Advertisement

BMC Medical Genetics

, 20:154 | Cite as

BRCA mutations in a cohort of Iraqi patients presenting to a tertiary referral center

  • Chantal Farra
  • Christelle Dagher
  • Lama Hamadeh
  • Nagi El Saghir
  • Deborah MukherjiEmail author
Open Access
Research article
  • 156 Downloads
Part of the following topical collections:
  1. Genetic epidemiology and genetic associations

Abstract

Background

Unique pathogenic mutations in BRCA1 and 2 genes have been reported in different populations of patients originating from the Middle East region. Limited data are available for the Iraqi population. For many reasons a large number of Iraqi patients present to Lebanon for medical care. This is the first report of BRCA full gene sequencing conducted in a cohort of high-risk patients originating from Iraq.

Methods

This is a retrospective review of Iraqi patients diagnosed with breast or ovarian cancer referred for BRCA mutation testing at the American University of Beirut from January 2012 to October 2018.

Results

Of the 42 Iraqi women who underwent genetic testing at our institution, 3 BRCA pathogenic variants were found. Two mutations in BRCA1 c.224_227delAAAG and c.5431C > T and one mutation in BRCA2 c.5576_5579delTTAA were identified. Three other patients had sequence changes considered as variants of undetermined significance.

Conclusion

In this cohort of high-risk patients, one out of the three pathogenic BRCA variants detected has not previously been reported in the Middle Eastern population. Further studies are required to delineate the spectrum of BRCA mutations in the Iraqi population.

Keywords

Iraq BRCA1 BRCA2 Breast cancer Middle East 

Abbreviations

AUBMC

American University of Beirut Medical Center

IRB

Institutional Review Board

MENA

Middle East and North Africa

Background

The American University of Beirut Medical Center (AUBMC) is a major tertiary referral center in the Middle East. For several reasons including conflict-related deficiencies in healthcare services, a significant number of Iraqi patients seek cancer care at AUBMC [1]. Breast cancer is reportedly the most common primary tumor diagnosed in Iraqi females, with around 3000 cases registered in 2009 [2]. We have recently reviewed the literature regarding BRCA mutations reported in the Middle East and North Africa (MENA) region [3], we found a certain number of mutations specific to each country. Very limited information is however available regarding BRCA mutations in the Iraqi population.

The aim of our study is to review the data from patients of Iraqi origin referred to our institution for BRCA genetic testing. As far as we are aware, this is the first report of whole BRCA1 and BRCA 2 genes sequencing performed in this population.

Methods

After Institutional Review Board (IRB) approval, we retrospectively reviewed all cases of Iraqi patients who presented for BRCA mutation testing at the Medical Genetics Unit at AUBMC between January 2012 and October 2018. All data were collected from medical charts available at our institution.

QiaAmp DNA Mini kit was used to extract DNA from blood samples. BRCA1 and 2 genes were amplified using exon-specific primers designed via Primer 3. PCR efficiency was confirmed by amplification on agarose gel 2%. The amplicons were then enzymatically purified using Exonuclease I and Shrimp Alkaline Phosphatase, sequenced and loaded on the Genetic Analyzer (Applied Biosystem ABI 3500). Obtained sequences were analyzed using Seqscape® v2.7 software and compared to the corresponding reference sequences ([4, 5]). ClinVar was used to determine the significance of the variations found [6].

