Aldo Prawira1, Prabhakaran Munusamy1, Jimin Yuan1, Claire Hian Tzer Chan1, Geok Ling Koh1, Timothy Wai Ho Shuen2, Jiancheng Hu1,3, Yoon Sim Yap2, Min Han Tan2,4,5, Peter Ang2,6, Ann Siew Gek Lee7,8,9. 1. Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore, Singapore. 2. Department of Medical Oncology, National Cancer Centre, Singapore, Singapore. 3. Cancer and Stem Cell Biology Programme, Duke-NUS Medical School, Singapore, Singapore. 4. Institute of Bioengineering and Nanotechnology, Singapore, Singapore. 5. Lucence Diagnostics Pte Ltd, Singapore, Singapore. 6. Oncocare Cancer Centre, Gleneagles Medical Centre, Singapore, Singapore. 7. Division of Cellular and Molecular Research, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore, Singapore. dmslsg@nccs.com.sg. 8. Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore. dmslsg@nccs.com.sg. 9. Office of Clinical and Academic Faculty Affairs, Duke-NUS Medical School, Singapore, Singapore. dmslsg@nccs.com.sg.
Abstract
PURPOSE: PARP4 has been proposed as a candidate breast cancer susceptibility gene. However, its function and involvement in breast carcinogenesis is unclear. We sought to determine the variant frequency of PARP4 in BRCA-negative women referred for genetic testing from Singapore and to perform functional analyses of PARP4. METHODS: Next-generation sequencing of PARP4 was conducted for 198 BRCA-negative cases from Singapore. Three independent case-control association analyses of PARP4 were performed for (1) our Singaporean cohort, (2) three dbGaP datasets, and (3) cases from TCGA, with controls from the Exome Aggregation Consortium (ExAC). PARP4 knockout cells were generated utilizing the CRISPR-Cas9 approach in MDA-MB-231 (breast cancer) and MCF10A (normal breast) cell lines, and colony formation, cell proliferation, and migration assays carried out. RESULTS: Candidate variants in PARP4 were identified in 5.5% (11/198) of our Singapore cohort. Case-control association studies for our cases and the dbGaP datasets showed no significant association. However, a significant association was observed for PARP4 variants when comparing 988 breast cancer cases from the TCGA provisional data and 53,105 controls from ExAC (ALL) (OR 0.249, 95% CI 0.139-0.414, P = 2.86 × 10-11). PARP4 knockout did not affect the clonogenicity, proliferation rate, and migration of normal breast cells, but appeared to decrease the proliferation rate and clonogenicity of breast cancer cells. CONCLUSIONS: Taken together, our results do not support that PARP4 functions as a cancer susceptibility gene. This study highlights the importance of performing functional analyses for candidate cancer predisposition genes.
PURPOSE:PARP4 has been proposed as a candidate breast cancer susceptibility gene. However, its function and involvement in breast carcinogenesis is unclear. We sought to determine the variant frequency of PARP4 in BRCA-negative women referred for genetic testing from Singapore and to perform functional analyses of PARP4. METHODS: Next-generation sequencing of PARP4 was conducted for 198 BRCA-negative cases from Singapore. Three independent case-control association analyses of PARP4 were performed for (1) our Singaporean cohort, (2) three dbGaP datasets, and (3) cases from TCGA, with controls from the Exome Aggregation Consortium (ExAC). PARP4 knockout cells were generated utilizing the CRISPR-Cas9 approach in MDA-MB-231 (breast cancer) and MCF10A (normal breast) cell lines, and colony formation, cell proliferation, and migration assays carried out. RESULTS: Candidate variants in PARP4 were identified in 5.5% (11/198) of our Singapore cohort. Case-control association studies for our cases and the dbGaP datasets showed no significant association. However, a significant association was observed for PARP4 variants when comparing 988 breast cancer cases from the TCGA provisional data and 53,105 controls from ExAC (ALL) (OR 0.249, 95% CI 0.139-0.414, P = 2.86 × 10-11). PARP4 knockout did not affect the clonogenicity, proliferation rate, and migration of normal breast cells, but appeared to decrease the proliferation rate and clonogenicity of breast cancer cells. CONCLUSIONS: Taken together, our results do not support that PARP4 functions as a cancer susceptibility gene. This study highlights the importance of performing functional analyses for candidate cancer predisposition genes.
Entities:
Keywords:
Breast cancer; Cancer predisposition; Germline variants; PARP4
Authors: Irene Malo Estepa; Haidee Tinning; Elton Jóse Rosas Vasconcelos; Beatriz Fernandez-Fuertes; José María Sánchez; Gregory W Burns; Thomas E Spencer; Pat Lonergan; Niamh Forde Journal: Int J Mol Sci Date: 2020-04-20 Impact factor: 5.923
Authors: Maria Skydt Lindgren; Philippe Lamy; Sia Viborg Lindskrog; Emil Christensen; Iver Nordentoft; Karin Birkenkamp-Demtröder; Benedicte Parm Ulhøi; Jørgen Bjerggaard Jensen; Lars Dyrskjøt Journal: Eur Urol Open Sci Date: 2021-11-05