Key Points

  • A new series of ceramide analogs were synthesized, identifying some that effectively inhibit PEL growth and progression in vitro and in vivo.

  • The mechanisms of new ceramide analogs, including novel sphingolipid-regulated cellular genes required for PEL survival, were identified.

Abstract

Primary effusion lymphoma (PEL) is an aggressive malignancy with poor prognosis even under chemotherapy. Kaposi sarcoma–associated herpesvirus (KSHV), one of the human oncogenic viruses, is the principal causative agent. Currently, there is no specific treatment for PEL; therefore, developing new therapies is of great importance. Sphingolipid metabolism plays an important role in determining the fate of tumor cells. Our previous studies have demonstrated that there is a correlation between sphingolipid metabolism and KSHV+ tumor cell survival. To further develop sphingolipid metabolism-targeted therapy, after screening a series of newly synthesized ceramide analogs, here, we have identified compounds with effective anti-PEL activity. These compounds induce significant PEL apoptosis, cell-cycle arrest, and intracellular ceramide production through regulation of ceramide synthesizing or ceramide metabolizing enzymes and dramatically suppress tumor progression without visible toxicity in vivo. These new compounds also increase viral lytic gene expression in PEL cells. Our comparative transcriptomic analysis revealed their mechanisms of action for inducing PEL cell death and identified a subset of novel cellular genes, including AURKA and CDCA3, controlled by sphingolipid metabolism, and required for PEL survival with functional validation. These data provide the framework for the development of promising sphingolipid-based therapies against this virus-associated malignancy.

REFERENCES

1.
Chang
Y
,
Cesarman
E
,
Pessin
MS
, et al
.
Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi’s sarcoma
.
Science
.
1994
;
266
(
5192
):
1865
-
1869
.
2.
Cesarman
E
,
Chang
Y
,
Moore
PS
,
Said
JW
,
Knowles
DM
.
Kaposi’s sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas
.
N Engl J Med
.
1995
;
332
(
18
):
1186
-
1191
.
3.
Oksenhendler
E
,
Boutboul
D
,
Galicier
L
.
Kaposi sarcoma-associated herpesvirus/human herpesvirus 8-associated lymphoproliferative disorders
.
Blood
.
2019
;
133
(
11
):
1186
-
1190
.
4.
Calabrò
ML
,
Sarid
R
.
Human herpesvirus 8 and lymphoproliferative disorders
.
Mediterr J Hematol Infect Dis
.
2018
;
10
(
1
):
e2018061
.
5.
Yarchoan
R
,
Uldrick
TS
.
HIV-associated cancers and related diseases
.
N Engl J Med
.
2018
;
378
(
22
):
2144
-
2145
.
6.
Chen
Y-B
,
Rahemtullah
A
,
Hochberg
E
.
Primary effusion lymphoma
.
Oncologist
.
2007
;
12
(
5
):
569
-
576
.
7.
Simonelli
C
,
Spina
M
,
Cinelli
R
, et al
.
Clinical features and outcome of primary effusion lymphoma in HIV-infected patients: a single-institution study
.
J Clin Oncol
.
2003
;
21
(
21
):
3948
-
3954
.
8.
Guillet
S
,
Gérard
L
,
Meignin
V
, et al
.
Classic and extracavitary primary effusion lymphoma in 51 HIV-infected patients from a single institution
.
Am J Hematol
.
2016
;
91
(
2
):
233
-
237
.
9.
Gopalakrishnan
R
,
Matta
H
,
Tolani
B
,
Triche
T
Jr.
,
Chaudhary
PM
.
Immunomodulatory drugs target IKZF1-IRF4-MYC axis in primary effusion lymphoma in a cereblon-dependent manner and display synergistic cytotoxicity with BRD4 inhibitors
.
Oncogene
.
2016
;
35
(
14
):
1797
-
1810
.
10.
Patil
A
,
Manzano
M
,
Gottwein
E
.
CK1α and IRF4 are essential and independent effectors of immunomodulatory drugs in primary effusion lymphoma
.
Blood
.
2018
;
132
(
6
):
577
-
586
.
11.
Lurain
K
,
Polizzotto
MN
,
Aleman
K
, et al
.
Viral, immunologic, and clinical features of primary effusion lymphoma
.
Blood
.
2019
;
133
(
16
):
1753
-
1761
.
12.
