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Role of Damage Response in Bone Marrow Failure in Fanconi Anemia

损伤反应在范可尼贫血骨髓衰竭中的作用

基本信息

批准号：
8113396
负责人：
JULIA SIDOROVA
金额：
$ 7.49万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-20 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8113396
关键词：
Affect Behavior Biological Assay Birth Bone Marrow Bone Marrow Cells CD34 gene Cell Line Cells Child Complement Congenital Abnormality DNA DNA Damage DNA Replication Damage Data Defect Development Disease Dissection Etiology Fanconi anemia protein Fanconi&apos s Anemia Fibroblasts Genome Stability Genomic Instability Goals Hematologic Neoplasms Hematopoietic Hematopoietic stem cells Hereditary Disease Human Lesion Link Maintenance Malignant Neoplasms Measures Metabolism Methodology Microfluidics Molecular Morbidity - disease rate Mutation Pancytopenia Pathway interactions Patients Phenotype Pilot Projects Predisposition Proteins Protocols documentation Public Health Research Personnel Risk Role S Phase Scheme Simian virus 40 Staging Stem cells Stress Stretching Syndrome System Technology Testing Work arm cell type clinically relevant established cell line gene cloning high risk improved in vivo insight lymphoblast mortality mutant novel progenitor response single molecule skeletal tool

项目摘要

DESCRIPTION (Provided by Applicant): The goal of this project is to gain mechanistic understanding of the functional defects of DNA metabolism associated with Fanconi anemia (FA). FA is a highly variable genetic disorder that manifests soon after birth with progressive pancytopenia and then bone marrow failure. Children with FA can also have congenital abnormalities of skeletal and other systems, and later in the course of the disease they are at high risk of developing certain cancers. FA pathway is implicated in genome stability maintenance and specifically in supporting replication of damaged DNA during S phase of the cell cycle. However, it is not known how exactly the pathway performs this function. This is in part due to the limited availability of direct, in vivo functional assays to measure replication fork metabolism in human cells. The investigators will use a novel, quantitative, and sensitive technology, microfluidics-assisted display of stretched DNA, to determine how deficiency in FA pathway affects the ability of cells to replicate lesion-containing DNA in vivo. The investigators will first analyze cell lines established from FA patient cells, and then apply their experimental scheme to the cell type that is clinically most relevant to the etiology of FA -hematopoietic stem cells. Selective depletion of this cell type is a leading cause of mortality and morbidity associated with FA, and it can be hypothesized that this phenotype is at least in part due to the increased sensitivity of these cells to DNA damage-induced replication stress, and/or increased requirement for the FA pathway in the response to this stress. By measuring replication in vivo in FA protein-deficient and control hematopoietic stem cells on the one hand, and comparing it with FA-deficient primary fibroblasts on the other, they will be able to test this hypothesis and set the stage for further mechanistic dissection of the FA pathway function in stem cells. As a first step towards this mechanistic insight, the investigators will establish a protocol for a screen for compounds that improve replication in the presence of DNA damage in hematopoietic stem cells. PROJECT NARRATIVE: This project uses a unique tool to query molecular defects of a human bone marrow failure and cancer predisposition disorder, Fanconi anemia. The results of this work will contribute to our understanding of the etiology of this disease and potentially of other heritable genomic instability syndromes.

描述（由申请人提供）：该项目的目标是获得与范可尼贫血（FA）相关的 DNA 代谢功能缺陷的机制理解。 FA 是一种高度可变的遗传性疾病，出生后不久就会出现进行性全血细胞减少症，然后出现骨髓衰竭。患有 FA 的儿童还可能有骨骼和其他系统的先天性异常，在疾病的后期，他们患某些癌症的风险很高。 FA 途径涉及基因组稳定性维持，特别是支持细胞周期 S 期受损 DNA 的复制。然而，尚不清楚该通路究竟如何执行此功能。部分原因是用于测量人类细胞复制叉代谢的直接体内功能测定的可用性有限。研究人员将使用一种新颖、定量且灵敏的技术，即微流体辅助显示拉伸 DNA，来确定 FA 途径的缺陷如何影响细胞在体内复制含有病变的 DNA 的能力。研究人员将首先分析从 FA 患者细胞建立的细胞系，然后将他们的实验方案应用于临床上与 FA 造血干细胞病因最相关的细胞类型。这种细胞类型的选择性耗竭是与 FA 相关的死亡和发病的主要原因，并且可以假设这种表型至少部分是由于这些细胞对 DNA 损伤诱导的复制应激的敏感性增加，和/或在响应这种应激时对 FA 途径的需求增加。通过一方面测量 FA 蛋白缺陷和对照造血干细胞的体内复制，另一方面将其与 FA 缺陷的原代成纤维细胞进行比较，他们将能够检验这一假设，并为进一步解析干细胞中 FA 途径功能奠定基础。作为实现这一机制洞察的第一步，研究人员将建立一个筛选化合物的方案，以改善造血干细胞 DNA 损伤时的复制。项目叙述：该项目使用一种独特的工具来查询人类骨髓衰竭和癌症易感性疾病（范可尼贫血）的分子缺陷。这项工作的结果将有助于我们了解这种疾病的病因学以及其他潜在的遗传性基因组不稳定综合征的病因学。