Upper motor neurones corticospinal, Methods and compositions for Differentiating neural stem cells by modulating the Signaling of the CB1 cannabinoid receptor and Uses Thereof

摘要

In vitro method for obtain a higher corticospinal motor Neuron from a neural stem cell.The said method comprises contacting a neural stem cell in appropriate conditions to increase the signal mediated by the CB1 cannabinoid receptor in the cell and conditionS to promote the dorsalizaciu00f3n this cell and the appropriate conditions to increase the signal mediated by the CB1 cannabinoid receptor implies an increase in the number of cells CTISATB2 P2 + - with regard to basal levels of at least 5% and involve: (i) contacting Said cell with a neural cannabinoid CB1 receptor agonist selected GroupConsists of anandamide and 2-arachidonoylglycerol (2-ag), N - or arachidonyl ethanolamide dopamine, - 2 '- cloroetilamida (ACEA), Hu - 210, jwh-073, WIN 55212 - 2, cannabinol, tetrahydrocannabinol (THC),11 Hydroxy (- Oh? 9 - 11 - tetrahydrocannabinol THC), levonantradol, dronabinol, am-2201 (1 - (5 - fluoropentil) - 3 - (1 - naftoil) indole), jwh-018 (1 - pentyl - 3 - (1 - naftoil) indole), AM - 678,2 - (2 - age or arachidonyl glyceryl ether, ether noladina), Palmitoiletanolamida (PEA) and CP 55940; and / or (ii) introduce a neural stem cell in the expression vector encoding the CB1 cannabinoid receptor to increase expression of the cannabinoid receptor CB1 above itS basal levels.And / or (iii) introduce a neural stem cell in the expression vector that encodes an enzyme Synthesis of endocannabinoids in the cell to increase the activity of an enzyme SynthesisEndocannabinoids above basal levels.And / or (iv) make this a neural stem cell is an agent that endocannabinoid degradation Enzyme is selected from the group consisting of a Plasmid, a arnip,A ribozyme and antisense nucleic acid and / or contacting Said cell with a neural inhibitor the Enzyme degradation of endocannabinoids.And the conditions for Promoting the dorsalizaciu00f3n that involve contacting Said cell with a neural Stem Cell dorsalizaciu00f3n Agent selected from: (a) an inhibitor of SoniC Hedgehog selected from the group consisting of 3 ', 5' - cyclic monophosphate sodium salt of N6 '- -, O2 dibutyryl adenosine, ay 9944, cyclopamine KAAD cyclopamine -, forskolin, gant58, gant61, jk184,HPI HPI - 1, - 3, - 1 jervina, Sant robotnikinina 0449 -, GDC, and IPI - 926; and (b) a Smad inhibitor selected from the group consisting nogina, dorsomorfina, sb431542, LDN - 193189, Lefty, Activin,TGF beta and combinations thereof.