Introduction: Abnormalities in splicing factors, such as mutations or deregulated expression, can lead to aberrant splicing of target genes, potentially contributing to the pathogenesis of acute myeloid leukemia (AML). Despite this, the precise mechanism underlying the abnormal alternative splicing (AS) induced by SRSF1, a splicing factor associated with poor AML prognosis, remains elusive. Methods: Using strict splicing criteria, we globally screened for AS events in NPMc-positive and NPMc-negative AML samples from TCGA. An AS network associated with AML prognosis was then established. Functional assays, including CCK-8, flow cytometry, and Western blot, were conducted on K562 and THP-1 cells overexpressing SRSF1. Cell viability following 72-h Omipalisib treatment was also assessed. To explore the mechanism of SRSF1-induced AS, we created a BCL2L11 miniGene with a site-specific mutation at its branch point. The AS patterns of both wild-type and mutant miniGenes were analyzed following SRSF1 overexpression in HEK-293T, along with the subcellular localization of different spliceosomes. Results: SRSF1 was significantly associated with AML prognosis. Notably, its expression was markedly upregulated in refractory AML patients compared to those with a favorable chemotherapy response. Overexpression of SRSF1 promoted THP-1 cell proliferation, suppressed apoptosis, and reduced sensitivity to Omipalisib. Mechanistically, SRSF1 recognized an aberrant branch point within the BCL2L11 intron, promoting the inclusion of a cryptic exon 3, which in turn led to apoptosis arrest. Conclusion: Overexpression of SRSF1 and the resulting abnormal splicing of BCL2L11 are associated with drug resistance and poor prognosis in AML.

1.
Marando
L
,
Huntly
BJP
.
Molecular landscape of acute myeloid leukemia: prognostic and therapeutic implications
.
Curr Oncol Rep
.
2020
;
22
(
6
):
61
.
2.
Thomas
D
,
Majeti
R
.
Biology and relevance of human acute myeloid leukemia stem cells
.
Blood
.
2017
;
129
(
12
):
1577
85
.
3.
Sung
H
,
Ferlay
J
,
Siegel
RL
,
Laversanne
M
,
Soerjomataram
I
,
Jemal
A
, et al
.
Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries
.
CA Cancer J Clin
.
2021
;
71
(
3
):
209
49
.
4.
Surveillance, epidemiology, and end results Program: cancer stat facts: leukemia – acute myeloid leukemia (AML)
.
Bethesda (MD)
:
National Cancer Institute, DCCPS, Surveillance Research Program
;
2020
. Available online. (Last accessed December 15, 2021).
5.
Grech
G
,
Pollacco
J
,
Portelli
M
,
Sacco
K
,
Baldacchino
S
,
Grixti
J
, et al
.
Expression of different functional isoforms in haematopoiesis
.
Int J Hematol
.
2014
;
99
(
1
):
4
11
.
6.
Larsson
CA
,
Cote
G
,
Quintas-Cardama
A
.
The changing mutational landscape of acute myeloid leukemia and myelodysplastic syndrome
.
Mol Cancer Res
.
2013
;
11
(
8
):
815
27
.
7.
Leggere
JC
,
Saito
Y
,
Darnell
RB
,
Tessier-Lavigne
M
,
Junge
HJ
,
Chen
Z
.
NOVA regulates Dcc alternative splicing during neuronal migration and axon guidance in the spinal cord
.
Elife
.
2016
;
5
.
8.
Inoue
D
,
Chew
GL
,
Liu
B
,
Michel
BC
,
Pangallo
J
,
D’Avino
AR
, et al
.
Spliceosomal disruption of the non-canonical BAF complex in cancer
.
Nature
.
2019
;
574
(
7778
):
432
6
.
9.
Anczukow
O
,
Rosenberg
AZ
,
Akerman
M
,
Das
S
,
Zhan
L
,
Karni
R
, et al
.
The splicing factor SRSF1 regulates apoptosis and proliferation to promote mammary epithelial cell transformation
.
Nat Struct Mol Biol
.
2012
;
19
(
2
):
220
8
.
10.
Pani
T
,
Rajput
K
,
Kar
A
,
Sharma
H
,
Basak
R
,
Medatwal
N
, et al
.
Alternative splicing of ceramide synthase 2 alters levels of specific ceramides and modulates cancer cell proliferation and migration in Luminal B breast cancer subtype
.
Cell Death Dis
.
2021
;
12
:
171
.
11.
Zhang
J
,
Manley
JL
.
