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Development and Application of Model Systems to Study Extrachromosomal DNA Generation in Glioblastoma

胶质母细胞瘤染色体外 DNA 生成研究模型系统的开发和应用

基本信息

批准号：
10647263
负责人：
Amit Gujar
金额：
$ 8.38万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-18 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10647263
关键词：
Biological Models CRISPR interference Cell Line Cells Circular DNA Communities Cryopreservation DNA Damage DNA Repair DNA Repair Gene DNA Repair Inhibition DNA Repair Pathway DNA amplification DNA analysis DNA biosynthesis Dependence Development Diagnosis Dihydrofolate Reductase Disease Drug resistance Etiology Future Gene Amplification Generations Genes Glioblastoma Goals Growth Heterogeneity Knowledge Malignant Neoplasms Malignant neoplasm of brain Methotrexate Modeling Molecular Monitor Normal Cell Normal tissue morphology Oncogenes Pathway interactions Patient-Focused Outcomes Patients Play Prevalence Process Proliferating Protocols documentation Repression Research Resistance Resources Role System Technology Testing Therapeutic Transcription Repressor Tumor Tissue Xenograft procedure aggressive therapy biobank brain cell cancer cell cancer therapy cancer type design extrachromosomal DNA gene induction/repression genetic manipulation improved insight interest loss of function mouse model neoplastic cell novel repaired resistance mechanism response therapeutic target therapeutically effective tool tumor tumor growth

项目摘要

PROJECT SUMMARY/ABSTRACT Glioblastoma (GBM) is the most common primary malignant brain tumor with little improvement in patient survival in past few decades despite aggressive treatment options. Better understanding of the mechanisms underlying GBM is necessary to design more effective therapeutic strategies. Extrachromosomal DNAs (ecDNAs) are a well-known mechanism of oncogene amplification that promotes rapid tumor growth. Although discovered decades ago, recent technological advances have characterized ecDNAs in finer detail and have shown that they are a common occurrence in many cancer types. In a pan-cancer analysis, patients whose tumors had ecDNA amplifications were found to have significantly shorter survival than patients whose tumors had other types of amplifications. A recent study using GBM patient tumor tissues, their derivative cell lines, and orthotopic xenograft mouse models generated from these lines, showed that the majority of oncogene amplifications in these systems are extrachromosomal. Indeed, studies have shown that around 60% of GBM tumors contain ecDNAs, making it the cancer with highest ecDNA prevalence. Mechanisms that enable ecDNA generation at such high rates in GBM are starting to be revealed; however, no model systems are currently available to causally analyze the role of specific genes in their generation. In Aim 1, we will therefore utilize a drug resistance mechanism that allows ecDNA generation to develop clonal lines from primary patient-derived GBM cells to model ecDNA generation. These systems will be isogenic (i.e., without/before and with/after ecDNA generation) and clonal to minimize intercellular heterogeneity in ecDNA levels. Procurement of clones before and after ecDNA generation, and knowledge of clones that are able to generate ecDNAs in this approach, combined with desired manipulations of genes of interest permits assessment of causality of those genes in the generation process. Using these model systems developed, we will test a hypothesis in Aim 2 that DNA repair machinery is necessary for ecDNA generation with specific DNA repair genes governing the process. There is increasing evidence implicating DNA repair in ecDNA generation. Moreover, DNA repair is actively being studied as a target for GBM treatment due to its well-documented role in resistance to standard therapy for GBM via repair of therapy-induced DNA damage. The causal role of key DNA repair genes in ecDNA generation will be analyzed by determining the effects of CRISPRi-induced repression of these genes on ecDNA generation. We expect that the findings from this project will help define the ecDNA generation process and the therapeutic potential of targeting ecDNA in GBM with implications for other cancers with high ecDNA prevalence.

项目总结/文摘