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基因中的双重突变引起fanconi贫血（FA），骨髓衰竭综合征与基因组不稳定性和癌症有关。 FA患者患有广泛的临床表现形式，包括白血病的高风险和其他导致终生发病率和的恶性肿瘤死亡。此外，急性白血病和非FA中的其他癌症发生了获得的FA基因突变患者，表明FA基因保护普通人群免受癌症的侵害。因此，对足总的理解基因组房屋保存功能对于公众的健康非常重要。在相间，检测 DNA损伤促进了策划遗传修复的多蛋白FA复合物的形成病变。我们以前的工作表明，FA信号传导可确保高保真有丝分裂染色体隔离通过调节主轴组件检查点（SAC）并保持中心体结构和功能（Nalepa等，JCI 2013）。这些发现以及他人的工作表明基因组维持 FA/BRCA途径的作用超出了相间的范围，以预防癌症。但是，它不是机械上的清除FA基因的破坏如何损害有丝分裂的SAC和其他阶段。此外，尽管我们已经证明了最常见的FA基因Fanca的丧失会增强相间和有丝分裂误差在人类造血（Abdul-Sater等人，Exp Hem 2015）期间，尚不清楚FA-分裂是否有故障缺乏细胞直接导致恶性转化。在初步研究中，我们进行了 Fanca中的合成致命性shRNA筛查 - / - 衍生的原代细胞，并确定了强烈的在有丝分裂过程中与FA信号相互作用的新型候选途径。在这里，我们建议机械剖析这些新发现的途径在体内和体内。此外，我们已经产生了一个新的 FA的转基因小鼠模型直接确定是否促进错误有丝分裂通过进一步的SAC遗传损害，在FA缺陷背景中的白血病发生。在一起，我们的工作将为基因组不稳定性和因损失而导致的恶性转化提供机械洞察力 FA信号传导，将缺陷的细胞分裂评估为FA - / - 白血病中的因子，并创建临床前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。