Therapy against HER2-positive Breast Cancerarchives.innovationinbreastcancer.com/files/... · · 2016-03-07Therapy against HER2-positive Breast Cancer JoaquínArribas ... PVR. LAMA3

Therapy against HER2-positiveBreast Cancer

Joaquín ArribasVall d’Hebron Institute of Oncology (VHIO)Institució Catalana de Recerca i Studis Avançats (ICREA)

Summary

• Introduction: HER2 and p95HER2

• Antibodies against p95HER2 and response to anti-HER2 therapies

• HER2-induced cellular senescence in breast cancer progression and treatment

• Immune-therapy based on p95HER2

Summary





ERBB receptor family

Yarden, Y and Pines, G (2012) Nature Review Cancer 12, 553-563

Introduction

The EGFR (HER, ErbB) signaling pathway in breast cancer

A17

NRG

Nucleus

HER3

HER2

NRG

GeneExpressionPrograms

Introduction

HER2HER2

HER2HER2HER2

p95HER2

HER2 fragments in human tumors

PM

HER2

Gene expression

S S

p95HER2(648-CTF)

p95HER2(611-CTF)

Arribas et al (2010) Clin Can Res 16, 4226-4235Morancho et al (2013) Oncogene 32, 1452-1459

Introduction

HER3

NRG

p95HER2 and breast cancer progression

Pedersen K et al (2009) Mol Cell Biol 29, 3319-3331

Introduction

Summary





611

NGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTS

624 634 642 653630626

MPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTS

600590580571

TM

N629 100-115 kDa p95HER2(611-CTF)HER2

Specific anti-p95HER2 antibodies (anti-100-115 kDa p95HER2 (611-CTF))

Parra-Palau et al (2010) Cancer Res 70, 8537-8546Sperinde J et al (2010) Clin Cancer Res 16:4226-4235

Analysis of p95HER2 expression in human breast tumor samples overexpressing HER2

Parra-Palau J L et al (2010) Cancer Res 70, 8537-8546

62 107

p95HER2 /CTFs

Lysates

CB11

CB11

32H2

62

CB11

32H2

107A B

147 147

CB11

32H2

HER2-positive breast cancer patient derived xenograftsPDX Subtype

173 HER2

67 HER2

118 Lum B

288 HER2

289 HER2

290 HER2

251 HER2Loading

control

75 -

100 -

150 -

250 -

kDa

PDX173 PDX67 PDX288 PDX289 PDX333 PDX144PDX251 PDX118PDX290

5 μm

PDX173 PDX67 PDX251 PDX118

Her

cept

est

anti-

p95H

ER2

H-Score+1 +2 +310 10 20

+1 +2 +310 10 20

+1 +2 +30 10 20

p95HER2 and response to trastuzumab + chemotherapy

******

***

PDX118

anti-HER2

anti-p95HER2

0

200

400

600

800

0 15 30 45 60

Tum

or v

olum

e (m

m3 )

Time (days)

Control Trastuzumab

***

0

250

500

750

1000

0 5 10 15 20 25 30 35 40 45

Tum

or v

olum

e (m

m3 )

Time (days)

Control

Trastuzumab

PDX118 (p95HER2-negative; lum B)

PDX67 (p95HER2-positive; HER2-enriched) PDX67

anti-HER2

anti-p95HER2

Parra-Palau J Ll et al (2014) J Natl Cancer Inst 106, dju291

Summary





Expression of p95HER2 may result in senescence

Dox + -

+ Dox

- Dox

-220

kDa

-120

-100

100 µm

Ponceau-S

Cel

l Num

ber

(rel

ativ

e to

day

0)

Time (days)

0

4

8

12

16

0 5 10

+Dox

-Dox

***

Angelini PD et al (2013) Cancer Res 73, 450-458

p95HER2-induced senescence secretome

MMP1 IL6 IL11 ANGPTL4


c

320 367

16316

-6

HER2p95HER2

Log2FC (-/+ Dox)