Ogretmen
B
,
Hannun
YA
.
Biologically active sphingolipids in cancer pathogenesis and treatment
.
Nat Rev Cancer
.
2004
;
4
(
8
):
604
-
616
.
13.
Cuvillier
O
,
Pirianov
G
,
Kleuser
B
, et al
.
Suppression of ceramide-mediated programmed cell death by sphingosine-1-phosphate
.
Nature
.
1996
;
381
(
6585
):
800
-
803
.
14.
Liu
J
,
Beckman
BS
,
Foroozesh
M
.
A review of ceramide analogs as potential anticancer agents
.
Future Med Chem
.
2013
;
5
(
12
):
1405
-
1421
.
15.
Saddoughi
SA
,
Ogretmen
B
.
Diverse functions of ceramide in cancer cell death and proliferation
.
Adv Cancer Res
.
2013
;
117
:
37
-
58
.
16.
Selzner
M
,
Bielawska
A
,
Morse
MA
, et al
.
Induction of apoptotic cell death and prevention of tumor growth by ceramide analogues in metastatic human colon cancer
.
Cancer Res
.
2001
;
61
(
3
):
1233
-
1240
.
17.
Qin
Z
,
Dai
L
,
Trillo-Tinoco
J
, et al
.
Targeting sphingosine kinase induces apoptosis and tumor regression for KSHV-associated primary effusion lymphoma
.
Mol Cancer Ther
.
2014
;
13
(
1
):
154
-
164
.
18.
Dai
L
,
Trillo-Tinoco
J
,
Bai
A
, et al
.
Ceramides promote apoptosis for virus-infected lymphoma cells through induction of ceramide synthases and viral lytic gene expression
.
Oncotarget
.
2015
;
6
(
27
):
24246
-
24260
.
19.
Cao
Y
,
Qiao
J
,
Lin
Z
,
Zabaleta
J
,
Dai
L
,
Qin
Z
.
Up-regulation of tumor suppressor genes by exogenous dhC16-Cer contributes to its anti-cancer activity in primary effusion lymphoma
.
Oncotarget
.
2017
;
8
(
9
):
15220
-
15229
.
20.
Dai
L
,
Plaisance-Bonstaff
K
,
Voelkel-Johnson
C
, et al
.
Sphingosine kinase-2 maintains viral latency and survival for KSHV-infected endothelial cells
.
PLoS One
.
2014
;
9
(
7
):
e102314
.
21.
Liu
J
,
Antoon
JW
,
Ponnapakkam
A
,
Beckman
BS
,
Foroozesh
M
.
Novel anti-viability ceramide analogs: design, synthesis, and structure-activity relationship studies of substituted (S)-2-(benzylideneamino)-3-hydroxy-N-tetradecylpropanamides
.
Bioorg Med Chem
.
2010
;
18
(
14
):
5316
-
5322
.
22.
Foroozesh
M
,
Goyal
N
,
Jackson
T
, et al
.
Optimization of scale-up synthesis of anti-cancer ceramide analog 315
.
J Undergrad Chem Res
.
2017
;
16
(
3
):
89
-
90
.
23.
Ponnapakam
AP
,
Liu
J
,
Bhinge
KN
, et al
.
3-Ketone-4,6-diene ceramide analogs exclusively induce apoptosis in chemo-resistant cancer cells
.
Bioorg Med Chem
.
2014
;
22
(
4
):
1412
-
1420
.
24.
Bielawski
J
,
Szulc
ZM
,
Hannun
YA
,
Bielawska
A
.
Simultaneous quantitative analysis of bioactive sphingolipids by high-performance liquid chromatography-tandem mass spectrometry
.
Methods
.
2006
;
39
(
2
):
82
-
91
.
25.
Qin
Z
,
Dai
L
,
Bratoeva
M
,
Slomiany
MG
,
Toole
BP
,
Parsons
C
.
Cooperative roles for emmprin and LYVE-1 in the regulation of chemoresistance for primary effusion lymphoma
.
Leukemia
.
2011
;
25
(
10
):
1598
-
1609
.
26.
Chaudhary
AK
,
Yadav
N
,
Bhat
TA
, et al
.
A potential role of X-linked inhibitor of apoptosis protein in mitochondrial membrane permeabilization and its implication in cancer therapy
.
Drug Discov Today
.
2016
;
21
(
1
):
38
-
47
.
