Modeling uniquely human developmental gene regulatory networks using humanized mice

使用人源化小鼠模拟独特的人类发育基因调控网络

基本信息

批准号：
10579859
负责人：
James P Noonan
金额：
$ 56.81万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-10 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10579859
关键词：
3-Dimensional ATAC-seq Acceleration Affect Animal Model Architecture Binding Biological Process CRISPR/Cas technology Cell physiology Cells Cerebral cortex ChIP-seq Chondrocytes Coupled Data Development Developmental Gene Developmental Process Digit structure Elements Embryo Enhancers Epigenetic Process Evolution Exhibits Experimental Models Funding Gene Expression Gene Expression Regulation Genes Genetic Transcription Genetic Variation Genome Goals Growth Human Human Development Human Genetics Length Limb Bud Limb Development Limb structure Macaca mulatta Maps Modeling Molecular Morphogenesis Morphology Mus Mutation Orthologous Gene Pan Genus Pattern Phenotype Play Regulator Genes Regulatory Element Regulatory Pathway Research Project Grants Resolution Role Specific qualifier value Time Walking Wild Type Mouse biochip cell type chromosome conformation capture cognitive ability data integration developmental disease epigenetic profiling gain of function gene regulatory network genome editing genome-wide human tissue humanized mouse in vivo innovation insight mouse model novel single-cell RNA sequencing skeletal temporal measurement tool trait transcription factor transcriptome

项目摘要

Physical adaptation is a hallmark of human evolution. Morphological changes in our limbs allowed us to make and use sophisticated tools and to walk upright, and the expansion of the human cortex is the origin of our advanced cognitive abilities. These traits are ultimately encoded in genetic changes that arose during human evolution, and which acted to alter molecular and cellular processes during development. The goal of this ongoing research project, which began in 2010, is to determine where in the genome those changes reside and to understand their biological functions. Our efforts focus on two classes of gene regulatory elements that may encode novel functions in humans. The first are Human Accelerated Regions (HARs), many of which encode transcriptional enhancers that are highly conserved across species but show multiple human-specific sequence changes. The second class of elements are Human Gain Enhancers (HGEs), which are transcriptional enhancers that show increased activity in developing human tissues based on comparisons of epigenetic marks associated with enhancer activity in human, rhesus macaque and mouse. These discoveries reveal that changes in developmental gene regulation played a central role in the evolution of uniquely human morphology and provide the means to experimentally model the evolution of human development. In this funding cycle, we will use humanized mouse models to study the biological function of HACNS1, which we identified as the first known HAR to encode human-specific regulatory activity. HACNS1 maintains its human-specific activity in the developing mouse limb and alters the expression of the nearby transcription factor Gbx2 in the developing embryo. We hypothesize that HACNS1 is acting within a larger network of human-specific regulatory changes that modified development. Using the HACNS1 mouse model and our maps of human-specific regulatory functions in the limb as entry points, we will identify transcriptional and regulatory changes downstream of HACNS1 at single-cell resolution, model additional human-specific regulatory functions that may interact with the regulatory changes driven by HACNS1, and determine how these changes influence the development of the limb.

身体适应是人类进化的标志。四肢的形态变化使我们能够并使用复杂的工具并直立行走，而人类皮质的扩展是我们的起源高级认知能力。这些特征最终在人类期间出现的遗传变化中编码进化，并在发育过程中改变了分子和细胞过程。目标的目标始于2010年的正在进行的研究项目是确定基因组中的这些变化位于哪里，并且了解他们的生物学功能。我们的努力专注于两类基因调节元素，可能编码人类中的新功能。第一个是人类加速区（HAR），其中许多编码转录增强子在物种之间高度保守但显示多个人类特异性序列更改。第二类元素是人力增强器（HGES），它们是转录的根据表观遗传标记的比较，增强剂显示出增强的人体组织活性与人类，恒河猴和小鼠的增强剂活性相关。这些发现揭示了变化在发育中的基因调节在独特的人类形态的进化中起着核心作用提供实验建模人类发展的进化的手段。在这个资金周期中，我们将使用人源化的小鼠模型研究HACNS1的生物学功能，我们将其确定为第一个已知用于编码人类特异性调节活动的HAR。 HACNS1保持其人类特异性活动发育中的小鼠肢体并改变了附近转录因子GBX2在开发胚胎。我们假设HACNS1在更大的人类特异性网络中起作用改变发展的监管变化。使用HACNS1鼠标模型和我们的地图肢体中人类特异性的调节功能作为入口点，我们将确定转录和 HACNS1下游的调节变化在单细胞分辨率下，建模其他人类特异性可能与HACNS1驱动的监管变化相互作用的调节功能，并确定这些变化如何影响肢体的发展。