Novel sources of endogenous DNA damage in hematopoietic stem progenitor cells

造血干祖细胞内源性 DNA 损伤的新来源

基本信息

批准号：
10400381
负责人：
Moonjung Jung
金额：
$ 23.55万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-20 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10400381
关键词：
Acceleration Acetaldehyde Acute Myelocytic Leukemia Affect Age Aldehydes Allogenic Attention Award Biological Assay Blood Bone Marrow CRISPR screen CRISPR/Cas technology Cell Death Cell Line Cells Clustered Regularly Interspaced Short Palindromic Repeats DNA Adducts DNA Damage DNA Interstrand Crosslinking DNA Repair Defect Disease Progression Drug Metabolic Detoxication Dysmyelopoietic Syndromes Engraftment Enzymes FANCD2 protein Fanconi Anemia Complementation Group A Protein Fanconi Anemia pathway Fanconi anemia protein Fanconi&apos s Anemia Formaldehyde Frequencies Functional disorder General Population Genes Genetic Genome Genomic Instability Goals Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Human Impairment In Vitro Inherited Knock-out Knowledge Lead Lesion Maintenance Metabolic Modernization Monitor Mus Pancytopenia Pathogenesis Pathway interactions Patients Physicians Play Prevention Reporting Research Ribonucleoproteins Risk Role Scientist Screening Result Solid Neoplasm Source Testing Training Transplantation Validation Variant Work acute T-cell lymphoblastic leukemia cell aldehyde dehydrogenases base bone marrow failure syndrome cell transformation cell type crosslink experience fitness genetic information genome integrity graft vs host disease hematopoietic tissue in vivo liquid chromatography mass spectrometry mortality mouse model new therapeutic target next generation sequencing novel novel therapeutics prevent repaired screening skills therapeutic target

项目摘要

Project Summary The overarching goal of this training award is to acquire the skills and experience necessary to become an independent physician scientist with research expertise on fundamental mechanisms of bone marrow failure, with a final goal of developing novel therapies. Maintaining genome integrity is an essential task for cells that need to pass correct genetic information to their progeny. Cells are equipped with a variety of mechanisms to protect their genome including robust DNA repair. Hematopoietic stem cells are especially vulnerable to DNA damage and rely on Fanconi anemia DNA repair pathway that removes DNA interstrand crosslinks. Patients lacking the Fanconi anemia proteins invariably develop bone marrow failures. Unfortunately, there is no therapy that prevents bone marrow failures in Fanconi anemia. While bone marrow failure in Fanconi anemia is thought to be caused by the inappropriate repair of the DNA interstrand crosslinks, the source of that DNA damage is poorly understood. Recent studies have implicated endogenous metabolic by-products, such as acetaldehyde and formaldehyde, in acceleration of disease progression in Fanconi anemia. To identify other sources of DNA damage, we have performed a CRISPR-Cas9 screen and identified a novel source of endogenous DNA damage that needs Fanconi anemia pathway for its proper repair. In this application, we propose to validate our impactful in vitro findings in the mouse hematopoietic stem cells using in vivo transplantation assays and genetically modified mouse models. We will also assess the type of DNA damage and subsequent cellular consequences, paying special attention to the genomic instability that is caused by this endogenous source of DNA damage. We will also test whether any other protective pathways play a role in genome maintenance of hematopoietic stem cells using an unbiased in vivo CRISPR screen approach. Knowledge gained through our studies may unveil novel therapeutic targets useful for prevention of bone marrow failures in Fanconi anemia and in the general population.

项目摘要该培训奖的总体目标是获得成为一个必要的技能和经验独立医师科学家具有骨髓衰竭基本机制的研究专业知识，以开发新疗法的最终目标。维持基因组完整性是细胞的重要任务需要将正确的遗传信息传递给其后代。细胞配备了多种机制保护其基因组，包括健壮的DNA修复。造血干细胞特别容易受到DNA的影响损坏并依赖于去除DNA链间交联的Fanconi贫血DNA修复途径。患者缺乏fanconi贫血蛋白总是会出现骨髓衰竭。不幸的是，没有治疗这可以防止范科尼贫血中的骨髓失败。虽然认为法科尼贫血的骨髓衰竭是由DNA间链交联的不适当修复引起的，该DNA损伤的来源是理解不佳。最近的研究暗示了内源性代谢副产品，例如乙醛和甲醛，在法科尼贫血中疾病进展的加速。识别其他DNA来源损坏，我们进行了CRISPR-CAS9屏幕，并确定了内源性DNA损伤的新来源这需要法科尼贫血途径才能进行适当的修复。在此应用程序中，我们建议验证我们有影响力的使用体内移植测定和遗传学的体外发现，在小鼠造血干细胞中发现修改的鼠标模型。我们还将评估DNA损伤的类型和随后的细胞后果，特别注意由DNA损伤的内源性来源引起的基因组不稳定性。我们还将测试任何其他保护途径是否在造血的基因组维持中起作用干细胞使用无偏的体内CRISPR筛选方法。通过我们的研究获得的知识可能揭示新型治疗靶标有助于预防范科尼贫血和中的骨髓衰竭一般人口。