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Breast Cancer Res Treat (2015) 150:7180

DOI 10.1007/s10549-015-3293-7

PRECLINICAL STUDY

Prevalence of BRCA1 and BRCA2 germline mutations in patients


with triple-negative breast cancer
Michelle W. Wong-Brown Cliff J. Meldrum Jane E. Carpenter
Christine L. Clarke Steven A. Narod Anna Jakubowska Helena Rudnicka
Jan Lubinski Rodney J. Scott

Received: 28 January 2015 / Accepted: 31 January 2015 / Published online: 15 February 2015
Springer Science+Business Media New York 2015

Abstract Triple-negative breast cancers (TNBC) lack


expression of oestrogen, progesterone and HER2 receptors.
The gene expression profiles of TNBCs are similar to those
of breast tumours in women with BRCA1 mutations. Reports to date indicate that up to 20 % of TNBC patients
harbour germline BRCA mutations; however, the prevalence
of BRCA mutations in TNBC patients varies widely between
countries and from study to study. We studied 774 women
with triple-negative breast cancer, diagnosed on average at
age 58.0 years. Samples of genomic DNA were provided by
the Australian Breast Cancer Tissue Bank (ABCTB) (439
patients) and by the Department of Genetics and Pathology

Electronic supplementary material The online version of this


article (doi:10.1007/s10549-015-3293-7) contains supplementary
material, which is available to authorized users.
M. W. Wong-Brown  R. J. Scott (&)
School of Biomedical Sciences & Pharmacy, Centre for
Information-Based Medicine, Hunter Medical Research Institute,
University of Newcastle, Lot 1 Kookaburra Circuit, New
Lambton Heights, Newcastle, NSW 2305, Australia
e-mail: rodney.scott@newcastle.edu.au
C. J. Meldrum  R. J. Scott
Division of Molecular Medicine, Pathology North, NSW
Pathology, Lookout Road, Newcastle 2305, NSW, Australia
J. E. Carpenter  C. L. Clarke
Australian Breast Cancer Tissue Bank, University of Sydney at
the Westmead Millennium Institute, Westmead, NSW, Australia
S. A. Narod
Familial Breast Cancer Research Unit, Womens College
Research Institute, Toronto, Canada
A. Jakubowska  H. Rudnicka  J. Lubinski
Department of Genetics and Pathology, Pomeranian Medical
University, Szczecin, Poland

of the Pomeranian Medical University (335 patients). The


entire coding regions and the exonintron boundaries of
BRCA1 and BRCA2 were amplified and sequenced by nextgeneration sequencing. We identified a BRCA1 or BRCA2
mutation in 74 of 774 (9.6 %) triple-negative patients. The
mutation prevalence was 9.3 % in Australia and was 9.9 %
in Poland. In both countries, the mean age of diagnoses of
BRCA1 mutation carriers was significantly lower than that of
non-carriers, while the age of onset of BRCA2 mutation
carriers was similar to that of non-carriers. In the Australian
cohort, 59 % of the mutation-positive patients did not have a
family history of breast or ovarian cancer, and would not
have qualified for genetic testing. The triple-negative phenotype should be added as a criterion to genetic screening
guidelines.
Keywords Triple-negative breast cancer  BRCA1 
BRCA2  Germline mutations  Prevalence  Genetic testing
List of abbreviations
BRCA Breast cancer susceptibility gene
DNA
Deoxyribonucleic acid
PARP Poly (adenosine diphosphate)-ribose polymerase
TNBC Triple-negative breast cancer
UV
Unclassified variant
pCR
Pathological complete response

Introduction
TNBC describes a subgroup of breast cancers that are
negative for the oestrogen, progesterone and HER2 receptors. They account for between 15 and 20 % of all
breast cancers diagnosed but are over-represented in young
women and in black women in the United States.

