ABORDAJE GENÉTICO DEL CÁNCER DE MAMA Y OVARIO HEREDITARIO
1. 1
Abordaje genético
del cáncer de mama y ovario hereditario
Dra. Sara Alvarez de Andrés
Dirección Médica NIMGentics, S.L
Conflicto de Intereses: Socio de NIMGenetics, SL
2. 2
Contenidos:
1. Selección de pacientes para el estudio del Cáncer de Mama y Ovario hereditario
2. Técnicas para la identificación de las Mutaciones
3. Tipos de Mutaciones en BRCA1 y BRCA2
4. Utilidad y Aplicaciones de los Paneles Multigen en el Ca de Mama y Ovario
hereditario
5. Evaluación de los Factores de riesgo que incrementan el riesgo
relativo de desarrollo de Ca de Mama
• Edad (>65a vs <65a, con un incremeto progresivo del riego hasta los 80a )
• Determinadas Mutaciones germinales heredadas (BRCA1 y/o BRCA2)
• Antecedentes personales de Patología mamaria
• Dos o más familiares de primer grado con Cáncer de mama diagnosticados a edades tempranas
• Historia Personal de Cáncer de Mama (>40 años)
• Niveles Hormonales elevados de estrógenos ó testosterona (postmenopaúsicas)
• Radioterapia torácica previa
• Un antecedente familiar de Cáncer de Mama de primer grado
• Historia menstrual (>12a y >55a)
• Historia reproductiva (Nuliparidad, 1ºemb>30ª, No lactancia, No embarazos a término)
• Antecedentes personales de herencia Ashkenazi (Europa del Este)
• Hº personal de cáncer de endometrio, ovario y colon.
• Terapias Hormonales sustitutivas ó contraceptivas
• Otros: Obesidad, Consumo Alcohol, status socioeconómico alto, estatura alta.
Breast Cancer. Facts and Figures 2013-2014
Riego Relativo
>4
2.1-4
1.1-2.0
6. - ELEVADA INCIDENCIA DEL CANCER DE MAMA- familias con múltiples casos pueden
agrupar casos de Cáncer hereditario y esporádico.
- PENETRANCIA VARIABLE del GEN CAUSAL dependiente del género, la edad, y de
factores no genéticos
- HETEROGENICIDAD DE LOCUS- Diferentes genes son responsables de la
predisposicíon al Cáncer de Mama en las diferentes familias con HBOC
- HETEROGENICIDAD ALÉLICA- Diferentes Alelos alterados en los diferentes genes
causales en las diferentes familias.
6
Factores que dificultan el diagnóstico clínico del
Cáncer de Mama y Ovario Hereditario
Mary-Claire King, Science 343, 1462-65 (2014)
7. Criterios de Selección de la SEOM+NCCN 2014
para el estudio de los genes BRCA1 y BRCA2 en pacientes con Cancer de Mama
1) Independientemente de la historia familiar si:
• Antecedente de una mutación conocida ó herencia Ashkenazi
• SEOM: Paciente con cáncer de mama y cáncer de ovario sincrónico o metacrónico vs
NCCN: Cáncer de ovario epitelial, cáncer tubárico o primario peritoneal
• Pacientes >60años con Cáncer de Mama triple negativo
• SEOM: Un caso de cáncer de mama en paciente de 30 años ó menor vs NCCN<45a
• Un caso de cáncer de mama bilateral en menor de 40 años.
2) Familias con dos miembros afectos con cáncer de mama y/o cáncer de ovario y al
menos una de las siguientes características:
• Varón con cáncer de mama.
• Uno de ellos es un Cáncer de ovario, cáncer tubárico o primario peritoneal.
• Ambos casos diagnosticados en menores de 50 años
3) Familias con 3 ó más individuos con cáncer de mama y/o cáncer de ovario en la misma
rama de la familia.
4) Familias con dos familiares (1º, 2º ó 3º grado) con Cáncer de páncreas y y/o prostata
(Gleason score 7)
Cáncer Hereditario II ed. Alonso A et al. Ed. SEOM, 2010; pp:428
8. 8
Criterios de NCCN 2014 para considerar la realización de Estudios Genéticos
1) En pacientes AFECTOS:
• Antecedente de una mutación conocida
• Cáncer de ovario, cáncer tubárico o primario peritoneal
• Cáncer de mama:
– En paciente de <45años
– Cáncer Triple Negativo
– Cáncer de Mama en varón
– Cáncer de mama bilateral ó dos tumores ipsilaterales sincrónicos ó asincrónicos
• Con Antecedentes familiares de:
– 1 familiar con Ca de mama<50 años
– 1 familiar con Ca de Ovario a cualquier edad
– 2 familiares con Ca de mama y/o cancer de páncreas de cualquier edad
– Pertenecer a una población de riesgo (pej: Ashkenazi)
– 1 familiar con Ca de mama y otro con Ca de pancreas, prostata, SNC, endometrio, Leucemia/linfoma,
Tiroides, alteraciones dermatológicas, macrocefalia, hamartomas GI ó cancer gástrico difuso.
