Mecanismos de resistencia a terapia anti-HER2: una visión ... · Cerv. EGFR HER2 HER3 HER4...
Transcript of Mecanismos de resistencia a terapia anti-HER2: una visión ... · Cerv. EGFR HER2 HER3 HER4...
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)