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Breast Cancer Res Treat (2015) 150:7180

Table 1 TNBC demographic and clinical data of the Australian and


Polish patient cohorts
Characteristic

Australian
patients,
N = 439 (%)

Polish patients, N = 335


(%)

57 15 years

59 10 years

Australian cohort
Age at diagnosis
B50 years

153 (34.9)

49 (14.6)

[50 years

286 (65.1)

286 (85.4)

Breast cancer only

80 (18.2)

Information only available


for mutation carriers (see
Table 2)

Breast and ovarian


cancer

11 (2.5)

Breast and prostate


cancer

2 (0.5)

Breast and other


cancer

43 (9.8)

Ovarian cancer
only

5 (1.1)

Ovarian and other


cancer

6 (1.3)

Other cancer

85 (19.4)

No recorded family
history of cancer

207 (47.2)

Family history of
cancer

Methods
Study cohort

Type of primary tumour


Ductal (Invasive/
in situ/not
otherwise
specified)

409 (93.2)

Papillary

3 (0.7)

2 (0.6)

Medullary

10 (2.3)

42 (12.5)

Apocrine

4 (1.2)

Lobular (Invasive/
in situ/not
otherwise
specified)

9 (2)

14 (4.2)

Metaplastic

7 (2.1)

3 (0.9)

8 (1.8)

36 (10.7)
N = 219

Others
Unknown
Tumour grade

227 (67.8)

6 (1.4)

1 (0.5)

48 (10.9)

54 (24.6)

374 (85.2)

143 (65.3)

Not known

11 (2.5)

21 (9.6)

BRCA1 and BRCA2 are breast cancer susceptibility genes that are part of the DNA repair pathway. Pathogenic
mutations in both genes confer a high risk of breast cancer
[1], and together they account for approximately 5 % of all
breast cancer cases [2]. A large proportion of tumours in
women with a BRCA1 mutation exhibited a triple-negative
phenotype. Not all women with breast cancer qualify for
BRCA1 and BRCA2 testing. Testing is currently based on

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the age of onset, the family history and in some cases (e.g.
Ashkenazi Jews) on the ethnic group. Currently, tumour
histology is not a clear criterion for the recommendation
for genetic testing, but some have suggested that all women
with triple-negative breast cancer be candidates for genetic
testing, regardless of age of onset or family history. With
the advent of next-generation DNA sequencing (NGS), it is
cost-effective to perform BRCA1 and BRCA2 testing to
women outside of a familial cancer setting. Several studies
have reported that up to 20 % of women with TNBC breast
cancer carry a BRCA mutation [312]. The aim of this
study was to define the prevalence of germline BRCA1 and
BRCA2 mutations in two independent populations of consecutively collected TNBC cases, in Poland and in Australia, unselected for family history and age of diagnosis.

439 patients with TNBC from Australia were included.


Cases were selected based on triple-negative status and not
on age of onset or family history. The demographic and
clinical data are described in Table 1. This study was approved by the Hunter New England Health Human Research Ethics Committee. Samples were provided by the
Australian Breast Cancer Tissue Bank (ABCTB).
DNA samples from 335 Polish patients with TNBC were
provided by the Department of Genetics and Pathology,
Pomeranian Medical University. The study was approved
by the local institutional ethics review committee for participation in this study. There are three common BRCA1
founder mutations in the Polish population. It is currently
recommended that all breast cancer patients in Poland be
tested for these three mutations and, for the purposes of the
current study, patients with one of these mutations were
excluded. 49 women were diagnosed under 50 years of age
and 286 were diagnosed over age 50. The average age of
diagnosis of these two patient groups is shown in Table 1.
BRCA1 and BRCA2 sequencing
Target-specific primers were designed by Fluidigm Corp.
(San Francisco, CA). Common sequence tags (CS1 and
CS2) were added to the forward and reverse primers for
Access Array amplicon tagging. 184 primer pairs were
designed to cover all coding exons of BRCA1 and BRCA2.
Genomic DNA from TNBC patients were normalised to
50 ng/ll concentration and 1 ll of the solution was loaded
onto a Fluidigm Access Array, (a microfluidic array in
which many concurrent PCR reactions were performed
with nested primer pairs). A two-primer protocol was used

