Patología molecular del carcinoma de mama

José PalaciosServicio de Anatomía PatológicaHHUU Virgen del Rocío, Sevilla

Xàtiva, 12 de enero de 2009

CK8, 18, 19

Cadherina-E

CK5/6, 14, 17

P-Cadherin

Actin

vimentin

Luminal epithelial

Myoepithelial

(Basal)

CELLULAR TYPES IN MAMMARY GLAND

Basal epithelialmarkers

Mesenchymal markers

CK5/6

CK14

CK17

P-CADHERIN

p63

S100

CD10

CD44

VIMENTIN

CAV1

OSTEONECTIN

SMA

CALPONIN

H-CALDESMON

CALPONINp63

MYOEPITHELIAL MARKERS

Limiting dilution transplantations studies

Retroviral-tagged mammary implants

Prospective isolation of putative stem cells using cell surface markers

IDENTIFICATION OF MAMMARY STEM CELLS

Multipotent epithelial cells (“stem cells”) with the ability to generate the entire

functional mammary epithelium

Visvader and Lindeman, Cancer Res 2006

MODEL OF THE EPITHELIAL CELL HIERARCHY IN THE MOUSE MAMMARY GLAND.

AEP/ADH/LG-DCIS

ALH/LCIS

+8q+17q-17pP53HER2MYC

-16q +11q

CCND1E-CD (-)

BREAST CANCER PROGRESION MODEL

Invasive Lobular Carcinoma

Low Grade (invasive ductal G1,

tubular, cribiforme...)

High grade (invasive ductal G3, apocrine,

medullary metaplastic...)

HG-CDIS

Breast epithelium

TDLU

CCC

Perou et al., Nature 2000

MOLECULAR CLASSIFICATION OF BREAST CANCER

c Luminal, ER+ (A and B)

d ERBB2e Basalf Normal

Sorlie et al; PNAS 2003

LUMINAL (ER+) PHENOTYPES

Invasive ductal carcinoma (grade 1-2/3)

Invasive lobular carcinoma

Tubular carcinoma

Invasive cribiforme carcinoma

Mucinous carcinoma

LUMINAL A/B PHENOTYPES

GRADE 1-2/3

ER LEVELS

PROLIFERATION MARKERS

HER2 EXPRESSION

•Invasive breast carcinoma composed by non-cohesive cells individually dispersed or arranged in single-file linear pattern in a fibrous stroma.

LOBULAR BREAST CANCER

INCREASING INCIDENCE (HRT?)

HIGHER AGE AT DIAGNOSIS

HIGHER SIZE AT DIAGNOSIS

DIFFUSE GROWTH PATERN

LOWER SENSITIVITY OF MAMMOGRAPHY TO DETECT ILC

HIGHER RATE OF FAILURE AFTER BCT?

METASTATIC PATTERN

Lobular carcinomas completely lack E-cadherin expression

Lobular carcinoma in situ

Infiltrating lobular carcinoma

Invasive Ductal carcinoma

Absence of E-cadherin >80% Reduced E-cadherin: 50%

Gamallo et al; Am J Pathol 1993

C-terminal

• CDH1 is a tumor and invasion suppressor gene

16q22.1

The E-cadherin/catenin adhesion complex

-catenin

CBD

JMD

E-CADHERIN

actin

p120

-catenin

Protein Mw Gene location

E-cadherin 120 kDa 16q22.1 -catenin 102 kDa 5q31 -catenin 92 kDa 3p21 -catenin 83 kDa 17q21 p120ctn 120 kDa 11q11

N

IS

IF

D16S398

Wt N T IF IS

E-CADHERIN INACTIVATION IS THE HALLMARK OF LN

E-CD E-CD

Sarrió et al; Int J Cancer 2004

D16S265

CDH1CDH1

D16S496

D16S398

D16S3057

D16S752

16q2

2.1

T

N

D16S496

-126 -21 +81 +144+1

M U M U M U M U M U M U M U M U

14 15 16 17 24 25 38 H2O

CDH1

Mechanisms of CDH1 inactivation in lobular tumors

CDH1 LOH (60-80%)

CDH1 promoter hypermethylation (40-60%)

6 6 Wt6 6 Wt

Exon 13Exon 13

Wt 5 5Wt 5 5

Exon 10Exon 10

Wt 19 19Wt 19 19

Exon 2Exon 2

1712 G > C

Wt 52 52Wt 52 52

Exon 11Exon 11

• CDH1 mutations (20-40%).

