Discovering how DNA replication errors cause cancer chromosomal instability

目的：采用一套新的最先进的DNA测序技术（包括单细胞和单分子方法）来剖析特定的DNA损伤修复过程对基因组的精确影响。理论基础：复制应激是已知的人类病理包括衰老和癌症的驱动因素，但涉及的确切细胞机制尚不清楚，靶向复制应激治疗仍然是一个挑战。与正常细胞相比，癌症携带更高的内在RS水平。这种增加理论上是可以利用的，但是这种方法需要达到一个精确的阈值，以避免对正常细胞产生毒性。因此，为了增加癌症特异性细胞杀伤的特异性，我们正在寻找癌症细胞和正常细胞在如何耐受或修复复制应激和随后的染色体不稳定性方面的特异性机制差异。存在各种DNA修复途径来处理或绕过RS，然而，这些途径在癌细胞中的相对用途通常在这些途径中携带突变尚不清楚。我们将通过结合单细胞CNA分析和新开发的单分子实验（靶向长读测序）来确定癌症RS修复中特定缺陷的遗传特征。

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).