A gene trap mutagenesis screen for genes underlying cellular response to the mood stabilizer lithium. Issue 5 (12th April 2013)

• Record Type: Journal Article
• Authors: Gow, Matthew; Mirembe, Dora; Longwe, Zaomba; Pickard, Benjamin S.
• Is Part Of: Journal of cellular and molecular medicine. Volume 17:Issue 5(2013)
• Page Start: 657
• Page End: 663
• Publication Date: 2013-04-12
• Subjects: Cytology; Medicine; Molecular Biology; Cytologie -- Périodiques; Médecine -- Périodiques; Biologie moléculaire -- Périodiques; Cytology -- Periodicals; Medicine -- Periodicals; Molecular biology -- Periodicals; 611.01805
• DOI: 10.1111/jcmm.12048
• Languages: English
• ISSNs: 1582-1838

Abstract

<abstract abstract-type="main" xml:lang="en" id="jcmm12048-abs-0001"> <title>Abstract</title> <p>Identifying the biological pathways mediating the action of a therapeutic compound may help the development of more specific treatments while also increasing our understanding of the underlying disease pathology. Salts of the metal lithium are commonly used as a front‐line mood stabilizing treatment for bipolar disorder. Lithium's action has been variously linked to inositol phosphate metabolism and the WNT/Glycogen Synthase Kinase 3β (GSK3β)/β‐Catenin signalling cascade, but, to date, little is known about which of these provides the principal therapeutic benefit for patients and, more specifically, which constituent genes, through presumed sequence variation, determine differences in patient response to treatment. Here, we describe a functional screen in which SH‐SY5Y neuroblastoma cells were randomly mutated through genomic integration of the pMS1 poly A 'gene trap' plasmid vector. Lithium normally induces differentiation of neuroblastoma cells, but a small proportion of mutated cells continued to proliferate and formed colonies. Rapid amplification of cDNA ends (RACE)‐PCR was used to identify the 'trapped' gene in each of these lithium‐resistant colonies. Heterozygous, gene trap integrations were identified within ten genes, eight of which are likely to produce loss‐of‐function mutations including <italic>MED10</italic>, <italic> MSI2</italic> and three long intergenic non‐coding (LINC) RNAs. Both <italic>MED10</italic> and <italic>MSI2</italic> have been previously linked with WNT/GSK3β/β‐Catenin pathway function suggesting that this is an important mediator of lithium action in this screen. The methodology applied here provides a rapid, objective and economic approach to define the genetic contribution to drug action, but could also be readily adapted to any desired <italic>in vitro</italic> functional selection/screening paradigm.</p> </abstract>