Beyond the CENTRIC Trial eyo d t e C C a Integrin Targeting for - - PowerPoint PPT Presentation

▶
beyond the centric trial eyo d t e c c a integrin
SMART_READER_LITE
LIVE PREVIEW

Beyond the CENTRIC Trial eyo d t e C C a Integrin Targeting for - - PowerPoint PPT Presentation

Jun 13, 2023 213 likes •535 views

OncoRay OncoRay National Center for National Center for Radiation Research in Oncology, Dresden Beyond the CENTRIC Trial eyo d t e C C a Integrin Targeting for Tumor Therapy Sensitization Nils Cordes Cell adhesion and Survival

slide-1
SLIDE 1

OncoRay National Center for OncoRay – National Center for Radiation Research in Oncology, Dresden

Beyond the CENTRIC Trial – eyo d t e C C a Integrin Targeting for Tumor Therapy Sensitization

Nils Cordes

slide-2
SLIDE 2

Cell adhesion and Survival

Cress

Paula Cordes, Easter, 2008

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-3
SLIDE 3

Functions of the ECM

Fertilized egg

n

Adulthood

Lifespan

Death

Rozario & DeSimone. Developmental Biology, 341, 1, 2010, 126–140 Fléchon et al. Reproductive Biology and Endocrinology 2003 1:124 Nishida et al., Invest. Ophthalmol. Vis. Sci. 29:1887–1890, 1988 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-4
SLIDE 4

Integrin heterodimers

Hynes, Cell, 2000 Legate et al., Genes Dev, 2009 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-5
SLIDE 5

Focal adhesion - SIGNALING

Extracellular space

ECM

Extracellular space Cell membrane

EGFR Integrin PINCH1 Nck2 ILK

P i

Talin

Cell membrane

PP1

MEK4/7

FAK Grb2

Parvin Paxillin

C t l

Akt

p130Cas

JNK

MEK1/2

Actin

Cytosol

Schaller and Parsons. Curr Opin Cell Biol. 1994 Burridge and Chrzanowska- Wodnicka Annu Rev Cell

ERK1/2

  • Wodnicka. Annu Rev Cell

Dev Biol. 1996 Legate et al., Nat Rev Mol Cell Biol. 2006 Eke and Cordes, Radiother Oncol, 2011 H hl t l BBA

Survival, Proliferation, Differentiation

Hehlgans et al., BBA reviews, 2007 Vehlow and Cordes, BBA reviews, 2013 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-6
SLIDE 6

Overexpression of (various) Integrins

HNSCC Lung Lung Pancreas Prostate Breast Breast Brain ….

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-7
SLIDE 7

Targeting beta1 integrin

Extracellular space

ECM

Extracellular space Cell membrane

EGFR beta1 Integrin PINCH1 Nck2 ILK

P i

Talin

Cell membrane

X

PP1

MEK4/7

FAK Grb2

Parvin Paxillin

C t l

X

Akt

p130Cas

JNK

MEK1/2

Actin

Cytosol

X X X X

ERK1/2

Radio- and Chemosensitization

X

Radio and Chemosensitization Anti-invasion

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-8
SLIDE 8

beta1 integrin targeting - clonogenicity

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013 Eke, Cordes et al., IJROBP, 2012

slide-9
SLIDE 9

3D procedure

3D lrECM

Co = IgG1a AIIB2 = anti-1 integrin IgG mAb

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-10
SLIDE 10

AIIB2 cytotoxicity

Co = IgG1a AIIB2 = anti-1 integrin IgG mAb

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-11
SLIDE 11

1 integrin inhibition and radiosensitivity

* P < 0.05 ** P < 0.01

Co = IgG1a AIIB2 = anti-1 integrin IgG mAb

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-12
SLIDE 12

Radiosensitization by 1 integrin inhibition

* P < 0.05 ** P < 0.01

Cal33 0 1 1 FaDu 0 1 1 HSC4 0 1 1 UTSCC5 0 1 1

* *

0 001 0.01 0.1 0 001 0.01 0.1 0 001 0.01 0.1 0 001 0.01 0.1

ction

* * ** * * ** **

2 4 6 0.001 2 4 6 0.001 2 4 6 0.001 2 4 6 0.001 UTSCC14 1 UTSCC45 1 UTSCC8 1

urviving frac

0.1 1 0 01 0.1 1 0 01 0.1 1

Su

IgG#1 (1 µg/ml) AIIB2 (1 µg/ml) IgG#1 (10 µg/ml) AIIB2 (10 µg/ml)

