Functional characterization of myo-inositol monophosphatase

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/55 (2006.01) C12N 9/16 (2006.01) C12Q 1/68 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2523441

This invention relates to the functional characterization of myo-inositol monophosphatase 2 (IMPA2), one of the enzymes acting in the phosphatidyl inositol signaling pathway. In particular, the present invention provides evidence that IMPA2 is associated with depression- and anxiety inducing conditions, in particular anxiety and affective disorders. In a first aspect the present invention provides the use of an IMPA2 enzyme in an assay to identify anti-anxiety or an anti-depression compounds. In particular to the use of an isolated polynucleotide encoding said IMPA2 protein, wherein said IMPA2 protein is preferably being selected from polynucleotides encoding the mouse, rat or human IMPA2 enzyme. It is thus an object of the present invention to provide a method for identifying anti-anxiety or anti-depression compounds wherein said compounds are capable of enhancing neuronal plasticity, said method comprising the steps of: a) providing a composition comprising an IMPA2 protein; b) contacting the IMPA2 protein with the test compound; and c) measuring the activity of the IMPA2 protein wherein a decrease in the IMPA2 activity in the presence of the test compound is an indicator of an anti- anxiety or anti-depression compound. In these assays the activity of the IMPA2 protein is assessed by measuring the hydrolysis of myo-inositol 1-phosphate to generate inositol and inorganic phosphate, in particular by measuring the accumulation of either myo-inositol monophosphate product in the form of radiolabeled inositol or inorganic phosphate (Pi) in the form of radiolabeled32Pi or in a colorimetric assay. The compositions comprising the IMPA2 protein could either be cellular extracts, whole cells or organisms expressing the IMPA2 proteins according to the invention.

L'invention se rapporte à la caractérisation fonctionnelle de myo-inositol monophosphatase 2 (IMPA2), une des enzymes agissant dans la voie de signalisation de phosphatidyle inositol. L'invention concerne plus particulièrement la preuve que IMPA2 est associée à la dépression et l'anxiété induisant des pathologies, plus particulièrement les troubles affectifs et l'anxiété. Selon une première variante, l'invention concerne l'utilisation d'une enzyme IMPA2 dans un essai biologique afin d'identifier des composants d'anti-anxiété ou d'anti-dépression. On se penche plus particulièrement sur l'utilisation d'un polynucléotide isolé codant ladite protéine IMPA2, qui est sélectionnée de préférence parmi des polynucléotides codant l'enzyme IMPA2 de la souris, du rat ou de l'humain. L'invention vise donc à fournir un procédé d'identification de composés anti-anxiété ou anti-dépression aptes à améliorer la plasticité neuronale et consistant à : a) fournir une composition comprenant une protéine IMPA2; b) mettre en contact la protéine IMPA2 avec le composé de l'essai; et c) mesurer l'activité de la protéine IMPA2, une diminution de l'activité de IMPA2 en présence du composé de l'essai indiquant la présence d'un composé anti-anxiété ou anti-dépression. Dans ces essais biologiques, l'activité de la protéine IMPA2 protéine est évaluée par la mesure de l'hydrolyse demyo-inositol 1-phosphate afin de générer un phosphate inositol et inorganique, plus particulièrement par la mesure de l'accumulation du produit de myo-inositol monophosphate sous forme de phosphate inositol ou inorganique radiomarqué (Pi) sous forme de ?32¿Pi radiomarqué ou dans un essai colorimétrique. Les compositions comprenant la protéine IMPA2 peuvent être soit des extraits cellulaires, des cellules entières, soit des organismes exprimant les protéines IMPA2 de cette invention.

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