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.

1 .- 8 . (canceled) 
     
     
         9 . An expression vector comprising a nucleic acid sequence that encodes for a MANF polypeptide, wherein the MANF polypeptide consists of a signal sequence and an amino acid sequence at least 95% identical to SEQ ID NO: 3. 
     
     
         10 . The expression vector of  claim 9 , wherein the amino acid sequence is at least 97% identical to SEQ ID NO: 3. 
     
     
         11 . The expression vector of  claim 9 , wherein the signal sequence is the first 21 amino acids of SEQ ID NO: 2. 
     
     
         12 . The expression vector of  claim 9 , further comprising a promoter. 
     
     
         13 . A genetically engineered cell comprising a recombinant nucleic acid that encodes for a MANF polypeptide, wherein the MANF polypeptide consists of a signal sequence and an amino acid sequence at least 95% identical to SEQ ID NO: 3. 
     
     
         14 . The genetically engineered cell of  claim 13 , wherein the amino acid sequence is at least 97% identical to SEQ ID NO: 3. 
     
     
         15 . The genetically engineered cell of  claim 13 , wherein the signal sequence is the first 21 amino acids of SEQ ID NO: 2. 
     
     
         16 . The genetically engineered cell of  claim 13 , wherein the recombinant nucleic acid further comprises a promoter. 
     
     
         17 . The genetically engineered cell of  claim 13 , wherein the genetically engineered cell is a eukaryotic cell. 
     
     
         18 . A method of manufacturing comprising:
 a. culturing the genetically engineered cell of  claim 13  in a culture medium; and   b. purifying a secreted MANF polypeptide from the culture medium.   
     
     
         19 . The method of  claim 18 , wherein the secreted MANF has an amino acid sequence that consists of a sequence at least 95% identical to SEQ ID NO: 3. 
     
     
         20 . The method of  claim 18 , wherein the secreted MANF has an amino acid sequence that consists of a sequence at least 97% identical to SEQ ID NO: 3. 
     
     
         21 . A secreted MANF polypeptide produced by the method of  claim 18 . 
     
     
         22 . A non-glycosylated MANF polypeptide, where the amino acid sequence of the MANF polypeptide consists of a sequence at least 95% identical to SEQ ID NO: 3. 
     
     
         23 . The non-glycosylated MANF polypeptide of  claim 22 , wherein the amino acid sequence of the MANF polypeptide consists of a sequence at least 97% identical to SEQ ID NO: 3. 
     
     
         24 . A formulation comprising:
 a. a MANF polypeptide, where the amino acid sequence of the MANF polypeptide consists of a sequence at least 95% identical to SEQ ID NO: 3;   b. a sterile liquid.   
     
     
         25 . The formulation of  claim 24 , wherein the amino acid sequence of the MANF polypeptide consists of a sequence at least 97% identical to SEQ ID NO: 3. 
     
     
         26 . A method of treating a neurodegenerative disease comprising administering to a subject in need thereof an effective amount of the formulation of  claim 24 . 
     
     
         27 . The method of  claim 26 , wherein the neurodegenerative disease comprises Parkinson's disease. 
     
     
         28 . The method of  claim 26 , wherein the neurodegenerative disease comprises Alzheimer's disease or amyotrophic lateral sclerosis.