** * * ** * *

2 4 6 0.001 0.01 2 4 6 0.001 0.01 2 4 6 0.001 0.01

Radiation dose [Gy]

( µg )

** ** *

Radiation dose [Gy]

Eke, Cordes et al., J Clin Invest, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-13
SLIDE 13

1 Integrin inhibition in vivo

UTSCC15 xenograft model g

(day 11; post 2.5 mg/kg mAb plus 20 Gy)

Time (d) to reach:

anti-1 Co-IgG

n = 129

( )

Co-Ab + 20 Gy anti-β1 Ab + 20 Gy ER 26 [VB:19; 26] 56 [VB:28; 108] 5-fold starting volume + 20 Gy 20 Gy ER 2.33

P < 0.025

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-14
SLIDE 14

DSB repair upon 1 integrin blocking

* P < 0,05 ** P < 0,01 Dickreuter et al., in prep. Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-15
SLIDE 15

1 Integrin in radioresistance: The mechanisms

PKA Calpha/beta T197 JNK T183/Y185

Phosphoproteome array analysis

XF354fl2 SAS UTSCC15

in vitro

AIIB2 IgG#2

UTSCC15

in vivo

FAK S843 FAK S722 FAK S910 FAK Y397 PKA Cbeta S338 PKCbeta1/2 T500 p

FAK pFAK Y397 AIIB2 IgG#2

GSK3beta S216 GSK3alpha Y279 GSK3alpha S21 Src Y529 PAK1/2/3 S144/S141/S154 PKCzeta/lambda T410/T403 FAK S843

pJNK2 T183/Y185 JNK2 pJNK1 T183/Y185 JNK1

Raf1 S259 ERK2 T185/Y187 GRK2 S670 GSK3beta S9 GSK3beta S216

0 0.5 1 1.5 2 2.5 3 3.5

-Actin JNK2 Fold change upon 1 integrin inhibition

(normalized to total protein expression)

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-16
SLIDE 16

1 integrin-FAK interaction

In situ Proximity ligation assay

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-17
SLIDE 17

1 Integrin inhibition affects FAK

Gabarra-Niecko et al., Biochem J. 2002 Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-18
SLIDE 18

1 Integrin inhibition affects FAK

* P < 0.05 ** P < 0.01

*

Increase of radioresistance Reduction of AIIB2 susceptibility

Co = IgG1a AIIB2 = anti-1 integrin IgG mAb Gabarra-Niecko et al., Biochem J. 2002 Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-19
SLIDE 19

1 Integrin inhibition affects FAK

* P < 0.05 ** P < 0.01

*

Increase of radioresistance Reduction of AIIB2 susceptibility

Co = IgG1a AIIB2 = anti-1 integrin IgG mAb Gabarra-Niecko et al., Biochem J. 2002 Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-20
SLIDE 20

1 Integrin inhibition affects FAK

* P < 0.05 ** P < 0.01

*

Increase of radioresistance Reduction of AIIB2 susceptibility

Co = IgG1a AIIB2 = anti-1 integrin IgG mAb Gabarra-Niecko et al., Biochem J. 2002 Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-21
SLIDE 21

Cell rounding upon 1 integrin blocking

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-22
SLIDE 22

FAK interacts with Cortactin

UTSCC15

IP: FAK IP: IgG

UTSCC15

FAK CTTN

-Actin CTTN pCTTN Y421

CTTN

Eke et al., J Clin Invest. 2012 Apr 2;122(4):1529-40 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-23
SLIDE 23

Modified Cortactin by 1 integrin inhibition

In situ Proximity ligation assay IP M S h IP – Mass Spec approach Cortactin : Cortical actin binding protein:

  • cytoplasmic expression
  • polymerization and rearrangement of

polymerization and rearrangement of actin cytoskeleton

Eke, Cordes et al., JCI, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-24
SLIDE 24

Proper FAK-Cortactin interaction critical for radiation cell survival

Kinase domain P873A/P876A/P879A FERM FAT GFP Kinase domain P371A/P374A FERM FAT GFP

FAK-Cort 3 FAK-Cort 1

Kinase domain P973A/P976A FERM FAT GFP Kinase domain P642A/P645A/P648A FERM FAT GFP

FAK-Cort 4 FAK-Cort 2

UTSCC15 UTSCC15

1

FAK CTTN

GFP wt FAK-Cort 1 FAK-Cort 2

* *

g fraction

0.6 0.8 1.0 1.2

  • el. CTTN

precipitation

** ** **

n.s.