Exon

BRCA1

p.Val233Asnfs*4
p.Leu502Alafs*2
p.Gln563*
p.Lys652Glufs*21

p.Lys654Serfs*47

c.213-11T[G

c.514delC

c.697_698del

c.1504_1508del

c.1687C[T

c.1952dup

c.1961del

int 5

11

p.Gln172Asnfs*62

Results in frameshift

Activates cryptic splice


acceptor site. Leads to
aberrant splicing and
insertion of 59 bp

p.Cys61Gly

Skip of exon 5 very likely

c.181T[G

Intronic retention of 58 bp

p.Cys24Serfs*13

c.70_80del

c.135-2A[G

p.Glu23Valfs*17

Amino acid change

c.68_69del

Nucleotide
change

int 3

Australian cohort

Gene

rs80357747

rs80357522

rs80357885

rs80356898

rs80357888

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

rs80358061

rs80357872

Y/class 5pathogenic

Y/UV

Y/class 5pathogenic

Y/class 5pathogenic

BIC
(Y/Nclass)

rs28897672

rs80358065

rs80359877

rs80357713

dbSNP

45

03-11-043

02-12-076

02-08-102

05-08-009

05-11-096

04-08-214

08-12-105

01-10-155

04-10-067

07-12-007

01-10-014

47

36

62

68

43

41

69

41

37

47

40

55

03-09-148

03-11-100

26

Age of
diagnosis

01-07-014

Patient

Mother, ovarian, 66

Paternal uncle, bowel, 60

Maternal grandmother, breast, unknown

Mother, breast cancer, 45

Paternal cousin, breast, 40 s

Paternal aunt, breast, 50 s

Paternal aunt, breast, 50 s

Maternal niece, breast, 39

Sister, colorectal, 55

Sister, ovarian, 53

Father, pancreatic, 76

Mother, breast, 73

Brother, bowel, unknown

Grandfather, prostate, unknown

Maternal aunt, stomach, 75

Mother, cervical, 38

Maternal cousin, breast, 50 s

Paternal grandmother, breast, 90 s

Mother, breast, 38

Maternal grandaunt, ovarian, 20 s

Maternal grandfather, brain, 30

Paternal grandaunt, stomach, 50

Paternal grandmother, pancreas, 77

Paternal grandfather, leukaemia, 52

Paternal aunt, breast, 36

Father, non-Hodgkins
lymphoma and colon, 75

Maternal 2nd cousin, breast, 45

Maternal aunt, breast, 47

Maternal cousin, ovarian, 45

Family history
(relationship, type of
cancer, age of diagnosis)

Table 2 Deleterious mutations detected in the Australian and Polish cohorts and any recorded family history of diseases associated with the mutation carriers

Breast Cancer Res Treat (2015) 150:7180


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123

123

BRCA2

Gene

c.5266dup

c.5272A[T

20

p.Cys419Trpfs*11
p.Phe620Leufs*24
p.Thr630Asnfs*14

c.1257del

c.1860del

c.1889del

10

Classified as pathogenic
according to kConFab

Skip of exon 7 is very likely

c.631?2T[G

int 7

p.Leu105*

c.314T[G

p.Leu1764*

p.His1732Phefs*5

p.Thr1677Ilefs*2

c.5289del

p.Arg1758*

c.5194-12G[A

int 19

21

p.Gln1756Profs*74

c.5030_5033del

17

p.Trp1508*

p.Leu1404*

p.Ser1253Argfs*10

c.3756_3759del

c.4523G[A

p.Gln1200*

c.3598C[T

c.4210del

p.Ile946Tyrfs*6

c.2835dup

15

Amino acid change

Nucleotide
change

13

Exon

Table 2 continued

rs80359315

rs80359272

rs81002899

rs80358561

rs80357906

rs80358079

rs80357862

rs80357765

rs80357868

rs62625307

rs80357519

dbSNP

Y/class 5pathogenic

Y/class 5pathogenic

Y/UV

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

BIC
(Y/Nclass)

