SHORT COMMUNICATION

The deletion of exons 3–5 of BRCA1 is the first founderrearrangement identified in breast and/or ovarian cancerSpanish families

Sarai Palanca • Inmaculada de Juan • Gema Perez-Simo •

Eva Barragan • Isabel Chirivella • Eduardo Martınez •

Oscar Fuster • Pascual Bolufer

Published online: 2 November 2012

� Springer Science+Business Media Dordrecht 2012

Abstract We recently described a novel g.8097_22733

del14637 deletion encompassing exons 3–5 in BRCA1 gene.

This rearrangement was detected in 3 of 15 (20 %) breast

and/or ovarian cancer families of Eastern Spain. This finding

made us suspect that the newly identified deletion could be a

founder mutation. To confirm this hypothesis we studied 18

subjects belonging to the three families under study, 11

deletion carriers and 7 non-carriers. We performed a hap-

lotype analysis using two BRCA1 intragenic microsatellite

markers and two markers surrounding the BRCA1 locus. The

segregation analysis showed one common particular haplo-

type established by D17S1325, D17S1323, D17S855 and

D17S1320 markers detected in the deletion carriers but

absent in the non-carriers. Our study sustain that the deletion

of exons 3–5 of BRCA1, g.8097_22733del14637, identified

in families of southeastern of the Valencian Community is

the first founder rearrangement until now reported in Spanish

population, confirming the hypothesis that this mutation

could have Iberian ancestry.

Keywords Familial breast and/or ovarian cancer �BRCA1/2 � Large genomic rearrangements �Founder mutation

Introduction

Large genomic rearrangements (LGRs) in BRCA1/2 genes

may account for up to 36 % of all BRCA1/2 disease causing

mutations identified in different populations [1, 2]. To date,

more than 120 different LGRs have been reported in

BRCA1/2 genes, and in 12 of them a founder effect have

been established (Table 1). The most striking examples have

been observed in Dutch population, for the deletions of exon

13 and exon 22, which represent the 31.7 % of all BRCA1

mutations identified in breast and/or ovarian cancer (BC/OC)

families [1, 3]. Recently, haplotype studies have confirmed a

common ancestor for two other deletions in exons 1, 2 in this

same population [4]. Furthermore, a duplication of exon 13,

first identified in one Portuguese and in three American

families [5], was subsequently found in 11 additional fami-

lies from Australia, Belgium, Canada, Great Britain, and the

United States [6]. These families likely derived from a

common ancestor of northern British origin. Indeed, the exon

13 duplication is the seventh most frequent BRCA1 mutation

reported in the Breast Cancer Information Core database

(http://research.nhgri.nih.gov/bic/) [7]. The high proportion

of LGRs detected in Danish population can also be explained

by a single founder deletion of exons 3–16 of BRCA1 gene.

Besides, other founder BRCA1 rearrangements have been

identified in France, Italian, Germany, etc. populations

(Table 1). In BRCA2 gene, an Alu insertion in exon 3

(c.156_157insAlu) has showed a founder effect in Portu-

guese population [8]. This LGR accounts for more than a

quarter (24.7–37 %) of deleterious BRCA1/2 mutations

This study was carried out on behalf of the Group for Assessment of

Hereditary Cancer of Valencia Community.

S. Palanca � I. de Juan � G. Perez-Simo � E. Barragan �O. Fuster � P. Bolufer (&)

Laboratorio de Biologıa Molecular, Escuela de Enfermerıa 7a

planta, Hospital Universitario La Fe, Avd. Campanar 21,

46009 Valencia, Spain

e-mail: bolufer_pas@gva.es

I. Chirivella

Unit of Genetic Counselling in Cancer (Medical Oncology),

University Clinical Hospital, Valencia, Spain

E. Martınez

Unit of Genetic Counselling in Cancer (Medical Oncology),

Consorcio Provincial Hospital, Castellon, Spain

123

Familial Cancer (2013) 12:119–123

DOI 10.1007/s10689-012-9579-6

detected in families with BC/OC from northern/central

Portugal [9].

