Mechanisms of genomic instability, tumor initiation and progression following the disruption of the RTF2-RNase H2 axis

RTF2-RNase H2 轴破坏后基因组不稳定、肿瘤发生和进展的机制

基本信息

批准号：
10728877
负责人：
Nicolas Johannes Blobel
金额：
$ 5.27万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10728877
关键词：
13q Alleles Atlases Automobile Driving Basic Science Biological Assay Biology Career Choice Cell Line Cell model Cells Chronic Lymphocytic Leukemia Clonal Evolution DNA Repair DNA Sequence Rearrangement DNA replication fork Data Disease Progression Doctor of Philosophy Enzymes Excision Excision Repair Exhibits Future Genes Genetic Genetic Materials Genome Genome Stability Genomic Instability Genomics Goals Hematologist Human Human Cell Line Hydrolysis In Vitro Institution Laboratories Light Loss of Heterozygosity Maintenance Malignant Neoplasms Measures Mediating Mentorship Methods Molecular Mutagenesis Mutate Mutation Oncologist Pathogenicity Pathway interactions Patients Pattern Peptides Physicians Poly(ADP-ribose) Polymerase Inhibitor Prediction of Response to Therapy Predisposition Process Proteins RNA RNase 2 Recombinants Recurrence Regulation Relapse Research Ribonuclease H Ribonucleases Ribonucleotides Role Running Sampling Scientist Stimulator of Interferon Genes Stress Tertiary Protein Structure Testing Therapeutic Intervention Topoisomerase Tumor Suppressor Proteins Universities University Hospitals Variant Work chromothripsis chronic lymphocytic leukemia cell doctoral student exome in vivo insight micronucleus mutant novel prevent programs protein purification recruit repaired replication stress response termination factor tumor initiation tumor progression whole genome

项目摘要

Project Summary/Abstract The aberrant incorporation or retention of ribonucleic acids (RNAs) in the genome is a common cause of genomic instability, rendering it susceptible to hydrolysis and downstream mutagenesis. The enzyme RNase H2 is one of the primary mechanisms protecting against this destabilization of the genome by removing these genome- embedded RNAs. Our lab recently uncovered a novel mechanism of regulation of RNase H2, by uncovering that replication termination factor 2 (RTF2) is involved in localizing and regulating the levels of RNase H2 at the replication fork. Further elucidation of this interaction is required to understand the basic biology behind the regulation and function of how RNase H2 prevents genome instability. Interestingly, copy number loss of RNase H2 is commonly found in Chronic Lymphocytic Leukemia (CLL), in over 30% of cases. In my preliminary work, I have developed various cellular models in which RNase H2 and RTF2 can be depleted, and I have expressed and purified recombinant RNase H2 and RTF2, allowing for both in vivo studies of which RNase H2 activities are regulated by RTF2 and in vitro studies of their interaction. Furthermore, I have developed a novel assay allowing a quantitative analysis of ribonucleotide incorporation in the genomes of human cells. This assay will be used to study the regulation of RNase H2 by RTF2, and will be assessed in its applicability to predict CLL responses to PARP-inhibitors. The direct mechanism behind tumor progression in the loss of RNase H2 has not been studied. In this proposal, building on my above preliminary work, I will test the hypothesis that RTF2 interacts directly with and regulates the activities of RNase H2 at the replication fork and examine the mechanism behind how loss of RNase H2 compromises genomic stability and leads to tumor progression. I am an MD/PhD student at the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional Program, where I am in the laboratory of Dr. Agata Smogorzewska at The Rockefeller University. My long-term goal is to become a physician scientist, practicing as a hematologist-oncologist as well as running an independent basic science lab as an academic university hospital. The plan outlined in this proposal, along with the support and mentorship of Dr. Agata Smogorzewska, my thesis research committee, and the Tri-Institutional MD-PhD program will help me achieve these career aspirations.

项目摘要/摘要基因组中核糖核酸（RNA）的异常掺入或保留是基因组的常见原因不稳定，使其容易受到水解和下游诱变的影响。酶RNase H2是通过去除这些基因组 - 嵌入的RNA。我们的实验室最近通过发现复制来揭示了RNase H2调节的新型机制终止因子2（RTF2）涉及在复制叉处定位和调节RNase H2的水平。需要进一步阐明这种相互作用，以了解调节背后的基本生物学和 RNase H2如何防止基因组不稳定性的功能。有趣的是，RNase H2的拷贝数损失通常为在慢性淋巴细胞性白血病（CLL）中发现，超过30％的病例。在我的初步工作中，我已经开发了 RNase H2和RTF2可以耗尽的各种细胞模型，我已经表达和纯化重组RNase H2和RTF2，允许调节RNase H2活性的两个体内研究通过RTF2及其相互作用的体外研究。此外，我已经开发了一种新颖的测定法人类细胞基因组中核糖核苷酸掺入的定量分析。该测定将习惯研究RTF2对RNase H2的调节，并将在其适用性中进行评估以预测CLL的响应 PARP抑制剂。尚未研究RNase H2丢失中肿瘤进展的直接机制。在此提案中，在我上述初步工作的基础上，我将测试RTF2直接相互作用的假设并调节RNase H2在复制叉处的活性，并检查损失如何丢失的机制 RNase H2损害了基因组稳定性并导致肿瘤进展。我是Weill Cornell/Rockefeller/Sloan Kettering Tri机构计划的医学博士/博士学位学生在洛克菲勒大学的Agata Smogorzewska博士实验室中。我的长期目标是成为一个医师科学家，作为血液学家对起医生的实践，并运行独立的基础科学实验室作为学术大学医院。该提案中概述的计划，以及 Agata Smogorzewska博士，我的论文研究委员会和三机构MD-PHD计划将帮助我实现这些职业愿望。