Dopaminergic neuronal survival-promoting factors and uses thereof

Abstract

In general, the invention features substantially purified MANF and substantially purified nucleic acids encoding the same. The invention also features a pharmaceutical composition that includes MANF and a pharmaceutically-acceptable excipient, methods for treatment of a neurodegenerative disease, methods for improving dopaminergic neuronal survival during or following cell transplantation, methods for production of neurons for transplantation, and methods for identifying compounds that modulate or mimic MANF's biological activity.

What we claim is:  
     
         1 . A substantially purified MANF polypeptide.  
     
     
         2 . The substantially purified MANF polypeptide of  claim 1 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6,or 7.  
     
     
         3 . The substantially purified polypeptide of  claim 2 , wherein said MANF polypeptide consists of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         4 . A substantially purified polynucleotide encoding a MANF polypeptide.  
     
     
         5 . The substantially purified polynucleotide of  claim 5 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         6 . The substantially purified polynucleotide of  claim 5 , wherein said MANF polypeptide consists of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         7 . A pharmaceutical composition comprising: (i) a substantially purified MANF polypeptide; and (ii) a carrier that is pharmaceutically acceptable for administration to the central nervous system.  
     
     
         8 . The pharmaceutical composition of  claim 7 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         9 . The pharmaceutical composition of  claim 8 , wherein said MANF polypeptide consists of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         10 . The pharmaceutical composition of  claim 7 , further comprising a neural cell.  
     
     
         11 . The pharmaceutical composition of  claim 10 , wherein said neural cell is a neuron, a neural stem cell, or a neuronal precursor cell.  
     
     
         12 . A method for increasing survival of dopaminergic neurons, said method comprising the step of contacting said dopaminergic neurons with a survival-promoting amount of a substantially purified MANF polypeptide.  
     
     
         13 . The method of  claim 12 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         14 . The method of  claim 13 , wherein said MANF polypeptide consists of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         15 . A method for growing dopaminergic neurons for transplantation, said method comprising the step of culturing said neurons, or progenitor cells thereof, with a survival-promoting amount of a substantially purified MANF polypeptide.  
     
     
         16 . The method of  claim 15 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         17 . The method of  claim 16 , wherein said MANF polypeptide consists of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         18 . A method of treating a patient having a disease or disorder of the nervous system, said method comprising the step of administering to said patient a dopaminergic neuronal survival-promoting amount of a substantially purified MANF polypeptide.  
     
     
         19 . The method of  claim 18 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         20 . The method of  claim 19 , wherein said MANF polypeptide consists of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         21 . A method for preventing dopaminergic neuronal cell death in a mammal, said method comprising administering to said mammal a dopaminergic neuronal survival-promoting amount of a substantially purified MANF polypeptide.  
     
     
         22 . The method of  claim 21 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         23 . The method of  claim 22 , wherein said MANF polypeptide consists of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         24 . A method of transplanting cells into the nervous system of a mammal, comprising (i) transplanting cells into the nervous system of said mammal; and (ii) administering a dopaminergic neuronal survival-promoting amount of a MANF polypeptide to said mammal in a time window from four hours before said transplanting of said cells to four hours after said transplanting of said cells.  
     
     
         25 . The method of  claim 24 , wherein said mammal is a human.  
     
     
         26 . The method of  claim 24 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         27 . The method of  claim 26 , wherein said MANF polypeptide consists of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         28 . The method of  claim 24 , wherein said time window is from two hours before said transplanting of said cells to two hours after said transplanting of said cells.  
     
     
         29 . A method of transplanting cells into the nervous system of a mammal, said method comprising the steps of: 
 (a) contacting said cells in vitro with a MANF polypeptide; and    (b) transplanting said cells into the nervous system of said mammal.    
     
     
         30 . The method of  claim 29 , wherein step (a) and step (b) are performed within four hours of each other.  
     
     
         31 . The method of  claim 29 , wherein said mammal is a human.  
     
     
         32 . The method of  claim 29 , wherein said MANF polypeptide consists essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         33 . The method of  claim 32 , wherein said MANF polypeptide consists of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         34 . An expression vector encoding a MANF polypeptide consisting essentially of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         35 . The expression vector of  claim 34 , wherein said MANF polypeptide consists of the sequence of SEQ ID NO: 2, 3, 4, 5, 6, or 7.  
     
     
         36 . The expression vector of  claim 34 , wherein said vector is an adenoviral vector or retroviral vector.