Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells

Hiroyuki Konishi, Morassa Mohseni, Akina Tamaki, Joseph P. Garay, Sarah Croessmann, Sivasundaram Karnan, Akinobu Ota, Hong Yuen Wong, Yuko Konishi, Bedri Karakas, Khola Tahir, Abde M. Abukhdeir, John P. Gustin, Justin Cidado, Grace M. Wang, David Cosgrove, Rory Cochran, Danijela Jelovac, Michaela J. Higgins, Sabrina ArenaLauren Hawkins, Josh Lauring, Amy L. Gross, Christopher M. Heaphy, Yositaka Hosokawa, Edward Gabrielson, Alan K. Meeker, Kala Visvanathan, Pedram Argani, Kurtis E. Bachman, Ben Ho Park

Research output: Contribution to journalArticle

Abstract

Biallelic inactivation of cancer susceptibility gene BRCA1 leads to breast and ovarian carcinogenesis. Paradoxically, BRCA1 deficiency in mice results in early embryonic lethality, and similarly, lack of BRCA1 in human cells is thought to result in cellular lethality in view of BRCA1's essential function. To survive homozygous BRCA1 inactivation during tumorigenesis, precancerous cells must accumulate additional genetic alterations, such as p53 mutations, but this requirement for an extra genetic "hit" contradicts the two-hit theory for the accelerated carcinogenesis associated with familial cancer syndromes. Here, we show that heterozygous BRCA1 inactivation results in genomic instability in nontumorigenic human breast epithelial cells in vitro and in vivo. Using somatic cell gene targeting, we demonstrated that a heterozygous BRCA1 185delAG mutation confers impaired homology-mediated DNA repair and hypersensitivity to genotoxic stress. Heterozygous mutant BRCA1 cell clones also showed a higher degree of gene copy number loss and loss of heterozygosity in SNP array analyses. In BRCA1 heterozygous clones and nontumorigenic breast epithelial tissues from BRCA mutation carriers, FISH revealed elevated genomic instability when compared with their respective controls. Thus, BRCA1 haploinsufficiency may accelerate hereditary breast carcinogenesis by facilitating additional genetic alterations.

Original languageEnglish
Pages (from-to)17773-17778
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume108
Issue number43
DOIs
Publication statusPublished - Oct 25 2011

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Genomic Instability
Neoplasm Genes
Carcinogenesis
Breast
Epithelial Cells
Alleles
Mutation
Clone Cells
Haploinsufficiency
Gene Dosage
Gene Targeting
Loss of Heterozygosity
DNA Repair
DNA Damage
Single Nucleotide Polymorphism
Hypersensitivity
Epithelium
Neoplasms

Keywords

  • Breast cancer 1, early onset
  • Human cell gene targeting
  • Somatic cell knock-in

ASJC Scopus subject areas

  • General

Cite this

Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells. / Konishi, Hiroyuki; Mohseni, Morassa; Tamaki, Akina; Garay, Joseph P.; Croessmann, Sarah; Karnan, Sivasundaram; Ota, Akinobu; Wong, Hong Yuen; Konishi, Yuko; Karakas, Bedri; Tahir, Khola; Abukhdeir, Abde M.; Gustin, John P.; Cidado, Justin; Wang, Grace M.; Cosgrove, David; Cochran, Rory; Jelovac, Danijela; Higgins, Michaela J.; Arena, Sabrina; Hawkins, Lauren; Lauring, Josh; Gross, Amy L.; Heaphy, Christopher M.; Hosokawa, Yositaka; Gabrielson, Edward; Meeker, Alan K.; Visvanathan, Kala; Argani, Pedram; Bachman, Kurtis E.; Park, Ben Ho.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, No. 43, 25.10.2011, p. 17773-17778.

