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.

抽象的 唐氏综合症（DS）或三体疾病与影响影响的多种发育异常有关 心血管，血液学，呼吸道，免疫学，内分泌，胃肠道和神经系统。 染色体（CHR）21的附加副本仍然存在这些系统问题的机制仍然存在 理解不佳。我们以前曾招募600例先天性心脏缺陷（CHD）和100例DS患者 无冠心病的DS患者。使用在手术修复期间获得的废弃CHD组织的分析，我们确定了 异倍体和DS CHD患者之间出乎意料的和不同的心脏基因表达差异。包括 其中我们发现，DS的SOST（在ChR 17上编码硬化蛋白）的表达比在 整倍体CHD组织。由于硬化蛋白抑制Wnt和BMP信号，我们假设失调可能 一些DS患者有助于CHD。硬化蛋白还会影响骨密度，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定义异常基因的遗传基础 影响DS表型的表达。 我们假设对三体第21三体的分子反应受到其他基因型的影响，这些基因型 导致患者特异性的DS表型。我们将定义这些基因型及其相关的监管网络 改变了细胞和器官的发育。我们将利用〜700 ds的整个基因组序列（WGS） 具有不同DS表型的患者将用于拟议研究。我们正在产生诱导 来自200名DS患者的多能干细胞（IPSC）。 WGS和IPSC推导得到其他资金的支持 机制。 我们建议分析WGS以定义与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 与IPSC区分开。通过将DS IPS细胞分化为心肌细胞（IPSC-CM）， 内皮，神经细胞和其他人将定义序列对基因表达的后果。我们将 还使用增强的CRISPR/ CAS9工程和IPSC的单细胞分析来研究该基因 与DS表型相关的调节机制。我们还将探讨DS-的后果是否 与整倍体细胞中的相关变体，以确定是否需要调节基因型。我们建议 使用DS患者IPSC和丢弃的Euploid和21心脏组织进行验证。 为了实现这些广泛的目标，我们将：1）使用整体确定与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Euploid IPSC，分化细胞和CHD组织的轮廓。 3）定义基因和调节途径 通过在DS IPSC和细胞谱系中扰动基因表达来调节DS表型。