Subtipos moleculares

del Cncer de Mama

Joseph Pinto
Investigacin Bsica &
translacional
Flujo de la Informacin
Gentica
Flujo de la Informacin
Gentica
Diferenciacin celular
Qu estudiamos en el perfil
gentico?
Tumor A Tumor B

Expresin: 4
Expresin: 21
Los MIcroarrays
Affymetrix Expression Arrays

http://www.affymetrix.com/technology/ge_analysis/index.affx
 Target Sequences and Probes
Example:
1415771_at:
Description: Mus musculus nucleolin mRNA, complete cds
LocusLink: AF318184.1 (NT sequence is 2412 bp long)
Target Sequence is 129 bp long
11 probe pairs tiling the target sequence
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
gagaagtcaaccatccaaaactctgtttgtcaaaggtctgtctgaggataccactgaagagaccttaaaagaatcatttgagggctctgttcgtgcaagaatagtcactgatcgggaaactggttctt
Affymetrix Chip Pseudo-image

*image created using dChip software

1415771_at on MOE430A

*image created using dChip software

1415771_at on MOE430A

PM
MM

*Note that PM, MM are always adjacent

*image created using dChip software

 Sample 1 Sample 2 Sample M

Gene 1
Gene 2

Expression Signature

Expression Profile

Gene N
 Squamous, small cell, and large cell lung tumors express a unique set of genes.

Garber M E et al. PNAS 2001;98:13784-13789

2001 by National Academy of Sciences
 Heat Map for 2880 Ontario Genes (39 Tissues)

Genes

Tissues
Los subtipos de cncer de
mama
 Human Breast Cancer Is Highly Heterogeneous

STAGE Lymph. infiltrate

invasive

In situ
low
Well-
Margins Nuclear Grade
Differentiation
Poorly-

high

infiltrating single-file
pushing
Can we decipher new molecular genetic information for
these complex and variable tumors and establish a new
classification with real therapeutic impact.
Perou et. al.,
2000

Sorlie et. al.,

Sorlie et. al.,
 Figure 1a.

Sorlie T, Tibshirani R, Parker J, et al: Repeated observation of

breast tumor subtypes in independent gene expression data
sets. Proc Natl Acad Sci U S A 100:8418-23, 2003
HER2-amplicon

basal epithelia

uminal epithelial

proliferation

Claudin-low
 Intrinsic Breast Cancer
Subtypes described by
Perou et al.

Luminal A
Luminal B
HER2+
Basal-like
Express amounts
Express levels of EGFR,
Of luminal cyto-
c-kit, & growth factors like
Keratins & genetic
hepatocyte growth factor
Markers of luminal
and IGF
Epithelial cells of
Normal tissue

Sotiriou, C. et al. NEJM, 2009.

CURRENT THERAPEUTIC BREAST CANCER
SUBTYPES

15-18%

20-25% 60-65%
Cmo se originan los subtipos
de cncer de mama?
 Modelos que explican el posible
origen de los subtipos de cncer de
mama
1. Evolucin lineal
 Modelos que explican el posible
origen de los subtipos de cncer de
mama
2. Alteracin de distintas vas

Chreighton,
Cmo funcionan los
receptores
hormonales?
Miller JCO 2014
Zabransky JCO 2014
Cmo funciona el
HER2?
 HER2 Overexpression in Breast
Cancer

HER2 is overexpressed in
~ 25% of breast cancers

Normal (1x)
~ 25,000-50,000 HER2
receptors

Overexpressed
HER2 (10-100x)
up to ~ 2,000,000
HER2 receptors

Pegram MD, et al. Cancer Treat Res. 2000;103:57-75.

