Understanding the complexity of gene dosage imbalance in Down syndrome

了解唐氏综合症基因剂量不平衡的复杂性

基本信息

批准号：
9894132
负责人：
STELLA T CHOU
金额：
$ 335.22万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-20 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9894132
关键词：
4 year old Acute Megakaryocytic Leukemias Address Alleles Amino Acids Aneuploidy Architecture Blood CRISPR/Cas technology Candidate Disease Gene Cardiac Cell Line Cell model Cells Child Childhood Leukemia Chromatin Chromosomes Chromosomes, Human, Pair 21 Clinical Clinical Trials Comorbidity Complex Complication Congenital Anemia Congenital Heart Defects DNA Methylation Diamond-Blackfan anemia Disease Dose Down Syndrome Epigenetic Process Erythroid Functional disorder GATA1 gene Gene Dosage Gene Expression Gene Expression Alteration Gene Mutation General Population Genes Genetic Genome Genomics Germ Layers Goals Hematological Disease Hematopoiesis Hematopoietic Human In Vitro Individual Intellectual functioning disability Intervention Knowledge Medical Methods Molecular Mus Mutation Myeloproliferative disease Neurons Newborn Infant Noise Pathologic Patients Pharmacology Pharmacology Study Phenotype Population Predisposition Presenile Alzheimer Dementia Production Proteins Relative Risks Research Role Signal Transduction Signaling Molecule Somatic Mutation Source Structure Syndrome System Technology Testing Tissues Translating United States National Institutes of Health Variant Viral actionable mutation clinical phenotype cohesion comparative dosage genome-wide human model induced pluripotent stem cell innovation innovative technologies insight interest leukemia leukemic transformation leukemogenesis mouse model mutational status neuropathology novel overexpression small hairpin RNA transcription factor transcriptome transcriptomics

项目摘要

ABSTRACT Patients with Down syndrome (DS, trisomy 21, T21) demonstrate a spectrum of clinical phenotypes due to an extra copy of chromosome 21 (HSA21). The clinical spectrum encompasses intellectual disability, early onset Alzheimer’s disease, congenital heart defects, and blood abnormalities including childhood leukemia. DS phenotypes are likely related to alteration of gene expression due to the extra copy of HSA21, and understanding the genomic determinants that contribute to the different phenotypes is a major objective in DS research. Our efforts will address several existing challenges that include: i. murine models do not recapitulate all of the human DS phenotypes, ii. manipulation of HSA21 gene dosage in murine and human DS cellular models have relied on methods that alter expression in non-physiologic doses, and iii. transcriptomic studies between normal and T21 tissues are challenged by considerable “noise” from inherent gene expression variation among individuals. In this application, we will address these challenges by using innovative technologies to create, genetically manipulate, and analyze patient-derived induced pluripotent stem cells (iPSCs). We will use a defined set of T21 iPSC lines and CRISPR-CAS9 gene editing to produce isogenic cell lines that differ only by copy number of HSA21 or specific candidate genes. Using isogenic T21 and euploid iPSCs, we will test whether T21 disrupts overall chromosomal architecture that results in genome-wide gene expression dysregulation. We will perform studies in gene expression, chromosome architecture, DNA methylation, and chromatin signatures to provide a comparative view of genome-wide transcriptome and chromatin contacts in isogenic T21 and euploid cells. Further studies will examine whether abnormalities are due to the overexpression of one or more specific HSA21 genes, or as a consequence of heterozygous mutations in CTCF or cohesion components identified in DS patients with leukemia. Our goal is to produce novel, medically relevant knowledge that advances our understanding of gene dosage imbalance in DS, with a particular interest in DS-associated leukemia and potential new treatments. We aim to not only provide insight into DS abnormalities, but may have broader implications for other diseases associated with aneuploidy.

抽象的唐氏综合症患者（DS，Trisome 21，T21）表现出由于临床表型 21染色体的额外副本（HSA21）。临床范围包括智力障碍，早期发作阿尔茨海默氏病，先天性心脏缺陷和血液异常，包括儿童白血病。 DS 表型可能与由于HSA21的额外副本和了解有助于不同表型的基因组决定者是DS的主要目标研究。我们的努力将应对包括：i的现有挑战。鼠模型不会概括所有人类DS表型，ii。在鼠和人类DS细胞中操纵HSA21基因剂量模型依赖于改变非生理剂量和III中表达的方法。转录组研究正常组织和T21组织之间的质疑是固有基因表达的“噪声”的挑战个体之间的差异。在此应用程序中，我们将通过使用创新来解决这些挑战创建，一般操纵和分析患者衍生的诱导多能干细胞的技术（IPSC）。我们将使用定义的T21 IPSC线和CRISPR-CAS9基因编辑来产生同源细胞仅由HSA21或特定候选基因的拷贝数不同的线。使用等源性T21和Euploid IPSC，我们将测试T21是否会破坏导致全基因组基因的总体染色体结构表达失调。我们将进行基因表达，染色体结构，DNA的研究甲基化和染色质特征，以提供全基因组转录组和等源性T21和多倍体细胞中的染色质接触。进一步的研究将检查异常是否是由于一个或多个特异性HSA21基因的过表达，或者由于杂合的结果 DS患者的CTCF或凝聚成分中的突变。我们的目标是生产新颖，医学上相关的知识，使我们对DS中基因剂量失衡的理解发展，对与DS相关的白血病和潜在的新疗法特别感兴趣。我们的目标不仅提供洞察力 DS异常，但可能对与非整倍性相关的其他疾病具有更广泛的影响。