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