Evaluating novel architectural proteins as therapeutic targets for cohesin dysfunction

摘要 Cornelia de Lange综合征(CDLS)是一种罕见的累及多个器官的发育障碍 包括中枢神经系统，导致可变的神经发育延迟。CDLS主要是由 通过粘附素复合体的调节或结构成分的突变，这是必不可少的结构 染色体(SMC)与染色质相互作用并调节的含蛋白质复合体的维持 基因组折叠。假设CDLS症状是转录失调的结果。 这是由于凝集素破坏时基因组结构的变化。到目前为止，还没有针对 直接纠正粘附素功能障碍或染色质错误折叠。这些问题在一定程度上是由于缺乏 已知的调节粘附素介导的染色质折叠的因子。我们进行了高通量荧光检测 基于原位杂交(Hi-FISH)的人细胞筛查，使用寡核苷酸探针技术检测 并测量大量样本中的染色质相互作用。我们瞄准了大约8000个人类基因， 代表“可用药的基因组”，并分离出五个候选基因，它们的耗尽可以抵消细胞 与粘附素功能障碍相关的表型。这项提案的目标是描述这些角色的特征 核结构和粘附素功能中的新的抗粘附素因子及其抑制能力 纠正CDLS模型中的基因错误表达。通过了解与粘连有关的分子因素 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治疗镉 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.