Site-specific control of human gene regulation for therapeutically applicable mechanistic insights

人类基因调控的位点特异性控制以获得治疗上适用的机制见解

• 批准号: 10282969
• 负责人: Isaac Hilton
• 金额: $ 37.71万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10282969
• 关键词: Affect; Cells; Chromatin; Chromatin Remodeling Factor; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Methylation; Disease; Equilibrium; Gene Expression; Gene Expression Regulation; Genes; Goals; Health; Histones; Human; Human Genome; Knowledge; Modification; Post-Translational Protein Processing; Process; Proteins; Proteomics; Regulator Genes; Regulatory Element; Research Personnel; Resolution; Site; System; Therapeutic; Widespread Disease; base; cell behavior; cell type; chromatin modification; epigenome editing; epigenomics; genomic locus; human disease; improved; insight; interdisciplinary approach; novel therapeutics; nuclease; programs; protein complex; spatiotemporal; tool; transcription factor

PROJECT SUMMARY Dysregulated gene expression is a widespread and disease-agnostic driver of human illness. Therefore, the ability to understand and precisely control gene expression has the potential to revolutionize the therapeutic landscape. The expression of human genes is naturally controlled by an elegant convergence of regulatory forces, including the physical compaction of chromatin, post-translational modifications (PTMs) to histone proteins, DNA methylation, and the dynamic engagement between transcription factors and chromatin modifying proteins and the human genome. Although this coordinated control safeguards the balance between health and disease, our mechanistic understanding of how these regulatory forces unite to drive human gene expression, and how they can be predictably redesigned for new therapies, remains limited. Programmable epigenome editing tools based upon nuclease null CRISPR/Cas-based systems have recently emerged and enable new ways to control endogenous human gene expression and covalent modifications to native chromatin. Despite this exciting progress, major technological and conceptual gaps remain. For instance, it is mechanistically unclear how the expression levels of specific genes can be precisely tuned over wide ranges in human cells. In addition, it is incompletely understood why the same transcription factors and chromatin modifiers have different effects when localized to specific regulatory elements in different human cell types. Further, the spatiotemporal stability and functional durations of transcription factors and chromatin modifiers at different genomic loci are not well defined. The goal of this MIRA project is to overcome these conceptual and technological gaps by developing and combining diverse CRISPR/Cas-based transcription factors and chromatin modifiers with bulk and single cell epigenomics and sensitive proteomics. We will use this multidisciplinary approach to answer fundamental questions about human gene regulatory mechanisms including: (1) How can human gene expression be site- specifically controlled at the resolution observed in health and disease? (2) What are the causal functions and operational stabilities of diverse chromatin modifications? (3) To what extent does chromatin compaction, modification state, protein complex composition, and spatial proximity affect the function of transcription factors and chromatin modifiers? Altogether, our proposal has great potential to uncover and enable control over pivotal mechanisms with broad and significant importance to human health.

项目概要 基因表达失调是人类疾病的一个广泛且与疾病无关的驱动因素。因此， 理解和精确控制基因表达的能力有可能彻底改变治疗方法 景观。人类基因的表达自然地受到监管的优雅融合的控制 力，包括染色质的物理压缩、组蛋白的翻译后修饰 (PTM) 蛋白质、DNA 甲基化以及转录因子和染色质修饰之间的动态结合 蛋白质和人类基因组。尽管这种协调控制保障了健康与健康之间的平衡 疾病，我们对这些调控力量如何联合起来驱动人类基因表达的机械理解， 以及如何针对新疗法进行可预测的重新设计仍然有限。 基于无核酸酶 CRISPR/Cas 系统的可编程表观基因组编辑工具最近已 的出现并实现了控制内源性人类基因表达和共价修饰的新方法 天然染色质。尽管取得了这些令人兴奋的进展，但主要的技术和概念差距仍然存在。例如， 从机制上讲，尚不清楚如何在大范围内精确调节特定基因的表达水平 在人体细胞中。此外，尚不完全理解为什么相同的转录因子和染色质 当修饰剂定位于不同人类细胞类型的特定调节元件时，会产生不同的效果。 此外，转录因子和染色质修饰剂的时空稳定性和功能持续时间 不同的基因组位点尚未明确定义。 MIRA 项目的目标是通过开发和克服这些概念和技术差距 将多种基于 CRISPR/Cas 的转录因子和染色质修饰剂与散装细胞和单细胞相结合 表观基因组学和敏感蛋白质组学。我们将使用这种多学科方法来回答基本问题 关于人类基因调控机制的问题包括：（1）人类基因表达如何进行位点- 具体控制在健康和疾病中观察到的分辨率？ (2) 因果函数是什么？ 不同染色质修饰的操作稳定性？ (3) 染色质压缩到什么程度， 修饰状态、蛋白质复合物组成和空间邻近性影响转录因子的功能 和染色质修饰剂？总而言之，我们的建议具有巨大的潜力，可以发现并实现对关键的控制 对人类健康具有广泛和重大意义的机制。