Evaluating novel architectural proteins as therapeutic targets for cohesin dysfunction

摘要 科尔内利亚·德兰格综合症（CdLS）是一种罕见的发育障碍，影响多种器官 包括中枢神经系统，引起可变的神经发育延迟。CDLS主要是由 通过在粘附素复合物的调节或结构组分中的突变，这是必不可少的 维持与染色质相互作用并调节的含有染色体（SMC）蛋白的复合物 基因组折叠据推测，CdLS症状是转录失调的结果 这是由于内聚蛋白破坏后基因组结构的变化。到目前为止，还没有针对 直接纠正粘连蛋白功能障碍或染色质错误折叠。这些问题的部分原因是， 已知调节粘着蛋白介导的染色质折叠的因子。我们进行了高通量的荧光检测， 在人类细胞中进行基于原位杂交（Hi-FISH）的筛选，使用Oligopaint探针技术检测 并测量大量样本中的染色质相互作用。我们瞄准了约8,000个人类基因， 代表“可药物化的基因组”，并分离出五个候选基因， 与粘附素功能障碍相关的表型。本提案的目标是描述这些机构的作用， 新的“抗粘连蛋白因子”在核结构和粘连蛋白功能中的作用，并测试它们抑制 在CdLS模型中纠正基因错误表达。通过了解参与粘附素的分子因素 生物学和染色质折叠，我们可以开始考虑有针对性的方法来CdLS治疗 发展

Abstract Cornelia de Lange Syndrome (CdLS) is a rare developmental disorder affecting a multitude of organs including the central nervous system, inducing a variable neurodevelopmental delay. CdLS is primarily caused by mutations in regulatory or structural components of the cohesin complex, which are essential Structural Maintenance of Chromosomes (SMC) protein-containing complexes that interact with chromatin and modulate genome folding. It is hypothesized that CdLS symptoms are a consequence of transcriptional dysregulation due to changes in genome architecture upon cohesin disruption. To date, there are no therapies aimed at correcting cohesin dysfunction or chromatin misfolding directly. These issues are due, in part, to the paucity of factors known to regulate cohesin-mediated chromatin folding. We performed a high-throughput fluorescent in situ hybridization (Hi-FISH)-based screen in human cells, which uses Oligopaint probe technology to detect and measure chromatin interactions in a large number of samples. We targeted ~8,000 human genes, which represent the “druggable genome” and isolated five candidate genes whose depletion can offset cellular phenotypes associated with cohesin dysfunction. The goal of this proposal is to characterize the role of these novel ‘anti-cohesin factors’ in nuclear architecture and cohesin function and test the ability of their inhibition to correct gene misexpression in CdLS models. Through understanding the molecular factors involved in cohesin biology and chromatin folding, we can begin to consider targeted approaches to CdLS therapeutic development.