Results

A total of 42 unrelated Iraqi women were referred for genetic testing performed at AUBMC. The mean age was 47.2 (27–69) years with 10.4 standard deviation. All of the 42 women had a diagnosis of breast cancer, 14 had a positive first-degree family history of breast or ovarian cancer (Table 1). Patients with a diagnosis of breast cancer without a positive first-degree family history were referred for testing for high-risk features in line with National Comprehensive Cancer Network (NCCN) guidelines [7].
Table 1

Patients’ demographics

Variable

Number

Number of patients

42

Mean age (years)

47.2 (Range 27–69)

Breast or ovarian cancer family history

14

Breast cancer personal history

42

Two disease causing variants (c.224_227delAAAG and c.5431C > T) in BRCA1 and one (c.5576_5579delTTAA) in BRCA2 was observed in the present study (Table 2) (Additional files 1, 2 and 3). Three variants of unknown significance (VUS) in BRCA1 (c.536A > G, c.1458 T > G, c.1648A > C) were observed in two patients whereas one patient had one VUS in BRCA2 (c.1075G > A) (Table 3) (Additional files 4, 5, 6, 7, 8, 9 and 10).
Table 2

Pathogenic BRCA gene mutations detected in the cohort

Gene

Nucleotide Change

AA change

Nomenclature protein

Population previously reported in

BRCA1

c.224_227delAAAG

p.Glu75Valfs

343del4

Kurdish Jewish

BRCA1

c.5431C > T

p.Gln1811Ter

Q1811X

Greek

BRCA2

c.5576_5579delTTAA

p.Ile1859Lysfs

5804del4

Lebanese

Chinese

Table 3

BRCA VUS detected in the cohort

Gene

Nucleotide Change

AA change

Nomenclature protein

Number of patients with mutation

BRCA1

c.536A > G

p.Tyr179Cys

Y179C

2

c.1458 T > G

p.Phe486Leu

F486 L

c.1648A > C

p.Asn550His

N550H (benign)

BRCA2

c.1075G > A

p.Glu359Lys

E359K

1

Discussion

Of the 958 Iraqi patients referred to AUBMC in the past 3 years for treatment of cancer, 14.5% presented for diagnosis, treatment or consultation exclusively for breast cancer [1].

Patients from Iraq receiving cancer care in Lebanon are not covered by third-party insurance resulting in considerable financial constraints related to diagnostic tests and treatment. For this reason, and decreased availability of genetic counseling, we suspect significant underutilization of BRCA testing for eligible patients.

We conducted a literature review of previously reported mutations in BRCA1 and 2 genes detected in patients of Iraqi origin. Very few data have been published on BRCA findings in the Iraqi population and of the studies available, none has conducted whole BRCA1 and 2 genes sequencing.

In 1997, a study investigating the presence of 5 specific BRCA mutations in Ashkenazi and non-Ashkenazi Jewish females with breast or ovarian cancer identified the presence of the Ashkenazi Jewish common mutation BRCA1 185delAG, for the first time in an Iraqi patient with no known Ashkenazi ancestry [8] The BRCA1 185delAG has been identified in several other ethnic groups [9]. One other study used PCR amplification with 3 specific primer pairs to investigate the prevalence of 3 specific mutations (BRCA1 185delAG, BRCA1 5382insC and BRCA2 6174delT) in a cohort of Patients in the Hilla region of Iraq however these results have not been validated [10].

In the present study 3 BRCA pathogenic variants were detected by whole gene sequencing performed for clinical reasons. The BRCA2 c.5576_5579delTTAA, which was previously identified in one patient of Lebanese origin [11], is also reportedly a founder mutation in the Chinese population [12]. The BRCA1 c.224_227delAAAG found in our study was previously identified in in 2 out of 53 patients with breast cancer of Kurdish Jewish descent [13].

The third pathogenic variant found in our cohort, BRCA1 c.5431C > T has been documented in 3 unrelated patients with breast cancer of Greek origin [14]. It has not to date been reported in the Middle East region.

Conclusion

In a cohort of high-risk patients for hereditary breast or ovarian cancer of Iraqi origins referred for BRCA testing at our institution, one out of three identified pathogenic variants has not previously been reported in the Middle East region. Because of the heterogeneity of the variants reported to date in the different populations that have been studied in the region, a unified and affordable, panel that would target specific variants cannot be recommended at this stage. Further studies utilizing whole-gene sequencing approaches are required to formulate evidence-based guidelines for BRCA and other cancer related genetic analyses in the region.

