During the last decade, PALB2 has been defined as a high-risk breast cancer susceptibility gene alongside BRCA1 and BRCA2. Heterozygous mutation in PALB2 increases the lifetime breast cancer risk of female carriers to an average of 53%, but the risk estimate is affected by family history, studied population, and the specific pathogenic variant.

We have previously shown that lymphoblastoid cell lines (LCLs) of persons heterozygous for PALB2 c.1592delT, a Finnish founder mutation, show defects in DNA replication and damage response. Now we examine how various heterozygous, protein-truncating PALB2 founder mutations from different populations affect cell function, and whether the observed phenotype correlates with disease risk. The study cohorts come from Canada, Australia, and Finland and consist of 80 LCLs of PALB2 mutation carriers (c.1592delT, c.2323C>T, c.3113G>A or c.3323delA) and suitable population controls. There is some evidence that these variants are associated with different magnitudes of breast cancer risk, and PALB2 c.3113G>A has been associated with an exceptionally high cancer risk of 91% (95% CI 44-100) to age 70.

Utilizing these LCLs, we have studied DNA damage response, cell cycle progression, and DNA replication with various methods. DNA fiber assay results show that PALB2 mutation carriers elongate their DNA more slowly during replication, likely due to increased fork stalling, but compensate this by increased dormant origin firing. The strongest replication phenotype was observed for PALB2 c.3113G>A carriers. High content analysis shows that most of the mutation carriers had compromised RAD51 foci formation after DNA damage, whereas 53BP1 foci levels were elevated. These results indicate that during DNA double-strand break repair, heterozygous PALB2 mutation carriers may rely more on alternative, error-prone pathways that are independent of RAD51, thus contributing to genomic instability. Additionally, we are currently performing RNA-Seq to determine if the LCLs show adaptation of their gene expression in response to pathogenic PALB2 variants.

Citation Format: Niina Laurila, Kerstin Borgmann, Muthiah Bose, Felix Meyer, Katri Pylkäs, Bing Xia, William Foulkes, Melissa Southey, Helmut Pospiech, Robert Winqvist. Heterozygous PALB2 mutations cause replication stress and DNA repair defects in carrier derived lymphoblastoid cell lines [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2335.