Development and Optimization of a High-Content Analysis Platform to Identify Suppressors of Lamin B1 Overexpression as a Therapeutic Strategy for Autosomal Dominant Leukodystrophy.
Development and Optimization of a High-Content Analysis Platform to Identify Suppressors of Lamin B1 Overexpression as a Therapeutic Strategy for Autosomal Dominant Leukodystrophy.
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DOI：
10.1177/2472555220915821
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发表时间：
2020-09
期刊：
影响因子：
--
通讯作者：
Vogt A
中科院分区：
文献类型：
--
作者：
Nmezi B;Vollmer LL;Shun TY;Gough A;Rolyan H;Liu F;Jia Y;Padiath QS;Vogt A
常染色体显性脑白质营养不良（ADLD）是一种致命的、进行性的成人发病疾病，其特征是广泛的中枢神经系统（CNS）脱髓鞘和显著的发病率。发病年龄晚，疾病进展相对缓慢，为该疾病提供了较大的治疗窗口。然而，ADLD没有治疗方法，这是一种迫切且未满足的临床需求。我们以前已经表明，ADLD是由重复的核纤层蛋白B1基因引起的核纤层蛋白B1蛋白，核纤层的主要成分的表达增加，并证明，少突胶质细胞特异性过度表达的核纤层蛋白B1的转基因小鼠表现出时间和组织病理学特征，让人联想到人类疾病。由于核纤层蛋白B1水平的增加是触发ADLD的致病事件，因此旨在降低核纤层蛋白B1水平和相关功能后果的方法代表了发现小分子ADLD治疗剂的有希望的策略。为此，我们建立了一个诱导型细胞培养模型的核纤层蛋白B1过表达和开发高含量的分析与多变量分析，以定义，分析和量化核纤层蛋白B1的表达及其相关的异常核表型在小鼠成纤维细胞（MEF）。已对该测定法进行优化，以满足多日变异性研究中的高通量筛选（HTS）标准。为了控制原代MEF的批次间差异，我们实施了一种筛选策略，该策略采用哨兵细胞以避免HTS期间的昂贵损失。我们认为，该检测将确定真正的抑制核纤层蛋白B1的病理生理作为候选人开发成潜在的治疗ADLD。
Autosomal Dominant Leukodystrophy (ADLD) is a fatal, progressive adult-onset disease characterized by widespread central nervous system (CNS) demyelination and significant morbidity. The late age of onset together with the relatively slow disease progression provides a large therapeutic window for the disorder. However, no treatment exists for ADLD, representing an urgent and unmet clinical need. We have previously shown that ADLD is caused by duplications of the lamin B1 gene causing increased expression of the lamin B1 protein, a major constituent of the nuclear lamina, and demonstrated that transgenic mice with oligodendrocyte specific over-expression of lamin B1 exhibit temporal and histopathological features reminiscent of the human disease. As increased levels of lamin B1 are the causative event triggering ADLD, approaches aimed at reducing lamin B1 levels and associated functional consequences represent a promising strategy for discovery of small molecule ADLD therapeutics. To this end, we have created an inducible cell culture model of lamin B1 overexpression and developed high-content analysis in connection with multivariate analysis to define, analyze, and quantify lamin B1 expression and its associated abnormal nuclear phenotype in mouse fibroblasts (MEF). The assay has been optimized to meet high throughput screening (HTS) criteria in multi-day variability studies. To control for batch-to-batch variation in the primary MEFs we have implemented a screening strategy that employs sentinel cells to avoid costly losses during HTS. We posit the assay will identify bona fide suppressors of lamin B1 pathophysiology as candidates for development into potential therapies for ADLD.
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影响因子：
3.1
作者：
Nmezi, Bruce;Giorgio, Elisa;Padiath, Quasar S.
通讯作者：
Padiath, Quasar S.
影响因子：
3.5
作者：
Giorgio, Elisa;Robyr, Daniel;Brusco, Alfredo
通讯作者：
Brusco, Alfredo
影响因子：
11.4
作者：
Barascu, Aurelia;Le Chalony, Catherine;Bertrand, Pascale
通讯作者：
Bertrand, Pascale
影响因子：
1.8
作者：
Agresti, A;Coull, BA
通讯作者：
Coull, BA
影响因子：
11.4
作者：
Butin-Israeli, Veronika;Adam, Stephen A.;Goldman, Anne E.;Goldman, Robert D.
通讯作者：
Goldman, Robert D.