Notes

Acknowledgements

Not applicable.

Authors’ contributions

CF, DM, NES designed the study, CD and LH collected data, CF, CD, LH, NES and DM were responsible for data analysis, interpretation and manuscript writing. All authors read and approved the final manuscript: CF, CD, LH, NES, DM.

Funding

No funding was required for this study.

Ethics approval and consent to participate

The Institutional Review Board (IRB) of the American University of Beirut Medical Center has approved this research. Due to the retrospective nature of data collection, a waiver of written consent was granted.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Supplementary material

12881_2019_885_MOESM1_ESM.jpg (170 kb)
Additional file 1: BRCA1 c.224_227delAAAG- 16-018 BC (JPG 170 kb)
12881_2019_885_MOESM2_ESM.jpg (239 kb)
Additional file 2: BRCA1 c.5431C > T-16-053 BC (JPG 238 kb)
12881_2019_885_MOESM3_ESM.jpg (178 kb)
Additional file 3: BRCA2 c.5576_5579delTTAA-13-046 BC (JPG 178 kb)
12881_2019_885_MOESM4_ESM.jpg (180 kb)
Additional file 4: BRCA1 VUS c.536A > G- 15-050 BC (JPG 179 kb)
12881_2019_885_MOESM5_ESM.jpg (175 kb)
Additional file 5: BRCA1 VUS c.536A > G- 14-072 BC (JPG 175 kb)
12881_2019_885_MOESM6_ESM.jpg (245 kb)
Additional file 6: BRCA1 VUS c.1458 T > G- 14-072 BC (JPG 244 kb)
12881_2019_885_MOESM7_ESM.jpg (223 kb)
Additional file 7: BRCA1 VUS c.1458 T > G- 15-050 BC (JPG 223 kb)
12881_2019_885_MOESM8_ESM.jpg (224 kb)
Additional file 8: BRCA1 VUS c.1648A > C-14-072 BC (JPG 223 kb)
12881_2019_885_MOESM9_ESM.jpg (220 kb)
Additional file 9: BRCA1 VUS c.1648A > C- 15-050 BC (JPG 220 kb)
12881_2019_885_MOESM10_ESM.jpg (244 kb)
Additional file 10: BRCA2 VUS c.1075G > A- 17-059 BC (JPG 244 kb)

