Therapeutic Use of the Encoding Sequence of the Carboxy-Terminal Domain of the Heavy Chain of the Tetanus Toxin

Abstract

The present invention relates to the therapeutic use of the encoding sequence of the carboxy-terminal domain of the heavy chain of the tetanus toxin and of the polypeptide encoded by said sequence, preferably for the treatment of amyotrophic lateral sclerosis (ALS).

1 - 24 . (canceled) 
     
     
         25 . A method for ameliorating the symptoms of motor neuron degeneration in a subject with Amyotrophic Lateral Sclerosis (ALS), comprising administering an isolated polynucleotide, wherein:
 (i) the polynucleotide is an HcTeTx polynucleotide encoding a HcTeTx polypeptide comprising the carboxy-terminal domain of the heavy subunit of the tetanus toxin, wherein the sequence of the HcTeTx polypeptide comprises the sequence of SEQ ID NO:2 or SEQ ID NO: 5 or a fragment thereof; and,   (ii) the HcTeTx polypeptide encoded by the HcTeTx polynucleotide increases the survival rate of transgenic SOD1G93A mice, compared to the survival rate of transgenic SOD1G93A mice that have not been administered the HcTeTx polynucleotide.   
     
     
         26 . The method according to  claim 25 , wherein the sequence of the HcTeTx polynucleotide comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 6 
     
     
         27 . The method according to  claim 25 , wherein the sequence of the HcTeTx polynucleotide consists of the sequence of SEQ ID NO: 1 or SEQ ID NO: 6. 
     
     
         28 . The method according to  claim 25 , wherein the HcTeTx polynucleotide is an RNA, a DNA, or a combination thereof. 
     
     
         29 . The method according to  claim 28 , wherein the RNA or DNA is mono-catenary o bi-catenary. 
     
     
         30 . The method according to  claim 25 , wherein the HcTeTx polynucleotide is a naked DNA. 
     
     
         31 . The method according to  claim 25 , wherein the HcTeTx polynucleotide is operably linked to a promoter, a terminator, a silencer sequence, or a combination thereof. 
     
     
         32 . The method according to  claim 25 , wherein the HcTeTx polynucleotide is inserted into a vector. 
     
     
         33 . The method according to  claim 32 , wherein the vector is a plasmid, a phage, a cosmid, a phagemid, a yeast artificial chromosome (YAC), a bacterial artificial chromosome (BAC), a human artificial chromosomes (HAC), a viral vector, or a combination thereof 
     
     
         34 . The method according to  claim 33 , wherein the viral vector is an adenovirus, a retrovirus, or a combination thereof. 
     
     
         35 . The method according to  claim 32 , wherein the vector is used to generate a transgenic cell. 
     
     
         36 . The method according to  claim 32 , wherein the HcTeTx polynucleotide in the vector is expressed in vivo. 
     
     
         37 . The method according to  claim 36 , wherein the HcTeTx polynucleotide in the vector is expressed in vivo in a muscle cell. 
     
     
         38 . The method according to  claim 31 , wherein the promoter is capable of expressing the HcTeTx polypeptide encoded by the HcTeTx polynucleotide. 
     
     
         39 . The method according to  claim 38 , wherein the promoter is a cytomegalovirus (CMV) promoter. 
     
     
         40 . The method according to  claim 39 , wherein the CMV promoter is a pCMV promoter. 
     
     
         41 . The method according to  claim 32 , wherein the vector is a plasmid. 
     
     
         42 . The method according to  claim 41 , wherein the vector is a  PC DNA™ plasmid. 
     
     
         43 . The method according to  claim 42 , wherein the  PC DNA™ plasmid is a  PC DNA™3.1 plasmid. 
     
     
         44 . The method according to  claim 25 , wherein the administration of the HcTeTx polynucleotide is oral, nasal, parenteral, or combinations thereof. 
     
     
         45 . The method according to  claim 44 , wherein the parenteral administration is intraperitoneal, intravenous, subcutaneous, intramuscular, or combinations thereof. 
     
     
         46 . The method according to  claim 45 , wherein the HcTeTx polynucleotide is administered intramuscularly to a single muscle group. 
     
     
         47 . The method according to  claim 45 , wherein the HcTeTx polynucleotide is administered intramuscularly to more that one muscle group. 
     
     
         48 . The method according to  claim 25 , wherein the HcTeTx polynucleotide has neuroprotectant activity. 
     
     
         49 . The method according to  claim 25 , wherein the HcTeTx polynucleotide is administered in a combination treatment. 
     
     
         50 . The method according to  claim 49 , wherein the combination treatment comprises polypeptides, polynucleotides, vectors, cells, or combinations thereof. 
     
     
         51 . The method according to  claim 25 , wherein HcTeTx polynucleotide is genetically fused to a polynucleotide encoding a non-HcTeTx therapeutic moiety, wherein the HcTeTx polypeptide encoded by the HcTeTx polynucleotide is not a targeting moiety for the non-HcTeTx therapeutic moiety. 
     
     
         52 . The method according to  claim 25 , wherein HcTeTx polynucleotide is genetically fused to a polynucleotide encoding a non-HcTeTx non-therapeutic moiety, wherein the HcTeTx polypeptide encoded by the HcTeTx polynucleotide is not a targeting moiety for the non-HcTeTx non-therapeutic moiety. 
     
     
         53 . The method according to  claim 25 , wherein the HcTeTx polynucleotide encodes a HcTeTx polypeptide consisting essentially of the carboxy-terminal domain of the heavy subunit of the tetanus toxin polypeptide, wherein the HcTeTx polypeptide comprises the sequence of SEQ ID NO:2 or SEQ ID NO: 5 or a fragment thereof. 
     
     
         54 . The method according to  claim 25 , wherein the HcTeTx polynucleotide encodes a HcTeTx polypeptide comprising the carboxy-terminal domain of the heavy subunit of the tetanus toxin polypeptide, wherein the HcTeTx polypeptide consists of the sequence of SEQ ID NO:2 or SEQ ID NO: 5.