Post on 17-Jan-2016
Combination of Vaccine-derived Measles and Mumps Virus Enhanced anti-tumor Effects: Special Focus on Acute Leukemia and Prostate Cancer
Sen. Colonel A/Pr. Dr. Nguyen Linh Toan1$, Ho Anh Son1#, LiFeng Zhang3#, Bui Khac Cuong1, Hoang Van Luong2, Naoki Yamamoto3$, et al.
1Vietnam Military Medical University, Vietnam.2Department of Microbiology, NUS, Singapore.
Nguyen Linh Toan - VMMU - Vietnam
2015 Asia Pacific Military Health Exchange
Da Nang - 2015
BACKGROUND
(Torre LA et al. CA Cancer J Clin. 2015).
14.1 million new cancer cases & 8.2 million cancer deaths, 2012
Cancer is major leading cause of death in worldwide.
(Torre LA et al. CA Cancer J Clin. 2015).
Lung and breast cancer are the most frequently and the leading causes
of cancer death in men and women worldwide .
However, prostate cancer is the second most frequently diagnosed
cancer in men worldwide, with 1.1mil. new cases to occurred in 2012
Despite the considerable progress accomplished in recently years, most advanced cancers remain incurable prompting the need for novel, less toxic and more targeted therapy.
Freidman et al., Pediatric Research (2012)
CSC: cancer stem cells
OLV is a novel treatment approach that exploits the ability of viruses to selectively infected and destroy cancer cells. OLV replication, cell killing, virus release and spread within cancer cells but not normal cells
Kirn D et al. (2001). Nat Med Liu T-C et al. (2007) Nat Clin Pract Oncol 5
Oncolytic virotherapy (OLV)
CD46Nectin 4
CD46Nectin 4
CD150: B,T, M, DC
Summay: phase I-III clinical trials of OLV
(Buijs PR et al. 2015; Lapp et al. 2014; Msaouel et al. 2013)
It is highly anticipated that the oncolytic activity of It is highly anticipated that the oncolytic activity of
combined OLVs should be improved quantitatively as combined OLVs should be improved quantitatively as
well as qualitatively. This situation is rather well as qualitatively. This situation is rather
reminiscent of drug therapy in the clinical settingsreminiscent of drug therapy in the clinical settings. .
1) Improved targeting 1) Improved targeting
2) Synergy and complementation of 2) Synergy and complementation of multiple virusesmultiple viruses
3) Produces enhanced anti-tumor effects3) Produces enhanced anti-tumor effects
Our strategy is to exploit Viral Our strategy is to exploit Viral SynergySynergy
Nguyen Linh Toan - VMMU - Vietnam 8
HK1(Nasopharyngeal)
HCT116(Colorectal)
H358(Lung)
EC109(Esohageal)
PC-3(prostate)
MCF-7(Breast)
HepG2(Liver)
Mock
MMR-Infected
Mock
MMR-Infected
Cytopathic effects (CPE) were observed in a wide range of human solid malignancy cell lines infected with the MMR viruses
MMR:MeaslesMumpsRubella
9
Cytopathic effects (CPE) was observed in a wide range of hematological malignancy cell lines infected with the MMR viruses
KI-4
Raji
Mock
MMR-Infected
U937
HuT 102
Lymphoma cell lines
Mock
MMR-Infected
Leukemia cell lines
HL-60
Jurkat
10(Zhang….. Toan. ..et al, Cancer Letter, 2014)
Post infection (P.I.)
MeV MuV MM
day 0 day 3 day 60
10
2020
70
120
day 0 day 3 day 60
10
2020
70
120Huc-Fm
PC-3
MCF-7
KATO III
HEPG2
HUH4
H358
H1299day 0 day 3 day 60
10
2020
70
120
HELA
SIHA
HK-1
HONE-1
EC109
KYSE70
HT29
HCT116day 0 day 3 day 6
0
10
2020
70
120
day 0 day 3 day 60
10
2020
70
120
day 0 day 3 day 60
10
2020
70
120
The MM targeted a wider range of solid cancer cell lines and displayed greater oncolytic effect on several cell lines compared with single virus
PC3:Prostate C., MCF-7: Breast C., KATO III: Gastric C., HepG2, HuH4: Liver C., H358, H1299: Lung C.,Hela and SiHa: cervix C. HK-1:NPC, HONE-2 EC109: Esohageal, KYSE70, HT29 and HCT166: Colorectal C.,
11
The MM targeted a wider range of hematological cancer cell lines and displayed greater oncolytic effect on several cell lines compared with single virus
day 0 day 3 day 60
5
10
THP-1
U937
HL-60
Jurkat
KI-1
KI-2
KI-4
HUT-102
Raji
Namalwa
MeV
10
80
150
day 0 day 3 day 60
5
10
THP-1
U937
HL-60
Jurkat
KI-1
KI-2
KI-4
HUT-102
Raji
Namalwa
MuV
10
80
150
day 0 day 3 day 60
5
10
THP-1
U937
HL-60
Jurkat
KI-1
KI-2
KI-4
HUT-102
Raji
Namalwa
MM
10
80
150
MeV MuV
MM
(Zhang….. Toan. ..et al, Cancer Letter, 2014)
Oncolytic effects between MeV, MuV and MM on human hematological malignant cell lines.