05-10-073

03-07-065

01-08-025

01-06-021

48

56

84

34

89
46

05-09-110

55

36

47

45

55

45

04-11-051

01-11-082

03-09-059

04-10-126

01-08-061

02-08-30S

01-12-021

29

33

04-11-088
03-06-041

30

43

44

Age of
diagnosis

02-08-124

08-10-069

02-08-049

Patient

Maternal grand-aunt, breast, 70

Paternal grandfather, unknown, 80

Mother, breast, 39

Maternal uncle, colorectal, 60

Maternal uncle, colorectal, 60

Maternal uncle, liver, 23

Maternal 2nd cousin, breast, 50 s

Maternal grandfather, colorectal, 68

Maternal cousin, breast, 33

Mother, breast, 58

Paternal cousin, breast, unknown

Paternal aunt, breast, unknown

Maternal aunt, ovarian, 40

Mother, breast, 40

Mother, bladder, 74

Maternal great-aunt, breast, 50

Maternal grandmother, breast, 60

Mother, breast, 60

Mother, breast, 47

Paternal aunt, breast, 50

Maternal cousin, breast, 35

Maternal aunt, breast, 40

Paternal aunt, breast, 60 s

Paternal great-grandmother, breast, 80 s

Maternal aunt, breast, unknown

Cousin, breast

Cousin, breast

Father, throat, 40 s

Maternal aunt, ovarian, 42

Family history
(relationship, type of
cancer, age of diagnosis)

74
Breast Cancer Res Treat (2015) 150:7180

Exon

11

16

BRCA1

p.Gln1273*

c.3817C[T

BRCA2

p.Gln1096*

c.3286C[T

c.4689C[A

c.2886dup

p.Tyr1563*

p.Ile963Tyrfs*19

p.Gln1396*

p.Gln1090*

c.3268C[T

c.4186C[T

p.Gly563*

c.1687C[T

13

p.Gly284*

c.80?2T[C

c.850C[T

i2

11

Amino acid change

p.Thr2766Asnfs*11

Nucleotide change

c.8297del

18

p.Ile2315Lysfs*12

p.Val2228Glyfs*5

c.6944_6947del

13

c.6682dup

p.Asn1784Thrfs*7

c.5351del
p.Leu1908Argfs*2

p.Tyr1710*

c.5130_5133del

p.Ser1970*

p.Asp1469Lysfs*11

c.4405_4409del

c.5909C[A

p.Gln1452*

c.4354C[T

c.5722_5723del

p.Ala938Profs*21

c.2808_2811del

11

Amino acid change

Nucleotide
change

Exon

BRCA1

BRCA1

Polish cohort

Gene

Gene

Table 2 continued

rs80357433

rs80357011

rs80357208

rs80357485

rs80357402

rs80356898

rs80358128

dbSNP

rs80359705

rs80359629

rs80359621

rs80358824

rs80359531

rs80359509

rs80359485

rs80359352

dbSNP

Y/class 5pathogenic

NUV

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

BIC (Y/N Class)

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

BIC
(Y/Nclass)

01-08-037

08-11-063

04-08-036

05-10-118

04-09-164

08-10-074

02-09-527

01-12-019

03-11-109

04-12-030

Patient

43

136,755

160,333#

220,693

161,467

36

41

54

53

62

55

349,099
192,682

33
59

342,887

40

Family history
of breast
and/or ovarian
cancer (Y/N)

Brother, melanoma, 35

Paternal cousin, breast, 40

Mother, multiple myeloma


& lung, 60

Maternal grandmother,
melanoma, 90 s

Brother, skin, 40 s

Father, skin, 60 s

Father, prostate, 65

Daughter, thyroid, 54

Maternal grandmother,
gastrointestinal, unknown

Brother, prostate, 59

Father, bowel, 54

Maternal aunt, breast, 50 s

Mother, breast, 50 s

Family history
(relationship, type of
cancer, age of diagnosis)

