-
62
.
33.
Zheng
L
,
Xu
L
,
Xu
Q
, et al
.
Utx loss causes myeloid transformation
.
Leukemia
.
2018
;
32
(
6
):
1458
-
1465
.
34.
Gozdecka
M
,
Meduri
E
,
Mazan
M
, et al
.
UTX-mediated enhancer and chromatin remodeling suppresses myeloid leukemogenesis through noncatalytic inverse regulation of ETS and GATA programs
.
Nat Genet
.
2018
;
50
(
6
):
883
-
894
.
35.
Ratajczak
MZ
,
Suszynska
M
.
Emerging Strategies to Enhance Homing and Engraftment of Hematopoietic Stem Cells
.
Stem Cell Rev Rep
.
2016
;
12
(
1
):
121
-
128
.
36.
Ntziachristos
P
,
Tsirigos
A
,
Welstead
GG
, et al
.
Contrasting roles of histone 3 lysine 27 demethylases in acute lymphoblastic leukaemia
.
Nature
.
2014
;
514
(
7523
):
513
-
517
.
37.
Nagamachi
A
,
Matsui
H
,
Asou
H
, et al
.
Haploinsufficiency of SAMD9L, an endosome fusion facilitator, causes myeloid malignancies in mice mimicking human diseases with monosomy 7
.
Cancer Cell
.
2013
;
24
(
3
):
305
-
317
.
38.
Demarest
RM
,
Ratti
F
,
Capobianco
AJ
.
It’s T-ALL about Notch
.
Oncogene
.
2008
;
27
(
38
):
5082
-
5091
.
39.
Vervoort
SJ
,
van Boxtel
R
,
Coffer
PJ
.
The role of SRY-related HMG box transcription factor 4 (SOX4) in tumorigenesis and metastasis: friend or foe?
Oncogene
.
2013
;
32
(
29
):
3397
-
3409
.
40.
Glass
C
,
Wilson
M
,
Gonzalez
R
,
Zhang
Y
,
Perkins
AS
.
The role of EVI1 in myeloid malignancies
.
Blood Cells Mol Dis
.
2014
;
53
(
1-2
):
67
-
76
.
41.
Sandoval
S
,
Kraus
C
,
Cho
EC
, et al
.
Sox4 cooperates with CREB in myeloid transformation
.
Blood
.
2012
;
120
(
1
):
155
-
165
.
42.
Zhang
H
,
Alberich-Jorda
M
,
Amabile
G
, et al
.
Sox4 is a key oncogenic target in C/EBPα mutant acute myeloid leukemia
.
Cancer Cell
.
2013
;
24
(
5
):
575
-
588
.
43.
Irvine
DA
,
Copland
M
.
Targeting hedgehog in hematologic malignancy
.
Blood
.
2012
;
119
(
10
):
2196
-
2204
.
44.
Bigas
A
,
Waskow
C
.
Blood stem cells: from beginning to end
.
Development
.
2016
;
143
(
19
):
3429
-
3433
.
45.
Wahlestedt
M
,
Pronk
CJ
,
Bryder
D
.
Concise review: hematopoietic stem cell aging and the prospects for rejuvenation
.
Stem Cells Transl Med
.
2015
;
4
(
2
):
186
-
194
.
46.
Liang
Y
,
Van Zant
G
,
Szilvassy
SJ
.
Effects of aging on the homing and engraftment of murine hematopoietic stem and progenitor cells
.
Blood
.
2005
;
106
(
4
):
1479
-
1487
.
47.
Sun
D
,
Luo
M
,
Jeong
M
, et al
.
Epigenomic profiling of young and aged HSCs reveals concerted changes during aging that reinforce self-renewal
.
Cell Stem Cell
.
2014
;
14
(
5
):
673
-
688
.
48.
Blank
U
,
Karlsson
S
.
TGF-β signaling in the control of hematopoietic stem cells
.
Blood
.
2015
;
125
(
23
):
3542
-
3550
.