2) En individuos NO AFECTOS con Hª familiar de:
– Mutación conocida
– Cáncer de mama bilateral ó dos tumores ipsilaterales sincrónicos ó asincrónicos
– Más de dos individuos con Cáncer de mama en la misma familia
– cancer de ovario
– Cáncer de mama en <45 años
– 1 familiar con Ca de mama y otro con Ca de pancreas, prostata, SNC, endometrio, Leucemia/linfoma,
Tiroides, alteraciones dermatológicas, macrocefalia, hamartomas GI ó cancer gástrico difuso.
– Cancer de mama en Varon
9. • Aspectos Legales
– Liberación de la Patente de Myriad (U.S Supreme court, Junio 2013)
– “The Genetic Information Nondiscrimination Act of 2008 (Public Law 110–233)”
• Desarrollo Científico/Tecnológico:
– Identificación de múltiples genes de predisposición al desarrollo del Cáncer de
Mama y Ovario hereditario.
– Desarrollo de la Secuenciación masiva
• Evidencias epidemiológicas*
– Más del 50% de las mutaciones en BRCA1 y BRCA2 son heredadas de
progenitores NO AFECTOS.
– El 50% de las mujeres con mutaciones en BRCA1 y BRCA2 tienen poca o ninguna
historia familiar de Ca de mama u ovario (familias NO INFORMATIVAS)
9
Pérdida de Barreras para el Estudio mutacional de BRCA1/2
“Hasta que no se erradique en Ca de Mama y Ovario en mujeres portadoras la carrera de los
BRCAs no se habrá finalizado”-Mary-Claire King, 2014
* M.-C. King, Science 302, 643–646 (2003).
11. 11
Se analizan simultáneamente todos los exones y las regiones
de splicing -20 a +20, al menos -10 a +10 de BRCA1/BRCA2
>1 nt
F1 F2
R2R1
+20
Los amplicones son solapantes
Aplicación de la Secuenciación masiva al Estudio mutacional
de BRCA1/2
12. Coverage:
• 100% ORF
• All, but 1 amplicon, >100x
• Mean per amplicon: 2100 ± 302
Análisis Simultáneo de 8 muestras en una sola carrera
BRCA1
cds
exon
Depthofcoverage
BRCA2
cds
exon
Depthofcoverage
Aplicación de la Secuenciación masiva al Estudio mutacional
de BRCA1/2
13. 13
Nonsense
Frameshift deletion
Frameshift insertion
Missense
Normal
mRNADNA
C
T
CA A G GCG C T A A C T
GU U C C GC GA U U GA
GU U C U U G A
CA A G A A C T
GU U U A G
C A A A C T
CA A G C G A A C T
GU U C G C U U GA
GU U C G A U U GA
CA A G C T A A C T
Clasificación de las Mutaciones Puntuales
PROTEINA TRUNCADA Y
DEGRADADA POR EL SISTEMA
NMD (Nonsense Mediated Decay)
PROTEINA CON UN AMINOACIDO
DISTINTO A LA PROTEINA WT
14. 14
Identificación de los Grandes Reordenamientos
Detección de las pérdidas y ganancias de regiones del genoma en una muestra de ADN
Hibridar y Escanear
MLPA
(Multiplex ligation dependent probe
amplification)
aCGH de Alta Resolución
(Comparative Genomic Hybridization)
15. Deteccion de grandes delecioness
Se ha detectado una deleción patogénica en la citobanda 13q12.2, coordeandas genómicas
chr13: 32,890,289-32,890,740. El análisis bioinformático con el Software Alamut ESES predice
que la deleción identificada implica una inactivación del gen BRCA2, debido al cambio del marco
de lectura generándose una proteína truncada.
