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 regeneration, comprising: 
 a gene activated matrix comprising a biocompatible matrix and at least one neuronal therapeutic encoding agent having an operably linked promoter.    
     
     
         2 . A device for promoting neuronal survival, comprising: 
 a gene activated matrix comprising a biocompatible matrix and at least one neuronal therapeutic encoding agent having an operably linked promoter.    
     
     
         3 . The device of either  claim 1  or  claim 2  wherein the promoter is an inducible promoter.  
     
     
         4 . The device of either  claim 1  or  claim 2  wherein the promoter is a tissue specific promoter.  
     
     
         5 . The device of either  claim 1  or  claim 2  wherein the promoter is selected from the group consisting of GAP43 promoter, GFAP promoter, neuron specific enolase promoter, FGF-receptor promoter, elastase I gene control region, immunoglobulin gene control region, alpha-1-antitrypsin gene control region, beta-globin gene control region, myelin basic protein gene control region, myosin light chain 2 gene control region, RSV promoter, vaccinia virus 7.5K promoter, SV40 promoter, HSV promoter, MLP adenovirus promoter, MMTV LTR promoter, CMV promoter, metallothionein promoter, a promoter having at least one cAMP response element, tie promoter, VCAM-1 promoter, alpha V-beta 3 integrin promoters, ICAM-3 promoter, CD44 promoter, CD40 promoter, notch 4 promoter, and an event type-specific promoter.  
     
     
         6 . The device of either  claim 1  or  claim 2  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 either  claim 1  or  claim 2  wherein the neuronal therapeutic encoding agent encodes a neurotrophic factor.  
     
     
         9 . The device of  claim 8  wherein the neurotrophic factor is a member of the neurotrophin family.  
     
     
         10 . The device of  claim 8  wherein the neurotrophic factor is a member of the FGF family.  
     
     
         11 . The device of  claim 8  wherein the neurotrophic factor is selected from the group consisting of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), cardiotrophin-1 (CT-1), choline acetyltransferase development factor (CDF), ciliary neurotrophic factor (CNTF), oncostatin M (OSM); fibroblast growth factor-1 (FGF-1), FGF-2, FGF-5, glial cell-line-derived neurotrophic factor (GDNF), insulin, insulin-like growth factor-1 (IGF-1), IGF-2, interleukin-6 (IL-6), leukemia inhibitor factor (LIF), neurite promoting factor (NPF), neurotrophin-3 (NT-3), NT-4, platelet-derived growth factor (PDGF), protease nexin-1 (PN-1), S-100, transforming growth factor-β (TGF-β), and vasoactive intestinal peptide (VIP).  
     
     
         12 . The device of either  claim 1  or  claim 2  wherein the neuronal therapeutic encoding agent encodes an inhibitor of an antagonist of axonal generation or regeneration.  
     
     
         13 . The device of  claim 12  wherein the inhibitor of an antagonist of axonal generation or regeneration is an inhibitor of TGF-beta.  
     
     
         14 . The device of  claim 13  wherein the inhibitor of TGF-beta is selected from the group consisting of decorin, a TGF-beta inhibitory chemokine, an anti-TGF-beta antibody, an antisense TGF-beta oligonucleotide, a TGF-beta gene specific ribozyme and a mutated TGF-beta.  
     
     
         15 . The device of  claim 14  wherein the TGF-beta inhibitory chemokine is an ELR containing member of the CXC chemokine family.  
     
     
         16 . The device of  claim 15  wherein the ELR containing member of the CXC chemokine family is selected from the group consisting of interleukin-8, ENA-78, GROα, GROβand GROγ.  
     
     
         17 . The device of  claim 13  wherein the inhibitor of TGF-beta is decorin.  
     
     
         18 . The device of  claim 13  wherein the inhibitor of TGF-beta is an anti-TGF-beta antibody.  
     
     
         19 . The device of  claim 13  wherein the inhibitor of TGF-beta is a mutated TGF-beta.  
     
     
         20 . The device of either  claim 1  or  claim 2  wherein the neuronal therapeutic encoding agent is non-covalently associated with the gene activated matrix.  
     