Ross JS, et al. Am J Clin Pathol. 1999;112(suppl 1):S53-S71.
Excessive cellular division
Slamon DJ, et al. Science. 1987;235:177-182.
 The human epidermal growth factor receptors
(HER) are a family of structurally-related cell
surface proteins

Extracellular ligand-binding
domain
HER1/EGFR HER2 HER3 HER4

Transmembrane
domain

Intracellular tyrosine kinase domain

42 Oncologist. 2003;8(suppl 3):5-17. Yarden et al. Nature Rev Mol Cell Biol. 2001;2:127-137.
Rowinsky.
 Con excepcin de HER2, las proteinas HER atraviesan
por un cambio conformacional sobre la unin de
lingandos esencial para la dimerizacin y sealizacin.
Ligand primes receptor Closed conformation
for activity

Open conformation

Cho et43
al. Science. 2002;297:1330-1333. Ferguson et al. Mol Cell. 2003;11:507-517. Ogiso et al. Cell. 2002;110:775-787.
 HER2 se encuentra siempre en una conformacin
abierta convirtindose en un compaero ideal
para la dimerizacin.

HER2 HER2 does not require a

ligand to be primed

Garrett44
et al. Mol Cell. 2003;11:495-505. Graus-Porta et al. EMBO J. 1997;16:1647-1655.
 De todos los posibles dimeros, el par HER2:HER3
presenta la sealizacin mitgena ms fuerte.

Homodmeros Heterodmeros
HER1:HER3 HER1:HER4
HER4:HER4 HER1:HER2
HER3:HER3 HER2:HER3
HER2:HER2 HER2:HER4
HER1:HER1
HER3:HER4

+ + + + + + + +
+ + + +
+ +
+
Signaling activity

Tzahar45
et al. Mol Cell Biol. 1996;16:5276-5287. Lenferink et al. EMBO J. 1998;17:3385-3397.
 La fosforilacin de los dominios tirosin
quinasa de los dmeros de HER es regulada
va interacciones alostricas.

Zhang46
et al. Cell. 2006;125:1137-1149.
 Biologic Effects of HER-2/neu
Amplification/Overexpression in Human Breast
Cancer Cells

DNA
Synthesis
Cell Growth

HER2- HER-2 HER2+
Breast Growth in

Cancer Cell Breast Cancer Soft Agar
Lines Transfect Cell Lines
ion Tumorigenicity

Metastatic

Potential
E2 Response,
Tam
Resist.
Los subtipos sustitutos
(inmunofenotipo)
 Intrinsic Breast Cancer
Subtypes described by
Perou et al.

Luminal A
Luminal B
HER2+
Basal-like
Express amounts
Express levels of EGFR,
Of luminal cyto-
c-kit, & growth factors like
Keratins & genetic
hepatocyte growth factor
Markers of luminal
and IGF
Epithelial cells of
Normal tissue

Sotiriou, C. et al. NEJM, 2009.

Cncer de mama basal/triple
negativo
 Relevance of Triple-Negative
Breast Cancer
Tumors that do not express the ER, PR, and
HER2/neu markers are more likely to be basal-
subtype tumors
Inherently aggressive biologic behavior
Triple-negativity used as a surrogate for basal subtype

Tumors that do not express the ER, PR, and

HER2/neu markers have fewer systemic therapy
options
Endocrine/hormonal therapy will be ineffective
Herceptin therapy will be ineffective
Chemotherapy IS effective
 Basal-like breast cancer
Gene expression profile
Most tightly clustered subgroup
in gene expression arrays
CK 5/6 and 17 expression
P53 mutations
EGFR overexpression
Mostly triple negative
BRCA1 connection
Gene expression similar
Morphology similar
Morphology
High grade
Mainly ductal or medullary
High mitotic count
Scant stroma
Central necrosis
Pushing border
Lymphocytic infiltrate
Apoptotic figures
A B
Analysis of Interobserver Agreeme

Mackay JNCI
Mas subtipos?
Los subgrupos integradores tienen distinto perfil de
nmero de copias
Examen
ESTROGEN PROGESTERONE

1
HER2 Ki 67
ESTROGEN PROGESTERONE

HER2 2 Ki 67
ESTROGEN PROGESTERONE

HER2 3 Ki 67
ESTROGEN PROGESTERONE

HER2 4 Ki 67
ESTROGEN PROGESTERONE

HER2
5 Ki 67