49.
Richardson
C
,
Yan
S
,
Vestal
CG
.
Oxidative stress, bone marrow failure, and genome instability in hematopoietic stem cells
.
Int J Mol Sci
.
2015
;
16
(
2
):
2366
-
2385
.
50.
Krivtsov
AV
,
Twomey
D
,
Feng
Z
, et al
.
Transformation from committed progenitor to leukaemia stem cell initiated by MLL-AF9
.
Nature
.
2006
;
442
(
7104
):
818
-
822
.
51.
Ivanova
NB
,
Dimos
JT
,
Schaniel
C
,
Hackney
JA
,
Moore
KA
,
Lemischka
IR
.
A stem cell molecular signature
.
Science
.
2002
;
298
(
5593
):
601
-
604
.
52.
Chambers
SM
,
Shaw
CA
,
Gatza
C
,
Fisk
CJ
,
Donehower
LA
,
Goodell
MA
.
Aging hematopoietic stem cells decline in function and exhibit epigenetic dysregulation
.
PLoS Biol
.
2007
;
5
(
8
):
e201
.
53.
Gekas
C
,
Graf
T
.
CD41 expression marks myeloid-biased adult hematopoietic stem cells and increases with age
.
Blood
.
2013
;
121
(
22
):
4463
-
4472
.
54.
Li
T
,
Zhou
ZW
,
Ju
Z
,
Wang
ZQ
.
DNA Damage Response in Hematopoietic Stem Cell Ageing
.
Genomics Proteomics Bioinformatics
.
2016
;
14
(
3
):
147
-
154
.
55.
Flach
J
,
Bakker
ST
,
Mohrin
M
, et al
.
Replication stress is a potent driver of functional decline in ageing haematopoietic stem cells
.
Nature
.
2014
;
512
(
7513
):
198
-
202
.
56.
Zhang
C
,
Hong
Z
,
Ma
W
, et al
.
Drosophila UTX coordinates with p53 to regulate ku80 expression in response to DNA damage
.
PLoS One
.
2013
;
8
(
11
):
e78652
.
57.
Gao
J
,
Zou
J
,
Li
J
, et al
.
Folate deficiency facilitates coordination of KDM6A with p53 in 1 response to DNA damage
.
bioRxiv
. March 28, 2019.
doi.org/10.1101/591768
.
58.
Purton
LE
,
Scadden
DT
.
Limiting factors in murine hematopoietic stem cell assays
.
Cell Stem Cell
.
2007
;
1
(
3
):
263
-
270
.
59.
Yang
YR
,
Bu
FT
,
Yang
Y
, et al
.
LEFTY2 alleviates hepatic stellate cell activation and liver fibrosis by regulating the TGF-β1/Smad3 pathway
.
Mol Immunol
.
2020
;
26
:
31
-
39
.
60.
Chang
H
,
Liu
Y
,
Xue
M
, et al
.
Synergistic action of master transcription factors controls epithelial-to-mesenchymal transition
.
Nucleic Acids Res
.
2016
;
44
(
6
):
2514
-
2527
.
61.
Oki
S
,
Ohta
T
,
Shioi
G
, et al
.
ChIP-Atlas: a data-mining suite powered by full integration of public ChIP-seq data
.
EMBO Rep
.
2018
;
19
(
12
):
e46255
.
62.
Placek
K
,
Hu
G
,
Cui
K
, et al
.
MLL4 prepares the enhancer landscape for Foxp3 induction via chromatin looping
.
Nat Immunol
.
2017
;
18
(
9
):
1035
-
1045
.
63.
Sun
Y
,
Zhou
B
,
Mao
F
, et al
.
HOXA9 Reprograms the Enhancer Landscape to Promote Leukemogenesis
.
Cancer Cell
.
2018
;
34
(
4
):
643
-
658.e5
.
64.
De
S
,
Wurster
AL
,
Precht
P
,
Wood
WH
III
,
Becker
KG
,
Pazin
MJ
.
