Molecular Causes of Down Syndrome Associated Congenital Heart Disease and Other Phenotypes

唐氏综合症相关先天性心脏病和其他表型的分子原因

• 批准号: 9894531
• 负责人: JONATHAN G SEIDMAN
• 金额: $ 372.43万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-19 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9894531
• 关键词: ATAC-seq; Affect; Attention; Blood; Bone Density; CRISPR/Cas technology; Cardiac; Cardiac Myocytes; Cardiovascular Abnormalities; Cardiovascular system; Cell Differentiation process; Cell Line; Cell Lineage; Cell Nucleus; Cells; Cessation of life; Child; Chromosomes; Chromosomes, Human, Pair 17; Chromosomes, Human, Pair 21; Clinical; Clinical Data; Complement; Congenital Heart Defects; DNA; Derivation procedure; Development; Diabetes Mellitus; Down Syndrome; Ear; Elements; Endocrine; Endothelium; Engineering; Epigenetic Process; Eye; Funding; Funding Mechanisms; Gene Abnormality; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genomics; Genotype; Goals; Grant; Health; Hematology; Immunologics; Individual; Infant; Infrastructure; Knowledge; Life; Link; Metabolic; Molecular; National Heart, Lung, and Blood Institute; Neurologic; Neurologic Deficit; Neurons; Online Systems; Operative Surgical Procedures; Participant; Pathway interactions; Patient Recruitments; Patients; Pediatrics; Phenotype; Procedures; Proteins; Protocols documentation; RNA analysis; Reagent; Regulator Genes; Regulatory Pathway; Research; Risk; Sampling; Secure; Sequence Analysis; Signal Transduction; Specimen; Stroke; Structure; System; Technology; Testing; Tissues; Transcript; Trisomy; Untranslated RNA; Variant; clinical phenotype; conditioning; congenital heart disorder; data hub; gastrointestinal; genetic variant; human subject; improved; induced pluripotent stem cell; innovation; insight; leukemia; obesity risk; organ growth; premature; recruit; relating to nervous system; repaired; respiratory; response; single cell analysis; skeletal; stem cell differentiation; transcriptome sequencing; whole genome

ABSTRACT Down syndrome (DS) or trisomy 21 is associated with multiple developmental anomalies affecting the cardiovascular, hematological, respiratory, immunological, endocrine, gastrointestinal and neurological systems. The mechanisms by which an additional copy of chromosome (Chr) 21 produces these systemic issues remains poorly understood. We have previously recruited 600 DS patients with congenital heart defects (CHD) and 100 DS patients without CHD. Using analyses of discarded CHD tissues obtained during surgical repair, we identified unexpected and distinct cardiac gene expression differences between euploid and DS CHD patients. Included among these we found that SOST (encoding sclerostin on Chr 17) expression was 10-fold higher in DS than in euploid CHD tissues. As sclerostin inhibits WNT and BMP signaling, we hypothesis that dysregulation may contribute to CHD in some DS patients. Sclerostin also impact bone density and may contribute to skeletal and other abnormalities in DS. We propose to expand these analyses to define genetic basis for abnormal gene expression that influence DS phenotype. We hypothesize that the molecular response to trisomy 21 are influenced by additional genotypes that result in patient-specific DS phenotypes. We will define these genotypes and their related regulatory networks that alter cell and organ development. We will capitalize on whole genome sequences (WGS) from ~700 DS patients with different DS phenotypes that will be available for the proposed studies. We are generating induced pluripotent stem cells (iPSCs) from 200 DS patients. WGS and iPSC derivation are supported by other funding mechanisms. We propose to analyze WGS to define sequence elements associated with DS phenotypes. We will explore the transcriptional and epigenetic consequences of these associations using DS iPSCs and cell lineages differentiated from iPSCs. Through the differentiation of DS iPS cells into cardiomyocytes (iPSC-CMs), endothelial, neural cells and others will define the consequences of sequences on gene expression. We will also use enhanced CRISPR/ Cas9 engineering and single cell analyses of iPSCs to investigate the gene regulatory mechanisms associated with DS phenotypes. We will also explore if the consequences of DS- associated variants with in euploid cells, to determine if trisomy is required conditioning genotype. We propose to validate finding using DS patient iPSCs and discarded euploid and trisomy 21 cardiac tissues. To achieve these broad goals, we will: 1) Identify genetic variants associated with DS phenotypes using whole genome sequence and RNA expression eQTLs. 2) Compare single cell/nucleus transcriptional and ATACseq profiles of DS and euploid iPSCs, differentiated cells, and CHD tissues. 3) Define genes and regulatory pathways that modulate DS phenotypes by perturbing gene expression in DS iPSCs and cell lineages.

摘要 唐氏综合征（DS）或21三体与多种发育异常有关， 心血管、血液、呼吸、免疫、内分泌、胃肠道和神经系统。 21号染色体的额外拷贝产生这些系统性问题的机制仍然存在 不太了解。我们之前招募了600名患有先天性心脏病（CHD）的DS患者和100名 无CHD的DS患者。通过对手术修复过程中获得的废弃CHD组织进行分析， 整倍体和DS CHD患者之间的意外和独特的心脏基因表达差异。包括 其中，我们发现SOST（编码Chr 17上的硬化蛋白）表达在DS中比在DGS中高10倍。 整倍体CHD组织。由于sclerostin抑制WNT和BMP信号传导，我们假设调节异常可能 在某些DS患者中导致CHD。硬化蛋白还影响骨密度，并可能有助于骨骼和 DS中的其他异常。我们建议扩展这些分析，以确定异常基因的遗传基础 表达影响DS表型。 我们假设对21三体的分子反应受到其他基因型的影响， 导致患者特异性DS表型。我们将定义这些基因型及其相关的调控网络 从而改变细胞和器官的发育。我们将利用全基因组序列（WGS）从约700 DS 可用于拟定研究的具有不同DS表型的患者。我们正在产生诱导 多能干细胞（iPSC）来自200名DS患者。WGS和iPSC衍生得到其他资金的支持 机制等 我们建议分析WGS来定义与DS表型相关的序列元件。我们将 使用DS iPSC和细胞谱系探索这些关联的转录和表观遗传后果 从iPSC分化而来。通过DS iPS细胞分化为心肌细胞（iPSC-CM）， 内皮细胞、神经细胞和其他细胞将定义基因表达序列的结果。我们将 我还使用增强的CRISPR/ Cas9工程和iPSC的单细胞分析来研究基因 与DS表型相关的调节机制。我们还将探讨如果DS的后果- 在整倍体细胞中的相关变体，以确定是否需要调节基因型的三体性。我们提出 使用DS患者iPSC和丢弃的整倍体和21三体心脏组织验证发现。 为了实现这些广泛的目标，我们将：1）使用全基因组测序技术鉴定与DS表型相关的遗传变异。 基因组序列和RNA表达eQTL。2)比较单细胞/核转录和ATACseq DS和整倍体iPSC、分化细胞和CHD组织的概况。3)定义基因和调控途径 其通过干扰DS iPSC和细胞谱系中的基因表达来调节DS表型。