Deleciónde 451 pb
(Exón 2 de BRCA2)
18. 18
Knudson “two-hit” Model
Genes Supresores de Tumores
Primera mutacion
(Portador)
GEN NORMAL
Regula el Crecimiento tumoral
Segunda Mutación
(Desarrollo del tumor)
Cáncer
Esporádico
Cáncer
Familiar
Mayor riesgo a desarrollar un Cáncer
Menor Edad de presentación
Asociación con múltiples tumores
Habitualmente con un patrón de Herencia AD
19. BRCA1 (Chr17)
BRCA2 (Chr13)
~1800 mutaciones
~2000 mutaciones
Splice-site
Nonsense/Frameshift
Missense
Díez O, et al . Hum Mutat 2003;22:301-312
≠ Locus y ≠ alelos = fenotipo
Mutaciones fundadoras-Estudios dirigidos
Más de 1M de individuos han sido testados en las últimas dos décadas
El panorama mutacional de BRCA1/2
BRCA1 (14% de las mutaciones)
BRCA2 (2.6%de las mutaciones)
GRANDES
REORDENAMIENTOS
MUTACIONES
20. 20
La pentrancia interfamilias depende del dominio proteíco
alterado en función de la localización de la mutación
Missense
Nonsense/Frameshift
>Riesgo de
Ca de Ovario
Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA)
Las mutaciones en BRCA1 y BRCA2 presentan una Penetrancia variable e incompleta
Mutaciones
en 3´y 5´
>Riesgo de
Ca de Mama
21. 21
1.- Variante Benigna
2.- Variante Probablemente Benigna
3.-Variante de Significado incierto
4.- Variante probablemente patogénica
5.- Variante Patogénica
Interpretación de las variantes genómicas en el informe
Clínico de diagnóstico Genético
VARIANTES
INFORMADAS
Son aquellas que NO pueden ser
clasificadas como
NEUTRALES ó PATOGÉNICAS
VUS: 2 al 10% de las variantes
Cheon et al. Genome Medicine 2014, 6:121
22. 22
Variantes de Significado Incierto (VUS)
1. Las VUS incluyen variantes missense, intronicas, y pequeños Indels in frame.
2. La clasificación de estas variantes se depura a través de bases de datos como
Clinvar (repositorios de los resultados de diversos laboratorios)
3. La evaluación de la patogenicidad de las variantes se realiza por sistemas de
predicción in sílico y el desarrollo de consorcios como el “Evidence-based
Network for the Interpretation of Germline Mutant Alleles (ENIGMA)”*.
4. La probabilidad de patogenicidad de cada variante en base a:
*N. M. Lindor et al., Hum. Mutat. 33, 8–21 (2012).
• Nivel de conservación evolutiva de la variante (Align-GVGD)
• Segregación familiar
• Características AP del tumor
• Efectos en el RNA splicing
• Ensayos “in vitro”
• Recurrencia de la variante ¿Problemas éticos y de Regulación?
23. 23
Variantes de Riesgo bajo Moderado (Hipomórficas)
Mutaciones en BRCA1/2 que REDUCEN la actividad de la Proteína pero NO la anulan. Se
asocian a un riesgo bajo ó moderado de Ca de Mama y Ovario
Inhibe la expresión del
microRNA-155
BRCA1 p.Arg1699Gln (R1699Q)
Afecta al dominio BRCT domain
Riesgo acumulado del 24% a los 70 años Vs el 12% de la Población general
Se estima que en el futuro una caracterización mas precisa de las
variantes BRCA1 y BRCA2 de riesgo moderado permitirá el
desarrollo de estrategias terapéuticas específicas y distintas de las
que se aplican a los pacientes de alto riesgo
S. Chang et al., Nat. Med. 17, 1275–1282 (2011).
24. Fergus J. Couch et al. Science (2014) 34; 1466-1477
Síndrome de Cáncer de Mama y Ovario Hereditario
CHECK2 1100delC:
•1.2% de los controles europeos
•4.2% de los casos de Ca de Mama familiar BRCA1/BRCA2-negativos
•Riesgo de Ca de Mama del del 35% en >70a
PALB2 1592delT:
•Riesgo de Ca de Mama del 14% a los 50a y del 35% en >70a
•Se asocia a T. triple negativos y de Mal pronóstico
BRCA1: Riesgo de Ca de Mama del 50-70% a los 70a
BRCA2: Riesgo de Ca de Mama del 40-60% a los 70a
Weischer M, J Clin Oncol. 2008 Feb 1;26(4):542-8
26. 26
1.- Síndrome de Li-Fraumeni
• Gen TP53
• Criterios de Chompret ó mujeres con Ca de Mama <35 años
BRCA negativas
• Se asocia a tumores HER2+/ER+/PR+
2.- Síndrome de Cowden/PTHS
• Gen PTEN y SDHB-D, PIK3A, AKT1
• Riesgo de desarrollar Ca de Mama del 85% con una penetrancia
del 50% a partir de los 50 as (Ca de mama criterio mayor)
3.- Síndrome de Peutz-Jeghers
• Gen STK11/LKB1
• Riesgo de Ca de mama 44-50% y de Ca de ovario 18-21%
4.- Síndrome Hereditario del Cáncer Gástrico Difuso
• Gen CDH1
• Riesgo de Ca de mama 39-52%
29. ….RESULTADOS DE UN PANEL NGS AMPLIADO (39 genes)
Estudio de 198 pacientes de Alto riesgo:
- 57 pacientes BRCA positivas
- 141 pacientes BRCA negativas
- Se identificaron 16 variantes patogénicas en los genes:
MLH1
ATM
SLX4
BLM
NBN
CDH1
MUTYH
CDKN2A
PRSS1
AllisonW. Kurian, et al J Clin Oncol (2014)32: pp1-9
Problema: 428 Variantes de Significado incierto eran identificadas en 39 genes
Rentabilidad Diagnóstica: 11,3% en mujeres con Ca de Mama e Historia familiar
Cx profiláctica salpingo-ooforectomía
Vigilancia Intensiva Mama y/o
Gastrointestinal
30. Paneles de NGS multigen
30
1. Los paneles de NGS han demostrado ser una herramienta coste/efectiva
para la identificación de individuos y familias de alto riesgo.