Misregulation of pre-mRNA alternative splicing in cancer
.
Cancer Discov
.
2013
;
3
(
11
):
1228
37
.
12.
Smith
PJ
,
Zhang
C
,
Wang
J
,
Chew
SL
,
Zhang
MQ
,
Krainer
AR
.
An increased specificity score matrix for the prediction of SF2/ASF-specific exonic splicing enhancers
.
Hum Mol Genet
.
2006
;
15
(
16
):
2490
508
.
13.
Karni
R
,
de Stanchina
E
,
Lowe
SW
,
Sinha
R
,
Mu
D
,
Krainer
AR
.
The gene encoding the splicing factor SF2/ASF is a proto-oncogene
.
Nat Struct Mol Biol
.
2007
;
14
(
3
):
185
93
.
14.
Gout
S
,
Brambilla
E
,
Boudria
A
,
Drissi
R
,
Lantuejoul
S
,
Gazzeri
S
, et al
.
Abnormal expression of the pre-mRNA splicing regulators SRSF1, SRSF2, SRPK1 and SRPK2 in non small cell lung carcinoma
.
PLoS One
.
2012
;
7
(
10
):
e46539
.
15.
Sinnakannu
JR
,
Lee
KL
,
Cheng
S
,
Li
J
,
Yu
M
,
Tan
SP
, et al
.
SRSF1 mediates cytokine-induced impaired imatinib sensitivity in chronic myeloid leukemia
.
Leukemia
.
2020
;
34
(
7
):
1787
98
.
16.
Nishimura
K
,
Yamazaki
H
,
Zang
W
,
Inoue
D
.
Dysregulated minor intron splicing in cancer
.
Cancer Sci
.
2022
;
113
(
9
):
2934
42
.
17.
de Necochea-Campion
R
,
Shouse
GP
,
Zhou
Q
,
Mirshahidi
S
,
Chen
CS
.
Aberrant splicing and drug resistance in AML
.
J Hematol Oncol
.
2016
;
9
(
1
):
85
.
18.
Zhang
J
,
Gu
Y
,
Chen
B
.
Mechanisms of drug resistance in acute myeloid leukemia
.
Onco Targets Ther
.
2019
;
12
:
1937
45
.
19.
Ku
J
,
Kim
R
,
Kim
D
,
Kim
D
,
Song
S
,
Lee
K
, et al
.
Single-cell analysis of AIMP2 splice variants informs on drug sensitivity and prognosis in hematologic cancer
.
Commun Biol
.
2020
;
3
(
1
):
630
.
20.
Makela
E
,
Pavic
K
,
Varila
T
,
Salmenniemi
U
,
Loyttyniemi
E
,
Nagelli
SG
, et al
.
Discovery of a novel CIP2A variant (NOCIVA) with clinical relevance in predicting TKI resistance in myeloid leukemias
.
Clin Cancer Res
.
2021
;
27
(
10
):
2848
60
.
21.
Sciarrillo
R
,
Wojtuszkiewicz
A
,
Assaraf
YG
,
Jansen
G
,
Kaspers
GJL
,
Giovannetti
E
, et al
.
The role of alternative splicing in cancer: from oncogenesis to drug resistance
.
Drug Resist Updat
.
2020
;
53
:
100728
.
22.
Wang
BD
,
Lee
NH
.
Aberrant RNA splicing in cancer and drug resistance
.
Cancers
.
2018
;
10
(
11
):
458
.
23.
Paronetto
MP
,
Passacantilli
I
,
Sette
C
.
Alternative splicing and cell survival: from tissue homeostasis to disease
.
Cell Death Differ
.
2016
;
23
(
12
):
1919
29
.
24.
Oltean
S
,
Bates
DO
.
Hallmarks of alternative splicing in cancer
.
Oncogene
.
2014
;
33
(
46
):
5311
8
.
25.
Zhang
B
,
Yang
L
,
Wang
X
,
Fu
D
.
Identification of survival-related alternative splicing signatures in acute myeloid leukemia
.
Biosci Rep
.
2021
;
41
(
7
).
26.
Meyers
J
,
Yu
Y
,
Kaye
JA
,
Davis
KL
.
Medicare fee-for-service enrollees with primary acute myeloid leukemia: an analysis of treatment patterns, survival, and healthcare resource utilization and costs
.
Appl Health Econ Health Policy
.
2013
;
11
(
3
):
275
86
.
27.
Xie
ZC
,
Gao
L
,
Chen
G
,
Ma
J
,
Yang
LH
,
He
RQ
, et al
.