FAK-Cort 2 FAK-Cort 3 FAK-Cort 4

** *

Surviving

0.0 0.2 0.4

re Co-p

** **

2 4 6 0.1

**

Radiation dose [Gy] S

Eke et al., J Clin Invest. 2012 Apr 2;122(4):1529-40 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-25
SLIDE 25

Proper FAK-Cortactin interaction critical for radiation cell survival

Kinase domain P873A/P876A/P879A FERM FAT GFP Kinase domain P371A/P374A FERM FAT GFP

FAK-Cort 3 FAK-Cort 1

Kinase domain P973A/P976A FERM FAT GFP Kinase domain P642A/P645A/P648A FERM FAT GFP

FAK-Cort 4 FAK-Cort 2

UTSCC15 UTSCC15

1

FAK CTTN

GFP wt FAK-Cort 1 FAK-Cort 2

* *

g fraction

0.6 0.8 1.0 1.2

  • el. CTTN

precipitation

** ** **

n.s.

FAK-Cort 2 FAK-Cort 3 FAK-Cort 4

** *

Surviving

0.0 0.2 0.4

re Co-p

** **

2 4 6 0.1

**

Radiation dose [Gy] S

Eke et al., J Clin Invest. 2012 Apr 2;122(4):1529-40

*p < 0.05; **p < 0.01

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-26
SLIDE 26

Proper FAK-Cortactin interaction critical for radiation cell survival

Kinase domain P873A/P876A/P879A FERM FAT GFP Kinase domain P371A/P374A FERM FAT GFP

FAK-Cort 3 FAK-Cort 1

Kinase domain P973A/P976A FERM FAT GFP Kinase domain P642A/P645A/P648A FERM FAT GFP

FAK-Cort 4 FAK-Cort 2

UTSCC15 UTSCC15

1

FAK CTTN

GFP wt FAK-Cort 1 FAK-Cort 2

* *

g fraction

0.6 0.8 1.0 1.2

  • el. CTTN

precipitation

** ** **

n.s.

FAK-Cort 2 FAK-Cort 3 FAK-Cort 4

** *

Surviving

0.0 0.2 0.4

re Co-p

** **

2 4 6 0.1

**

Radiation dose [Gy] S

Eke et al., J Clin Invest. 2012 Apr 2;122(4):1529-40

*p < 0.05; **p < 0.01

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-27
SLIDE 27

Our model

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-28
SLIDE 28

Additional Data

  • No effects of beta1 integrin inhibition in normal
  • No effects of beta1 integrin inhibition in normal

cells

Eke, Cordes et al., JCI, 2012

  • No irritation of mice parameters such as weight,

behavior

Eke, Cordes et al., JCI, 2012

  • No signs of hypoxia, modified perfusion, vascular

d it d t t i i i density and metastasis in vivo

  • Modulation of radiation induced oral mucositis

Eke, Cordes et al., JCI, 2012

  • Modulation of radiation-induced oral mucositis

(mouse) by selective inhibition of beta1 integrin

Albert, Cordes, Dörr, et al., Radiother Oncol 2012 Radiother Oncol, 2012 Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-29
SLIDE 29

Head & neck cancer, Lung cancer Lung cancer, Pancreatic cancer, Breast cancer, L k i Leukemia, Glioblastoma multiforme Glioblastoma multiforme …

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-30
SLIDE 30

Take home messages

 Cell-ECM interactions essentially impact on tumor cell survival mechanisms and therapy resistance  beta1 integrin mediated radioresistance is conferred  beta1 integrin-mediated radioresistance is conferred by FAK-Cortactin-JNK1 signaling in HNSCC  beta1 integrins seem to be potential targets in various tumor entities – adjuvant targeting to radiochemotherapy might be beneficial

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013

slide-31
SLIDE 31

Acknowledgements

Molecular and Cellular Radiobiology Ellen Dickreuter Iris Eke Claudia Förster MPI Dresden (GER) Anna Shevchenko Institute of Pathology, UKD (GER) Julia Hennig Inga Lange Mandy Poschau Andre Sagerer Christina Schorradt gy ( ) Gustavo Baretton MPI of Biochemistry (GER) Reinhard Fässler Fabio Stanchi Ahmed Soffar Anne Steglich Katja Storch Anne Vehlow Nicole Wachtel UCSF (USA) David Schlaepfer West Virginia University (USA) Nicole Wachtel Suse Wiedemann Elisa Zienert Katja Zscheppang Nils Cordes West Virginia University (USA) Michael D. Schaller NIH-NIDCR (USA) Kenneth M. Yamada OncoRay Research Groups Michael Baumann, Mechthild Krause Turku University Central Hospital (Finland)

  • R. Grenman

Molecular and Cellular Radiobiology | Cordes | NCT Symposium 2013