     
         21 . The device of either  claim 1  or  claim 2  wherein the neuronal therapeutic encoding agent is adsorbed to the gene activated matrix.  
     
     
         22 . The device of either  claim 1  or  claim 2  wherein the neuronal therapeutic encoding agent is absorbed in the gene activated matrix.  
     
     
         23 . The device of either  claim 1  or  claim 2  wherein the neuronal therapeutic encoding agent is capable of inducing neuronal axonal generation or regeneration.  
     
     
         24 . A device for promoting neuronal 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 . A device for promoting neuronal survival, comprising: 
 a gene activated matrix;    at least one support cell; and    at least one neuronal therapeutic encoding agent having an operably linked promoter.    
     
     
         26 . The device of either  claim 24  or  claim 25  wherein the support cell is a Schwann cell.  
     
     
         27 . The device of either  claim 24  or  claim 25  wherein the support cell is an oligodendrocyte.  
     
     
         28 . The device of either  claim 24  or  claim 25  wherein the support cell is an astrocyte.  
     
     
         29 . The device of either  claim 24  or  claim 25  wherein the support cell is a microglial cell.  
     
     
         30 . The device of either  claim 24  or  claim 25  wherein the support cell is a fibroblast.  
     
     
         31 . The device of either  claim 24  or  claim 25  wherein the support cell is a macrophage.  
     
     
         32 . The device of either  claim 24  or  claim 25  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 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the neuronal therapeutic encoding agent is capable of maintaining axonal generation or regeneration.  
     
     
         34 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the gene activated matrix is an implant for a neuronal injury site.  
     
     
         35 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the gene activated matrix is formed upon administration.  
     
     
         36 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the gene activated matrix is administered to a neuronal injury site.  
     
     
         37 . The device of any one of claims  1 ,  2 ,  24  or  25  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 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the gene activated matrix is a paste.  
     
     
         39 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the neuronal therapeutic encoding agent is selected from the group consisting of a nucleic acid molecule, a vector, an antisense nucleic acid molecule and a ribozyme.  
     
     
         40 . The device of any one of claims  1 ,  2 ,  24  or  25 , further comprising a targeting agent, wherein said targeting agent is complexed with the neuronal therapeutic encoding agent and is capable of binding a neuronal cell surface receptor.  
     
     
         41 . The device of any one of claims  1 ,  2 ,  24  or  25 , further comprising a targeting agent, wherein said targeting agent is conjugated to the neuronal therapeutic encoding agent and is capable of binding a neuronal cell surface receptor.  
     
     
         42 . The device of any one of claims  1 ,  2 ,  24  or  25 , further comprising a targeting agent, wherein said targeting agent is complexed with the neuronal therapeutic encoding agent and is capable of binding a repair cell surface receptor.  
     
     
         43 . The device of any one of claims  1 ,  2 ,  24  or  25 , further comprising a targeting agent, wherein said targeting agent is conjugated to the neuronal therapeutic encoding agent and is capable of binding a repair cell surface receptor.  
     
     
         44 . The device of any one of claims  1 ,  2 ,  24  or  25 , further comprising a targeting agent, wherein said targeting agent is complexed with the neuronal therapeutic encoding agent and is capable of binding extracellular matrix.  
     
     
         45 . The device of any one of claims  1 ,  2 ,  24  or  25 , further comprising a targeting agent, wherein said targeting agent is conjugated to the neuronal therapeutic encoding agent and is capable of binding extracellular matrix.  
     
     
         46 . The device of any one of claims  1 ,  2 ,  24  or  25 , 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 any one of claims  1 ,  2 ,  24  or  25 , 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 ,  2 ,  24  or  25  wherein the device contains sub-physiologic amounts of a neuronal therapeutic agent.  
     
     
         49 . The device of any one of claims  1 ,  2 ,  24  or  25  wherein the device contains physiologic amounts of a neuronal therapeutic agent.  
     
     
         50 . A device according to any one of claims  1 ,  2 ,  24  or  25 , further comprising a conduit having a lumen.  
     
     
         51 . The device of  claim 50  wherein the conduit comprises the gene activated matrix.  
     
