DNA Replication Control and Its Application to Selective Killing of Cancer Cells

基因组稳定性的维持在细胞周期中是极其重要的，基因组不稳定性被认为是细胞周期中最重要的因素。 癌细胞的标志DNA再复制是基因组不稳定性的一种，它伴随着复制起点的启动 在同一个细胞周期中多次发生。尽管调控DNA再复制的几个基因已经被发现， 虽然已经查明，但其背后的机制仍有待确定。此外，DNA复制是否 控制可用于癌症治疗是未知的。本提案的目的是调查 Cdk 1在人类癌症和乳腺癌中阻止G2/M期DNA再复制的机制 正常细胞，筛选在癌细胞中选择性地引起DNA再复制的小分子， 进行DNA再复制控制的全基因组研究。这项研究不仅有助于了解 我们的细胞如何维持稳定的基因组的一个方面，但也为我们提供了有用的治疗靶点， 以及治疗癌症的潜在化合物。因此，这项建议与联合国的使命有关， NIH/NCI在其对生命系统的基础知识和支持研究，培训和研究的追求， 关于癌症的病因、诊断、预防和治疗的其他方案。 该提案的具体目标是：目标1。研究Cdkl在调节Orel活性中的作用， 在G2/M期阻止DNA再复制。特别是，我们将研究Orel磷酸化的作用， G2/M期Cdk 1激酶的作用以及Orel是否参与了G2/M期的再复制调控。目标2. 通过HTS试验筛选小分子，以鉴定选择性引起DNA再复制的小化合物 在癌细胞中。目标3：进行基于细胞的DNA再复制控制的全基因组研究， 通过siRNA转染的HTS测定以鉴定参与DNA再复制控制的新基因。 我的最终职业目标是成为癌症研究领域的独立研究人员。到 为了实现这一目标，目前我正在研究导致癌细胞基因组不稳定的机制 及其在癌症治疗中的应用，导致申请该奖项。

Maintenance of genome stability is extremely important in the cell cycle, with genomic instability being the hallmark of cancer cells. DNA re-replication is one type of genomic instability with replication origins firing multiple times during the same cell cycle. Although several genes that regulate DNA re-replication have been identified, the mechanisms behind them remain to be determined. Moreover, whether DNA re-replication control can be used for cancer therapy is unknown. The aim of this proposal is to investigate the mechanisms by which Cdk1 prevents DNA re-replication during G2/M phase in both human cancer and normal cells, to screen for small molecules that selectively cause DNA re-replication in cancer cells, and to perform genome-wide studies of DNA re-replication control. This study will be useful not only to understand one facet of how our cells maintain a stable genome, but also to provide us with useful therapeutic targets and potential compounds for the treatment of cancer. Therefore, this proposal is relevant to the mission of NIH/NCI in its pursuit of fundamental knowledge about living systems and supporting research, training, and other programs with respect to the cause, diagnosis, prevention, and treatment of cancer. The Specific Aims of this proposal are: Aim 1. Investigate the role of Cdkl in regulating Orel activity to prevent DNA re-replication during G2/M. In particular, we will investigate the role of phosphorylation of Orel by Cdk1 kinases during G2/M and whether Orel is involved in re-replication regulation during G2/M. Aim 2. Screen for small molecules by a HTS assay to identify small compounds that selectively cause DNA rereplication in cancer cells. Aim 3. Perform cell-based genome-wide studies of DNA re-replication control via a HTS assay by siRNA transfection to identify new genes that involve in DNA re-replication control. My ultimate career goal is to become an independent researcher in the field of cancer research. To accomplish this, currently I am investigating the mechanisms that cause genome instability in cancer cells and its application for cancer therapy, resulting in the application for this award.