CCNN

SIGSIG PREPRE CaCa2+ 2+ binding domain TMTM CP CP

60-70% produced truncated proteins.

Mechanisms of CDH1 inactivation in lobular tumors

Sarrió et al, Int J Cancer 2003

HDGC is caused by germline mutation of CDH1.

The lifetime risk of developing gastric cancer by age 80 is 65%-80%.

Female CDH1 mutation carriers are at increased risk for developing lobular breast cancer.

Penetrance studies have shown that female CDH1 germline mutation carriers have an additional risk of breast cancer, particularly lobular breast cancer, in about 39% of patients.

Some families have been misclassified as breast cancer families due to clustering of breast cancer cases.

This suggests that families with negative test results for BRCA1/2 in whom the proband has lobular breast cancer should be reevaluated for HDGC and screened for CDH1 mutations.

BREAST CANCER IN HDGC PATIENTS

Perou et al., Nature 2000

MOLECULAR CLASSIFICATION OF BREAST CANCER

c Luminal, ER+ (A and B)

d ERBB2e Basalf Normal

BREAST CANCER AND THE BASAL-LIKE PHENOTYPE

Palacios et al; Am J Pathol 1995 Tsuda et al; Am J Surg Pathol 2000

ER/HER2-negative,

CK5- and/or EGFR-positive

Nielsen et al., Clin Cancer Res 2004

BASAL-LIKE PHENOTYPE

VIMENTIN, P-CADHERIN, EGFR, CK5/6, FASCIN

p63, CD10, OSTEONECTIN, SMA, CALPONIN, H-CALDESMON

BASAL-LIKE PHENOTYPE (ER/HER2-negative, CK5- and/or EGFR-positive)

Rodríguez-Pinilla el al., Clin Cancer Res 2006

Medullary carcinoma

IBCMF IDCG3 P

ERPositiveNegative

2/35 (5.7%)33/35 (94.3%)

24/39 (61.5%)15/39 (38.5%)

<0.001

HER2PositiveNegative

0/35 (0.0%)35/35 (100.0%)

9/38 (23.7%)29/38 (76.3%)

0.002

Ck5/6PositiveNegative

21/35 (60.0%)14/35 (40.0%)

7/39 (17.9%)32/39 (82.1%)

<0.001

EGFRPositiveNegative

9/35 (25.7%)26/35 (74.3%)

9/37 (24.3%)28/37 (75.7%)

0.553

Ck19PositiveNegative

13/35 (37.1%)22/35 (62.9%)

26/38 (68.4%)12/38 (31.6%)

0.007

P-CadherinPositiveNegative

14/35 (40.0%)21/35 (60.0%)

3/38 (7.9%)35/38 (92.1%)

0.001

Basal-like phenotypePositiveNegative

22/35 (62.9%)13/35 (37.1%)

7/37 (18.9%)30/37 (81.1%)

<0.001

Differences between invasive breast carcinomas with medullary features (IBCMF) and grade 3 invasive ductal carcinoma of no

special type (IDCG3).

Rodríguez-Pinilla el al., Am J Surg Pathol 2007

Medullary carcinoma Metaplastic carcinomaPoorly differentiated carcinoma with central

acellular zones

MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL HETEROGENEITY OF BREAST CARCINOMAS WITH BASAL-LIKE PHENOTYPE

VIMENTIN, P-CADHERIN, EGFR, CK5/6, FASCIN

p63, CD10, OSTEONECTIN, SMA, CALPONIN, H-CALDESMON

SOX2 EXPRESSION IN BREAST CANCER

SOX2 Ki67

CK5/6VIMENTIN

SOX2 (-) SOX2 (+) P

Rodríguez-Pinilla et al., Mod Pathol 2007

Perou et al., Nature 2000

MOLECULAR CLASSIFICATION OF BREAST CANCER

c Luminal, ER+ (A and B)

d ERBB2e Basalf Normal

HER2 EXPRESSION IN BREAST CANCER

BASAL+ HER2+ BASAL/HER2-

RE (-) 95% 60% 20%

RP (-) 93% 58% 37%

GRADO 3 98% 55% 32%

CK5 57% 10% 6%

CK8 50% 98% 95%

P53 (>30%) 55% 42% 16%

Ki67 (>15%) 72% 46% 32%

EGFR (+) 35% 10% 5%

DIFERENCIAS ENTRE TUMORES CON FENOTIPO BASAL+ Y HER2+

Laakso et al., Clin Cancer Res 2006

HER2 EXPRESSION IN BREAST CANCER

Oncogene 2005

Apocrine histology

Non-basal ER-negative

AR-positive

HER2-positive (50%)

