Epigenome Editing Technologies for Treating Diverse Disease

用于治疗多种疾病的表观基因组编辑技术

• 批准号: 10438803
• 负责人: Charles A. Gersbach
• 金额: $ 39.85万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-08 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10438803
• 关键词: Address; Adoption; Algorithms; Angelman Syndrome; Animal Disease Models; BCAR1 gene; Bacterial Genome; Bioinformatics; Biological Assay; Biology; CRISPR/Cas technology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Custom; DNA; DNA Methylation; DNA Repair; DNA Sequence; DNA Sequence Alteration; Disease; Disease Progression; Enzymes; Epigenetic Process; Gene Expression; Gene Expression Regulation; Gene Transduction Agent; Genes; Genetic; Genetic Transcription; Genome; Genome engineering; Genomics; Goals; Guide RNA; Health; Hereditary Disease; Heritability; Higher Order Chromatin Structure; Histones; Human; Immunity; Individual; Inherited; Modification; Mutation; Neuromuscular Diseases; Nucleic Acids; Open Reading Frames; Outcome; Pathology; Population; Prader-Willi Syndrome; Primary Lateral Sclerosis; Property; Proteins; RNA; Rare Diseases; Risk; Safety; Sampling; Site; Somatic Cell; Specificity; Structure; System; Technology; Testing; Therapeutic; Translating; Variant; Viral; Viral Vector; Work; adeno-associated viral vector; base; biological research; biomaterial compatibility; clinical translation; epigenetic regulation; epigenome; epigenome editing; epigenomics; experimental study; gene therapy; genome editing; genome wide association study; genome-wide; histone modification; human disease; human pathogen; immunogenic; immunogenicity; improved; in vivo; mouse model; next generation; novel; nuclease; off-target mutation; preclinical development; programs; somatic cell gene editing; therapeutic target; tool

The recent revolution in novel nucleic acid-targeting systems has generated incredible opportunities for treating disease by targeted manipulation of DNA and RNA. While the emphasis thus far has largely been on genome editing to treat rare, inherited disorders, this represents only one mechanism by which these DNA-targeting tools can be applied to improve human health. In fact, a significantly broader set of pathologies can be addressed by modulating gene regulation and epigenetic states, in contrast to altering underlying DNA sequences. Moreover, this approach has a number of advantages with respect to efficiency, safety, and reversibility. While several studies have demonstrated proof-of-principle that in vivo somatic cell epigenome editing can be used to program cell phentoypes and modulate therapeutic targets, there are a number of challenges that must be overcome to prepare this technology for treatment of human disease. First, an ideal DNA-targeting system that is facile and broadly applicable has yet to be developed. While CRISPR-Cas9 systems have dramatically transformed genome engineering, their application for human epigenome editing is limited by specificity, incompatibility with size-restricted viral vectors, and pre-existing immunity in the human population. Therefore, we will mine bacterial genomes for novel small CRISPR-Cas9/Cas12 systems that meet these criteria for in vivo epigenome editing. We will examine genome-wide specificity of epigenomic modifications with unbiased assays and assess both induced immunity in mouse models and pre-existing immunity in human samples. Second, it remains unclear in the field of epigenome editing which epigenetic modifications are necessary and sufficient to achieve desired outcomes in gene expression and genome structure. We will complete a comprehensive analysis of the relationship between epigenetic states and epigenome editing activity to develop a set of rules for achieving corresponding changes in gene expression. Finally, we will validate these epigenome editing tools in vivo in a set of pilot experiments in mouse models of neuromuscular disease encompassing a representative set of epigenomic states. In close collaboration with the Somatic Cell Genome Editing Consortium, this work will prepare epigenome editing technology for human clinical translation in which it may have a transformative effect on a broad array of both rare and common disease.

新的核酸靶向系统的最新革命为治疗癌症创造了令人难以置信的机会。 通过靶向操纵DNA和RNA来治疗疾病。虽然迄今为止的重点主要是基因组 编辑来治疗罕见的遗传性疾病，这只代表了这些DNA靶向治疗的一种机制。 工具可以用来改善人类健康。事实上，可能存在更广泛的病理学 通过调节基因调控和表观遗传状态来解决，而不是改变潜在的DNA 序列的此外，这种方法在效率、安全性和可持续性方面具有许多优点。 可逆性虽然一些研究已经证明了体内体细胞表观基因组 编辑可用于编程细胞表型和调节治疗靶点， 这些挑战必须克服，以准备这项技术用于治疗人类疾病。首先，一个理想 简单易行且广泛适用的DNA靶向系统尚待开发。CRISPR-Cas9 系统极大地改变了基因组工程，它们在人类表观基因组编辑中的应用 受特异性、与大小受限的病毒载体的不相容性以及人中预先存在的免疫力的限制， 人口因此，我们将挖掘细菌基因组，以获得满足以下条件的新型小CRISPR-Cas9/Cas 12系统： 这些标准用于体内表观基因组编辑。我们将研究表观基因组特异性， 使用无偏试验进行修改，并评估小鼠模型中诱导的免疫力和预先存在的免疫力。 人体样本的免疫力其次，在表观基因组编辑领域， 修饰对于在基因表达和基因组修饰中实现期望的结果是必要的和足够的。 结构我们将完成对表观遗传状态与 表观基因组编辑活动，以制定一套规则，以实现基因表达的相应变化。 最后，我们将在小鼠模型中的一组先导实验中在体内验证这些表观基因组编辑工具。 神经肌肉疾病，包括一组代表性的表观基因组状态。的密切协作下 体细胞基因组编辑联盟，这项工作将准备表观基因组编辑技术，为人类 临床翻译，其中它可能对广泛的罕见和常见的 疾病