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The role of microhomology-mediated end joining in Fanconi anemia pathogenesis

微同源介导的末端连接在范可尼贫血发病机制中的作用

基本信息

批准号：
10367981
负责人：
Neil Johnson
金额：
$ 45.41万
依托单位：
RESEARCH INST OF FOX CHASE CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10367981
关键词：
Address Adult Alleles Amino Acids Anemia Animal Model Ashkenazim BRCA1 gene BRCA2 gene Biological Biological Assay Bone marrow failure CRISPR/Cas technology Cell Line Cells Chemicals Child Coiled-Coil Domain Complex DNA DNA Double Strand Break DNA Repair DNA Repair Gene DNA Repair Pathway DNA-Directed DNA Polymerase Data Defect Development Disease Embryo Embryonic Development Equilibrium Etiology Excision Fanconi&apos s Anemia Flow Cytometry Functional disorder Genomic Instability Health Hematology Hematopoiesis Hematopoietic stem cells Hereditary Breast and Ovarian Cancer Syndrome Human Hyperactivity Immunofluorescence Microscopy Incidence Individual Knockout Mice Laboratories Life Measures Mediating Molecular Mus Musculoskeletal Mutation Oncogenes Organism PALB2 gene Pathogenesis Pathologic Pathway interactions Patients Phenotype Population Process Proteins Reporter Resected Role Severities Single-Stranded DNA Subgroup Symptoms Testing Transgenic Animals Transplantation Work cancer predisposition developmental disease experimental study genetic approach homologous recombination inhibitor insight leukemia mouse model mutant mouse model neonatal mice nuclease p53-binding protein 1 pre-clinical protein expression recombinational repair recruit repaired therapeutic development tool

项目摘要

PROJECT SUMMARY Children with Fanconi anemia (FA) demonstrate developmental disorders that include short stature, musculoskeletal defects, cancer predisposition, bone marrow failure (BMF) and anemia. In a significant proportion of cases, FA is associated with biallelic mutations in the hereditary breast and ovarian cancer (HBOC) genes. Pre-clinical FA studies have largely relied on transgenic animal models. However, currently available FA mouse models are born without developmental defects and hematological abnormalities have to be chemically induced. Our laboratory has developed a new Brca1CC mutant mouse model with a 3-amino acid deletion in the coiled-coil (CC) domain of Brca1 that specifically disrupts the Brca1-Palb2 association, resulting in loss of Rad51 loading and HR deficiency. Notably, Brca1CC homozygous mice are born at sub-Mendelian ratios, and neo-natal mice demonstrate a range of phenotypes analogous to FA in humans, including short stature, BMF with severe anemia, and adult mice develop leukemia. Therefore, Brca1CC mice closely resemble human FA and provide a new tool to gain unprecedented insight into biological pathways that underpin FA etiology. Both homologous recombination (HR) and microhomology-mediated end joining (MMEJ) are double stranded DNA break (DSB) repair pathways that are initiated by DNA end resection, a process where DSBs are resected by nucleases to form single stranded (ss)DNA regions. In preliminary data, we examined human FA patient cells, as well as Brca1CC MEFs, for HR and MMEJ activity. Interestingly, while HR was lowered, MMEJ was hyperactivated in FA cell lines. We now hypothesize that hyperactive DNA end resection and MMEJ promote the molecular pathogenesis of FA. We will address the following Specific Aims: 1) identify DNA repair pathways that are hyperactive in FA; 2) uncover mechanisms that promote FA embryonic development and pathogenesis; and 3) examine the effects of MMEJ inhibition on FA pathogenesis. Collectively, the proposed experiments will yield new insight into DNA repair mechanisms that promote genome instability and FA.

项目总结