2. Pueden dirigir decisiones clínicas y estrategias terapéuticas (pej:
identificación de mutaciones en genes de reparación)
A favor..
1. La interpretación clínica de los resultados es complicada. Así, la
penetrancia del cáncer de mama y el riesgo de desarrollo de otros tumores
no ha sido todavía establecido para las mutaciones patogénicas.
2. En la mayoría de los paneles hay un elevado número de VUS, cuya
interpretación causa ansiedad en la paciente y en el médico
3. Diversos paneles contienen genes no claramente asociados al Cáncer
de mama (pej: APC ó VHL)
En contra..
En cuanto a la Tasa de detección general, solo comentarte lo que probablemente ya sepas y es que BRCA1 y BRCA2 representan aproximadamente el 20-25% de los cánceres hereditarios de Mama (1) y aproximadamente un 5-10% del total de los cánceres de Mama (2). Además, estas mutaciones representan el 15% de los casos de Cáncer de Ovario(3). Estos números resultan difíciles de estimar desde el laboratorio dado que en muchos casos la historia de la paciente es insuficiente y no hacemos seguimiento de posterior de los casos.
Paología mamaria que incrementa el riesgo en >4
Cáncer de Mama (<40 años)
Carcinoma Lobular in situ
hiperplasia atípica confirmada
Tejido mamario de alta densidad
Terapias Hormonales como el Diethylstilbestrol
El Dx clínico es complejo PORQUE el HBOC es altamente heterogéneo a nivel biológico y genético debido a:
The discovery of BRCA1 and its sister
genes illustrates that the degree of biological complexity
underlying a phenotype is an excellent
predictor of its genetic heterogeneity (30).
Antecedente de una mutación conocida
Comentarios:
NCCN- National Comprehensive Cancer Network
Evidentemente sin antecedentes familiares de mutación previa
Según el US el antecedente de un cáncer de mama bilateral ya es criterio para hacer el test
En familias judías:
un antecedente de familiar de primer grado con mama u ovario
dos familiares de segundo grado en la misma rama de la familia con cáncer de mama u ovario.
- En el grupo 2) también se incluye el antecedente de bilateralidad
Dadas la implicaciones terapeúticas de la identificación de estas mutaciones, el estudio mutacional de BRCA1/2 debería ser incluido en el cribado de salud de mujeres adultas jóvenes…y como cualquier screening la frecuencia de resultados positivos sería baja, pero para aquellas mujeres en las que se detectase las consecuencias serían una mesurables e implicarían salvar una vida.
So what next? Given that 50% of BRCA1 and
BRCA2 mutations are inherited from unaffected
fathers, and given the small size of modern families,
almost 50% of women with BRCA1 and
BRCA2 mutations have little or no family history
of breast or ovarian cancer. Yet, cancer risks to
mutation carriers with no cancer family history are as high as risks to mutation carriers from severely
affected families (36). Identification of
cancer-causing mutations in BRCA1 and BRCA2
has clear and actionable implications for prevention.
BRCA1 and BRCA2 screening as part of
routine health care for young adult women is sensible
and feasible. As in any population-screening
program, genetic or otherwise, few participants
will prove positive, but for women who learn
that they carry mutations in BRCA1 or BRCA2,
the consequences are enormous, addressable, and
life-saving.
Until there are no more breast or ovarian cancers
among women with BRCA1 or BRCA2 mutations,
the real race is not over.
36. M.-C. King, J. H. Marks, J. B. Mandell; The New York
Breast Cancer Study Group, Science 302, 643–646 (2003).
2013, the U.S. Supreme Court ruled unanimously
that genes are products of nature and
therefore cannot be patented (31), nullifying the
Myriad patents on BRCA1 and BRCA2. The ruling
was a victory for science and for patients and led
immediately to broader availability of clinical genetic
testing.
