Project 2: Targeting DNA replication in BRCA-associated breast cancer

项目 2：针对 BRCA 相关乳腺癌中的 DNA 复制

基本信息

批准号：
10599900
负责人：
Bing Xia
金额：
$ 37.12万
依托单位：
RUTGERS BIOMEDICAL AND HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10599900
关键词：
Ablation Address Aftercare BRCA deficient BRCA1 gene BRCA2 gene Binding Breast Breast Cancer Cell Breast Cancer Risk Factor Breast Cancer cell line Cancer Etiology Cell Proliferation Cells ChIP-seq Clinical Clone Cells DNA DNA Damage DNA Double Strand Break DNA Repair DNA Replication Factor DNA biosynthesis DNA lesion DNA replication fork Defect Development Disease Double Strand Break Repair Doxorubicin Drug resistance Engineering Fiber Generations Genes Genome Stability Genomic Instability Germ-Line Mutation Kinetics Knockout Mice MCM10 gene Malignant Neoplasms Mammary Neoplasms Mammary Tumorigenesis Measures Modeling Molecular Mutation Ovarian PALB2 gene Pancreas Phenocopy Phenotype Platinum Play Poly(ADP-ribose) Polymerase Inhibitor Prostate Proteins Public Health Regulation Relapse Research Resistance Risk Role S phase SLC19A1 gene Salts Speed Testing Time Topoisomerase Topoisomerase Inhibitors Topoisomerase-I Inhibitor Tumor Suppressor Proteins Type I DNA Topoisomerases Xenograft procedure cancer cell cancer therapy chemotherapy combat comparative efficacy conditional knockout efficacy evaluation gene product genome sequencing high risk histone methyltransferase homologous recombination inhibitor irinotecan malignant breast neoplasm mouse model mutant radioresistant rational design recombinational repair repaired replication stress response restraint side effect targeted treatment treatment response triple-negative invasive breast carcinoma tumor tumorigenesis whole genome

项目摘要

PROJECT SUMMARY Germline mutations in BRCA1 and BRCA2 and PALB2 predispose to breast cancer and also increase the risks of ovarian, pancreatic, prostate, and other cancers. The products of the 3 genes constitute a BRCA1-PALB2- BRCA2 axis that plays essential roles in the DNA damage response, particularly in DNA double strand break repair by homologous recombination (HR). In addition, BRCA1 and BRCA2 have also been implicated in the regulation of DNA replication after DNA damage, as their mutant cells are defective in the intra-S phase checkpoint, which slows down DNA synthesis after the damage presumably to allow time for DNA repair and minimize the generation of mutations. However, the mechanisms by which the two proteins regulate DNA synthesis have remained elusive, and the impact of such “unrestrained” DNA synthesis on replication fidelity and genome stability after DNA damage remains unknown. In our preliminary studies, we found that BRCA2 interacts with an essential DNA replication factor, MCM10, and that either loss of BRCA2 or disruption of the BRCA2- MCM10 interaction results in unrestrained replication fork speed, reduced fork stalling, and reduced origin firing after DNA damage and replication stress. Moreover, we also found that loss of BRCA1 leads to reduced replication fork speed both before and after DNA damage. In this proposal, we will determine how BRCA2 and BRCA1 regulate DNA replication kinetics, the impact of unrestrained DNA synthesis on replication fidelity and therapy response, and the potential of replication stress inducers as a targeted therapy for BRCA and PALB2 mutant cancers. In Aim 1, we will define the mechanisms of BRCA2 and BRCA1 in the regulation of replication fork speed, fork stalling and origin firing after replication stress induced by topoisomerase inhibitors. In Aim 2, we will engineer triple negative breast cancer (TNBC) cell lines to assess the impact of replication kinetics dysregulation on replication fidelity, cancer cell response to topoisomerase inhibitors and potential relapse after treatment. In Aim 3, we will use conditional knockout and syngeneic mouse models to determine the efficacy of replication stress inducers on Brca1, Brca2 and Palb2 null mammary tumors.

项目摘要 BRCA1和BRCA2和PALB2中的种系突变易感乳腺癌，也增加了风险卵巢，胰腺，前列腺和其他癌症。 3个基因的产物构成BRCA1-PALB2- BRCA2轴在DNA损伤响应中起着重要作用，尤其是在DNA双链断裂中通过同源重组（HR）修复。此外，BRCA1和BRCA2也与 DNA损伤后DNA复制的调节，因为它们的突变细胞在Intra-S相中有缺陷检查点，损伤后会减慢DNA合成的减慢，大概可以花费时间进行DNA修复和最小化突变的产生。但是，两种蛋白质调节DNA的机制综合仍然难以捉摸，这种“不受限制”的DNA合成对复制保真度的影响 DNA损伤后的基因组稳定性仍然未知。在我们的初步研究中，我们发现BRCA2相互作用具有必不可少的DNA复制因子MCM10，并且BRCA2的丢失或BRCA2-的破坏 MCM10相互作用会导致不受约束的复制叉速度，减少叉子停滞和降低的起点射击 DNA损伤和复制应力后。此外，我们还发现BRCA1的损失导致减少 DNA损伤之前和之后的复制叉速度。在此提案中，我们将确定BRCA2和 BRCA1调节DNA复制动力学，不受约束的DNA合成对复制保真度和治疗反应以及复制应力影响者作为BRCA和PALB2的靶向疗法的潜力突变癌。在AIM 1中，我们将在复制调节中定义BRCA2和BRCA1的机制拓扑异构酶抑制剂引起的复制应力后，分叉的速度，叉子失速和起源发射。在AIM 2中，我们将设计三重阴性乳腺癌（TNBC）细胞系以评估复制动力学的影响关于复制保真度的失调，癌细胞对拓扑异构酶抑制剂的反应以及在治疗。在AIM 3中，我们将使用条件基因敲除和合成小鼠模型来确定 BRCA1，BRCA2和PALB2无效乳腺肿瘤上的复制应力诱导剂。