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

基本信息

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
项目类别：
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

项目摘要

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 21 三体症与多种发育异常有关，影响心血管、血液、呼吸、免疫、内分泌、胃肠道和神经系统。额外的 21 号染色体 (Chr) 副本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 名患有先天性心脏病 (CHD) 的 DS 患者。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，我们确定整倍体和 DS CHD 患者之间意外且明显的心脏基因表达差异。包括其中，我们发现 DS 中的 SOST（编码 Chr 17 上的硬化蛋白）表达量比在 DS 中高 10 倍整倍体CHD组织。由于硬化素抑制 WNT 和 BMP 信号传导，我们假设失调可能导致一些 DS 患者的 CHD。硬化蛋白也会影响骨密度，并可能有助于骨骼和骨骼健康 DS 的其他异常。我们建议扩大这些分析以确定异常基因的遗传基础影响 DS 表型的表达。我们假设对 21 三体性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 DS 表型。我们将定义这些基因型及其相关的调控网络改变细胞和器官的发育。我们将利用约 700 DS 的全基因组序列 (WGS) 具有不同 DS 表型的患者可用于拟议的研究。我们正在产生感应来自 200 名 DS 患者的多能干细胞 (iPSC)。 WGS 和 iPSC 衍生得到其他资金支持机制。我们建议分析 WGS 来定义与 DS 表型相关的序列元素。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 不同。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序列对基因表达的影响。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 表型相关的调控机制。我们还将探讨 DS 的后果是否- 与整倍体细胞中的相关变体，以确定三体性是否需要调节基因型。我们建议使用 DS 患者 iPSC 和废弃的整倍体和 21 三体心脏组织来验证发现。为了实现这些广泛的目标，我们将： 1) 使用整体鉴定与 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/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通过干扰 DS iPSC 和细胞谱系中的基因表达来调节 DS 表型。