slide1
Download
Skip this Video
Download Presentation
in vivo animal model studies in biological science

Loading in 2 Seconds...

play fullscreen
1 / 50

in vivo animal model studies in biological science - PowerPoint PPT Presentation


  • 101 Views
  • Uploaded on

in vivo animal model studies in biological science. Cancer 2. Neuroscience. Cancer research. 2. Neuroscience. Lung Cancer. Cure rate for all patients: 15%. Male. Female. EGFR Expression in NSCLC. Tumours showing high EGFR expression.

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' in vivo animal model studies in biological science' - maja


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
slide1

in vivo animal model studies in biological science

  • Cancer
  • 2. Neuroscience
  • Cancer research

2. Neuroscience

slide2

Lung Cancer

  • Cure rate for all patients: 15%

Male

Female

slide3

EGFR Expression in NSCLC

Tumours showinghigh EGFR expression

High expression associated with

Poor outcome

NSCLC 60-80%

slide4

EGFR signaling pathways

HER

HER

g

PLC

PI3K

PKC

PDK-1

Ras

Sos

AKT

PTEN

Grb2

Raf

p70

S6K

Shc

Bad/Bcl2

JAK

Mek

GSK3

STAT

Erk

Myc

p27

KIP1

E2F

Jun/Fos

Sp1

FKHR-L1

PEA3

Elk1

CyclinD1

Citri A, et al. Exp Cell Res 2003

slide5

Molecular Targeting of EGF Receptor

Cetuximab

Gefitinib (IRESSA)

Erlotinib (Tarceva)

slide6

Response to EGFR-TKI in NSCLC patients

Initial diagnosis

2 months after gefitinib

9 months later

Yu CJ. (2005) PloS Med

slide7

Primary activating mutations are mainly found in EGFR tyrosine kinase domain exons 18-21

EGF Ligand Binding

TM

Tyrosine Kinase

L858R (~40%)

S768I (2%)

R776C (2%)

G719A/C (5%)

L861Q (4%)

719

768

776

858

861

GxGxxG

18

K

19

20

R

L

21

R

22

23

24

E746-A750

766-768

Deletions (~45%)

Insertions (3%)

slide8

Modeling EGFR-TKI responses in pre-clinical model system

PC9, HCC827

TKI

Response(+)

Response(-)

( Acquired resistance )

EGFR

Mutation(+)

TKI

Response(-)

Primary mutation(s)

NSCLC

TKI

Response(+)

EGFR

Mutation(-)

TKI

Response(-)

slide9

Clinical response to EGFR-TKI in NSCLC patients

Initial diagnosis

2 months after gefitinib

9 months later

Yu CJ. (2005) PloS Med

slide10

Mechanisms of the Acquired Resistance

to EGFR tyrosine kinase inhibitors in NSCLC (2008)

Unknown Mechanisms

T790M secondary mutation

MET amplification

slide11

Modeling EGFR-TKI responses in pre-clinical model system

PC9, HCC827

TKI

Response(+)

Response(-)

( Acquired resistance )

EGFR

Mutation(+)

TKI

Response(-)

Primary mutation(s)

NSCLC

TKI

Response(+)

EGFR

Mutation(-)

TKI

Response(-)

erlotinib treatment in pc9 orthotopic lung cancer model
Erlotinib treatment in PC9 orthotopic lung cancer model

vehicle

treated

erlotinib (50mg/kg/day)

treated

Before administration

1 week

2 weeks

3 weeks

slide15

Multi-cycle resistance test

Inoculate previously obtained in vivo drug resistant cells to

second animals and subject the animals to repeated drug cycle

PC9TR

PC9

slide16

1st generation EGFR-TKI

gefitinib, erlotinb : reversible EGFR blocker

2nd generation EGFR-TKI

e.g. pan-erbB blocker, multi-target EGFR blocker,

irreversible EGFR blocker

(BIBW2992)

slide18

The Effect on Cell Viability of si-EGFR in PC-BR clones

scram

si-EGFR

si-EGFR

si-EGFR

si-EGFR

scram

scram

scram

EGFR

Actinin

PC9

#1

#6

#10

EGFR dependent

EGFR independent

slide19

Sequencing in PC9 & BR1,6,10

BR#1

PC9

I

T(790)

L

Q

I

T(790)

L

Q

BR#6

BR#10

I

T(790)

L

Q

I

T(790)

L

Q

slide20

Protein expression and phosphorylation profile

in PC9-BR clones

#6

#1

#10

PC9

0 0.2 2 20 200 BIBW2992 for24H

0 0.2 2 20 200

0 0.2 2 20 200

0 0.2 2 20 200

p-EGFR

EGFR

p-Her3

p-MET

MET

p-STAT3

STAT3

p70 S6K

p-AKT

AKT

p-ERK

ERK

BIM

Actinin

slide21

#1

#6

#10

PC

BIBW2992 treatment in inoculated mouse in vivo

slide23

Primary activating mutations are mainly found in EGFR tyrosine kinase domain exons 18-21

EGF Ligand Binding

TM

Tyrosine Kinase

L858R (~40%)

S768I (2%)

R776C (2%)

G719A/C (5%)

L861Q (4%)

719

768

776

858

861

GxGxxG

18

K

19

20

R

L

21

R

22

23

24

E746-A750

766-768

Deletions (~45%)

Insertions (3%)

slide24

EGFR Mutations

Gefitinib Responders 8/9

Non-responders 0/7 p= 0.00075

Lynch et al, NEJM 2004

slide28

EGFR mutation is oncogenic

Wong et al. (2006) Cancer Cell

slide29

EGFR expression is required

for the maintenance of tumor

Wong et al. (2006) Cancer Cell

slide31

Lung cancer originated from mutant EGFR

respond to various EGFR inhibitors

Wong et al. (2006) Cancer Cell

slide32

Transgenic mutant EGFR animal model study

1. mutant EGFR is oncogenic

2. continued expression of EGFR is required for the maintenance

of tumor

3. mutant EGFR is a therapeutic target

slide33

Factors controlling tumorigenesis

Tumor suppressor genes

Oncogenes

metastasis

Immune

Stroma

Angiogenesis

WT cells

slide35

Utility of genetically-engineered

mouse models of cancer

early detection

Genetically

engineered

mouse

prevention

Progression

analysis

Tumor development

chemotherapy

slide36

Genetic engineering of mouse genome

: knock-out and knock-in via homologous recombination

slide41

Confirmation of germ line transmission

and generation of knock-out(in) mouse

slide42

Conditional activation of p53

Advantages:

p53

Native promoters

Temporal, spatial

Regulation of gene expression

unbiased

slide44

P53 LSL/LSL is a phenocopy of p53-/-

Cre-recombinase-Oestrogen-Receptor-T2

slide50

p53 reactivation mouse model study with

conditional gene expression mouse

  • p53 inactivation is required for the maintenance of p53 mutant tumors
  • p53 gene delivery or other ways to reactivate p53 in p53 mutant tumor
  • could be a therapeutic option
ad