27.
Hannun
YA
,
Obeid
LM
.
Principles of bioactive lipid signalling: lessons from sphingolipids
.
Nat Rev Mol Cell Biol
.
2008
;
9
(
2
):
139
-
150
.
28.
Stiban
J
,
Tidhar
R
,
Futerman
AH
.
Ceramide synthases: roles in cell physiology and signaling
.
Adv Exp Med Biol
.
2010
;
688
:
60
-
71
.
29.
Hannun
YA
,
Obeid
LM
.
Many ceramides
.
J Biol Chem
.
2011
;
286
(
32
):
27855
-
27862
.
30.
Ichikawa
S
,
Sakiyama
H
,
Suzuki
G
,
Hidari
KIPJ
,
Hirabayashi
Y
.
Expression cloning of a cDNA for human ceramide glucosyltransferase that catalyzes the first glycosylation step of glycosphingolipid synthesis
.
Proc Natl Acad Sci USA
.
1996
;
93
(
10
):
4638
-
4643
.
31.
Song
M
,
Zang
W
,
Zhang
B
,
Cao
J
,
Yang
G
.
GCS overexpression is associated with multidrug resistance of human HCT-8 colon cancer cells
.
J Exp Clin Cancer Res
.
2012
;
31
(
1
):
23
-
28
.
32.
Liu
YY
,
Han
TY
,
Giuliano
AE
,
Cabot
MC
.
Expression of glucosylceramide synthase, converting ceramide to glucosylceramide, confers adriamycin resistance in human breast cancer cells
.
J Biol Chem
.
1999
;
274
(
2
):
1140
-
1146
.
33.
Dai
L
,
Trillo-Tinoco
J
,
Bai
L
, et al
.
Systematic analysis of a xenograft mice model for KSHV+ primary effusion lymphoma (PEL)
.
PLoS One
.
2014
;
9
(
2
):
e90349
.
34.
Antoon
JW
,
Liu
J
,
Ponnapakkam
AP
,
Gestaut
MM
,
Foroozesh
M
,
Beckman
BS
.
Novel D: -erythro N-octanoyl sphingosine analogs as chemo- and endocrine-resistant breast cancer therapeutics
.
Cancer Chemother Pharmacol
.
2010
;
65
(
6
):
1191
-
1195
.
35.
Ponnapakkam
T
,
Saulsberry
T
,
Hill
T
, et al
.
Inhibition of breast tumor growth in mice after treatment with ceramide analog 315
.
Anticancer Drugs
.
2018
;
29
(
9
):
898
-
903
.
36.
Samols
MA
,
Skalsky
RL
,
Maldonado
AM
, et al
.
Identification of cellular genes targeted by KSHV-encoded microRNAs
.
PLoS Pathog
.
2007
;
3
(
5
):
e65
.
37.
Haecker
I
,
Gay
LA
,
Yang
Y
, et al
.
Ago HITS-CLIP expands understanding of Kaposi’s sarcoma-associated herpesvirus miRNA function in primary effusion lymphomas
.
PLoS Pathog
.
2012
;
8
(
8
):
e1002884
.
38.
Lei
X
,
Zhu
Y
,
Jones
T
,
Bai
Z
,
Huang
Y
,
Gao
SJ
.
A Kaposi’s sarcoma-associated herpesvirus microRNA and its variants target the transforming growth factor β pathway to promote cell survival
.
J Virol
.
2012
;
86
(
21
):
11698
-
11711
.
39.
DiMaio
TA
,
Gutierrez
KD
,
Lagunoff
M
.
Kaposi’s sarcoma-associated herpesvirus downregulates transforming growth factor β2 to promote enhanced stability of capillary-like tube formation
.
J Virol
.
2014
;
88
(
24
):
14301
-
14309
.
40.
Park
SI
,
Lin
CP
,
Ren
N
, et al
.
Inhibition of aurora A kinase in combination with chemotherapy induces synthetic lethality and overcomes chemoresistance in Myc-overexpressing lymphoma
.
Target Oncol
.
2019
;
14
(
5
):
563
-
575
.
41.
Dauch
D
,
Rudalska
R
,
Cossa
G
, et al
.
A MYC-aurora kinase A protein complex represents an actionable drug target in p53-altered liver cancer
.
Nat Med
.
2016
;
22
(
7
):
744
-
753
.
42.