Natrajan et al, 2008

Natrajan et al, 2008

n HER2 (+)

Anderson (2003) 20 16 (80%)

Fu (2001) 14 14 (100%)

Schelfhout (2000) 30 26 (85%)

Meissner (1990) 23 23 (100%)

ENFERMEDAD DE PAGET y HER2/neu

HER2/neu Y CARCINOMA INFLAMATORIO

HER2 (+)

CDI-NOS 15-20%

CLA 30%

CI 35-55%

DSS in non-treated patients

Basal-like tumors (n=12)

Non Basal-like tumors (n=95)

Log Rank P=0.001

Survival Time (Months)

% S

urv

iva

l

DSS in CMF-treated

Survival Time (Months)

Log Rank P=0.633

% S

urv

iva

lBasal-like tumors (n=13)

Non Basal-like tumors (n=85)

ER/HER2-negative, CK5- and/or EGFR-positive

BASAL-LIKE PHENOTYPE AND CMF RESPONSE IN BREAST CANCER

Rodríguez-Pinilla el al., Clin Cancer Res 2006

Breast cancer Ovarian cancer Male BC

BRCA1 65% 40% -

BRCA2 45% 11% 8%

BCLC Meeting. Madrid, June 2003

BRCA1/2: 30% BRCA (-): 70%

Hereditary breast cancer

Histological type and grade

BRCA BRCA2 Sporadic

IDC 74% 71% 69%

Medullary features 18% 3% 3%

Grade 3 66% 41% 36%

Immunohistochemical markers

BRCA1 BRCA2 Sporadic

ER+ 20% 66% 65%

PR+ 20% 49% 66%

PRER

Probability of carrying a BRCA1 mutation by age, ER status and grade

(Lakhani et al, J Clin Oncol 2002)

Immunohistochemical markers

BRCA1 BRCA2 Sporadic

Ki67 high 56% 21% 22%

Cyclin D1 30% 56% 79%

Cyclin E 47% 35% 27%

P53+ 45% 27% 12%

HER2+ 0-7% 0-6% 18%

HER2 amplification

BRCA1 BRCA2 Sporadic

(n=14) (n=10) (n=54)

HER2 (FISH) 0 0 22%

Morpholgical and immunohistochemical features of BRCA1 and BRCA2 breast carcinomas

GRADE ER PR BCL2 Ki67 p53 HER2

BRCA1 3 - - - ++ ++ -

BRCA2 2/3 + + + + - -

Ki67 p53BCL2PRER

Sorlie et al. Proc Natl Acad Sci U S A. 2003

BASAL PHENOTYPE IN BRCA1 BREAST CARCINOMAS

BRCA1 BRCA2 Sporadic

Cytokeratin 5/6 46% 9% 8.5%

Vimentin 80% 15% 23%

Fascin 84% 17% 25%

Laminin 75% 7% 39%

Caveolin 1 22% 0 4.2%

Cytokeratin 5/6

BASAL CELL MARKERS

Palacios et al, J Nat Cancer Inst 2004Rodríguez-Pinilla et al, Clin Cancer Res 206; Breat Cancer Res and Treat 2006; J Clin Pathol 2007

BRCA1/2: 30% BRCA (-): 70%

Hereditary breast cancer

GRADE ER PR BCL2 Ki67 p53 HER2

BRCA1 3 - - - ++ ++ -

BRCA2 2/3 + + + + - -

BRCA(-) 1/2 + + + - - +/-

Dontu et al., 2004

BASAL PHENOTYPE

BASAL AND LUMINAL

MARKERS

LUMINAL PHENOTYPE

LUMINAL MARKERS

ONLY

HER2

PIK3CA

RAS

-16q

CDH1

IDC-G1/2

MUCINOUS

CRIBIFORME

TUBULAR

LOBULAR

METAPLASTIC

MEDULLARY

IDC-G3

IDC-G2/3

APOCRINE

BRCA1

EGFR

P53

MYC

SMA

VIMENTIN

C-KIT

EGFR

P-CADHERIN

CK5/6

CK8/18/19

HER2

AR

CK8/18/19

ER

PR

GATA3

CK8/18/19

BASAL

BASOLUMINAL

HER2

APOCRINE

LUMINAL B

LUMINAL A

LPER-

LPER+

Luminal cells

Luminal Progenitors

Common Progenitor

Stem Cell

ER-

ER-