TY - JOUR
T1 - Identification of deleterious germline CHEK2 mutations and their association with breast and ovarian cancer
AU - Kleiblova, Petra
AU - Stolarova, Lenka
AU - Krizova, Katerina
AU - Lhota, Filip
AU - Hojny, Jan
AU - Zemankova, Petra
AU - Havranek, Ondrej
AU - Vocka, Michal
AU - Cerna, Marta
AU - Lhotova, Klara
AU - Borecka, Marianna
AU - Janatova, Marketa
AU - Soukupova, Jana
AU - Sevcik, Jan
AU - Zimovjanova, Martina
AU - Kotlas, Jaroslav
AU - Panczak, Ales
AU - Vesela, Kamila
AU - Cervenkova, Jana
AU - Schneiderova, Michaela
AU - Burocziova, Monika
AU - Burdova, Kamila
AU - Stranecky, Viktor
AU - Foretova, Lenka
AU - Machackova, Eva
AU - Tavandzis, Spiros
AU - Kmoch, Stanislav
AU - Macurek, Libor
AU - Kleibl, Zdenek
N1 - Funding Information:
We thank Prof. Yves-Jean Bignon who kindly provided the CHEK2 plasmid, Prof. Martin Haluzik and Dr. Petra Kavalkova for their help with FluoroStar analysis, Jan Flemr for language editing. We thank all patients and their families for their participation in our study. The study was supported by grants of the Ministry of Health of the Czech Republic NV15-28830A, NV15-27695A, NV16-29959A, NV19-03-00279 and the grants of Charles University PROGRES Q28/LF1 and Q26/LF1, GAUK 762216, SVV2019/260367, PRIMUS/17/MED/9 and UNCE/MED/016. We thank The National Center for Medical Genomics (LM2015091) for providing allelic frequencies in ethnically matched populations (project CZ.02.1.01/0.0/0.0/16_013/0001634) and NPU II LQ1604. LM was partially supported by the Academy of Sciences of the Czech Republic (Strategie AV21, Qualitas).
Funding Information:
We thank Prof. Yves-Jean Bignon who kindly provided the CHEK2 plasmid, Prof. Martin Haluzik and Dr. Petra Kavalkova for their help with FluoroStar analysis, Jan Flemr for language editing. We thank all patients and their families for their participation in our study. The study was supported by grants of the Ministry of Health of the Czech Republic NV15-28830A, NV15-27695A, NV16-29959A, NV19-03-00279 and the grants of Charles University PROGRES Q28/LF1 and Q26/LF1, GAUK 762216, SVV2019/260367, PRIMUS/17/MED/9 and UNCE/MED/016. We thank The National Center for Medical Genomics (LM2015091) for providing allelic frequencies in ethnically matched populations (project CZ.02.1.01/ 0.0/0.0/16_013/0001634) and NPU II LQ1604. LM was partially supported by the Academy of Sciences of the Czech Republic (Strategie AV21, Qualitas).
Publisher Copyright:
© 2019 UICC
PY - 2019
Y1 - 2019
N2 - Germline mutations in checkpoint kinase 2 (CHEK2), a multiple cancer-predisposing gene, increase breast cancer (BC) risk; however, risk estimates differ substantially in published studies. We analyzed germline CHEK2 variants in 1,928 high-risk Czech breast/ovarian cancer (BC/OC) patients and 3,360 population-matched controls (PMCs). For a functional classification of VUS, we developed a complementation assay in human nontransformed RPE1-CHEK2-knockout cells quantifying CHK2-specific phosphorylation of endogenous protein KAP1. We identified 10 truncations in 46 (2.39%) patients and in 11 (0.33%) PMC (p = 1.1 × 10−14). Two types of large intragenic rearrangements (LGR) were found in 20/46 mutation carriers. Truncations significantly increased unilateral BC risk (OR = 7.94; 95%CI 3.90–17.47; p = 1.1 × 10−14) and were more frequent in patients with bilateral BC (4/149; 2.68%; p = 0.003), double primary BC/OC (3/79; 3.80%; p = 0.004), male BC (3/48; 6.25%; p = 8.6 × 10−4), but not with OC (3/354; 0.85%; p = 0.14). Additionally, we found 26 missense VUS in 88 (4.56%) patients and 131 (3.90%) PMC (p = 0.22). Using our functional assay, 11 variants identified in 15 (0.78%) patients and 6 (0.18%) PMC were scored deleterious (p = 0.002). Frequencies of functionally intermediate and neutral variants did not differ between patients and PMC. Functionally deleterious CHEK2 missense variants significantly increased BC risk (OR = 3.90; 95%CI 1.24–13.35; p = 0.009) and marginally OC risk (OR = 4.77; 95%CI 0.77–22.47; p = 0.047); however, carriers low frequency will require evaluation in larger studies. Our study highlights importance of LGR detection for CHEK2 analysis, careful consideration of ethnicity in both cases and controls for risk estimates, and demonstrates promising potential of newly developed human nontransformed cell line assay for functional CHEK2 VUS classification.
AB - Germline mutations in checkpoint kinase 2 (CHEK2), a multiple cancer-predisposing gene, increase breast cancer (BC) risk; however, risk estimates differ substantially in published studies. We analyzed germline CHEK2 variants in 1,928 high-risk Czech breast/ovarian cancer (BC/OC) patients and 3,360 population-matched controls (PMCs). For a functional classification of VUS, we developed a complementation assay in human nontransformed RPE1-CHEK2-knockout cells quantifying CHK2-specific phosphorylation of endogenous protein KAP1. We identified 10 truncations in 46 (2.39%) patients and in 11 (0.33%) PMC (p = 1.1 × 10−14). Two types of large intragenic rearrangements (LGR) were found in 20/46 mutation carriers. Truncations significantly increased unilateral BC risk (OR = 7.94; 95%CI 3.90–17.47; p = 1.1 × 10−14) and were more frequent in patients with bilateral BC (4/149; 2.68%; p = 0.003), double primary BC/OC (3/79; 3.80%; p = 0.004), male BC (3/48; 6.25%; p = 8.6 × 10−4), but not with OC (3/354; 0.85%; p = 0.14). Additionally, we found 26 missense VUS in 88 (4.56%) patients and 131 (3.90%) PMC (p = 0.22). Using our functional assay, 11 variants identified in 15 (0.78%) patients and 6 (0.18%) PMC were scored deleterious (p = 0.002). Frequencies of functionally intermediate and neutral variants did not differ between patients and PMC. Functionally deleterious CHEK2 missense variants significantly increased BC risk (OR = 3.90; 95%CI 1.24–13.35; p = 0.009) and marginally OC risk (OR = 4.77; 95%CI 0.77–22.47; p = 0.047); however, carriers low frequency will require evaluation in larger studies. Our study highlights importance of LGR detection for CHEK2 analysis, careful consideration of ethnicity in both cases and controls for risk estimates, and demonstrates promising potential of newly developed human nontransformed cell line assay for functional CHEK2 VUS classification.
KW - CHEK2
KW - KAP1
KW - VUS
KW - breast cancer
KW - functional assay
KW - germline mutations
KW - ovarian cancer
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U2 - 10.1002/ijc.32385
DO - 10.1002/ijc.32385
M3 - Article
C2 - 31050813
AN - SCOPUS:85066107734
SN - 0020-7136
VL - 145
SP - 1782
EP - 1797
JO - International journal of cancer
JF - International journal of cancer
IS - 7
ER -