Interrogating genome folding trajectories in health and disease

探究健康和疾病中的基因组折叠轨迹

• 批准号: 10473744
• 负责人: Stavros Lomvardas
• 金额: $ 62.27万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10473744
• 关键词: 3-Dimensional; Affect; Alleles; Alzheimer disease screening; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Anosmia; Architecture; Bar Codes; Biological Assay; Cells; Chemicals; Chromatin; Chromosomes; Complement; Complex; DNA; Detection; Development; Developmental Process; Disease; Down-Regulation; Enhancers; Environment; Environmental Risk Factor; Epigenetic Process; Essential Genes; Etiology; Exhibits; Future; Gene Activation; Gene Expression; Gene Expression Regulation; Gene Silencing; Generations; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genome; Genomics; Health; Heterochromatin; Human; Human body; Image; Individual; Label; Link; Maps; Molecular; Mus; Natural regeneration; Nerve Degeneration; Neurons; Nose; Nuclear; Odors; Olfactory Epithelium; Olfactory Pathways; Olfactory dysfunction; Organism; Output; Pharmacologic Substance; Prognosis; Property; Receptor Gene; Research; Resolution; Role; Route; Specificity; Symptoms; System; Technology; Transcriptional Activation; Transcriptional Regulation; Variant; Viral; base; cell type; diagnostic tool; experimental study; genetic manipulation; humanized mouse; hyposmia; in vivo; insight; interdisciplinary approach; molecular imaging; mouse model; mutant; nerve stem cell; neurotoxicity; novel; olfactory receptor; olfactory sensory neurons; prognostic tool; receptor expression; segregation; single molecule; tool

Project Summary Detection and identification of the astronomical number of volatile chemicals that we perceive as odors depends upon the monogenic and monoallelic expression of olfactory receptor (OR) genes in olfactory sensory neurons (OSNs). An intricate network of OSN-specific interchromosomal interactions coordinates the transcriptional activation of only 1 OR allele out of >2000 OR alleles distributed across 18 different chromosomes. Genomic interactions between silent OR genes assemble heterochromatic multi-chromosomal compartments that keep OR genes transcriptionally inactive, whereas genomic interactions between intergenic OR enhancers result in a multi-chromosomal enhancer hub that activates singular OR transcription. Here, we propose to combine Dip-C, a variation of single cell HiC, with viral-based cell tagging technologies, towards the identification of genome folding intermediates across OSN differentiation lineages. This “cradle to crate” genomic analysis will follow individual OSN progenitors and their barcoded progeny, allowing a complete cartography of genomic interactions made by every OR allele en route to transcriptional activation. Single molecule DNA FISH experiments will complement the proposed genomic studies, providing high resolution insight to the genomic choreography that orchestrates singular OR gene choice during development. Finally, we seek to explore how the trajectories of OSN genomic folding become altered and eventually disrupted in a humanized model for Alzheimer’s disease (AD). Olfactory dysfunction, hyposmia, and anosmia constitute well- established prodromal symptoms of AD, but the molecular etiology of this intriguing connection is not known. Using a humanized model for AD we discovered that interchromosomal OR compartments dissipate prior to the onset of neurodegeneration, resulting in strong downregulation of OR transcription. Thus, we propose to apply our single cell interrogation of genome folding transition in the context of AD and to establish the baseline of nuclear architecture in human OSNs. Deciphering how OR compartments assemble in health, and how they become disrupted in disease, may provide the basis for novel prognostic and diagnostic tools for AD, and molecular assays for in vivo screening of AD therapeutics.

项目摘要 我们认为是气味的挥发性化学物质的天文数量的检测和鉴定 取决于嗅觉感官中嗅觉受体（OR）基因的单相表达 神经元（OSN）。 OSN特异性互色体相互作用的复杂网络坐标 在> 2000中仅1或等位基因的转录激活或分布在18个不同的等位基因 染色体。静音或基因之间的基因组相互作用组装异颜色多染色体 保持或基因转录无活性的隔室，而基因组之间的基因组相互作用 或增强剂会导致一个多染色体增强子枢纽激活单数或转录。在这里，我们 提出将DIP-C与基于病毒的细胞标记技术相结合的DIP-C的建议 识别基因组折叠中间体跨OSN分化谱系。这个“摇篮” 基因组分析将遵循单个OSN祖细胞及其条形码后代，允许完整 每个或等位基因进行转录激活的基因组相互作用的制图。单身的 分子DNA鱼实验将完成提出的基因组研究，提供高分辨率 洞悉开发过程中精心策划单数或基因选择的基因组编排。最后， 我们试图探索OSN基因组折叠的轨迹如何改变并最终在 阿尔茨海默氏病（AD）的人源化模型。嗅觉功能障碍，低质量和厌食症构成很好的良好 已建立的AD前驱症状，但是这种有趣的联系的分子病因尚不清楚。 使用人源化模型，我们发现染色体或隔室在 神经变性的发作，导致强烈的下调或转录。那我们建议 在AD的背景下应用我们对基因组折叠过渡的单细胞询问，并建立 人类OSN核结构的基线。破译如何或隔间在健康中组装，以及 它们如何破坏疾病，可能为AD的新型预后和诊断工具提供基础， 和用于体内筛查AD疗法的分子测定。