Methods of using polynucleotides encoding truncated glial cell line-derived neurotrophic factor

Abstract

Disclosed are novel proteins, referred to as truncated glial cell line-derived neurotrophic factor (truncated GDNF) proteins, that promote dopamine uptake by dopaminergic cells and promote the survival of nerve cells. Also disclosed are processes for obtaining the truncated GDNF proteins by recombinant genetic engineering techniques.

1 . A method for promoting the survival or function of dopaminergic neurons comprising administering a polynucleotide encoding a truncated glial cell line-derived neurotrophic factor (GDNF) protein product comprising an amino acid sequence of Cys 41  through Cys 133  of SEQ ID NO:2. 
     
     
         2 . A method according to  claim 1  wherein said polynucleotide encodes a truncated GDNF protein product of the formula:
   X-[Cys41-Cys133]-Y 
 
       wherein
 [Cys41-Cys133] is Cys41 through Cys133 of SEQ ID NO: 2, 
 Y is absent or a carboxy-terminus amino acid residue of Ile134, and 
 X represents a methionylated or nonmethionylated amine group of Cys41 or amino-terminus amino acid residue(s) selected from the group: 
 
       
         
           
                 
                 
                 
               
                                                            G 
                     
                     
                 
                     
                 
                                                           RG 
                     
                 
                     
                 
                                                          NRG 
                     
                 
                     
                 
                                                         KNRG 
                   (SEQ ID NO: 3) 
                 
                     
                 
                                                        GKNRG 
                   (SEQ ID NO: 4) 
                 
                     
                 
                                                       RGKNRG 
                   (SEQ ID NO: 5) 
                 
                     
                 
                                                      QRGKNRG 
                   (SEQ ID NO: 6) 
                 
                     
                 
                                                     GQRGKNRG 
                   (SEQ ID NO: 7) 
                 
                     
                 
                                                    RGQRGKNRG 
                   (SEQ ID NO: 8) 
                 
                     
                 
                                                   RRGQRGKNRG 
                   (SEQ ID NO: 9) 
                 
                     
                 
                                                 G RRGQRGKNRG 
                   (SEQ ID NO: 10) 
                 
                     
                 
                                                KG RRGQRGKNRG 
                   (SEQ ID NO: 11) 
                 
                     
                 
                                               GKG RRGQRGKNRG 
                   (SEQ ID NO: 12) 
                 
                     
                 
                                              RGKG RRGQRGKNRG 
                   (SEQ ID NO: 13) 
                 
                     
                 
                                             SRGKG RRGQRGKNRG 
                   (SEQ ID NO: 14) 
                 
                     
                 
                                            NSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 15) 
                 
                     
                 
                                           ENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 16) 
                 
                     
                 
                                          PENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 17) 
                 
                     
                 
                                         SPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 51) 
                 
                     
                 
                                         NPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 18) 
                 
                     
                 
                                        ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 19) 
                 
                     
                 
                                      A ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 20) 
                 
                     
                 
                                     AA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 21) 
                 
                     
                 
                                    AAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 22) 
                 
                     
                 
                                   QAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 23) 
                 
                     
                 
                                  RQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 24) 
                 
                     
                 
                                 NRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 25) 
                 
                     
                 
                                RNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 26) 
                 
                     
                 
                               ERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 27) 
                 
                     
                 
                              RERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 28) 
                 
                     
                 
                             RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 29) 
                 
                     
                 
                           P RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 30) 
                 
                     
                 
                          LP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 31) 
                 
                     
                 
                         VLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 32) 
                 
                     
                 
                        AVLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 33) 
                 
                     
                 
                       MAVLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 34) 
                 
                     
                 
                      QMAVLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 35) 
                 
                     
                 
                     KQMAVLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 36) 
                 
                     
                 
                    DKQMAVLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 37) 
                 
                   and 
                     
                 
                     
                 
                   PDKQMAVLP RRERNRQAAA ANPENSRGKG RRGQRGKNRG 
                   (SEQ ID NO: 38) 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         3 . A method according to  claim 1  or  2  wherein X is selected from the group consisting of SEQ ID NOS: 3, 7, 8, 14, 17, 18, and 24. 
     
     
         4 . A method according to  claim 1  or  2  wherein X is G, RG or NRG. 
     
     
         5 . A method according to  claim 1  or  2  wherein said GDNF protein product has the amino acid sequence of SEQ ID NO:42. 
     
     
         6 . A method according to  claim 1  or  2  wherein said GDNF protein product has the amino acid sequence of SEQ ID NO:44. 
     
     
         7 . A method according to  claim 1  or  2 , wherein said GDNF protein product has the amino acid sequence of SEQ ID NO:46. 
     
     
         8 . A method according to  claim 1  or  2  wherein administering a polynucleotide is ex vivo administering. 
     
     
         9 . A method according to  claim 4  wherein administering a polynucleotide is ex vivo administering.