For nearly 20 years, while Myriad was the
only commercial source in the United States for
genetic testing of BRCA1 and BRCA2, cost was a
major deterrent to widespread screening. The
cost to women of BRCA1 and BRCA2 testing is
now dropping, due both to the end of the monopoly
and to two scientific developments that have
changed the landscape. First, there are now enough
genes identified with mutations predisposing to
breast and ovarian cancer that multigene screening
panels can be developed and effectively implemented.
Second, genomic technology now offers the opportunity
to sequence at costs orders of magnitude
lower than the cost of Sanger sequencing (32).
Previously, clinical genetic testingwas carried out
gene by gene, based on specific clinical indications
and family histories, with each test costing thousands
of dollars. With the advent of massively
parallel sequencing, large panels of genes are now
screened simultaneously at far lower cost (33).
There was another barrier to genetic testing
for inherited breast and ovarian cancer. Some
patients and physicians worried that a positive
finding would lead to loss of health care coverage.
In consequence, mutations were not identified
in some womenwho could have been saved
by risk-reducing surgery. Clinical guidelines have
been established for women harboring damaging
mutations in BRCA1 and BRCA2, including increased
surveillance, surgical removal of ovaries
and fallopian tubes (salpingo-oophorectomy) by
age 40 years or younger, and the possibility of
risk-reducing mastectomy (34, 35). The Genetic
Information Nondiscrimination Act of 2008 (Public
Law 110–233), which protects mutation carriers
against loss of health care coverage, should
have removed fear as a barrier to testing, so that
women with mutations in BRCA1 and BRCA2
can be identified without economic reprisal.
es un mecanismo celular de vigilancia del ARN mensajero para detectar mutaciones terminadoras y evitar la expresión de proteínas truncadas o erróneas. En mamíferos, la NMD se inicia por el complejo de unión de exones (EJC) que se depositan durante el splicing del pre ARNm. Normalmente, un EJC es retirado por el ribosoma durante la primera ronda de traducción del ARNm. No obstante, si se da un EJC en un punto de la secuencia más allá ("downstream") del codón "sin sentido" (también llamado de parada o terminación), entonces este EJC aún permanecerá unido al ARMm cuando el ribosoma llegue al codón de terminación, y de ese modo servirá para desencadenar la NMD, ya que los factores proteicos implicados en esta función identifican la presencia de un EJC "corriente abajo" como un problema, de modo que el ARNm erróneo se degrada, por ejemplo mediante el exosoma.1 Con raras excepciones, un EJC no se deposita corriente abajo de un codón terminador.
Multiplex ligation-dependent probe amplification (MLPA)[1] is a variation of the multiplex polymerase chain reaction that permits multiple targets to be amplified with only a single primer pair.[1] Each probe consists of two oligonucleotides which recognize adjacent target sites on the DNA. One probe oligonucleotide contains the sequence recognised by the forward primer, the other the sequence recognised by the reverse primer. Only when both probe oligonucleotides are hybridised to their respective targets, can they be ligated into a complete probe. The advantage of splitting the probe into two parts is that only the ligated oligonucleotides, but not the unbound probe oligonucleotides, are amplified. If the probes were not split in this way, the primer sequences at either end would cause the probes to be amplified regardless of their hybridization to the template DNA, and the amplification product would not be dependent on the number of target sites present in the sample DNA. Each complete probe has a unique length, so that its resulting amplicons can be separated and identified by (capillary) electrophoresis. This avoids the resolution limitations of multiplex PCR. Because the forward primer used for probe amplification is fluorescently labeled, each amplicon generates a fluorescent peak which can be detected by a capillary sequencer. Comparing the peak pattern obtained on a given sample with that obtained on various reference samples, the relative quantity of each amplicon can be determined. This ratio is a measure for the ratio in which the target sequence is present in the sample DNA.
Genetics revealed that
BRCA1 is a tumor suppressor gene following the
two-hit model (25): Cancer develops as the result
of one inherited loss-of-function mutation followed
by a somatic mutation causing loss of the
remaining wild-type allele in a vulnerable cell
type. The central puzzle is why complete loss of
function of BRCA1 leads to cancer. Solving this
puzzle has been especially challenging, because
the BRCA1 protein is involved inmultiple essential
biological functions (26).
As part of a multiprotein complex, BRCA1
repairs double-strand DNA breaks via the homologous
recombination repair pathway. The
C-terminal BRCT domain interacts with histone
deacetylase complexes and is involved in transcriptional
regulation. The N-terminal RING domain
heterodimerizes with a sister domain of BARD1
and acts as a ubiquitin ligase of the estrogen receptor
(27). Missense mutations that abrogate the
function of the RING domain lead to breast cancer.
