Myelination of congenitally dysmyelinated forebrains using oligodendrocyte progenitor cells

Abstract

One form of the present invention is directed to a method of remyelinating demyelinated axons by treating the demyelinated axons with oligodendrocyte progenitor cells under conditions which permit remyelination of the axons. Another aspect of the present invention relates to a method of treating a subject having a condition mediated by a loss of myelin or a loss of oligodendrocytes by administering to the subject oligodendrocyte progenitor cells under conditions effective to treat the condition mediated by a loss of myelin or a loss of oligodendrocytes. A further aspect of the present invention relates to an in vitro method of identifying and separating oligodendrocyte progenitor cells from a mixed population containing other mammalian brain or spinal cord cell types. This further aspect of the present invention involves removing neurons and neuronal progenitor cells from the mixed population to produce a treated mixed population. Oligodendrocyte progenitor cells are then separated from the treated mixed population to form an enriched population of oligodendrocyte progenitor cells.

What is claimed:  
     
         1 . A method of remyelinating demyelinated axons comprising: 
 treating the demyelinated axons with oligondendrocyte progenitor cells under conditions which permit remyelination of the axons.    
     
     
         2 . The method according to  claim 1 , wherein said treating is carried out by transuterine fetal intraventricular injection, intraventricular or intraparenchymal injections, intraparenchymal injections into adult and juvenile subjects, or intravascular administration.  
     
     
         3 . A method of treating a subject having a condition mediated by a loss of myelin or a loss of oligodendrocytes comprising: 
 administering to the subject oligondendrocyte progenitor cells under conditions effective to treat the condition mediated by a loss of myelin or a loss of oligodendrocytes.    
     
     
         4 . The method according to  claim 3 , wherein the method is used to treat a subject with a condition mediated by a loss of oligodendrocytes.  
     
     
         5 . The method according to  claim 3 , wherein the method is used to treat a subject with a condition mediated by a loss of myelin.  
     
     
         6 . The method according to  claim 5 , wherein the condition is an ischemic demyelination condition.  
     
     
         7 . The method according to  claim 6 , wherein the ischemic demyelination condition is selected from the group consisting of cortical stroke, Lacunar infarct, post-hypoxic leukoencephalopathy, diabetic leukoencephalopathy, and hypertensive leukoencephalopathy.  
     
     
         8 . The method according to  claim 5 , wherein the condition is an inflammatory demyelination condition.  
     
     
         9 . The method according to  claim 8 , wherein the inflammatory demyelination condition is selected from the group consisting of multiple sclerosis, Schilder's Disease, transverse myelitis, optic neuritis, post-vaccination encephalomyelitis, and post-infectious encephalomyelitis.  
     
     
         10 . The method according to  claim 5 , wherein the condition is a pediatric leukodystrophy.  
     
     
         11 . The method according to  claim 10 , wherein the pediatric leukodystrophy condition is selected from the group consisting of a lysosomal storage disease, Cavavan's Disease, Pelizaeus-Merzbacher Disease, and Crabbe's Globoid body leukodystrophy.  
     
     
         12 . The method according to  claim 10 , wherein the pediatric leukodystrophy condition is Tay-Sachs Disease.  
     
     
         13 . A method according to  claim 5 , wherein the condition is mucopolysaccharidosis.  
     
     
         14 . The method according to  claim 13 , wherein the condition is Sly's Disease.  
     
     
         15 . The method according to  claim 5 , wherein the condition is perinatal germinal matrix hemorrhage, periventricular leukoinalacia, or cerebral palsy.  
     
     
         16 . The method according to  claim 5 , wherein the condition is a radiation-induced condition.  
     
     
         17 . The method according to  claim 16 , wherein the radiation-induced condition is radiation-induced leukoencephalopathy or radiation-induced myelitis.  
     
     
         18 . The method according to  claim 5 , wherein the condition is an etiology causing sub-cortical leukoencephalopathy, said etiology being HIV/AIDS, head trauma, or multi-infarct states.  
     
     
         19 . The method according to  claim 3 , wherein said treating is carried out after administering radiation to the subject and before demyelination has occurred.  
     
     
         20 . The method according to  claim 3 , wherein the subject is a human.  
     
     
         21 . The method according to  claim 20 , wherein the subject is a post-natal human.  
     
     
         22 . The method according to  claim 20 , wherein the subject is an adult human.  
     
     
         23 . The method according to  claim 3 , wherein the method is carried out in the brain.  
     
     
         24 . The method according to  claim 3 , wherein the method is carried out in the spinal cord.  
     
