Devices containing dna encoding neurotrophic agents and related compositions and methods

Abstract

Devices useful in the delivery of DNA encoding neurotrophic agents, anti-fibrotic agents, and related compositions are disclosed herein for use in the treatment of central and/or peripheral nervous system injury. Methods of making and using the disclosed devices and DNA are also described. In various embodiments, the invention also discloses compositions and devices that may further include a targeting agent, such as a polypeptide that is reactive with an FGF receptor (e.g., bFGF), or another ligand that binds to cell surface receptors on neuronal cells, or a support cell. The invention also discloses methods of promoting neuronal survival and regeneration via transfection of an axon as it grows through a device or composition of the present invention, or via transfection of a repair cell.

1 . A device for promoting neuronal survival or regeneration, comprising:
 a gene activated matrix comprising a biocompatible matrix and at least one neuronal therapeutic encoding agent having an operably linked promoter.   
     
     
         2 .- 5 . (canceled) 
     
     
         6 . The device of  claim 1  wherein the promoter is a neuronal cell specific promoter. 
     
     
         7 . The device of  claim 6  wherein the promoter is selected from the group consisting of GAP43 promoter, FGF receptor promoter and neuron specific enolase promoter. 
     
     
         8 . The device of  claim 1  wherein the neuronal therapeutic encoding agent encodes a neurotrophic factor. 
     
     
         9 .- 11 . (canceled) 
     
     
         12 . The device of  claim 1  wherein the neuronal therapeutic encoding agent encodes an inhibitor of an antagonist of axonal generation or regeneration. 
     
     
         13 .- 19 . (canceled) 
     
     
         20 . The device of  claim 1  wherein the neuronal therapeutic encoding agent is non-covalently associated with the gene activated matrix. 
     
     
         21 . The device of  claim 1  wherein the neuronal therapeutic encoding agent is absorbed in or adsorbed to the gene activated matrix. 
     
     
         22 .- 23 . (canceled) 
     
     
         24 . A device for promoting neuronal survival or regeneration, comprising:
 a gene activated matrix;   at least one support cell; and   at least one neuronal therapeutic encoding agent having an operably linked promoter.   
     
     
         25 . (canceled) 
     
     
         26 . The device of  claim 24  wherein the support cell is selected from the group consisting of: a Schwann cell an oligodendrocyte an astrocyte, a microglial cell and a fibroblast. 
     
     
         27 .- 31 . (canceled) 
     
     
         32 . The device of  claim 24  wherein the support cell is an inflammatory cell selected from the group consisting of a macrophage, a neutrophil, a monocyte, a granulocyte and a lymphocyte. 
     
     
         33 . (canceled) 
     
     
         34 . The device of  claims 1  or  24  wherein the gene activated matrix is an implant for a neuronal injury site. 
     
     
         35 .- 36 . (canceled) 
     
     
         37 . The device of  claims 1  or  24  wherein the gene activated matrix is a composition selected from the group consisting of a solution, a paste, a suspension, a powder, a semisolid, an emulsion and a gel. 
     
     
         38 .- 39 . (canceled) 
     
     
         40 . The device of  claims 1  or  24 , further comprising a targeting agent, wherein said targeting agent is complexed with or conjugated to the neuronal therapeutic encoding agent and is capable of binding a neuronal cell surface receptor a repair cell surface receptor, or extracellular matrix. 
     
     
         41 .- 45 . (canceled) 
     
     
         46 . The device of  claims 1  or  24 , further comprising a nucleic acid binding domain, wherein said nucleic acid binding domain binds to a nucleic acid sequence that forms a portion of the neuronal therapeutic encoding agent. 
     
     
         47 . The device of  claims 1  or  24 , further comprising at least one linker that is selected from the group consisting of a cleavable linker, a linker that provides an intracellular protein sorting peptide sequence, a linker that reduces steric hindrance, a linker that provides a nuclear translocation signal and a linker that possesses a nucleic acid condensing ability. 
     
     
         48 . The device of any one of  claims 1  or  24  wherein the device contains sub-physiologic amounts of a neuronal therapeutic agent. 
     
     
         49 . (canceled) 
     
     
         50 . A device according to  claims 1  or  24  or  25 , further comprising a conduit having a lumen. 
     
     
         51 .- 58 . (canceled) 
     
     
         59 . A method for transferring a neuronal therapeutic encoding agent into a neuronal cell, comprising:
 contacting a neuronal cell with the device of  claims 1  or  24  to effectively transfer the neuronal therapeutic encoding agent into the neuronal cell.   
     
     
         60 .- 61 . (canceled) 
     
     
         62 . The method of  claim 59  wherein the device is contacted with a neuronal cell at a neuronal injury site. 
     
     
         63 .- 67 . (canceled) 
     
     
         68 . A method for transferring a neuronal therapeutic encoding agent into a repair cell, comprising:
 contacting a repair cell with the device of  claims 1  or  24  to effectively transfer the neuronal therapeutic encoding agent into the repair cell.   
     
     
         69 . The method of  claim 68  wherein the device is contacted with a repair cell at a neuronal injury site. 
     
     
         70 .- 76 . (canceled) 
     
     
         77 . A method of preparing a gene activated matrix for promoting neuronal regeneration and survival, comprising contacting a neuronal therapeutic encoding agent with a biocompatible matrix such that the neuronal therapeutic encoding agent associates non-covalently with the matrix. 
     
     
         78 .- 80 . (canceled)