Age of
diagnosis

285,146

228,532

146,990

Patient

44

44

39

63

57

83

60

78

58

59

Age of
diagnosis

Breast Cancer Res Treat (2015) 150:7180


75

123

123

c.9097dup

c.9253dup

24

p.Glu2089Aspfs*2

c.6267_6269delinsC

c.8946del

p.Tyr1894*

c.5682C[G

23

p.Ser1882*

c.5645C[A

22

p.Asn1784Lysfs*7

c.5352del

c.7558C[T

p.Asn1747*

c.5237dup

15

p.Lys1025Asn;Lys1026*

c.3075_3016delinsTT

11

p.Thr3085Asnfs*26

p.Thr3033Asnfs*11

p.Asp2983Ilefs*5

p.Arg2520*

p.Asn433Glnfs*18

p.Val220Ilefs*4

c.1296_1297del

p.Arg1835*

c.658_659del

c.5503C[T

24

p.Thr1677Ilefs*2

10

c.5278-2A[T

i20

Intronic retention of 65 bp

c.5030_5033del

17

p.Met1663Valfs*14

p.Glu1661*

c.4981G[T

c.4986?3G[C

Amino acid change

Nucleotide change

i16

Exon

Patient 160,333 harbours both BRCA1 and BRCA2 mutations

BRCA2

Gene

Table 2 continued

rs80359752

rs80359747

rs80358981

rs41293497

rs80358785

rs80359499

rs80358552

rs80359276

rs80359604

rs41293465

rs80357862

rs80358023

rs80357401

dbSNP

Y/class 5pathogenic

Y/class 5pathogenic

NUV

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic
COSMICdistribution in
ovary and endometrium

N/UV

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

Y/class 5pathogenic

COSMICdistribution in
ovary and large intestine

Y/UV

NUV

Y/class 5pathogenic

Y/Class 4likely pathogenic

Y/class 5pathogenic

BIC (Y/N Class)

33
41

73,032
353,587

53

174,157

192,325

285,224

266,895

99,173

151,271

45

68

73

62

52

58
50

195,860

68

349,466

346,635

63

273,641

49

52

171,190
87,035

58

71

77

58

66

38

54

135,383

119,568

334,861

173,119

156,407

166,662

94,836

62

53

101,115

136,084

Age of
diagnosis

Patient

Family history
of breast
and/or ovarian
cancer (Y/N)

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Breast Cancer Res Treat (2015) 150:7180

Breast Cancer Res Treat (2015) 150:7180

Distribution of age of diagnosis of BRCA mutation carriers


40

% of patients within mutation status

Fig. 1 Distribution of age of


diagnosis of A. BRCA mutation
carriers (Australian
average = 50.3 15 years,
Polish average = 53.6
11.5 years). The average age of
Australian and Polish BRCA1
mutation carriers is
47.2 11.8 years and average
age of BRCA2 mutation carriers
is 58.8 13.2. B. The
distribution of the age of
diagnosis of the non-mutation
carriers (average age of nonmutation carriers is 58.7 12.7
years)

77

35

BRCA1 mutation carriers

30

BRCA2 mutation carriers

25
20
15
10
5
0

25-34

35-44

45-54

55-64

65-74

75-84

85

Age of diagnosis (years)

% of patients within non-mutation


status

Distribution of age of diagnosis of patients without mutations


40
35

Non-mutation carriers

30
25
20
15
10
5
0

25-34

35-44

45-54

55-64

65-74

75-84

85

Age of diagnosis (years)

