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Investigation of human DNA polymerase epsilon variants

人类 DNA 聚合酶 epsilon 变异体的研究

基本信息

批准号：
10367753
负责人：
Zachary F Pursell
金额：
$ 38万
依托单位：
TULANE UNIVERSITY OF LOUISIANA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-15 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10367753
关键词：
Address Alleles BRCA2 Mutation BRCA2 gene Biochemistry Biological Models Bypass Catalytic Domain Cell model Cells Clinical Data Clustered Regularly Interspaced Short Palindromic Repeats Cytosine DNA Damage DNA Polymerase II DNA Repair DNA Sequence Alteration DNA biosynthesis DNA lesion DNA replication fork DNA-Directed DNA Polymerase Data Defect Development Disease Environment Exonuclease Failure Functional disorder Gene Rearrangement Genes Genetic Genetic Engineering Genetically Engineered Mouse Genome Stability Genomic Instability Genomics Goals Health Human Human Cell Line In Vitro Intervention Investigation Knowledge Laboratories Left Link Malignant Neoplasms Measures Mission Modeling Mouse Cell Line Mus Mutagenesis Mutate Mutation Normal Cell Nuclear Organoids PTEN gene Pathway interactions Phenotype Polymerase Positioning Attribute Publishing Research Short Interspersed Nucleotide Elements System Testing Therapeutic Therapeutic Intervention Time United States National Institutes of Health Variant Work base burden of illness cancer genome cancer type checkpoint therapy driver mutation gene repair genome database human DNA immune checkpoint blockade innovation insight loss of function man mutant neoantigens neoplastic cell novel prevent repaired replication stress response tumor tumorigenesis tumorigenic

项目摘要

SUMMARY Understanding how cells fail to protect against the mutations and structural rearrangements that are found in all cancers is critical to ultimately treating the disease. Mutant replication DNA polymerases are found in some of the most highly mutated tumors known. These tumors also have a very unique mutation spectrum. In our published work we generated and began to characterize mouse and human cell models of these tumors. While it was originally thought that this mutagenesis was both necessary and sufficient for tumor development, our published work and preliminary data strongly implicates additional factors as being necessary for POLE tumor development. The main goal of this project is to test our central hypothesis that while POLE tumors are some of the most highly mutagenic tumors observed, the essential tumorigenic feature of POLE mutants is a failure to stabilize stalled/collapsed replication forks in response to damaged DNA via inactivation of certain homology-directed repair genes like BRCA2 and/or loss of a nuclear PTEN activity that both normally stabilize stalled replication forks and thus suppress genome instability. POLE mutagenesis is concurrently enhanced by promoting acquisition of mutations in these pathways This hypothesis is based on our published work and unpublished preliminary evidence. Specifically, this project will 1) Define the mechanistic origins of POLE mutational signatures; 2) Characterize the cooperation between POLE mutations and homology-directed repair genes, in particular BRCA2; and 3) Characterize the interactions between a noncanonical PTEN mutant and POLE mutant alleles. The proposed work is significant because these studies linking POLE dysfunction to other dysregulated pathways will provide novel insight into both basic mechanisms of genome instability, but also new possible therapeutic approaches to treating replication-defective cancers. The work is also significant because it will advance our understanding of synthetic lethality and neo-epitopes in immune checkpoint therapies. The proposed work is innovative because 1) it is the first to identify genetic interactions between POLE mutant alleles and other pathways in tumors and then characterize these interactions in a mammalian system; 2) we will be the first to examine the effects of these interactions on genome stability and then compare to sequencing information from large, publicly available cancer genome databases; 3) the proposed studies are innovative in their use of CRISPR-based genetic engineering of multiple systems (mice, human cells, organoids) for comparing effects on phenotype and mutagenesis.

总结