权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Characterization of POLQ's Function in Replication Rescue

POLQ 在复制救援中的功能特征

基本信息

批准号：
10642910
负责人：
Susanna Jane Stroik
金额：
$ 7.63万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
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：
2021-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10642910
关键词：
Acute Camptothecin Cancerous Cell Cycle Checkpoint Cell Cycle Checkpoint Genes Cell Cycle Stage Cell Line Cell division Cells Cicatrix Compensation DNA DNA Damage DNA Double Strand Break DNA Repair Pathway DNA biosynthesis DNA replication fork Development Engineering Enzymes Gatekeeping Genes Genome Genomic Instability Genomics Goals Individual Kinetics Left Link Malignant Neoplasms Measures Mediating Microscopy Mitosis Molecular Monitor Mutate Mutation Neoplasm Metastasis Nonhomologous DNA End Joining Null Lymphocytes Optics Outcome PALB2 gene Pathway interactions Pharmaceutical Preparations Play Poison Polymerase Proteins Radiation therapy Recovery Reporter Resistance Role S phase Sequence Homologs Site Source Speed System Techniques Therapeutic Therapeutic Intervention Tumor Suppressor Genes Tumor Suppressor Proteins Type I DNA Topoisomerases Work brca gene cancer cell cancer diagnosis cancer genome cancer prevention cancer therapy cell injury chemotherapeutic agent deep sequencing genetically modified cells genome sequencing genome-wide homologous recombination individualized medicine insight malignant breast neoplasm neoplastic cell novel reconstruction recruit repaired replication stress response single molecule standard of care superresolution microscopy targeted treatment tumor tumorigenesis whole genome

项目摘要

Project Summary Genomic instability is a hallmark of cancer known to generate genetic alterations during cell division. It alone can drive oncogenesis and evolve cancer cells that no longer “play by the rules”. This is accomplished via introduction of key mutations in tumor suppressor and cell cycle checkpoint genes that allow cancer cells to resist many therapeutic interventions. Notably, the standard of care for many cancers is treatment with DNA damaging agents – chemotherapeutics and radiation therapy – with the goal of damaging cancer cells beyond repair. Thus, understanding the mechanisms by which DNA damage is repaired under these conditions is of the upmost importance for both cancer prevention and its treatment. Amongst DNA damage, the most severe threat to the genome are DNA double strand breaks (DSBs). DSBs have 3 major repair mechanisms which can be deployed in response to their accumulation – homologous recombination (HR), classical non-homologous end- joining, and Theta-mediated end-joining (TMEJ). The latter is upregulated in many cancers, specifically those in which HR genes are mutated (BRCA1/2, PALB2, etc.). However, little is known about the role this pathway plays in repairing breaks which occur during DNA replication, a cell cycle stage in which many cancer therapeutics induce DNA damage. I propose to study the precise role the central protein in TMEJ, POLQ, plays during DNA replication and the genomic consequences of such a role. In my preliminary work, I have established that POLQ deficiency renders cells sensitive to both acute and prolonged Camptothecin (CPT) treatment, a Topoisomerase I poison which induces replication fork collapse. Further, POLQ-null cells display high levels of terminally stalled replication forks after CPT exposure, implying POLQ is capable of repairing forks for replication restart. In this proposal, I aim to study how POLQ contributes to replication fork progression, protection, and recovery after the induction of replicative breaks. Further, I have engineered an inducible broken replication fork reporter system, with which I can measure the kinetics and repair signatures of individual replicative DSBs. Using these experimental approaches, I can dissect the molecular requirements and outcomes of POLQ-mediated repair of broken forks. Finally, I seek to elucidate the role POLQ/TMEJ plays during DNA replication in both HR-proficient and deficient cancers which would provide mechanistic insight into cancer treatments and tumor dynamics. Genome duplication is essential for cancer cell division and thus defining how POLQ repairs replicative breaks and facilitates subsequent replication is essential.

项目总结