Discovering how DNA replication errors cause cancer chromosomal instability

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AIM: To employ a novel set of state-of-the-art DNA sequencing technologies (including single cell, and single molecule approaches) to dissect the precise impact on the genome of specific deregulated DNA damage repair processes.Rationale:Replication stress is a known driver of human pathologies including aging and cancer, but exact cellular mechanisms involved remain unclear and targeting replication stress therapeutically is still a challenge. Cancer carries a higher intrinsic level of RS compared to normal cells. This increase can theoretically be exploited, however this approach is subject to attaining a precise threshold to avoid toxicity to normal cells. Therefore, to increase specificity for cancer-specific cell killing we are searching for specific mechanistic differences between cancer and normal cells in how they tolerate or repair replication stress and the ensuing chromosomal instability. Various DNA repair pathways exist to deal with, or bypass RS. However the relative use of these in cancer cells which often carry mutations in these pathways is not known. We will determine the genetic signature of specific defects in RS repair in cancer by using a combination of single cell CNA analysis, and newly developed single molecule experiments (targeted long-read sequencing).