Virtually all other cancer-causing mutations of
BRCA1 are truncations: nonsense mutations, frameshifts,
or large genomic deletions or duplications
leading to stops and loss of the C-terminal domain
Las Mutaciones Pathogenicas representan aproximadamente el 30%of high-risk breast cancer families y explican el ~15% de los breast cancer familial relative risk (the ratio of the risk of disease for a relative of an affected individual to that for the general population) (Fig. 1)
Genetic testing for mutations in BRCA1, BRCA2, and other breast cancer susceptibility genes has served as a model for the integration of genomics into the practice of personalized medicine, with proven efficacy as a tool to determine eligibility for enhanced screening and prevention strategies,as well as a marker for targeted therapy.
Cambios intronicos no representados en el dibujo
Todas las mutaciones registradas
due to the large
number of Alu repeats in the genomic region
containing the BRCA1 gene
Founder mutations
(common mutations in a population arising
from a small number of individuals) in BRCA1
and BRCA2 have been described in almost every
population studied. The best known are in the
Ashkenazi Jewish population, with 3% of individuals
carrying one of the three founder mutations,
namely BRCA1 c.68_69delAG [185delAG]
(1%), BRCA1 c.5266dupC [5382insC] (0.13%),
or BRCA2 c.5946delT [6174delT] (1.52%) (14, 15).
Other examples are the BRCA1 c.548-?_4185+?del
[ex9-12del] mutation found in ~10% of Hispanic
BRCA carriers and deletions of BRCA1 seen in
Dutch founder populations (16, 17). Thus, targeted
screening for specific BRCA1 and BRCA2
mutations before full gene testing is warranted in
a number of populations.
The BRCA1 amino terminus contains a RING domain that associates with BRCA1-associated RING domain protein 1 (BARD1) and a nuclear localization sequence (NLS). The central region of BRCA1 contains a CHK2 phosphorylation site on S988 (Ref. 25). The carboxyl terminus of BRCA1 contains: a coiled-coil domain that associates with partner anbind RAD51. The BRCA2 DNA-binding domain contains a helical domain (H), three oligonucleotide binding (OB) folds and a tower domain (T), which may facilitate BRCA2 binding to both single-stranded DNA and double-stranded DNA46. This region also associates with deleted in split-hand/split-foot syndrome (DSS1)42, 44, 45. The C terminus of BRCA2 contains an NLS and a cyclin-dependent kinase (CDK) phosphorylation site at S3291 that also binds RAD51 (Ref. 53).
d localizer of BRCA2 (PALB2); a SQ/TQ cluster domain (SCD) that contains approximately ten potential ataxia-telangiectasia mutated (ATM) phosphorylation sites and spans amino acid residues 1280–1524; and a BRCT domain that binds ATM-phosphorylated abraxas, CtBP-interacting protein (CtIP) and BRCA1-interacting protein C-terminal helicase 1 (BRIP1). The BRCA1–abraxas complex is associated with BRCA1 recruitment to sites of DNA damage19, 20, 108, 109. The BRCA1–BRIP1 complex, which also contains DNA topoisomerase 2-binding protein 1 (TOPBP1), is associated with DNA repair during replication110. The BRCA1–CtIP complex promotes ataxia-telangiectasia and Rad3-related (ATR) activation and homologous recombination (HR) by associating with the MRN complex (which is comprised of MRE11, RAD50 and Nijmegen breakage syndrome protein 1 (NBS1)) and facilitating DNA double-strand break resection22. The central region of BRCA1, which contains the SCD, is phosphorylated by ATM. This phosphorylation is important for BRCA1-mediated G2/M and S-phase checkpoint activation, as expression of a BRCA1 mutant that lacks three of the phosphorylation sites (S1387, S1423 and S1524) fails to rescue defective checkpoint activation and ionizing radiation hypersensitivity in a BRCA1-deficient cell line111, 112. b | The N terminus of BRCA2 binds PALB2 at amino acids 21–39 (Ref. 68). BRCA2 contains eight BRC repeats between amino acid residues 1009 and 2083 that
More than 1800 distinct rare variants—in the form of
intronic changes, missense mutations, and small
in-frame insertions and deletions—have been reported
inBRCA1and2000 inBRCA2 (BreastCancer
Information Core; www.research.nhgri.nih.gov/bic).
In BRCA1, missense mutations that are pathogenic
and highly penetrant (i.e., confer a high
risk of cancer) are located primarily in the RING
finger and BRCT domains (2, 9, 10), which are
critical for the DNA repair activity of BRCA1. In
BRCA2, highly penetrant, pathogenic missense
mutations are located predominantly in the DNA
binding domain (11, 12).
