Rho family antagonists and their use to block inhibition of neurite outgrowth
AssigneeUniversite de Montreal
InventorsLisa McKerracher, Maxime Lehmann
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Abstract
The invention relates to an antagonist of one or more of Rho family members having ability to elicit neurite outgrowth from cultured neurons in an assay method which includes culturing neurons on a substrate that incorporates a growth-inhibiting amount of Rho family member and exposing the cultured neurons to a candidate Rho family member antagonist agent to permit neuron growth. Candidates which elicit neurite outgrowth from the cultured neurons are thus identified as Rho family antagonists.
Claims21
An antagonist of one or more of Rho family members characterized by the ability to elicit neurite outgrowth from cultured neurons in an assay method, comprising the steps of:
- (a) culturing neurons on a growth permissive substrate that incorporates a growthinhibiting amount of Rho family member;
- and
- (b) exposing the cultured neurons of step a) to a candidate Rho family member antagonist agent in an amount and for a period sufficient prospectively to permit growth of the neurons;
- thereby identifying as Rho family antagonists the candidates of step b) which elicit neurite outgrowth from the cultured neurons of step a).
The use of antagonists of one or more Rho family members to promote neural growth by inhibiting Rho family members in the central nervous system.
The use of ADP-ribosyl transferase C3, or other closely related toxins, to promote neural growth by inhibiting one or more Rho family members in the central nervous system.
The use of a GTPase activating protein that is specific to Rho to convert GTP-bound active Rho to GDP-bound inactive Rho.
The use of biologically active fragments of ADP-ribosyl transferase C3, analogs and derivatives thereof, to promote neural growth by inhibiting one or more Rho family members in the central nervous system.
The use of Y27632, or related compounds, to promote neural growth by inhibiting Rho2 associated kinase in the central nervous system.
The use of genetically mutated forms of Rho, to promote neural growth by inhibiting one or more Rho family members in the central nervous system.
The use of dominant negative Rho to inactivate Rho, to foster axon growth in the central nervous system.
The use of GDP dissociation inhibitors, or stimulation thereof, to inhibit the dissociation of GDP from Rho and thereby prevent the binding of GTP necessary for the activation of Rho.
A method for producing Rho antagonists from Rho family members, fragments, analogs of derivatives by peptide synthesis or by recombinant DNA expression of either a truncated domain of Rho family members, incorporating one or more L- or D-amino acid substitutions, or of intact Rho family members using standard recombinant procedures and selecting antagonist characterized by the ability to elicit neurite outgrowth from cultured neurons in an assay method, comprising the steps of:
- (a) culturing neurons on a growth permissive substrate that incorporates a growth-inhibiting amount of a Rho family member;
- and
- (b) exposing the cultured neurons of step a) to a candidate Rho family member antagonist agent in an amount and for a period sufficient prospectively to permit growth of the neurons;
- thereby identifying as Rho family antagonists the candidates of step b) which elicit neurite outgrowth from the cultured neurons of step a).
An antagonist of one or more of Rho family members, characterized by the following properties:
- (a) blocks growth inhibition of neurites by myelin or myelin proteins;
- and
- (b) interferes with Rho family members-mediated growth inhibition as competitive but non-functional mimics of endogenous Rho family members.
An assay method useful to identify Rho family member antagonist agents that suppress inhibition of neuron growth, comprising the steps of:
- (a) culturing neurons on a growth permissive substrate that incorporates a growthinhibiting amount of a Rho family member;
- and
- (b) exposing the cultured neurons of step a) to a candidate Rho family member antagonist agent in an amount and for a period sufficient prospectively to permit growth of the neurons;
- thereby identifying as Rho family antagonists the candidates of step b) which elicit neurite outgrowth from the cultured neurons of step a).
A method to suppress the inhibition of neuron, comprising the steps of delivering, to the nerve growth environment, a Rho family antagonist in an amount effective to reserve myelin inhibition.
Dependencies are inferred from the claim text.