In this regard, in the Program of Genetic Counselling on

Cancer of the Valencian Community (Eastern Spain) which

hosts 745 families with high risk of BC/OC, we have

identified 133 carrier families of point mutations and 15

carrier families of LGRs (148/745, 19.9 %). The percent-

age of LGR among mutation carriers of our population is of

10.1 % (15/148). Of all rearrangements, we have identified

a novel one which consists in the g.8097_22733del14637

deletion affecting the exons 3–5 of BRCA1 gene [10]. This

rearrangement was detected in 3 of 15 families (20 %)

carrier of LGRs in BRCA1/2 genes. The relatively high

incidence of the rearrangement prompted us to assess the

hypothesis of the possible founder effect of this mutation.

Materials and methods

Patients

We performed a segregation analysis in 18 relatives of the

three families with the rearrangement. All the patients and

subjects were recruited by the units of genetic counselling

in cancer (UGCC) and all of them signed an informed

consent, following the guidelines of the institution’s ethics

committee in accordance with the Helsinki Declaration

(1964, amended in 1975 and 1983).

The first family (Fig. 1a) showed an index patient (IP)

N-41 affected with BC at age of 45 and OC 2 years later,

and her mother affected with BC had died at age of 78. In

other family (Fig. 1b), the IP N-80 was affected with

bilateral BC (BBC) at age of 43 and 53 years, respectively.

In a third family (Fig. 1c), the IP N-53 was affected with

BC at age of 43 and her mother (N-61) at age of 49 years.

All IP were found to bear exactly the same breakpoints

(g.8097_22733del14637).

Methods

In order to establish whether this recurrent deletion of the

exons 3–5 is a founder mutation, we performed a haplotype

analysis employing two BRCA1 intragenic microsatellite

markers (D17S855, D17S1323) and two markers (D17S1320,

D17S1325) surrounding the BRCA1 locus on the centromeric

and telomeric side, respectively. Amplification of microsat-

ellite markers was performed using 100 ng of genomic DNA

and FAM-labelled forward primers (0.4 lM). The final con-

centration of the other chemicals in PCR was: MgCl2(2.5 mM), dNTP (0.6 mM of each), 10 9 PCR Buffer (19)

and AmpliTaq Gold (0.03 U/ll). PCR cycling conditions were

94 �C for 10 min, 35 cycles of 94 �C 50 s, 50 �C 1 min,

72 �C 1 min, and a final extension at 72 �C for 45 min. The

sizes of PCR products were analysed by capillary electro-

phoresis on a 3130 Genetic Analyzer (Applied Biosystems)

using a 3130 POP-7 polymer matrix (Applied Biosystems).

ROX-labeled GeneScan 500 (Applied Biosystems) was used

as a molecular marker ladder. Data output was analyzed with

GeneMapper software (Version 4.0, Applied Biosystems) to

determine fragment lengths.

Results and discussion

Eleven (4 patients with cancer and 7 subjects unaffected) of

the 18 relatives studied were carriers of the deletion of the

Table 1 Founder BRCA1 and BRCA2 large genomic rearrangements

Gene Ex. involved LGR with breakpoints (ref seqa) No of families Population References

BRCA1 1, 2 AC060780: g.38037_46204delins9 4 Dutch [4]

3–16 IVS2 ? 7220_IVS16 ? 717del 12 Danish [19]

3–5 L78833: g.8097_22733del 3 Spanish [10]

8, 9 L78833: g.25302_32389del 3 American [11]

8–13 L78833: g.26967_50729del 3 French [20, 21]

9–12 L78833: g.29624_44280del 4 Hispanic [22]

9–19 L78833: g.29197_65577del 2 Italian [23]

11 c.1739_1740insAlu 2 Belgian [24]

