Mitotic failure in Fanconi anemia: mechanisms and role in carcinogenesis

范可尼贫血的有丝分裂失败：机制和在致癌作用中的作用

基本信息

批准号：
10001741
负责人：
David W Clapp
金额：
$ 4.66万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
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：
2020-02-28 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10001741
关键词：
Acetylation Acute Myelocytic Leukemia Acute leukemia Alleles Animal Model Animals BRCA2 gene Biochemistry CDC2 gene Cell division Cells Centrosome Child Chromosomal Instability Chromosome Segregation Clinical Complex DNA Damage Detection Dissection Dysmyelopoietic Syndromes Ensure Failure Fanconi Anemia Complementation Group A Protein Fanconi Anemia pathway Fanconi Anemia-BRCA Pathway Fanconi anemia protein Fanconi&apos s Anemia Future General Population Genes Genetic Genetic Diseases Genome Genomic Instability Goals Health Hematopoiesis Hematopoietic Housekeeping Human Impairment Individual Interphase Kinetochores Laboratories Lesion Link Maintenance Malignant - descriptor Malignant Neoplasms Maps Microscopy Mitosis Mitotic Mitotic Chromosome Molecular Morbidity - disease rate Mus Mutation PCAF gene Pathogenesis Pathway interactions Patients Pattern Phosphorylation Phosphotransferases Precision therapeutics Prometaphase Resolution Role Signal Transduction Stress Structure Testing Therapeutic Studies Tumor Suppression Tumor Suppressor Genes Tumor Suppressor Proteins Work bone marrow failure syndrome brca gene cancer cell cancer therapy carcinogenesis chromosome missegregation genome integrity high risk improved in vivo insight leukemia leukemogenesis mortality mouse model multidisciplinary novel novel strategies personalized therapeutic pre-clinical prevent repaired response small hairpin RNA small molecule

项目摘要

Abstract Biallelic mutations in the FA/BRCA genes cause Fanconi anemia (FA), a bone marrow failure syndrome associated with genomic instability and cancer. Individuals with FA suffer from a wide range of clinical manifestations, including high risk of leukemia and other malignancies that cause major lifelong morbidity and mortality. Furthermore, acquired mutations of FA genes occur in acute leukemia and other cancers in non-FA patients, indicating that FA genes protect the general population from cancer. Thus, understanding of the FA genome-housekeeping function is highly significant for health of the general public. In interphase, detection of the DNA damage promotes formation of the multiprotein FA complex that orchestrates the repair of genetic lesions. Our previous work revealed that FA signaling ensures high-fidelity mitotic chromosome segregation through regulating the spindle assembly checkpoint (SAC) and maintaining centrosome structure and function (Nalepa et al, JCI 2013). These findings, together with the work of others, suggest that the genome maintenance role of the FA/BRCA pathway extends beyond interphase to prevent cancer. However, it is not mechanistically clear how disruption of the FA genes impairs the SAC and other stages of mitosis. Furthermore, although we have demonstrated that loss of the most common FA gene, FANCA, enhances both interphase and mitotic errors during human hematopoiesis (Abdul-Sater et al, Exp Hem 2015), it is not known whether faulty division of FA- deficient cells directly contributes to malignant transformation. In preliminary studies, we have conducted a synthetic lethal kinome-wide shRNA screen in FANCA-/- patient-derived primary cells and have identified strong novel candidate pathways that functionally interact with FA signaling during mitosis. Here, we propose to mechanistically dissect these newly found pathways ex vivo and in vivo. Further, we have generated a new genetically modified mouse model of FA to directly determine whether erroneous mitosis promotes leukemogenesis in the FA-deficient background through further genetic impairment of the SAC. Together, our work will provide mechanistic insights into genomic instability and malignant transformation resulting from loss of FA signaling, evaluate defective cell division as a causative factor in FA-/- leukemia, and create a preclinical platform to test personalized therapeutic strategies against FA-/- cancers in future studies.

抽象的 FA/BRCA 基因的双等位基因突变导致范可尼贫血 (FA)，这是一种骨髓衰竭综合征与基因组不稳定和癌症有关。患有 FA 的个体患有多种临床疾病表现，包括白血病和其他恶性肿瘤的高风险，导致终身发病率和死亡。此外，FA 基因的获得性突变发生在非 FA 的急性白血病和其他癌症中。患者，表明 FA 基因可以保护普通人群免受癌症侵害。因此，对FA的理解基因组管家功能对于公众的健康非常重要。在间期，检测 DNA 损伤促进多蛋白 FA 复合物的形成，该复合物协调遗传修复病变。我们之前的工作表明 FA 信号传导确保高保真有丝分裂染色体分离通过调节纺锤体组装检查点（SAC）并维持中心体结构和功能（Nalepa 等人，JCI 2013）。这些发现与其他人的工作一起表明基因组维护 FA/BRCA 途径的作用超出了间期，可预防癌症。然而，这并不是机械地清楚 FA 基因的破坏如何损害 SAC 和有丝分裂的其他阶段。此外，虽然我们已经证明，最常见的 FA 基因 FANCA 的缺失会增加间期错误和有丝分裂错误在人类造血过程中（Abdul-Sater 等人，Exp Hem 2015），目前尚不清楚 FA- 的错误分裂是否细胞缺陷直接导致恶性转化。在初步研究中，我们进行了在 FANCA-/- 患者来源的原代细胞中进行合成致死性全激酶组 shRNA 筛选，并鉴定出强在有丝分裂过程中与 FA 信号传导功能相互作用的新候选途径。在此，我们建议机械地剖析这些新发现的离体和体内途径。此外，我们还生成了一个新的转基因小鼠 FA 模型可直接确定错误的有丝分裂是否促进通过 SAC 的进一步遗传损伤，FA 缺乏背景下的白血病发生。在一起，我们的这项工作将为基因组不稳定性和因丢失而导致的恶性转化提供机制见解 FA 信号传导，评估细胞分裂缺陷作为 FA-/- 白血病的致病因素，并创建临床前研究在未来的研究中测试针对 FA-/- 癌症的个性化治疗策略的平台。