     
         25 . An in vitro method of identifying and separating oligodendrocyte progenitor cells from a mixed population containing other mammalian brain or spinal cord cell types, said method comprising: 
 removing neurons and neuronal progenitor cells from the mixed population to produce a treated mixed population and    separating the oligodendrocyte progenitor cells from the treated mixed population to form an enriched population of oligodendrocyte progenitor cells.    
     
     
         26 . The method according to  claim 25 , wherein said separating oligodendrocyte progenitor cells comprises: 
 selecting a promoter which functions only in oligodendrocyte progenitor cells and not in the other cell types;    introducing a nucleic acid molecule encoding a fluorescent protein under control of said promoter into all cell types of said treated mixed population;    allowing only the oligodendrocyte progenitor cells, but not the other cell types, within said treated mixed population to express said fluorescent protein;    identifying cells of said treated mixed population of cell types that are fluorescent, which are restricted to oligodendrocyte progenitor cells; and    separating the fluorescent cells from said treated mixed population to form the enriched population of oligodendrocyte progenitor cells.    
     
     
         27 . The method according to  claim 26 , wherein said introducing comprises viral mediated transformation of all cell types of said treated mixed population containing other mammalian brain or spinal cord cell types.  
     
     
         28 . The method according to  claim 27 , wherein said viral mediated transformation comprises adenovirus mediated transformation, retrovirus-mediated transduction, lentivirus-mediated transduction, or adeno-associated virus-mediated transduction.  
     
     
         29 . The method according to  claim 26 , wherein said introducing comprises electroporation.  
     
     
         30 . The method according to  claim 26 , wherein said introducing comprises liposomal mediated transformation of all cell types of said treated mixed population containing other mammalian brain or spinal cord cell types.  
     
     
         31 . The method according to  claim 26 , wherein said separating the fluorescent cells comprises fluorescence activated cell sorting.  
     
     
         32 . The method according to  claim 26 , wherein said promoter is selected from the group consisting of a cyclic nucleotide phosphorylase I promoter, a myelin basic protein promoter, a JC virus minimal core promoter, a proteolipid protein promoter, a qk1 promoter, and a cyclic nucleotide phosphorylase II promoter.  
     
     
         33 . The method according to  claim 25 , wherein said separating oligodendrocyte progenitor cells comprises: 
 immunoseparating the oligodendrocyte progenitor cells.    
     
     
         34 . The method according to  claim 33 , wherein said immunoseparating is carried out by removing cells from the treated mixed population having an A2B5 antigen.  
     
     
         35 . The method according to  claim 33 , wherein said immunoseparating is carried out with a fluorescently labelled antibody which recognizes an antigen on the oligodendrocyte progenitor cells.  
     
     
         36 . The method according to  claim 35 , wherein said separating oligodendrocyte progenitor cells further comprises: 
 fluorescence activated cell sorting.    
     
     
         37 . The method according to  claim 25 , wherein said removing comprises: 
 selecting a promoter which functions only in neurons and neuronal progenitor cells;    introducing a nucleic acid molecule encoding a marker protein under control of said promoter into the mixed population;    allowing the neurons and neuronal progenitor cells to express the marker protein; and    separating the cells expressing the marker protein from the mixed population of cells, wherein said separated cells are the neurons and neuronal progenitor cells.    
     
     
         38 . The method according to  claim 37 , wherein said introducing comprises viral mediated transduction of the mixed population of cells.  
     
     
         39 . The method according to  claim 38 , wherein said viral mediated transduction comprises adenovirus-mediated transduction, retrovirus-mediated transduction, lentivirus-mediated transduction, or adeno-associated virus-mediated transduction.  
     
     
         40 . The method according to  claim 37 , wherein said introducing comprises electroporation.  
     
     
         41 . The method according to  claim 37 , wherein said introducing comprises biolistic transformation.  
     
     
         42 . The method according to  claim 37 , wherein said introducing comprises liposomal mediated transformation.  
     
     
         43 . The method according to  claim 37 , wherein the marker protein is a fluorescent protein and said separating comprises fluorescence activated cell sorting.  
     
     
         44 . The method according to  claim 37 , wherein the promoter is a Tα1 tubulin promoter, a MAP-1B promoter, an NCAM promoter, An HES-5 HLH promoter, an α-internexin promoter, or a GAP-43 promoter.  
     
     
         45 . The method according to  claim 25 , wherein the cell type is human.  
     
     
         46 . The method according to  claim 45 , wherein the human is an adult.  
     
     
         47 . The method according to  claim 45 , wherein the human is postnatal.  
     
     
         48 . The enriched population of oligodendrocyte progenitor cells produced according to the process of  claim 25.