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

基本信息

批准号：
8578098
负责人：
THOMAS W GLOVER
金额：
$ 37.96万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
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/8578098
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

项目摘要

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）：镰状细胞疾病（SCD）是人类发现的第一个分子遗传疾病，影响了全世界5万名美国人和数百万的人。羟基脲（HU）目前是唯一经认可的成人SCD疾病疗法，正在接受晚期临床试验，用于治疗受影响的婴儿和儿童。 HU刺激胎儿血红蛋白的产生，因此有效地改善了氧气传输，同时降低了血管结合性危机的发生率和严重程度。然而，HU也是已知的复制抑制剂，诱变剂和clastogen，但是尚未研究HU在后代和治疗受试者的孙子中的遗传作用，尤其是在基因组水平上，对于复制应激的影响，例如拷贝数变化（CNV）突变。 CNV是正常遗传变异和进化的关键因素，是遗传疾病中常见且重要的突变类别，包括智力低下，自闭症，精神分裂症等。我们已经开发了人类细胞培养模型系统，以研究CNV突变的遗传和环境风险因素。我们发现，HU的浓度与治疗患者的血清浓度相同，可显着诱导正常人细胞中的CNV。这些发现对接受HU治疗的SCD和其他疾病的患者的儿童和孙子对有害的，从头的CNV风险具有重要的遗传意义。我们建议将这些发现扩展到在体内动物模型中的直接研究。我们将通过使用高分辨率基因组微阵列检查F1和F2世代治疗的小鼠，评估HU对男性和雌性种系中CNV诱导的遗传作用。我们将确定从头CNV的父母起源，检查其基因组结构以推断细胞和机械起源，并将发现与研究组动物的表型相比。此外，我们将评估用HU和对照组治疗的小鼠细胞和组织中CNV的体细胞镶嵌。这些研究将使用复制模型抑制剂为CNV的哺乳动物提供第一个体内测试。获得的结果将对人类的种系和躯体有害CNV定义遗传和环境风险因素具有重要意义。此外，对于大量的SCD和后代HU治疗的患者，它们将具有重要且直接的临床意义。