Zullo
KM
,
Guo
Y
,
Cooke
L
, et al
.
Aurora A kinase inhibition selectively synergizes with histone deacetylase inhibitor through cytokinesis failure in T-cell lymphoma
.
Clin Cancer Res
.
2015
;
21
(
18
):
4097
-
4109
.
43.
Venkatakrishnan
K
,
Kim
TM
,
Lin
CC
, et al
.
Phase 1 study of the investigational Aurora A kinase inhibitor alisertib (MLN8237) in East Asian cancer patients: pharmacokinetics and recommended phase 2 dose
.
Invest New Drugs
.
2015
;
33
(
4
):
942
-
953
.
44.
Durlacher
CT
,
Li
ZL
,
Chen
XW
,
He
ZX
,
Zhou
SF
.
An update on the pharmacokinetics and pharmacodynamics of alisertib, a selective Aurora kinase A inhibitor
.
Clin Exp Pharmacol Physiol
.
2016
;
43
(
6
):
585
-
601
.
45.
Casey
NP
,
Fujiwara
H
,
Ochi
T
,
Yasukawa
M
.
Novel immunotherapy for adult T-cell leukemia/lymphoma: targeting aurora kinase A
.
OncoImmunology
.
2016
;
5
(
11
):
e1239006
.
46.
Chen
J
,
Zhu
S
,
Jiang
N
,
Shang
Z
,
Quan
C
,
Niu
Y
.
HoxB3 promotes prostate cancer cell progression by transactivating CDCA3
.
Cancer Lett
.
2013
;
330
(
2
):
217
-
224
.
47.
Adams
MN
,
Burgess
JT
,
He
Y
, et al
.
Expression of CDCA3 is a prognostic biomarker and potential therapeutic target in non-small cell lung cancer
.
J Thorac Oncol
.
2017
;
12
(
7
):
1071
-
1084
.
48.
Li
S
,
Liu
X
,
Liu
T
, et al
.
Identification of biomarkers correlated with the TNM staging and overall survival of patients with bladder cancer
.
Front Physiol
.
2017
;
8
:
947
-
954
.
49.
Zhang
W
,
Lu
Y
,
Li
X
, et al
.
CDCA3 promotes cell proliferation by activating the NF-κB/cyclin D1 signaling pathway in colorectal cancer
.
Biochem Biophys Res Commun
.
2018
;
500
(
2
):
196
-
203
.
50.
Zhang
Y
,
Yin
W
,
Cao
W
,
Chen
P
,
Bian
L
,
Ni
Q
.
CDCA3 is a potential prognostic marker that promotes cell proliferation in gastric cancer
.
Oncol Rep
.
2019
;
41
(
4
):
2471
-
2481
.
51.
Bigi
R
,
Landis
JT
,
An
H
,
Caro-Vegas
C
,
Raab-Traub
N
,
Dittmer
DP
.
Epstein-Barr virus enhances genome maintenance of Kaposi sarcoma-associated herpesvirus
.
Proc Natl Acad Sci USA
.
2018
;
115
(
48
):
E11379
-
E11387
.
52.
McHugh
D
,
Caduff
N
,
Barros
MHM
, et al
.
Persistent KSHV infection increases EBV-associated tumor formation in vivo via enhanced EBV lytic gene expression
.
Cell Host Microbe
.
2017
;
22
(
1
):
61
-
73
.
53.
Li
F
,
Zhang
N
.
Ceramide: therapeutic potential in combination therapy for cancer treatment
.
Curr Drug Metab
.
2015
;
17
(
1
):
37
-
51
.
54.
Liu
YY
,
Yu
JY
,
Yin
D
, et al
.
A role for ceramide in driving cancer cell resistance to doxorubicin
.
FASEB J
.
2008
;
22
(
7
):
2541
-
2551
.
55.
Petre
CE
,
Sin
SH
,
Dittmer
DP
.
Functional p53 signaling in Kaposi’s sarcoma-associated herpesvirus lymphomas: implications for therapy
.
J Virol
.
2007
;
81
(
4
):
1912
-
1922
.
56.
Muñoz-Fontela
C
,
Marcos-Villar
L
,
Hernandez
F
, et al
.
Induction of paclitaxel resistance by the Kaposi’s sarcoma-associated herpesvirus latent protein LANA2
.
J Virol
.
2008
;
82
(
3
):
1518
-
1525
.
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