De novo CNV formation in vivo with sickle cell anemia therapy

镰状细胞性贫血治疗体内从头形成 CNV

• 批准号: 8415873
• 负责人: THOMAS W GLOVER
• 金额: $ 36.43万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-30 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8415873
• 关键词: Adult; Affect; American; Animal Model; Animal Testing; Animals; Autistic Disorder; Biological Models; Cell Culture Techniques; Cell Therapy; Cells; Child; Clinical Treatment; Clinical Trials; Country; Data; Disease; Environmental Risk Factor; Female; Fetal Development; Fetal Hemoglobin; Fetus; Frequencies; Future Generations; Generations; Genes; Genetic; Genetic Polymorphism; Genetic Risk; Genetic Variation; Genomics; Germ-Line Mutation; Goals; Health; Hereditary Disease; Human; Inbred Strain; Incidence; Individual; Infant; Inherited; Location; Mammals; Mental Retardation; Methods; Mitotic; Modeling; Molecular Genetics; Monitor; Mosaicism; Multiple Anatomic Sites; Mus; Mutagens; Mutation; Oogenesis; Oogonia; Parents; Partner in relationship; Patients; Phenotype; Pregnancy; Production; Replication Error; Resolution; Risk; Risk Factors; Schizophrenia; Serum; Severities; Sickle Cell; Sickle Cell Anemia; Somatic Cell; Somatic Mutation; Spermatogenesis; Staging; Stress; Structure; Testing; Time; Tissue Sample; Tissues; Variant; Weaning; clastogen; clinically significant; genetic evolution; grandchild; high risk; hydroxyurea; improved; in vivo; inhibitor/antagonist; male; mouse model; offspring; oxygen transport; pregnant; public health relevance; risk variant

DESCRIPTION (provided by applicant): Sickle cell disease (SCD) is the first molecular genetic disorder identified in humans, affecting over 50,000 Americans and millions of people worldwide. Hydroxyurea (HU) is currently the only approved disease- modifying therapy for adult SCD and is in late-stage clinical trials for treatment of affected infants and children. HU stimulates fetal hemoglobin production and is therefore effective in improving oxygen transport while reducing the incidence and severity of vaso-occlusive crises. However, HU is also a known replication inhibitor, mutagen and clastogen, yet the genetic effects of HU in the offspring and grandchildren of treated subjects have not been studied, particularly at the genomic level for effects of replication stress such as copy number variant (CNV) mutations. CNVs are a key factor in normal genetic variation and evolution and are a common and important class of mutation in genetic disorders, including mental retardation, autism, schizophrenia and many others. We have developed a human cell culture model system to investigate the genetic and environmental risk factors for CNV mutations. We have found that HU, at concentrations identical to serum concentrations in treated patients, significantly induces CNVs in normal human cells. These findings have important and direct genetic implications for deleterious, de novo CNV risk in the children and grandchildren of patients treated with HU for SCD and other disorders. We propose to extend these findings to direct studies in animal models in vivo. We will evaluate the genetic effects of HU on CNV induction in both the male and female germlines by examining F1 and F2 generations of treated mice using high-resolution genomic microarrays. We will determine parental origins of de novo CNVs, examine their genomic structures to infer cellular and mechanistic origins and compare findings with those from phenotyping of study group animals. In addition, we will assess somatic mosaicism for CNVs in cells and tissues of mice treated prenatally with HU and controls. These studies will provide the first in vivo test in mammals of the somatic mutation/replication stress hypothesis for CNVs using a model inhibitor of replication. The results obtained will have important implications for defining genetic and environmental risk factors for both germline and somatic deleterious CNVs in humans. Moreover they will have important and immediate clinical significance to a large number of individuals with SCD and the future generations of HU-treated patients.

描述（由申请人提供）： 镰状细胞病（SCD）是第一个在人类中发现的分子遗传疾病，影响了超过50，000名美国人和全世界数百万人。羟基脲（HU）是目前唯一获批用于成人SCD的疾病缓解疗法，并处于治疗受影响婴儿和儿童的后期临床试验中。HU刺激胎儿血红蛋白的产生，因此有效地改善氧转运，同时降低血管闭塞性危象的发生率和严重程度。然而，HU也是一种已知的复制抑制剂、诱变剂和断裂剂，但尚未研究HU在治疗受试者的后代和孙辈中的遗传效应，特别是在基因组水平上对复制应激的影响，如拷贝数变异（CNV）突变。CNV是正常遗传变异和进化的关键因素，是遗传性疾病中常见且重要的一类突变，包括精神发育迟滞、自闭症、精神分裂症和许多其他疾病。我们已经开发了一个人类细胞培养模型系统来研究CNV突变的遗传和环境风险因素。我们已经发现，在与治疗患者的血清浓度相同的浓度下，HU显著诱导正常人细胞中的CNV。这些发现对于用HU治疗SCD和其他疾病的患者的子女和孙辈中有害的、新发CNV风险具有重要和直接的遗传意义。我们建议将这些发现扩展到体内动物模型的直接研究。我们将通过使用高分辨率基因组微阵列检查治疗小鼠的F1和F2代来评估HU对雄性和雌性生殖系中CNV诱导的遗传效应。我们将确定从头CNVs的亲本起源，检查其基因组结构以推断细胞和机制起源，并将结果与研究组动物的表型分析结果进行比较。此外，我们将评估产前用HU和对照处理的小鼠的细胞和组织中CNV的体细胞嵌合现象。这些研究将提供第一个在哺乳动物体内测试的体细胞突变/复制应激假说的CNVs使用模型的复制抑制剂。所获得的结果将具有重要的意义，为确定遗传和环境的危险因素，在人类生殖细胞和体细胞有害的CNVs。此外，它们将对大量患有SCD的个体和未来几代接受HU治疗的患者具有重要和直接的临床意义。