Log2

FC (-

/+ D

ox) p

95H

ER2

–Lo

g2FC

(-/+

Dox

) HER

2

b

-6

-4

-2

0

2

4

6

a

Proliferation Senescence

HER2 p95HER2

- Dox

HER2

p95HER2Transcriptome

Secretome p95HER2

MMP1

TFPI

KRT81

STC1

EPHA2

VEGFC

FSTL3

PODXL

SPTBN1 IL18

CLIP1

B4GALT1

TUBA4A

PLAUR

CDH3

AGR2

PVR

LAMA3

SPOCK1

IL11

PDLIM7

DSC2

LAMB3

EPPK1

HOMER3

ANGPTL4

LUMLAMC2

DST

ITGA2

ZN185

LAMB1

CD44

KIAA1609

MYH9

QSOX1

SERPINB1

IL6

CORO1C

PLOD2

CD55

LDLR

CD59

PRSS22

TSC22D2

CHST11

JAG1

B3GNT3

HS6ST2

MET

IL1RL1

FHL2

LOXL2

OLFML3

DBN1

ULBP2

p95HER2-induced senescence promotes metastasis cell non-autonomously

+ Dox

- Dox

MCF7 Tet-Off/p95HER2MDA-MB-231/Luc (1:1) -/+ Dox

Removal of primary tumor

Nor

mal

ized

pho

ton

flux

(x10

5 )

0.5

2.5

1.0

1.5

2.0

+ Dox - Dox + Dox - Dox

Nor

mal

ized

pho

ton

flux

22.2% 77.8%

*

MCF7 Tet-Off/p95HER2

MDA-MB-231/Luc


Purification of naturally occurring HCC1954 senescent cells

HCC1954CFSE lo HCC1954CFSE hi

40µm

0

100

200

300

400

1 100 10000

FL 1 Log

Cou

nts

0 days 7 days

0

100

200

300

400

1 100 10000

CSFE hi(5.3%)

0

10

20

30**

SA-b-gal

p21

gH2AX

53BP1 40µm

100µm

40µm

0

10

20

30

40SA-b-gal

p21

gH2AX / 53BP1

***

05

101520253035

Posi

tive

cells

(%)

**

Zacarias-Fluck M et al (2015) J Natl Cancer Inst 106, dju291

Secretory phenotype of naturally occurring HCC1954 senescent cells

02468

pg/m

l**IL-6

0

10

20

30

pg/m

l (X

10) **IL-8

0

10

20

30

pg/m

l

**AREG


Naturally occurring senescent cells from a HER2-PDX secrete IL-6

A

HER2 SA-ß-gal

0

20

40

60

80

100Po

sitiv

e ce

lls (%

)IL6 (-)

IL6 (+)

p<0.0001 p<0.0001

1

10001

20001

30001

Fluo

resc

ence

(AU

)

4 105

3 105

2 105

1 105

0

p21IL6

HER2-PDX (culture)B

40µm


IL-6 secreted by senescent cells is required for tumor growth

moIL-6 huIL-6

anti-moIL-6 anti-huIL-6

mIL-6

hIL-6 mIL-6

mIL-6

0,00,51,01,52,02,5

- +

pSTA

T3/S

TAT3 *

anti-huIL-6

anti-moIL-6+ anti-huIL-6anti-moIL-6

pSTAT3

STAT3

kDa-120

-120

0

100

200

300

400

0 10 20 30 40 50 60 70 80 90 100 110

Tum

or V

olum

e (m

m3)

Time (days)

anti-moIL-6

anti-moIL-6 + anti-huIL-6

* * *

*

Zacarias-Fluck et al (2015) J Natl Cancer Inst 107 (5): djv020

Senescent cells may contribute to tumor progression

MMP1

ICAM

IGFBP

AREG

ADAM9

IL8

IL6

Zacarias-Fluck et al (2015) J Natl Cancer Inst 107 (5): djv020

Summary





ERBB2 as a target in breast cancer therapy

Kroemer, G. et al (2015) Nature Medicine 10, 1128-11388

Inhibition of MHC expression

Humanized PDX models: strategies

Breast cancer

PBMC (healthy donors)PBMCs (matched patients)

TILs

CD34+ (cord blood)CD34+ (matched patients)

SubcutaneousOrthotopic

Vicario, R. et al (2015) unpublished

THANK YOU

Documents

Therapy against HER2-positive Breast Cancerarchives.innovationinbreastcancer.com/files/... · · 2016-03-07Therapy against HER2-positive Breast Cancer JoaquínArribas ... PVR. LAMA3