The role of nucleases in interstrand crosslink repair

核酸酶在链间交联修复中的作用

• 批准号: 9208149
• 负责人: Agata Smogorzewska
• 金额: $ 42.38万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9208149
• 关键词: ALS patients; Acute; Affect; Anemia; Attention; Binding; Biochemical; Blood Platelets; Bone Marrow; Cell Cycle; Cell Cycle Stage; Cell Death; Cell Line; Cell physiology; Cells; Chronic Kidney Failure; Complex; Cruciform DNA; DNA; DNA Damage; DNA Repair; DNA Repair Pathway; DNA lesion; Data; Development; Dialysis procedure; Disease; Enzymes; Erythrocytes; Event; Failure; Fanconi Anemia pathway; Fanconi anemia protein; Fanconi' s Anemia; Functional disorder; Genes; Genetic; Genetic Materials; Genetic Recombination; Genetic Transcription; Genetic study; Genome Stability; Goals; Grant; Growth; Head and Neck Cancer; Health; Hemorrhage; Hereditary Disease; Holliday Junction Resolvases; Human Genetics; In Vitro; Infection; Insect Proteins; International; Interstitial Nephritis; Kidney; Kidney Failure; Kidney Transplantation; Knowledge; Laboratory Study; Lead; Lesion; Leukocytes; Link; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Modeling; Molecular; Morphology; Mutate; Mutation; Myelogenous; Organ; Outcome; Oxygen; Pancytopenia; Pathogenesis; Pathway interactions; Patients; Predisposition; Process; Proteins; Rare Diseases; Registries; Regulation; Role; S Phase; Scaffolding Protein; Site; Substrate Specificity; Testing; Therapeutic; Toxic Environmental Substances; Travel; United States; Work; age related; cancer diagnosis; cancer therapy; chemotherapy; crosslink; disorder prevention; experimental study; fighting; genetic approach; in vivo; nuclease; prevent; public health relevance; repaired; response; scaffold; skeletal; tool

DESCRIPTION (provided by applicant): Damage to the genetic material of our cells can have many undesired consequences. It may lead to cell death, growth arrest, inappropriate growth or mutations. The outcomes of these on the organismal levels are failure of essential organ function or cancer. Rare human genetic diseases have enlightened us about how lack of DNA repair and thus persistence of DNA damage in our cells leads to these problems. Our laboratory studies two DNA repair diseases, Fanconi anemia (FA) and Karyomegalic Interstitial Nephritis (KIN). Patients with FA have developmental abnormalities including skeletal anomalies and bone marrow failure, which leaves them unable to produce enough red blood cells to carry oxygen, platelets to prevent bleeding or white blood cells to fight off infections. FA patients als have a very high predisposition to developing cancer including acute myelogenous anemia that occurs paradoxically in the setting of the bone marrow failure, head and neck cancers, and gynecologic cancers. KIN patients develop kidney failure and need dialysis and kidney transplantation. Although rare, these diseases can be used as powerful models for understanding how bone marrow and kidneys fail, and how cancer develops when the DNA is not repaired. We strive to understand the molecular underpinnings of these diseases, connections and differences between them. Even though the patients with the two diseases show different health problems, the cells from the patients lack the ability to repair a very particular type of DNA damage, interstrand crosslink, which links the two strands of DNA together precluding their separation. This kind of damage may be caused by environmental toxins, metabolites from cellular processes or by chemotherapy during cancer treatment. In this grant, we propose to concentrate our attention on the nucleases involved in processing of the interstrand crosslinks. We have identified SLX4 mutations in three patients with Fanconi anemia in the International Fanconi anemia registry. With our collaborators, we have described FAN1 mutations in KIN patients. In the first two aims we propose to use the patient cell lines to understand the pathogenesis of the two diseases. Using molecular approaches we want to understand the interaction of SLX4- bound nucleases as well as FAN1, their different requirements across cell cycle and across different DNA lesions and how they genetically interact with other DNA repair pathways in the cell. In the third aim, we will take a biochemical approach to understand these nucleases. Performing in vitro experiments, we want to study how they work on damaged DNA. Our goal is to have a detailed picture of how the cell deals with crosslinks in hopes of manipulating the repair pathways for therapeutic applications.

描述（由申请人提供）：细胞遗传物质的损伤可能会产生许多不良后果。它可能导致细胞死亡、生长停滞、不适当的生长或突变。这些在有机体水平上的结果是基本器官功能衰竭或癌症。罕见的人类遗传疾病让我们认识到细胞中缺乏 DNA 修复以及持续存在的 DNA 损伤是如何导致这些问题的。我们的实验室研究两种 DNA 修复疾病：范可尼贫血 (FA) 和巨核型间质性肾炎 (KIN)。 FA 患者存在发育异常，包括骨骼异常和骨髓衰竭，这使他们无法产生足够的红细胞来携带氧气、血小板来防止出血或白细胞来抵抗感染。 FA 患者也非常容易患上癌症，包括在骨髓衰竭、头颈癌和妇科癌症的情况下矛盾地发生的急性骨髓性贫血。 KIN 患者出现肾衰竭，需要透析和肾移植。虽然这些疾病很少见，但可以作为强大的模型来了解骨髓和肾脏如何衰竭，以及当 DNA 未修复时癌症如何发展。我们努力了解这些疾病的分子基础、它们之间的联系和差异。尽管患有这两种疾病的患者表现出不同的健康问题，但患者的细胞缺乏修复一种非常特殊类型的 DNA 损伤的能力，即链间交联，它将两条 DNA 链连接在一起，阻止它们分离。这种损害可能是由环境毒素、细胞过程的代谢物或癌症治疗期间的化疗引起的。在这笔资助中，我们建议将注意力集中在参与链间交联处理的核酸酶上。我们在国际范可尼贫血登记处发现了三名范可尼贫血患者的 SLX4 突变。我们与合作者一起描述了 KIN 患者中的 FAN1 突变。在前两个目标中，我们建议使用患者细胞系来了解这两种疾病的发病机制。使用分子方法，我们希望了解 SLX4 结合核酸酶和 FAN1 的相互作用、它们在细胞周期和不同 DNA 损伤中的不同需求，以及它们如何与细胞中的其他 DNA 修复途径进行遗传相互作用。在第三个目标中，我们将采用生化方法来了解这些核酸酶。通过进行体外实验，我们想要研究它们如何对受损的 DNA 起作用。我们的目标是详细了解细胞如何处理交联，以期操纵修复途径进行治疗应用。