to first amplify the target sequence followed by a second PCR


reaction that included the sequencing tags such that ultimately
each amplified product contained the target-specific sequence,
a tag sequence and a sample-specific unique barcode.
The exons were amplified and the PCR products were
harvested from each sample, pooled, and purified using
Ampure XP beads. The libraries were then sequenced
on an Illumina MiSeq instrument.
The sequencing data were analysed with NextGENe 2nd
Generation Sequencing Software v.2.3.3. (SoftGenetics,
Philadelphia). Raw data were converted into FASTA files
and aligned to BRCA1 and BRCA2 reference sequences from
the human reference sequences; GRCh37 Primary Assembly, BRCA1 NC_000017.10 and BRCA2 NC_00013.10
(http://www.ncbi.nih.gov). The CS1 and CS2 sequences
were trimmed, as well as the primer sequences to eliminate
the dilution of base calling from those regions.
The mutations were described using the nomenclature
guidelines of the Human Genome Variation Society (http://
www.hgvs.org). The DNA sequence numbering is based on

the cDNA sequences for BRCA1 (NM_ 007294.3) and


BRCA2 (NM_ 000059.3).
BRCA1 or BRCA2 mutations were only included in the
analyses if classified as deleterious or possibly deleterious
according to Breast Cancer Information Core (BIC) criteria
or if present in the BIC database [13]. The criteria are all
frameshift and nonsense variants (except those resulting in
neutral or polymorphic stop codons), all intronic variants in
the consensus splice acceptor or donor sites (within 2 bp of
exonintron junctions or if shown to result in aberrant
mRNA transcript processing, or non-synonymous variants
shown to have deleterious effect on known functional regions (based on functional or biochemical assays, or linkage analysis of high-risk families) [14].

Results
We identified 74 mutations in the 774 triple-negative breast
cancer patients (9.6 %). Mutations were present in 9.3 %

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78

40.0

Proportion of BRCA mutationpositive patients (%)

Fig. 2 The proportion of


patients with or without
recorded family histories of
breast and/or ovarian cancer in
each age group in those with
germline BRCA mutations. In
patients with family histories of
disease: \40 years17.1 %,
4060 years22 %,
C60 years2.4 %. In patients
without recorded family
histories of disease:
\40 years4.9 %,
4060 years36.6 %,
C60 years17.1 %

Breast Cancer Res Treat (2015) 150:7180

Distribution of age of diagnosis of patients with versus without


recorded family history of breast and/or ovarian cancer

35.0
30.0

With family history

25.0

Without recorded family


history

20.0
15.0
10.0
5.0
0.0
<40

40-60

>60

Age of diagnosis (years)

Table 3 Percentage of patients with BRCA1 or BRCA2 mutations compared to non-mutation carriers diagnosed with breast cancer
Age of diagnosis (years)

BRCA1 mutation carriers

BRCA2 mutation carriers

Non-mutation carriers

Australian breast cancer cohort

(%, n = 26)

(%, n = 15)

(%, n = 398)

\40

26.9

13.3

12.6

4060

57.7

60

43.7

C60

15.4

26.7

43.7

(%, n = 18)

(%, n = 15)

(%, n = 302)

\40

22.2

2.3

4060

61.1

60

54

C60

16.7

40

43.7

Polish breast cancer cohort

of the Australian patients and in 9.9 % of the Polish patients (see Table 2 for a complete list of deleterious mutations and Supplementary Table 1 for all other alterations
identified in BRCA1 and BRCA2). In Australia, 63 % of
the mutations were in BRCA1; in Poland 54.5 % of the
mutations were in BRCA1. The distribution of the age of
diagnoses is shown in Fig. 1.
In the Australian cohort, four of the 41 mutations are
novel and have not been reported in BIC, Leiden Open
(source) Variation Database (LOVD), Clinvar, or any
published literature. Of these, two are in BRCA1
(c.4523G[A, c.5272A[T) and two in BRCA2 (c.1860del,
c.4354C[T).
Among the 41 Australian patients with a mutation, 17
(41.5 %) had a family history of breast or ovarian cancer
(Table 2). In the Australian cohort, the mean age of disease
onset for BRCA1 mutation carriers is 46.0 years of age and
for BRCA2 mutation carriers is 57.6 years of age, and for
non-carriers, it was 57.8 years of age (p = 0.004).
A deleterious mutation was detected in 33 of 335 patients in Poland (9.9 %). Two mutations have not been
reported in BIC, Leiden Open (source) Variation Database
(LOVD) or Clinvar. These include one in BRCA1
(c.80?2T[C) and one in BRCA2 (c.2886dup).

