Functional roles of genetic risk factors for brain disorders in neurogenesis and neurodevelopment

脑部疾病遗传危险因素在神经发生和神经发育中的功能作用

• 批准号: 10065021
• 负责人: Guo-li Ming
• 金额: $ 80.5万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10065021
• 关键词: Adult; Affect; Animal Model; Behavior; Biological; Biology; Brain; Brain Diseases; Cell Culture System; Cell physiology; Cells; Copy Number Polymorphism; DNA Sequence Alteration; Development; Diagnosis; Disease; Environmental Risk Factor; Event; Generations; Genetic; Genetic Risk; Genetic Variation; Goals; Human; Human Development; Induced pluripotent stem cell derived neurons; Intervention; Knowledge; Measures; Molecular; Mus; Neurodevelopmental Disorder; Neurons; Neurophysiology - biologic function; Patients; Process; Research; Resolution; Resources; Risk; Risk Factors; Role; Technology; Testing; Therapeutic Intervention; Tissues; Transgenic Mice; adult neurogenesis; base; cell behavior; design; differentiation protocol; genetic risk factor; human stem cells; induced pluripotent stem cell; mouse model; nerve stem cell; nervous system disorder; neural model; neurodevelopment; neurogenesis; neuromechanism; neuron development; novel; prevent; programs; relating to nervous system; risk variant; stem cell biology; synaptogenesis; targeted treatment; technological innovation; three dimensional cell culture; tool; transcriptome

The overarching goal of this program is to define cellular and molecular events during neural development vulnerable to genetic perturbations that increase risk for neurodevelopmental and neurological disorders. Currently, our knowledge of human brain development is largely inferred from animal models, indirect measures of human development, and limited access to human neural tissue. All of these are valid tools to piece together the sequential processes of human neural development but are not sufficient to describe the dynamics with enough temporal or molecular resolution to understand mechanistically how genetic risk factors can affect brain formation and function. Technological advances in cellular reprogramming have now made it possible to derive induced pluripotent stem cells (iPSCs) from adult patients, which are a renewable resource for the generation of human neurons with disease-relevant genetic features. This long-term research program is designed to incorporate human iPSC-based studies with animal models to provide a comprehensive and longitudinal understanding of neural development, from neural stem cell behavior to neuronal development, synapse formation and circuit integration. As a proof-of-principle, these studies will use a prominent copy number variation (CNV) risk factor for multiple neurological disorders, 15q11.2CNVs, to illustrate how multifaceted interrogations of the basic biology of neural development in the context of genetic variation can reveal new targets for testing mechanism-based intervention in relevant subtypes of human neurons, as well as animal models of neural function and behavior. Building on significant scientific discoveries we have made in the fields of stem cell biology, adult neurogenesis, and patient-specific iPSCs, and technological innovations we have developed to meet critical challenges in each of these fields, our primary research focus is to integrate multiple levels of analysis to provide a high-resolution description of the cellular processes and molecular mechanisms of neural development that can be used to probe genetic or environmental risk for neurological disorders. Three interlinked projects will be pursued. Project 1 will focus on adult mouse neurogenesis as a model for neural development and use clonal analysis of neural stem cells and their development, single-cell transcriptome analysis, and transgenic mouse models to dissect molecular, cellular, and circuit level effects of genetic mutations on neural development; Project 2 will use human iPSCs with known genetic risk factors, and targeted differentiation protocols, to interrogate human neural development in 2D and 3D cultures; and Project 3 will focus on identifying the molecular mechanisms and targets of risk genes in both animal models and human iPSC-derived neurons and the rescue of observed deficits through rational therapeutic intervention. This is an opportune moment to synthesize recently developed technologies and build a novel translational platform to study underlying mechanisms of neurological disorders, and facilitate the identification of strategies to diagnose, treat, and prevent the often debilitating consequences of dysregulated neural development.

该计划的总体目标是定义神经发育过程中易受遗传扰动影响的细胞和分子事件，这些扰动会增加神经发育和神经系统疾病的风险。目前，我们对人类大脑发育的了解主要是从动物模型、人类发育的间接测量以及对人类神经组织的有限获取中推断出来的。所有这些都是将人类神经发育的顺序过程拼凑在一起的有效工具，但不足以以足够的时间或分子分辨率来描述动态，从而从机制上理解遗传风险因素如何影响大脑的形成和功能。细胞重编程技术的进步现在使得从成年患者中提取诱导多能干细胞（iPSC）成为可能，这是一种可再生资源，用于产生具有疾病相关遗传特征的人类神经元。这项长期研究计划旨在将基于人类 iPSC 的研究与动物模型相结合，以提供对神经发育的全面纵向理解，从神经干细胞行为到神经元发育、突触形成和电路整合。作为原理验证，这些研究将使用多种神经系统疾病的重要拷贝数变异 (CNV) 风险因素 15q11.2CNV，以说明如何在遗传变异的背景下对神经发育的基础生物学进行多方面的探究揭示了测试人类神经元相关亚型以及神经功能和行为动物模型的基于机制的干预的新目标。基于我们在干细胞生物学、成体神经发生和患者特异性 iPSC 领域取得的重大科学发现，以及我们为应对这些领域的关键挑战而开发的技术创新，我们的主要研究重点是整合多个层面分析提供神经发育的细胞过程和分子机制的高分辨率描述，可用于探测神经系统疾病的遗传或环境风险。将开展三个相互关联的项目。项目1将重点关注成年小鼠神经发生作为神经发育模型，并利用神经干细胞及其发育的克隆分析、单细胞转录组分析和转基因小鼠模型来剖析基因突变对神经发生的分子、细胞和回路水平的影响。神经发育；项目 2 将使用具有已知遗传风险因素的人类 iPSC 和有针对性的分化方案，来探究 2D 和 3D 培养物中的人类神经发育；项目3将重点确定动物模型和人类iPSC衍生神经元中风险基因的分子机制和靶点，并通过合理的治疗干预来挽救观察到的缺陷。这是综合最近开发的技术并建立一个新颖的转化平台来研究神经系统疾病的潜在机制，并促进确定诊断、治疗和预防神经发育失调所带来的经常衰弱后果的策略的好时机。