Dynamic BRG1 recruitment during T helper differentiation and activation reveals distal regulatory elements
.
Mol Cell Biol
.
2011
;
31
(
7
):
1512
-
1527
.
65.
Shi
J
,
Whyte
WA
,
Zepeda-Mendoza
CJ
, et al
.
Role of SWI/SNF in acute leukemia maintenance and enhancer-mediated Myc regulation
.
Genes Dev
.
2013
;
27
(
24
):
2648
-
2662
.
66.
Bossen
C
,
Murre
CS
,
Chang
AN
,
Mansson
R
,
Rodewald
HR
,
Murre
C
.
The chromatin remodeler Brg1 activates enhancer repertoires to establish B cell identity and modulate cell growth
.
Nat Immunol
.
2015
;
16
(
7
):
775
-
784
.
67.
Hohmann
AF
,
Martin
LJ
,
Minder
JL
, et al
.
Sensitivity and engineered resistance of myeloid leukemia cells to BRD9 inhibition
.
Nat Chem Biol
.
2016
;
12
(
9
):
672
-
679
.
68.
Li
G
,
So
AY-L
,
Sookram
R
, et al
.
Epigenetic silencing of miR-125b is required for normal B-cell development
.
Blood
.
2018
;
131
(
17
):
1920
-
1930
.
69.
Beerman
I
,
Rossi
DJ
.
Epigenetic Control of Stem Cell Potential during Homeostasis, Aging, and Disease
.
Cell Stem Cell
.
2015
;
16
(
6
):
613
-
625
.
70.
Goodell
MA
,
Rando
TA
.
Stem cells and healthy aging
.
Science
.
2015
;
350
(
6265
):
1199
-
1204
.
71.
Geiger
H
,
de Haan
G
,
Florian
MC
.
The ageing haematopoietic stem cell compartment
.
Nat Rev Immunol
.
2013
;
13
(
5
):
376
-
389
.
72.
Kramer
A
,
Challen
GA
.
The epigenetic basis of hematopoietic stem cell aging
.
Semin Hematol
.
2017
;
54
(
1
):
19
-
24
.
73.
Wątroba
M
,
Szukiewicz
D
.
The role of sirtuins in aging and age-related diseases
.
Adv Med Sci
.
2016
;
61
(
1
):
52
-
62
.
74.
Rimmelé
P
,
Bigarella
CL
,
Liang
R
, et al
.
Aging-like phenotype and defective lineage specification in SIRT1-deleted hematopoietic stem and progenitor cells
.
Stem Cell Reports
.
2014
;
3
(
1
):
44
-
59
.
75.
Brown
K
,
Xie
S
,
Qiu
X
, et al
.
SIRT3 reverses aging-associated degeneration
.
Cell Rep
.
2013
;
3
(
2
):
319
-
327
.
76.
Wang
H
,
Diao
D
,
Shi
Z
, et al
.
SIRT6 Controls Hematopoietic Stem Cell Homeostasis through Epigenetic Regulation of Wnt Signaling
.
Cell Stem Cell
.
2016
;
18
(
4
):
495
-
507
.
77.
Mohrin
M
,
Shin
J
,
Liu
Y
, et al
.
Stem cell aging. A mitochondrial UPR-mediated metabolic checkpoint regulates hematopoietic stem cell aging
.
Science
.
2015
;
347
(
6228
):
1374
-
1377
.
78.
Sen
P
,
Shah
PP
,
Nativio
R
,
Berger
SL
.
Epigenetic Mechanisms of Longevity and Aging
.
Cell
.
2016
;
166
(
4
):
822
-
839
.
79.
Schulz
WA
,
Lang
A
,
Koch
J
,
Greife
A
.
The histone demethylase UTX/KDM6A in cancer: Progress and puzzles
.
Int J Cancer
.
2019
;
145
(
3
):
614
-
620
.
80.
