Mecanismos de resistencia a terapia anti-HER2: una visión desde el laboratorio

Amplification/Overexpression

HER2 mRNA

HER2 protein

Terapias dirigidas a HER2

Pertuzumab

Trastuzumab

I

II

III

IV

Anticuerpos

Trastuzumab-emtansine (T-DM1)

Inhibidores

Lapatinib

Neratinib

ATP

Resistencia a esquemas

terapéuticos en uso

¿Cómo estudiamos esto en el laboratorio?

¿Cómo estudiamos esto en el laboratorio?

Núcleo

HER2 en la membrana

Control Trastuzumab

¿Cómo estudiamos esto en el laboratorio?

BTRH

Trastuzumab

BTRL

Lapatinib

BT474

BTRN

Neratinib

BT-TDM1R

T-DM1

BTRH/RL

Lapatinib

HER2TDM1

Ciclina B

Lisosoma

Núcleo

TRAIL-R

TRAIL

HER3

EV20/MMAF

HER2TDM1

Lisosoma

DM1: inhibitor of tubulin polymerization and microtubuledynamics.

HIGHLY CYTOTOXIC: 25-500 fold more potent thantaxane in cytotoxic assays

Systemic toxicity is limited due to low HER2 expression in normal tissues

1. T-DM1 binds HER2

2. T-DM1-HER2 complex

is internalized

Early endosome

Microtubule

4. Inhibition of microtubule

polymerization

3. Lysosomal degradation

and DM1 release

Lysosome

HER2

T-DM1

DM1

BT474 C#1 C#3 C#6

0 min

T-DM1, 3d

DAPIT-DM1 DAPIT-DM1 DAPIT-DM1 T-DM1 DAPI

2017 - Vol 77(17): 4639 - 4651

HER2TDM1

Ciclina B

Núcleo

InterphaseG1

S

G2

M

Cyclin B induction

Cyclin B induction upon T-DM1 treatment ocurred in

parental breast cancer cells, but not in their counterparts

with acquired resistance to T-DM1

TRAIL-R

TRAIL

BT474

Cellcycle

Transcription

Celldeath

Celladhesion

DNA damage

responseCell

differentiation

Metabolism

Cell migration

Sig

nif

ican

ce

(-1

0*l

og

10[p

va

lue])

Log2(MDE)

BT474 vs BT-RH

425 DEGs240 185

20

40

60

80

100

120

2 4 6-4-6 -2 00

140

BTRH

Trastuzumab

Perfil génico A Perfil génico B Perfil génico C Perfil génico D

BTRL

Lapatinib

BTRH/RL

Trastuzumab

+

Lapatinib

BT-TDM1R

T-DM1

-5

-4

-3

-2

-1

0

1

2

3

BIR

C3

BC

L2

BN

IP3

LD

R6

OP

GD

R5

HR

KT

RA

F3

TN

FS

F8

RL

PO

CA

SP

5IG

F1

RB

NIP

3D

R4

CA

SP

14

HP

RT

1N

AIP

BIR

C5

FA

SL

GB

CL

2L

2B

2M

MC

L1

CD

27

TP

73

BA

G3

CA

SP

7B

FA

RT

RA

IL/A

PO

2C

D4

0L

GB

IRC

2A

IFM

1R

IPK

2C

AS

P6

CA

SP

9B

IDT

NF

AC

TB

BR

AF

CA

SP

8D

IAB

LO

BC

L1

0F

AD

DD

FF

AC

IDE

BC

RA

DD

CA

SP

2B

CL

2L

11

TR

AD

DA

BL

1C

YC

SF

AS

TN

F-R

1G

AD

D4

5A

AP

O3

NO

D1

BIR

C6

BA

XN

KB

1C

AS

P3

LT

BR

BN

IP2

TR

AF

2A

PA

F1

NO

L3

AK

T1

BA

G1

XIA

PT

P5

3B

P2

CF

LIP

TP

53

BC

L2

L1

PY

CA

RD

DA

PK

1B

IKC

AS

P1

0C

AS

P4

Dif

fere

nti

alm

RN

Ae

xp

ress

ion

Hig

her

in B

T-R

HH

igh

er

in B

T474

Extrinsic

apoptotic

pathway

E

C

Figure 2

BT474

BT-RH

DR4

DR5

FADD

Control TRAIL

0

20

40

60

80

100

120

BT474 BT-RH

MT

T m

eta

bo

liza

tio

n

(% o

f u

ntr

ea

ted

co

ntr

ol)

Cell Death Cell Death

BT474 BT-RH

+ TRAIL+ TRAIL

FLIP

DR4/5

trimerHER2

FADD

CASP8

HER3

EV20/MMAF

HER2 –

IP: a-NRG endo

WB: a-NRG endo

HER3 –

HER4 –

EGFR –IP: a-EGFR

WB: a-EGFR

IP: a-HER2

WB: a-HER2

IP: a-HER3

WB: a-HER3

IP: a-HER4

WB: a-HER4

SS

C2

SS

C8

SS

C9

SS

C24B

SS

C29

SS

C34

SS

C38

SS

C40

SS

C42B

Hep

G2

Hu

h7

A2780

IGR

OV

1

OV

CA

R3

OV

CA

R8

SK

OV

3

A498

AC

NH

Caki2

769P

786O

Cabeza y Cuello Hígado Ovário Riñón

IP: a-TGFα

WB: a-TGFα

–

–proTGFα

–

–

proNRG

CA

PA

N1

IMIM

-PC

1

IMIM

-PC

2

NP

9

NP

18

NP

29

NP

31

PA

NC

1

SK

PC

1

DU

145

PC

3

HT

1080

Páncreas Próstata

HC

T116

HT

29

BT

549

HC

C70

HC

C1187

HC

C1937

HC

C3153

HS

578T

MD

AM

B-2

31

MD

AM

B-4

68

MC

F7

T47D

Mama TNBC

Mama ER/PR

CólonTC

BT

474

HC

C202

HC

C1419

HC

C1569

HC

C1954

HC

C2218

MD

AM

B-1

75

MD

AM

B-3

61

Mama HER2+

SK

BR

3

HB

L100

?

IMR

5

IMR

32

NB

59

SK

NB

E

SK

NM

C

Sy5y

T98G

U138

A431

A2085

SK

ME

L2

WM

115

Cerebro Piel

A549

DM

S92

GL

C8

H23

H441

H460

H727

H820

Pulmón

H1299

Hela

Cerv.

H1975

SW

1573

EGFR

HER2

HER3

HER4

Calnexin

0

20

40

60

80

100

120

Control Trastuzumab

EFGR

HER2

HER3

HER4

Calnexin

0

20

40

60

80

100

120

0,001 0,01 0,1 1 10

Pro

life

rati

on

rate

(%

co

ntr

ol)

[EV20-MMAF], nM

BT474BTRHHCC1419HCC1569HCC1954MDA-MB-361SKBr3

EV20/MMAF

C T C T C T C T C T C T C T

C T C T C T C T C T C T

Calnexin

EV20/MMAF

Calnexin

BA

0

200

400

600

800

1000

1200

1400

1600

1800

0 7 14 21

Days of treatment

Control

EV20 10 mg/kg

EV20/MMAF 3,3 mg/kg

EV20/MMAF 10 mg/kg

0

100

200

300

400

500

600

700

800

0 50 100 150 200 250 300 350

Me

an

tu

mo

r vo

lum

e (

mm

³)

Days

EV20/MMAF 3,3 mg/kg

EV20/MMAF 10 mg/kg

EV20/MMAFEV20/MMAF

(RTICC, CIBERONC)