A gene trap mutagenesis screen for genes underlying cellular response to the mood stabiliser lithium

Matthew Gow, Dora Mirembe, Zaomba Longwe, Benjamin S Pickard

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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 MED10, MSI2 and three long intergenic non-coding (LINC) RNAs. Both MED10 and MSI2 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 in vitro functional selection/screening paradigm.

LanguageEnglish
Pages657-663
Number of pages7
JournalJournal of Cellular and Molecular Medicine
Volume17
Issue number5
Early online date12 Apr 2013
DOIs
Publication statusPublished - May 2013

Fingerprint

Lithium
Mutagenesis
Glycogen Synthase Kinases
Genes
beta Catenin
Neuroblastoma
Long Noncoding RNA
Therapeutics
Poly A
Inositol Phosphates
Bipolar Disorder
Cell Differentiation
Plasmids
Complementary DNA
Salts
Metals
Economics
Pathology
Polymerase Chain Reaction
Mutation

Keywords

  • gene trap
  • mutagenesis screen
  • underlying cellular response
  • mood stabiliser lithium

Cite this

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A gene trap mutagenesis screen for genes underlying cellular response to the mood stabiliser lithium. / Gow, Matthew; Mirembe, Dora; Longwe, Zaomba; Pickard, Benjamin S.

In: Journal of Cellular and Molecular Medicine, Vol. 17, No. 5, 05.2013, p. 657-663.

Research output: Contribution to journalArticle

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