Research output: Contribution to journalArticle

Konishi, H, Mohseni, M, Tamaki, A, Garay, JP, Croessmann, S, Karnan, S, Ota, A, Wong, HY, Konishi, Y, Karakas, B, Tahir, K, Abukhdeir, AM, Gustin, JP, Cidado, J, Wang, GM, Cosgrove, D, Cochran, R, Jelovac, D, Higgins, MJ, Arena, S, Hawkins, L, Lauring, J, Gross, AL, Heaphy, CM, Hosokawa, Y, Gabrielson, E, Meeker, AK, Visvanathan, K, Argani, P, Bachman, KE & Park, BH 2011, 'Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells', Proceedings of the National Academy of Sciences of the United States of America, vol. 108, no. 43, pp. 17773-17778. https://doi.org/10.1073/pnas.1110969108
Konishi, Hiroyuki ; Mohseni, Morassa ; Tamaki, Akina ; Garay, Joseph P. ; Croessmann, Sarah ; Karnan, Sivasundaram ; Ota, Akinobu ; Wong, Hong Yuen ; Konishi, Yuko ; Karakas, Bedri ; Tahir, Khola ; Abukhdeir, Abde M. ; Gustin, John P. ; Cidado, Justin ; Wang, Grace M. ; Cosgrove, David ; Cochran, Rory ; Jelovac, Danijela ; Higgins, Michaela J. ; Arena, Sabrina ; Hawkins, Lauren ; Lauring, Josh ; Gross, Amy L. ; Heaphy, Christopher M. ; Hosokawa, Yositaka ; Gabrielson, Edward ; Meeker, Alan K. ; Visvanathan, Kala ; Argani, Pedram ; Bachman, Kurtis E. ; Park, Ben Ho. / Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells. In: Proceedings of the National Academy of Sciences of the United States of America. 2011 ; Vol. 108, No. 43. pp. 17773-17778.
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T1 - Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells

AU - Konishi, Hiroyuki

AU - Mohseni, Morassa

AU - Tamaki, Akina

AU - Garay, Joseph P.

AU - Croessmann, Sarah

AU - Karnan, Sivasundaram

AU - Ota, Akinobu

AU - Wong, Hong Yuen

AU - Konishi, Yuko

AU - Karakas, Bedri

AU - Tahir, Khola

AU - Abukhdeir, Abde M.

AU - Gustin, John P.

AU - Cidado, Justin

AU - Wang, Grace M.

AU - Cosgrove, David

AU - Cochran, Rory

AU - Jelovac, Danijela

AU - Higgins, Michaela J.

AU - Arena, Sabrina

AU - Hawkins, Lauren

AU - Lauring, Josh

AU - Gross, Amy L.

AU - Heaphy, Christopher M.

AU - Hosokawa, Yositaka

AU - Gabrielson, Edward

AU - Meeker, Alan K.

AU - Visvanathan, Kala

AU - Argani, Pedram

AU - Bachman, Kurtis E.

AU - Park, Ben Ho

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N2 - Biallelic inactivation of cancer susceptibility gene BRCA1 leads to breast and ovarian carcinogenesis. Paradoxically, BRCA1 deficiency in mice results in early embryonic lethality, and similarly, lack of BRCA1 in human cells is thought to result in cellular lethality in view of BRCA1's essential function. To survive homozygous BRCA1 inactivation during tumorigenesis, precancerous cells must accumulate additional genetic alterations, such as p53 mutations, but this requirement for an extra genetic "hit" contradicts the two-hit theory for the accelerated carcinogenesis associated with familial cancer syndromes. Here, we show that heterozygous BRCA1 inactivation results in genomic instability in nontumorigenic human breast epithelial cells in vitro and in vivo. Using somatic cell gene targeting, we demonstrated that a heterozygous BRCA1 185delAG mutation confers impaired homology-mediated DNA repair and hypersensitivity to genotoxic stress. Heterozygous mutant BRCA1 cell clones also showed a higher degree of gene copy number loss and loss of heterozygosity in SNP array analyses. In BRCA1 heterozygous clones and nontumorigenic breast epithelial tissues from BRCA mutation carriers, FISH revealed elevated genomic instability when compared with their respective controls. Thus, BRCA1 haploinsufficiency may accelerate hereditary breast carcinogenesis by facilitating additional genetic alterations.

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