IGF-1R: A key linker between chemoresistance and cancer stem cells in epithelial ovarian cancer cells

IGF-1R: A key linker between chemoresistance and cancer stem cells in epithelial ovarian cancer cells Ram Kumar Singh, Ankit Jinager, Ajit Dhadwe, Abh...
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IGF-1R: A key linker between chemoresistance and cancer stem cells in epithelial ovarian cancer cells Ram Kumar Singh, Ankit Jinager, Ajit Dhadwe, Abhijit De and Pritha Ray*

Advanced Centre for Treatment Research and Education in Cancer, Tata Memorial Centre, Navi Mumbai, India 5th Asia Pacific Summit on Cancer Therapy, 20th – 22nd July 2015

Ovarian Cancer  Ovarian cancer (OC) is the 4th leading cause of gynecological deaths around the world. (Siegel et al., Cancer J Clin, 2012)  As per the Tata Memorial Center, Mumbai’s registry India, it is the third most lethal cancer amongst women. Types  Germ cell OC (3-5%)  Epithelial OC (85-90%)  Sex-cord stromal cell OC (5-7%) Subtypes: Epithelial Ovarian Carcinoma

Chemotherapy  Platinum (cisplatin and carboplatin)  Taxane (paclitaxel)

Though the patients show response to 1 st line therapy, 50-60 % ultimately exhibit recurrence and finally succumb to the disease.

Key players behind drug resistance

There are subset of cells within the heterogeneous tumor designated as cancer stem cells. These CSCs have high DNA repair efficiency and chemo resistance property.

Ovarian cancer and chemoresistance

 Ovarian cancer-stem-like side population cells are tumorigenic and chemoresistant. (Hu et. al., British Journal of cancer, 2010)  In vivo and in vitro studies have showed the role of Insulin Growth Factor -1Receptor (IGF-1R) cross-talk in chemoresistance and have proven that IGF-1R inhibitors can overcome cytotoxic resistance. (Wong et. al., Gynecologic Oncology, 2012).

Neoadjuvent therapy Debulking Surgery Adjuvant Chemotherapy

Tumor Relapse

Tumor Relapse

Chemo resistance

Early detection of Chemo resistance

Targeting Right population

Chemo resistance

Cancer Stem Cells

IGF-1R Axis

Schematic representation of development of cellular resistant models in A2780 cells

Understanding the role of the biological and molecular players during induction of drug resistance CSC characterization • SP assay • Biomarker assay • Spheroid assay • Tumor xenograft assay

Cell line used • A2780/APFT • OAW42 • SKOV3 • Hey

Side population assay Paclitaxel Model

25

Combination Model

p = 0 .0 0 0 2

P e r c e n t S id e P o p u la tio n

Cisplatin Model

20

p = 0 .0 0 5 4

15

p < 0 .0 0 0 1 p = 0 .0 0 0 4

10

p = 0 .0 0 0 2 p < 0 .0 0 0 1

5

0

A

P

F

T C

IS

-E

R C

IS

-L

R P

A

C

-E

R P

A

C

-L C

R

O

M

B

I-

E

C

R

O

M

B

I-

L

R

Self Renewal and Chemoresistance Property B

110

* * * = P < 0 .0 0 1

***

110

***

90

100

80

90

P=0.0005 P=0.0004

70

***

60

Number of spheroids

N u m b e r o f s p h e r o id s

100

***

*** ***

50 40 ***

30 20

P=0.0008

80 70

P=0.0082

60

P=0.0014

50 P=0.0093

40 30 20

10 10

C

P

P

I

L

R

S

S

R L

7 A2

ER

S CI

LR

S CI

I

B

80

M

B

M C

O

O C

M O

0

N

P

P N

R

S

S

R E

I

E

I

B

B

M O C

P

C

A

C

A P

P

P

P

S

S L

C

C C A

A

N

R

L

R

P

P E

R

N

S

S

E

R

P

P

S

S

N P

C

IS

IS

L

R

L

R

S

P

100

*

80

*

*

40

20

R

R

I-

L

-L

B

C

C

O

M

A

C

IS

-L

F

R

T

0

P

SP NSP

60

A

P e r c e n t V ia b ilit y

MP

*

P

P R

N C

S

R

E

IS

IS

C

C

7 2 A

E

N

0

0

8

8

7 2

S

S

P

P

0

A

A

ER

C PA

LR

C PA

ER

LR

B MB OM C CO

Stemness gene expression across the resistant models 0.020 *** ***

0.015

***

0.010

** **

** *

0.005 * *

** * *

ns ns

N A

N O

G

2 S O X

T4 A

0.000

O C

Relative gene expression (2

-ct

)

APFT CIS-ER CIS-LR PAC-ER PAC-LR COMBI-ER COMBI-LR

Pluripotent Genes

ns

Biomarker analysis across the resistant models

Knock down of Oct4 gene using pll3.7 lentilox vector +1 Oct4 target Sequence adapted from Zaheres et al, Stem Cells, 2005

I

Sense

Loop

antisense

Terminator

TAACATGTGTAAGCTGCGGCCCTTCAAGAGAGGGCCGCAGCTTACACATGTTTTTTTTGC ATTGTACACATTCGACGCCGGGAAGTTCTCTCCCGGCGTCGAATGTGTACAAAAAAAACGCCGG

Effect of oct4 KD on its self renewal and drug resistance A Spheroid formation assay

8

60

**

N u m b e r o f s e r ia l p a s s a g e

***

* P

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