As studies of BRCA1 and BRCA2 unfolded, it
became apparent that the estimates of penetrance
(risk) of breast and ovarian cancer varied by the
ascertainment criteria for studies, with populationbased
studies showing much lower risks than
family-based studies (18). In clinical practice,
BRCA1 mutation carriers are generally estimated
to have a 57% chance of developing breast cancer
and a 40% chance of developing ovarian
cancer by age 70, whereas BRCA2 mutation carriers
are estimated to have a 49% chance of breast
cancer and an 18%chance of ovarian cancer (19).
Interindividual variability in the risk of breast and
ovarian cancer has been attributed to modifying
environmental and genetic effects, including the
location and type of mutations in BRCA1 and
BRCA2. Specifically, early reports focused on the
location of mutations in BRCA1/2 suggested that
nonsense and frameshift mutations located in the
central regions of either coding sequence, termed
ovarian cancer cluster regions (OCCR), were associated
with a greater risk of ovarian cancer than
similar mutations in the proximal and distal regions
of each gene (20–22). More recently, a Consortium
of Investigators of Modifiers of BRCA1/2
(CIMBA) study of 19,581 BRCA1 and 11,900
BRCA2 mutation carriers confirmed relative increases
in ovarian cancer and decreases in breast
cancer risk for mutations in the central region of
each gene and higher risk of breast cancer for
mutations in the 5′ and 3′ regions of each gene.
Variability in risk is also partly explained by common
genetic modifiers of breast and ovarian cancer
risk in BRCA1 and BRCA2 mutation carriers
that have been identified through genome-wide
association studies (23–29). Accounting for these
modifiers suggests that the BRCA1 mutation carriers
in the highest risk category may have an
81%or greater chance of breast cancer and a 63%
or greater chance of ovarian cancer by age 80,
whereas BRCA2 mutation carriers at greatest risk
may have more than an 83% chance of breast
cancer by age 80 (27, 28). In conjunction with other
variables modifying risk in BRCA1 and BRCA2
mutation carriers, these data offer the potential
for more precise personalized risk estimates.
The classification of VUS may be further
complicated by hypomorphic mutations in both
BRCA1 and BRCA2, which retain partial protein
activity and may be associated with moderate to
low risks of breast and ovarian cancer. The best
characterized of thesemutations is the p.Arg1699Gln
(R1699Q) missense mutation in the BRCT domain
of BRCA1 that abrogates the repression of
microRNA-155 (38) and is associated with a cumulative
risk of breast cancer of 24% by age 70
(30). This risk is lower than that associated with
other BRCA1 mutations but substantially greater
than the 12% risk of breast cancer in the general
population. In contrast, the well-known polymorphic
stop codon in BRCA2, p.Lys3326X, is associated
with only a modest increase in breast
cancer risk [odds ratio (OR) = 1.26] (39) and appears
to have little clinical relevance. As more
moderate risk variants in BRCA1 and BRCA2 are
validated, risk management strategies distinct from
those applied to carriers of high-risk mutations
must be developed.
Fig. 1. Genetic variants that predispose to breast cancer. The pie chart
on the left shows the estimated percentage contribution of mutations in highpenetrance
(BRCA1/2, TP53, CDH1, LKB1, and PTEN) and moderate-penetrance
(e.g., CHEK2, ATM, and PALB2) genes and common low-penetrance genetic
variants to familial relative risk.
“Known SNPs” are SNPs associated with breast cancer through GWAS, as listed
on the right. The odds ratios refer to the increase (or, in some cases, the
reduction) in risk conferred by the rare allele of the variants. “Other predicted
SNPs” refers to the estimated contribution of all SNPs, other than known loci,
that were selected for replication of breast cancer GWAS (5, 39).
Women with an abnormal PALB2 gene have a 14% risk of developing breast cancer by age 50 and a 35% risk of developing breast cancer by age 70. In comparison, women with an abnormal BRCA1 gene have a 50%-70% risk of developing breast cancer by age 70. Women with an abnormal BRCA2 gene have a 40%-60% risk of developing breast cancer by age 70.
The other breast and ovarian cancer genes
also harbor many different rare, recent damaging
mutations with effect sizes ranging from twofold
increased risk for CHEK2 to 10-fold for TP53.
Of the seven families in our 1990 linkage
analysis with young-onset breast cancer (3), six
families harbor mutations in BRCA1, and one
harbors a mutation in BRCA2. Of the 16 families
in that analysis that we predicted would not carry
mutations in BRCA1, six are explained by BRCA2;
one each is explained by PALB2,CDH1, andSLX4;
and seven remain unsolved. There are more breast
cancer genes to be found.