     
         52 . The device of  claim 50  wherein the lumen contains the gene activated matrix.  
     
     
         53 . The device of  claim 50  wherein the conduit comprises a bioabsorbable material.  
     
     
         54 . The device of  claim 53  wherein the bioabsorbable material comprises material selected from the group consisting of gene activated matrix, type I collagen, laminin, polyglycolic acid, glycolide trimethylene carbonate (GTMC), poly (L-lactide-co-6-caprolactone), glycoproteins, proteoglycans, heparan sulfate proteoglycan, nidogen, glycosaminoglycans, fibronectin, epidermal growth factor, fibroblast growth factor, nerve growth factor, cytokines, and DNA encoding growth factors and cytokines.  
     
     
         55 . The device of  claim 50  wherein the conduit comprises a non-bioabsorbable material.  
     
     
         56 . The device of  claim 55  wherein the non-bioabsorbable material is selected from the group consisting of polyamide, polyimide, polyurethane, segmented polyurethane, polycarbonate, and silicone.  
     
     
         57 . The device of  claim 55  wherein the non-bioabsorbable material comprises an etched microporous synthetic polymer surface.  
     
     
         58 . The device of  claim 50  wherein the conduit is tubular.  
     
     
         59 . A method for transferring a neuronal therapeutic encoding agent into a neuronal cell, comprising: 
 contacting a neuronal cell with the device of any one of claims  1 - 58  to effectively transfer the neuronal therapeutic encoding agent into the neuronal cell.    
     
     
         60 . The method of  claim 59  wherein transfer of the neuronal therapeutic encoding agent comprises retrograde axonal transport of the neuronal therapeutic encoding agent.  
     
     
         61 . The method of  claim 59 , further comprising expression of the neuronal therapeutic encoding agent at a neuronal cellular site distinct from a site of contact between the device and the neuronal cell.  
     
     
         62 . The method of  claim 59  wherein the device is contacted with a neuronal cell at a neuronal injury site.  
     
     
         63 . The method of  claim 59  wherein the device is contacted with a neuronal cell in a manner such that axonal generation or regeneration occurs.  
     
     
         64 . The method of  claim 63  wherein axonal regeneration occurs without axonal entrapment.  
     
     
         65 . The method of  claim 59  wherein the device is contacted with a neuronal cell in a manner that promotes neuronal survival.  
     
     
         66 . The method of  claim 65  wherein neuronal survival is promoted without axonal entrapment.  
     
     
         67 . The method of any one of claims  62 ,  63 ,  64 ,  65  or  66  wherein a neural connection is established or reestablished.  
     
     
         68 . A method for transferring a neuronal therapeutic encoding agent into a repair cell, comprising: 
 contacting a repair cell with the device of any one of claims  1 - 58  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 . The method of  claim 68  wherein the device is contacted with a repair cell in a manner such that axonal generation or regeneration occurs.  
     
     
         71 . The method of  claim 70  wherein axonal generation or regeneration occurs without axonal entrapment.  
     
     
         72 . The method of  claim 68  wherein the device is contacted with a repair cell in a manner that promotes neuronal survival.  
     
     
         73 . The method of  claim 72  wherein neuronal survival is promoted without axonal entrapment.  
     
     
         74 . The method of any one of claims  69 ,  70 ,  71 ,  72  or  73  wherein a neural connection is established or reestablished.  
     
     
         75 . The method of either  claim 59  or  claim 68  wherein the device contains sub-physiologic amounts of a neuronal therapeutic agent.  
     
     
         76 . The method of either  claim 59  or  claim 68  wherein the device contains physiologic amounts of a neuronal therapeutic agent.  
     
     
         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 . The method of  claim 77  wherein the neuronal therapeutic encoding agent is adsorbed to the gene activated matrix.  
     
     
         79 . The method of  claim 77  wherein the neuronal therapeutic encoding agent is absorbed in the gene activated matrix.  
     
     
         80 . The method of  claim 77  wherein the neuronal therapeutic encoding agent is selected from the group consisting of a nucleic acid molecule, a vector, an antisense molecule and a ribozyme.