Benyoucef
A
,
Palii
CG
,
Wang
C
, et al
.
UTX inhibition as selective epigenetic therapy against TAL1-driven T-cell acute lymphoblastic leukemia
.
Genes Dev
.
2016
;
30
(
5
):
508
-
521
.
81.
Van der Meulen
J
,
Sanghvi
V
,
Mavrakis
K
, et al
.
The H3K27me3 demethylase UTX is a gender-specific tumor suppressor in T-cell acute lymphoblastic leukemia
.
Blood
.
2015
;
125
(
1
):
13
-
21
.
82.
Mochizuki-Kashio
M
,
Aoyama
K
,
Sashida
G
, et al
.
Ezh2 loss in hematopoietic stem cells predisposes mice to develop heterogeneous malignancies in an Ezh1-dependent manner
.
Blood
.
2015
;
126
(
10
):
1172
-
1183
.
83.
Sashida
G
,
Wang
C
,
Tomioka
T
, et al
.
The loss of Ezh2 drives the pathogenesis of myelofibrosis and sensitizes tumor-initiating cells to bromodomain inhibition
.
J Exp Med
.
2016
;
213
(
8
):
1459
-
1477
.
84.
Komai
T
,
Inoue
M
,
Okamura
T
, et al
.
Transforming Growth Factor-β and Interleukin-10 Synergistically Regulate Humoral Immunity via Modulating Metabolic Signals
.
Front Immunol
.
2018
;
9
:
1364
.
85.
Zaiatz-Bittencourt
V
,
Finlay
DK
,
Gardiner
CM
.
Canonical TGF-β Signaling Pathway Represses Human NK Cell Metabolism
.
J Immunol
.
2018
;
200
(
12
):
3934
-
3941
.
86.
Liu
L
,
Cheung
TH
,
Charville
GW
, et al
.
Chromatin modifications as determinants of muscle stem cell quiescence and chronological aging
.
Cell Rep
.
2013
;
4
(
1
):
189
-
204
.
87.
Faralli
H
,
Wang
C
,
Nakka
K
, et al
.
UTX demethylase activity is required for satellite cell-mediated muscle regeneration
.
J Clin Invest
.
2016
;
126
(
4
):
1555
-
1565
.
88.
Schwörer
S
,
Becker
F
,
Feller
C
, et al
.
Epigenetic stress responses induce muscle stem-cell ageing by Hoxa9 developmental signals
.
Nature
.
2016
;
540
(
7633
):
428
-
432
.
89.
Mertens
J
,
Paquola
ACM
,
Ku
M
, et al
.
Directly Reprogrammed Human Neurons Retain Aging-Associated Transcriptomic Signatures and Reveal Age-Related Nucleocytoplasmic Defects
.
Cell Stem Cell
.
2015
;
17
(
6
):
705
-
718
.
90.
Rinnerthaler
M
,
Streubel
MK
,
Bischof
J
,
Richter
K
.
Skin aging, gene expression and calcium
.
Exp Gerontol
.
2015
;
68
:
59
-
65
.
91.
Mattson
MP
,
Arumugam
TV
.
Hallmarks of Brain Aging: Adaptive and Pathological Modification by Metabolic States
.
Cell Metab
.
2018
;
27
(
6
):
1176
-
1199
.
92.
Andricovich
J
,
Perkail
S
,
Kai
Y
,
Casasanta
N
,
Peng
W
,
Tzatsos
A
.
Loss of KDM6A Activates Super-Enhancers to Induce Gender-Specific Squamous-like Pancreatic Cancer and Confers Sensitivity to BET Inhibitors
.
Cancer Cell
.
2018
;
33
(
3
):
512
-
526.e8
.
93.
Kaneko
S
,
Li
X
.
X chromosome protects against bladder cancer in females via a KDM6A-dependent epigenetic mechanism
.
Sci Adv
.
2018
;
4
(
6
):
eaar5598
.
You do not currently have access to this content.

Sign in via your Institution

Sign In