Download Diapositiva 1 - 11 Simposio Oncología Traslacional

El papel de la anatomía patológica en
oncología: presente y futuro
The role of pathological anatomy in
oncology: present and future
Federico Rojo
Fundación Jiménez Díaz, Madrid
All patients with same diagnosis:
Potential role for biomarker-based diagnostics
The genetic basis for cancer
treatment decisions
Garraway, LA. J Clin Oncol 2012
Evolution of NSCLC subtyping
Li, T et al. J Clin Oncol 2013
Sparing cost for the health insurance
Example of gefitinib treatment in lung cancer
€ 1.7M
15 000 patients (gefinitib treatment:
8 weeks DFS; Mok
2009)
€ 69M
Spared cost of gefitinib treatment
EGFR
testing for
lung cancer
patients
1 724 patients +
(gefinitib treatment:
38 weeks DFS; Mok 2009)
€ 35M
Cost of gefitinib treatment
Sources of variability in molecular
testing
Allred, DC. Mod Pathol 2010
Delay to formalin fixation (cold ischemia)
modifies DNA quality
• The longer duration of fixation adversely affects the quality of tissue DNA
• The size of DNA extracted from FFPE decreases with increasing fixation time
• Tissues fixed in buffered formalin for 3 to 6 hours yield greater amounts of high-molecular
weight DNA
1h
A
B
2h
C
A
B
6h
C
A
B
12h
C
A
B
24h
C
A
B
C
Sample
Preservation of DNA in FFPE archieved tissue samples
• Buffered 4% Formaldehyde / 10% Formalin
– minimal prefixation time lag, 2 hours
– duration of fixation (3 to 6 hours)
– absolutely avoid low pH
• No Bouin, No Acid Formaline, No B5, No AZF, No decalcified samples (bone biopsies)
Lab A
Lab B
Lab C
Lab E
Lab D
Rojo, F. SPMM 2011
DNA sequence alterations due to
formalin fixation in archival specimens
FFPE tissues exhibit a high
frequency of nonreproducible
sequence alteration because of
cross-linking cytosine nucleotides on
DNA strands.
Taq-DNA polymerase fails to
recognize the cytosine and
incorporates an adenine in the place
of a guanosine, creating an artificial
C-T or G-A mutation.
Error rate is ~12%.
Williams, C et al. Am J Pathol 1999
Hofreiter, M et al. Nucleic Acids Res 2001
Akbari, M et al. J Mol Diag 2005
Lamy, A et al. Mod Pathol 2011
Recommendations for mutation testing
Su, KY. Et al. J Clin Oncol 2012
Querings, S. et al. PLOS One 2011
RAS mutations by pyrosequencing
KRAS WT
Codon 12
(GG
T)
NRAS WT
Codon 13
(GG
T)
Codon12
(GG
Codon13
T) (GG
T)
KRAS MUT
(GG
T)
(GA
T)
NRAS MUT
(G A T)
KRAS WT
Codon 146
(G C
A)
NRAS MUT
34
(T
KRAS MUT
(G C T)
G
T)
(G A T)
Importance of selection of tissue sample for
molecular diagnosis
Adenocarcinoma
Normal tissue
Liver
Necrosis
Heterogeneidad en cáncer: distinta
enfermedad en distintos pacientes, en
distintas localizaciones y variabilidad
intratumoral
Heterogeneidad en cáncer de mama:
distinta enfermedad en distintas mujeres
Heterogeneidad en cáncer de mama:
distinta enfermedad en distintas mujeres
Reis-Filho, JS. AACR 2012
Weigelt, B. et al. Breast Can Res 2010
Lee, JK. et al. Clin Cancer Res 2010
Heterogeneidad en cáncer de mama:
distinta enfermedad en distintas localizaciones
Tumor primario
Falk, AK. 2013
Martinez de
Dueñas, E.
2013
(ConvertHER)
Metástasis
ganglionar
Metástasis
visceral
(lab.
central)
Luminal A
(%)
Luminal B
(%)
HER2 (%)
TN (%)
Luminal A
85
0
9
0
Luminal B
11
100
91
0
HER2
0
0
0
8
TN
4
0
0
92
Luminal
74 (90)
21 (0)
22 (18)
HER2
16 (3)
76 (100)
7 (6)
TN
10 (7)
3 (0)
71 (76)
Falck, AK et al. BMC Cancer 2013
Martinez de Dueñas, E et al. Breast Cancer Res Treat 2013
Heterogeneidad en cáncer de mama:
distinta enfermedad en distintas localizaciones
Niikura, N. et al. J Clin Oncol 2012
Heterogeneity of mutations in colorectal
cancer
WT in both samples
MUT in both samples
WT in primary, MUT in mx
Baldus, SE. et al. Clin Cancer Res 2010
Voutsina, A et al. Mod Pathol 2013
MUT in primary, WT in mx
Intratumor heterogeneity: impact on biomarker
detection
Gerlinger, M. et al. NEJM 2012
Heterogeneity of mutations in cancer:
Clonal dynamics and resistance
Heterogeneidad en cáncer de mama:
variabilidad intratumoral
Heterogeneidad en cáncer de mama: variabilidad
intratumoral
Navin, N et al. Genome Res 2010
Heterogeneidad en cáncer de mama:
variabilidad intratumoral
Mosoyan, G et al. PLOS One 2013
Heterogeneidad en cáncer de mama:
variabilidad intratumoral
Mosoyan, G et al. PLOS One 2013
Heterogeneidad en cáncer de mama:
variabilidad intratumoral
Heterogeneous distribution of PIK3CA mutations in breast primary vs metastasis in 030%
Jensen, JD et al. Clin Can Res 2011
Gonzalez-Angulo, AM et al. Mol Cancer Ther 2011
Cancer genome:
Routine diagnostic in next years
Cancer genome:
Routine diagnostic in next years
Cancer genome:
Dynamic modulation of genetic background
Cancer genome:
Dynamic modulation of genetic background
Mensajes finales
• Estandarización en los procesos de laboratorio: fase
preanalítica y ensayos
• Elección del ensayo más apropiado a las características
de nuestro centro
• Inversión en tecnología y formación específica y continua
de los especialistas
• Alta complejidad de la enfermedad