The critical genes known
thus far encode proteins in the same and related
pathways.
Genetic variants that predispose to breast cancer. The pie chart
on the left shows the estimated percentage contribution of mutations in highpenetrance
(BRCA1/2, TP53, CDH1, LKB1, and PTEN) and moderate-penetrance
(e.g., CHEK2, ATM, and PALB2) genes and common low-penetrance genetic
variants to familial relative risk. Common genetic variants are denoted as SNPs.
“Known SNPs” are SNPs associated with breast cancer through GWAS, as listed
on the right. The odds ratios refer to the increase (or, in some cases, the
reduction) in risk conferred by the rare allele of the variants. “Other predicted
SNPs” refers to the estimated contribution of all SNPs, other than known loci,
that were selected for replication of breast cancer GWAS (5, 39).
Li-Fraumeni syndrome (caused by germline
mutations in TP53), Cowden disease (caused
by germline mutations in PTEN), and Peutz-Jeghers
syndrome (caused by germlinemutations in STK11)
These panels have proven effective in identifying
individuals and family members at elevated risk
of breast and other cancers. However, clinical
interpretation of results from the panels is complicated
by several factors. In particular, breast
cancer penetrance and risk of other cancers has
not yet been established for pathogenicmutations
in most of the panel genes, and guidelines for
clinical management of individuals found to carry
these mutations have not been developed (77).
Additionally, as is true for BRCA1/2, there is a
high rate of VUS in the panel genes, the interpretation
of which causes anxiety for both the
patient and the physician. Furthermore, several
commercial panels contain genes such as APC
and VHL, which have not been clearly associated
with susceptibility to breast cancer (78). Although
continued clinical research is needed to responsibly
integrate panel testing to practice, such approaches
may provide guidance for critical clinical
decisions such as whether a patient is at high risk
of contralateral breast cancer and/or should undergo
risk reduction surgeries. Conceivably, panel
testing also may prove useful for selecting patients
for treatment with PARP inhibitors, because
several of the genes in current panels encode
proteins involved in double-strand break repair,
which may influence responsiveness to platinum
and potentially PARPi (79).
BRCA1 y BRCA2 representan el 20-25% de los cánceres de Mama
INCIDENCIAS
BRCA 1/500-2500
PTEN 1/200.000
LYNCH 1/200-1000
PJ 1/120.000
LI FRAMUNENI Ycancer gástrico difuso RARO
Cowden syndrome tb llamado PTHS (PTEN hamartona tumor Syndrome)
Fijar en Dermatológico (Hamartomas)/circunferencia craneal/Tiroides
El Ca de Ovario de PJ es habitualmente cordoma
El ca en mutaciones CDH1 es lobular
INCIDENCIA DE LYNCH 1/200-1000
Asi mismo, los paneles que contienen genes implicados en la reparación del DNA de doble cadena pueden identificar pacientes potencialmente resistente a platinos y candidatos a tratamientos específicos como los inhibidores PARP
These panels have proven effective in identifying
individuals and family members at elevated risk
of breast and other cancers. However, clinical
interpretation of results from the panels is complicated
by several factors. In particular, breast
cancer penetrance and risk of other cancers has
not yet been established for pathogenicmutations
in most of the panel genes, and guidelines for
clinical management of individuals found to carry
these mutations have not been developed (77).
Additionally, as is true for BRCA1/2, there is a
high rate of VUS in the panel genes, the interpretation
of which causes anxiety for both the
patient and the physician. Furthermore, several
commercial panels contain genes such as APC
and VHL, which have not been clearly associated
with susceptibility to breast cancer (78). Although
continued clinical research is needed to responsibly
integrate panel testing to practice, such approaches
may provide guidance for critical clinical
decisions such as whether a patient is at high risk
of contralateral breast cancer and/or should undergo
risk reduction surgeries. Conceivably, panel
testing also may prove useful for selecting patients
for treatment with PARP inhibitors, because
several of the genes in current panels encode
proteins involved in double-strand break repair,
which may influence responsiveness to platinum
and potentially PARPi (79).
As part of personalizing risk
assessment, these genomic insights may soon
form a rational and cost-effective basis for selection
of women for breast cancer screening (91, 92).
Going forward, the reduced cost and increased
access to genomic profiling of breast tumors will
likely identify new therapeutic targets. However,
the anticipated increased uptake of sequencing
will require new approaches for communication
to patients of findings from germline DNA that
suggest increased risk for treatment toxicities or
risk for disorders other than breast cancer (90, 93, 94).
Two decades after the cloning of the BRCA genes,
clinical application of findings of breast cancer
genetic research continues to drive new paradigmsparadigms
of “personalized” genomics and precision
medicine.