Lukas Jan, Knospe Anne-Marie, Seemann Susanne, Citro Valentina, Cubellis Maria V, Rolfs Arndt
Albrecht-Kossel-Institute, University Rostock Medical Center;
Albrecht-Kossel-Institute, University Rostock Medical Center.
J Vis Exp. 2017 Dec 20(130):56550. doi: 10.3791/56550.
The use of personalized medicine to treat rare monogenic diseases like lysosomal storage disorders (LSDs) is challenged by complex clinical trial designs, high costs, and low patient numbers. Hundreds of mutant alleles are implicated in most of the LSDs. The diseases are typically classified into 2 to 3 different clinical types according to severity. Moreover, molecular characterization of the genotype can help predict clinical outcomes and inform patient care. Therefore, we developed a simple cell culture assay based on HEK293H cells heterologously over-expressing the mutations identified in Fabry and Pompe disease. A similar assay has recently been introduced as a preclinical test to identify amenable mutations for Pharmacological Chaperone Therapy (PCT) in Fabry disease. This manuscript describes an amended cell culture assay which enables rapid phenotypic assessment of allelic variants in Fabry and Pompe disease to identify eligible patients for PCT and may aid in the development of novel pharmacochaperones.
使用个性化医疗来治疗诸如溶酶体贮积症(LSDs)等罕见单基因疾病面临着复杂的临床试验设计、高昂成本和患者数量少等挑战。大多数溶酶体贮积症涉及数百个突变等位基因。这些疾病通常根据严重程度分为2至3种不同的临床类型。此外,基因型的分子特征有助于预测临床结果并为患者护理提供信息。因此,我们基于异源过表达在法布里病和庞贝病中鉴定出的突变的HEK293H细胞开发了一种简单的细胞培养检测方法。最近引入了一种类似的检测方法作为临床前测试,以鉴定法布里病中适合药物伴侣疗法(PCT)的突变。本手稿描述了一种改良的细胞培养检测方法,该方法能够快速对法布里病和庞贝病中的等位基因变体进行表型评估,以确定适合接受PCT的患者,并可能有助于新型药物伴侣的开发。