Epigenome Editing Technologies for Treating Diverse Disease

新型核酸靶向系统的最新革命为治疗创造了难以置信的机会 通过对DNA和RNA进行有针对性的操作而导致的疾病。虽然到目前为止，重点主要放在基因组上 编辑以治疗罕见的遗传性疾病，这只代表了这些DNA靶向的一种机制 工具可以用来改善人类健康。事实上，更广泛的病理学集合可以是 通过调节基因调节和表观遗传状态来解决，而不是改变潜在的DNA 序列。此外，这种方法在效率、安全性和 可逆性。虽然一些研究已经证明了体内体细胞表观基因组的原理证明 编辑可用于编程细胞表型和调节治疗靶点，有许多 为治疗人类疾病准备这项技术必须克服的挑战。第一，一个理想 简便易行、应用广泛的DNA靶向系统尚未开发出来。而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是 受限于特异性，与大小受限的病毒载体不相容，以及人类先前存在的免疫 人口。因此，我们将挖掘满足以下条件的新的小CRISPR-Cas9/Cas12系统的细菌基因组 这些是在体表基因组编辑的标准。我们将研究表观基因组学的全基因组特异性 用无偏试验进行修改，并评估小鼠模型和预先存在的诱导免疫 人体样本中的免疫力。其次，在表观基因组编辑领域，尚不清楚哪些表观遗传学 为了在基因表达和基因组方面达到预期的结果，修饰是必要的且充分的 结构。我们将完成表观遗传状态和表观遗传学之间关系的全面分析 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，我们将在活体内验证这些表观基因组编辑工具，在小鼠模型上进行一系列先导实验 一种神经肌肉疾病，包括一组典型的表观基因组状态。与……密切合作 体细胞基因组编辑联盟，这项工作将为人类准备表观基因组编辑技术 在临床翻译中，它可能会对一系列稀有和常见的 疾病。

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.