HuT 102KI-4
U937 HL-60
MeV MuV MM
Jurkat
THP-1
0
1
210
40
70
100 ****
0
25
50
75
100
*****
0
25
50
75
100
125**
**
0
25
50
75
100 **
**
0
10
20
30***
***
0
3
6
9 ***
***
MeV MuV MM MeV MuV MM
MeV MuV MM MeV MuV MM MeV MuV MM
The oncolytic effect of the MM compared with single virus was significantly enhanced in U937, THP-1, HL-60, Jurkat, KI-4 and HuT102 cell lines.
14(Zhang….. Toan. ..et al, Cancer Letter, 2014)
Nude Mice bearing human solid tumors
Colon cancer (HT
29)
Larynx cancer (Hep2)
Liver cancer (Hep 3B)
Lung cancer (H211)
MusclesMusclesPC tissue with PC tissue with prostate tubesprostate tubes
PC PC invaded venous system
Prostate Cancer (PC-3)
15
Groups Virus concentration
Frequency Inoculation site
Negative control
M199 medium 1 time or
2 time/wk x 3wks
Intra-tumor
Me 106pfu/100ul 1 time or
2 time/wk x 3wks
Intra-tumor
Mu 106pfu/100ul 1 time or
2 time/wk x 3wks
Intra-tumor
Me+Mu 106pfu/100ul + 106pfu/100ul
1 time or
2 time/wk x 3wks
Intra-tumor
Dose of intra-tumoral injections
OLV were inoculated intra-tumor when PC-3 tumors reached 0.5 mm3 volume
16
Multi-dose
MM enhances killing human PC-3 tumor xenograft
Time (day)T
um
or
size
(m
m3)
*
*
*
* P<0.05
Single dose
Tu
mo
r si
ze (
mm
3)
Time (day)
*
* P<0.05
17
MM combination prolonging survival time
Survival time (day)
Per
cen
t S
urv
ival
Per
cen
t S
urv
ival
Multi-doseSingle dose
18
Enhanced killing of U937 cells and prolonged mice survival by MM treatment in nude mice bearing U937 tumors
a. Tumor volume
b. Survival Days post treatment
Days post treatment
0 2 4 6 8 10 12 14 160.0
2.5
5.0
7.5
10.0
12.5Control
MeV
MuV
MM
*
0 10 20 30 400
30
60
90
120Control
MeV
MuV
MM
Days post treatment
19(Zhang….. Toan. ..et al, Cancer Letter, 2014)
MMKp7-6 (1mg/ml)
Mock
Annexin V
PI
Kp7-6 (1mg/ml)
Medium Kp7-6 MM MM + Kp7-60
10
20
30 **MM
6.9%
21.7%
8.6%
27.9%
1.6%
2.3%
1.2%
2.9%
THP-1
U937
Mechanistic studies: Involvement in apoptosis of U937 and Mechanistic studies: Involvement in apoptosis of U937 and THP-1 cells induced by MM infection.THP-1 cells induced by MM infection.
7.9%
31.1%
6.9%
41.1%
1.0%
2.7%
1.4%
2.2%
Medium Kp7-6 MM MM + Kp7-60
15
30
45 **
PI
(Zhang….. Toan. ..et al, Cancer Letter, 2014)
SUMMARY
In vitro MM displayed a more potent oncolytic effect than single virus on human solid and hematological cancer cell lines
In vivo, MM displayed significant tumor suppression and prolonged survival, compared to single virus, in a PC-3 and U937 xenograft tumor models.
Our results demonstrate that MM is a promising candidate for the treatment human cancers.
ACKNOWLEDGMENTS
Centre Biomedical & Pharmaceutical Lab.,
VMMU, Vietnam
Department of Microbiology, NUS, Singapore
NUS-VMMU collaboration project;
Vietnam NAFOSTED project, Viet Nam
VIETNAM MILITARY MEDICAL UNIVERSITYVIETNAM MILITARY MEDICAL UNIVERSITY