Prognostic alternative splicing regulatory network of splicing events in acute myeloid leukemia patients based on SpliceSeq data from 136 cases
.
Neoplasma
.
2020
;
67
(
3
):
623
35
.
28.
Chen
XX
,
Zhu
JH
,
Li
ZP
,
Xiao
HT
,
Zhou
H
.
Comprehensive characterization of the prognosis value of alternative splicing events in acute myeloid leukemia
.
DNA Cell Biol
.
2020
;
39
(
7
):
1243
55
.
29.
Chen
SL
,
Dai
YJ
,
Hu
F
,
Wang
Y
,
Li
H
,
Liang
Y
.
Effects of alternative splicing events on acute myeloid leukemia
.
DNA Cell Biol
.
2020
;
39
(
11
):
2040
51
.
30.
Rivera
OD
,
Mallory
MJ
,
Quesnel-Vallieres
M
,
Chatrikhi
R
,
Schultz
DC
,
Carroll
M
, et al
.
Alternative splicing redefines landscape of commonly mutated genes in acute myeloid leukemia
.
Proc Natl Acad Sci USA
.
2021
;
118
(
15
).
31.
Ng
KP
,
Hillmer
AM
,
Chuah
CT
,
Juan
WC
,
Ko
TK
,
Teo
AS
, et al
.
A common BIM deletion polymorphism mediates intrinsic resistance and inferior responses to tyrosine kinase inhibitors in cancer
.
Nat Med
.
2012
;
18
(
4
):
521
8
.
32.
Liu
J
,
Bhadra
M
,
Sinnakannu
JR
,
Yue
WL
,
Tan
CW
,
Rigo
F
, et al
.
Overcoming imatinib resistance conferred by the BIM deletion polymorphism in chronic myeloid leukemia with splice-switching antisense oligonucleotides
.
Oncotarget
.
2017
;
8
(
44
):
77567
85
.
33.
Ko
TK
,
Chin
HS
,
Chuah
CT
,
Huang
JW
,
Ng
KP
,
Khaw
SL
, et al
.
The BIM deletion polymorphism: a paradigm of a permissive interaction between germline and acquired TKI resistance factors in chronic myeloid leukemia
.
Oncotarget
.
2016
;
7
(
3
):
2721
33
.
34.
O’Connor
L
,
Strasser
A
,
O’Reilly
LA
,
Hausmann
G
,
Adams
JM
,
Cory
S
, et al
.
Bim: a novel member of the Bcl-2 family that promotes apoptosis
.
EMBO J
.
1998
.
35.
U
M
,
Miyashita
T
,
Shikama
Y
,
Tadokoro
K
,
Yamada
M
.
Molecular cloning and characterization of six novel isoforms of human Bim, a member of the proapoptotic Bcl-2 family
.
FEBS Lett
.
2001
;
509
(
1
):
135
41
.
36.
Dlamini
Z
,
Shoba
B
,
Hull
R
.
Splicing machinery genomics events in acute myeloid leukaemia (AML): in search for therapeutic targets, diagnostic and prognostic biomarkers
.
Am J Cancer Res
.
2020
;
10
(
9
):
2690
704
.
37.
Liu
J
,
Huang
B
,
Xiao
Y
,
Xiong
HM
,
Li
J
,
Feng
DQ
, et al
.
Aberrant expression of splicing factors in newly diagnosed acute myeloid leukemia
.
Onkologie
.
2012
;
35
(
6
):
335
40
.
38.
Hahn
CN
,
Venugopal
P
,
Scott
HS
,
Hiwase
DK
.
Splice factor mutations and alternative splicing as drivers of hematopoietic malignancy
.
Immunol Rev
.
2015
;
263
(
1
):
257
78
.
39.
Kuroda
J
,
Puthalakath
H
,
Cragg
MS
,
Kelly
PN
,
Bouillet
P
,
Huang
DC
, et al
.
Bim and Bad mediate imatinib-induced killing of Bcr/Abl+ leukemic cells, and resistance due to their loss is overcome by a BH3 mimetic
.
Proc Natl Acad Sci U S A
.
2006
;
103
(
40
):
14907
12
.
40.
Miao
J
,
Chen
GG
,
Yun
JP
,
Chun
SY
,
Zheng
ZZ
,
Ho
RL
, et al
.
Identification and characterization of BH3 domain protein Bim and its isoforms in human hepatocellular carcinomas
.
Apoptosis
.
2007
;
12
(
9
):
1691
701
.
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