References

  1. 1.
    Mukherji D, Skelton M, Alameddine R, Saifi O, Hammoud MS, Daher M, Charafeddine M, Faraj W, Temraz SN, Shamseddine A, Mula-Hussain LYI, Saleem M, Namiq KF, Dewachi O, Sitta GA. Financial toxicity associated with conflict-induced cross-border travel for cancer care: experience of Iraqi patients in Lebanon. J Clin Oncol. 2018;36(15_suppl):6562.  https://doi.org/10.1200/JCO.2018.36.15_suppl.6562.CrossRefGoogle Scholar
  2. 2.
    Iraqi Cancer Registry 2009 (2009). Iraqi Cancer board, Baghdad-Iraq.Google Scholar
  3. 3.
    Farra C, Dagher C, Badra R, Hammoud MS, Alameddine R, Awwad J, Seoud M, Abbas J, Boulos F, El Saghir N, Mukherji D. BRCA mutation screening and patterns among high-risk Lebanese subjects. Hered Cancer Clin Pract. 2019;17(1):4.  https://doi.org/10.1186/s13053-019-0105-9.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Phelan CM, Kwan E, Jack E, Li S, Morgan C, Aube J, Hanna D, Narod SA. A low frequency of non-founder BRCA1 mutations in Ashkenazi Jewish breast-ovarian cancer families. Hum Mutat. 2002;20(5):352–7.  https://doi.org/10.1002/humu.10123.CrossRefPubMedGoogle Scholar
  5. 5.
    Diop JPD, Diallo RN, Bourdon-Huguenin V, Dem A, Diouf D, Dieng MM, Ba SA, Dia Y, Ka S, Mbengue B, Thiam A, Faye O, Diop PA, Sobol H, Dieye A. Novel BRCA2 pathogenic variant c.5219 T > G; p.(Leu1740Ter) in a consanguineous Senegalese family with hereditary breast cancer. BMC Med Genet. 2019;20(1):73.  https://doi.org/10.1186/s12881-019-0814-y.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    ClinVar. https://www.ncbi.nlm.nih.gov/clinvar/. Accessed September 2018.
  7. 7.
    NCCN Guidelines for detection, prevention, & risk reduction. https://www.nccn.org/professionals/physician_gls/default.aspx#detection.
  8. 8.
    Abeliovich D, Kaduri L, Lerer I, Weinberg N, Amir G, Sagi M, Zlotogora J, Heching N, Peretz T. The founder mutations 185delAG and 5382insC in BRCA1 and 6174delT in BRCA2 appear in 60% of ovarian cancer and 30% of early-onset breast cancer patients among Ashkenazi women. Am J Hum Genet. 1997;60(3):505–14.PubMedPubMedCentralGoogle Scholar
  9. 9.
    Laitman Y, Feng BJ, Zamir IM, Weitzel JN, Duncan P, Port D, Thirthagiri E, Teo SH, Evans G, Latif A, Newman WG, Gershoni-Baruch R, Zidan J, Shimon-Paluch S, Goldgar D, Friedman E. Haplotype analysis of the 185delAG BRCA1 mutation in ethnically diverse populations. Eur J Hum Genet. 2013;21(2):212–6.  https://doi.org/10.1038/ejhg.2012.124.CrossRefPubMedGoogle Scholar
  10. 10.
    Razzak MS, Muhsin MA, Alshibly IK, Bearem WN. Prevalence of BRCA1/2 gene mutation carriage rate among local Iraqi population. Med J Babylon. 2014;11:2.Google Scholar
  11. 11.
    El Saghir NS, Zgheib NK, Assi HA, Khoury KE, Bidet Y, Jaber SM, Charara RN, Farhat RA, Kreidieh FY, Decousus S, Romero P, Nemer GM, Salem Z, Shamseddine A, Tfayli A, Abbas J, Jamali F, Seoud M, Armstrong DK, Bignon YJ, Uhrhammer N. BRCA1 and BRCA2 mutations in ethnic Lebanese Arab women with high hereditary risk breast cancer. Oncologist. 2015;20(4):357–64.  https://doi.org/10.1634/theoncologist.2014-0364.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Kim YC, Zhao L, Zhang H, Huang Y, Cui J, Xiao F, Downs B, Wang SM. Prevalence and spectrum of BRCA germline variants in mainland Chinese familial breast and ovarian cancer patients. Oncotarget. 2016;7(8):9600–12.  https://doi.org/10.18632/oncotarget.7144.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Zick A, Cohen S, Hamburger T, Goldberg Y, Zvi N, Sagi M, Perretz T. A BRCA1 frame shift mutation in women of Kurdish Jewish descent. Open Med J. 2015;2:31–6.CrossRefGoogle Scholar
  14. 14.
    Konstantopoulou I, Tsitlaidou M, Fostira F, Pertesi M, Stavropoulou AV, Triantafyllidou O, Tsotra E, Tsiftsoglou AP, Tsionou C, Droufakou S, Dimitrakakis C, Fountzilas G, Yannoukakos D. High prevalence of BRCA1 founder mutations in Greek breast/ovarian families. Clin Genet. 2014;85(1):36–42.  https://doi.org/10.1111/cge.12274.CrossRefPubMedGoogle Scholar

Copyright information

© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors and Affiliations

  1. 1.Medical Genetics Unit, Pathology DepartmentAmerican University of Beirut Medical CenterBeirutLebanon
  2. 2.Division of Hematology Oncology, Department of Internal MedicineAmerican University of Beirut Medical CenterBeirutLebanon

Personalised recommendations