123

Among the 33 Polish patients with a mutation, 19


(57.5 %) had a family history of breast or ovarian cancer
(Table 2). In the Polish cohort, the mean age of onset for
BRCA1 mutation carriers is 48.9 years of age, whereas for
BRCA2 mutation carriers, it is 59.9 years of age and for
non-mutation carriers, it is 60 9.8 years of age
(p = 0.004). The distribution of age of diagnosis of our
patient cohort according to age groups in those with and
without BRCA mutations is shown in Fig. 2.

Discussion
In both Poland and in Australia, approximately 10 % of the
patients with triple-negative phenotype harbour a germline
BRCA1 or BRCA2 mutation. Our study highlights the
importance of genetic testing for patients with TNBC regardless of age and in the absence of a family history. In
Poland, genetic testing for three founder mutations is already recommended for all breast cancer patients. We
recommend that in the absence of one of the founder mutations, Polish patients with triple-negative breast cancer be
considered for additional genetic testing, which should
include full sequencing of the coding regions of both genes.

Breast Cancer Res Treat (2015) 150:7180

In both countries, BRCA2 mutations were less common


than BRCA1 mutations, in agreement with other studies [3,
8, 15, 16]. These results may reflect the propensity for
BRCA1 carriers to develop primarily triple-negative cancers, whereas among BRCA2 carriers, the majority of
breast cancers are oestrogen-receptor positive. BRCA2
mutation carriers were shown in our study to be diagnosed
with TNBC at an older age than BRCA1 mutation carriers
and similar to the age of onset of non-carriers.
Recent guidelines by the National Comprehensive
Cancer Network (NCCN Guidelines in Oncology Version
3.2013 Genetic/Familial High-Risk Assessment: Breast and
Ovarian) recommends that TNBC diagnosed before or at
age 60 is considered to be sufficient to meet the threshold
of risk assessment for genetic testing [17]. In contrast, the
Cancer Institute New South Wales guidelines recommend
that patients with TNBC be offered testing if diagnosed
under the age of 40 years. [18]. From our results, 33.3 % of
BRCA2 mutation carriers in both the Australian and Polish
cohorts are over 60 years of age, therefore according to the
NCCN guidelines, they would not qualify for genetic
testing (see Table 3). Our results suggest that the age of
diagnosis should not be a limitation to genetic testing
eligibility.
The purposes of genetic testing of breast cancer patients
are to identify additional family members who might
benefit from preventive interventions and to guide the
choice of treatment of the breast cancer patients. Under
optimal circumstances, the genetic test results would be
made available within 2 weeks of diagnosis. BRCA1 mutations carriers have been shown to have enhanced benefit
from chemotherapy, in particular to neoadjuvant cisplatin
chemotherapy. In a recent study, we found that 61 % of
107 patients with a BRCA1 mutation had a complete
pathologic response when given cis-platinum [19, 20] and
all who experienced a pCR are currently alive. In addition,
women with a BRCA1 mutation have been shown to
benefit from bilateral mastectomy [21] and from
oophorectomy [22].
Acknowledgments DNA samples were received from the Australian Breast Cancer Tissue Bank, which is generously supported by
the National Health and Medical Research Council of Australia
(NHMRC), the Cancer Institute NSW (CINSW) and the National
Breast Cancer Foundation (NBCF). The tissues and samples are made
available to researchers on a non-exclusive basis. This work was
supported by the National Breast Cancer Foundation (NBCF), Australia. Dr Michelle Wong-Brown is supported by the Hunter Translational Cancer Research Centre with funding from the Cancer
Institute New South Wales.
Conflict of interest The authors of this article declare no competing
interests related to the study and no commercial associations that may
pose a conflict of interest.

79

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