13 L78833: g.44513_48347del 6 Dutch/German [1, 25]

13 L78833: g.44369_50449dup 91 Amer./Austr./Eur. [3, 5, 6]

17 L78833: g.58759_61875del 6 Italian/German [26, 27]

22 L78833: g.79505_80014del 25 Dutch [1]

BRCA2 3 c.156_157insAlu 32 Portuguese [8, 9, 24]

The bold identifies the founder LGR of the families of Valencian Communitya Reference sequences utilized for named the BRCA1/2 LGRs breakpoints: AC06078, U14680, L78833 and U43746

120 S. Palanca et al.

123

exons 3–5 in BRCA1 and the remaining seven (all of them

healthy) non carriers. The segregation analysis showed one

common specific haplotype defined by D17S1325,

D17S1323, D17S855 and D17S1320 markers in the dele-

tion carriers (Fig. 1) which was absent in non-carriers. This

finding strongly supports the existence of a common

ancestral mutant chromosome. In fact, the common origin

of the LGR for the three families was demonstrated by the

presence of different wild-type haplotypes with the char-

acteristic haplotype in all carriers studied (Fig. 1).

The deletion of exons 3–5 of BRCA1 affects the RING

domain (N-terminal zinc finger domain) of the BRCA1

protein and relevant interaction domains for multiple pro-

teins, compromising its function as tumor suppressor gene

[10, 11]. This fact supports that the carrier families have a

risk conferred for BC/OC. The estimate of the BRCA1/2

mutation carrier risk by the BRCAPRO and BOADICEA

models of genetic susceptibility [12–14], varied broadly

from 43 % (IP N-53), to 71 % (IP N-80) or 93 % (IP N-41)

in the families studied (Fig. 1). The low probability of

mutation carrier risk for IP N-53 supports the recommen-

dation of the systematic study of LGRs in all the patients

selected for BRCA1/2 genetic testing.

To our knowledge the deletion of exon 3–5 of BRCA1

has only been described in the population of the Valencian

Community, province of Alicante, Eastern Spain. Although

several studies in Spanish families with high risk of BC/OC

have identified cases of LGRs in the BRCA1/2 genes [15–

18], however the deletion of exons 3–5 of BRCA1 has not

yet been reported in any other population study to date. By

contrast, this rearrangement is relatively common among

the LGRs identified in our population (3/15 cases; 20 %);

and, furthermore the surnames of the mutation carriers

have all of them a Valencian ancestry.

In conclusion, our study show that the deletion of exons

3–5 of BRCA1 identified in three families of southeastern of

Fig. 1 Segregation analysis performed in families of IP N-41 (a),

N-80 (b) and N-53 (c). Pedigrees of breast and/or ovarian cancer

families with detected deletion of BRCA1 exons 3–5. Circles indicate

females, squares indicate males. The proband is indicated by an

arrow. The symbols filled in black denote individuals with cancer.

Type of cancer (BC breast cancer, BBC bilateral breast cancer, OCovarian cancer) and age at onset are indicated below each affected

individual. Haplotype of the genetic markers are shown as differently

filled bars. Mutation status: ? carrier, - no carrier (wt)

Deletion of exons 3–5 de BRCA1 is a founder mutation 121

123

the Valencian Community is the first founder LGR until now

reported in Spanish population, confirming the hypothesis

that this mutation could have Iberian origin. Although the

scientific and clinical impact of the finding here reported is

limited we thing that all the knowledge related for these rare

mutations as the LGR could be of interest for the scientific

community and also might provide future clinical benefits.

Acknowledgments We should express our gratitude to the ‘‘IIS La

Fundacion para la Investigacion del Hospital Universitario La Fe’’

for giving economical support to Gema Perez Simo, what made

possible their participation in the present study, and the ‘‘Fundacionde Investigacion de la Asociacion Espanola contra el Cancer’’.

Conflict of interest The authors declared that they have no conflict

of interest.

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