Regenerating functional neurons for treatment of disease and injury in the nervous system

Abstract

Methods for producing new neurons in the brain in vivo are provided according to aspects of the present invention which include introducing NeuroD1 into a glial cell, particularly into a reactive astrocyte or NG2 cell, thereby “converting” the reactive glial cell to a neuron. Methods of producing a neuronal phenotype in a glial cell are provided according to aspects of the present invention which include expressing exogenous NeuroD1 in the glial cell, wherein expressing exogenous NeuroD1 includes delivering an expression vector, such as a viral expression vector, including a nucleic acid encoding the exogenous NeuroD1 to the glial cell.

1 - 44 . (canceled) 
     
     
         45 . A method of treating a neurological condition in a subject in need thereof, wherein the neurological condition is characterized by loss of neurons and presence of reactive astrocytes, comprising:
 administering a therapeutically effective dose of transcription factor NeuroD1 to a reactive astrocyte of the subject,   wherein administering the therapeutically effective dose of NeuroD1 comprises administering an expression vector comprising a nucleic acid sequence encoding a NeuroD1 protein, thereby producing a neuronal phenotype in the reactive astrocyte to ameliorate the neurological condition of the subject.   
     
     
         46 . The method of  claim 45 , wherein the nucleic acid sequence comprises a nucleic acid having a sequence that has at least 70% identity to the DNA sequence set forth in SEQ ID NO: 1, SEQ ID NO: 3, a complement thereof, or a fragment thereof. 
     
     
         47 . The method of  claim 45 , wherein the NeuroD1 protein comprises a polypeptide having a sequence that has at least 70% identity to the polypeptide sequence set forth in SEQ ID NO: 2, SEQ ID NO: 4, or a functional fragment thereof. 
     
     
         48 . The method of  claim 45 , wherein the neuronal phenotype comprises one or more characteristics selected from the group consisting of: neuronal morphology, expression of one or more neuronal markers, electrophysiologic characteristics of neurons, synapse formation, and release of neurotransmitters. 
     
     
         49 . The method of  claim 45 , wherein the subject is a human. 
     
     
         50 . The method of  claim 45 , wherein the neurological condition is an injury to the central or peripheral nervous system. 
     
     
         51 . The method of  claim 45 , wherein the neurological condition is selected from the group consisting of: Alzheimer disease, Parkinson disease, amyotrophic lateral sclerosis (ALS), epilepsy, and stroke. 
     
     
         52 . The method of  claim 45 , wherein the therapeutically effective dose of NeuroD1 is administered via a systemic administration. 
     
     
         53 . The method of  claim 45 , wherein the therapeutically effective dose of NeuroD1 is administered via a local administration. 
     
     
         54 . The method of  claim 45 , wherein the expression vector is a retrovirus. 
     
     
         55 . The method of  claim 45 , wherein the expression vector is an adeno-associated virus. 
     
     
         56 . The method of  claim 45 , wherein the neuronal phenotype comprises an increase in expression of Neuronal Nuclei (NeuN) compared to before the administration. 
     
     
         57 . The method of  claim 45 , wherein the neuronal phenotype comprises a reduction in secretion of Chondroitin Sulfate Proteoglycan (CSPG) compared to before the administration. 
     
     
         58 . The method of  claim 45 , wherein the neuronal phenotype comprises a decrease in expression of Apolipoprotein E (ApoE) compared to before said administration. 
     
     
         59 . The method of  claim 45 , wherein the expression vector further comprises a regulatory element. 
     
     
         60 . The method of  claim 59 , wherein the regulatory element is selected from the group consisting of an internal ribosome entry site (IRES), a 2A domain, an intron, an origin of replication, a polyadenylation signal (pA), a transcription termination sequence, and an upstream regulatory domain. 
     
     
         61 . A method of treating a neurological condition in a subject in need thereof, wherein the neurological condition is characterized by loss of neurons and presence of reactive astrocytes, comprising:
 administering a therapeutically effective dose of transcription factor NeuroD1 to a reactive astrocyte of the subject,   wherein administering the therapeutically effective dose of NeuroD1 comprises administering an expression vector comprising (i) a human promoter selected from the group consisting of a GFAP promoter, an ALDH1L1 promoter, and an LCN2 promoter, and (ii) a nucleic acid sequence encoding a NeuroD1 protein, thereby producing a neuronal phenotype in the reactive astrocyte to ameliorate the neurological condition of the subject.   
     
     
         62 . The method of  claim 61 ,
 wherein the nucleic acid sequence comprises a nucleic acid having a sequence that has at least 70% identity to a DNA sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, a complement thereof, and a fragment thereof, or   wherein the nucleic acid sequence encodes a protein comprising a polypeptide having a sequence that has at least 70% identity to a polypeptide sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, and a functional fragment thereof.   
     
     
         63 . A method of treating a neurological condition in a subject in need thereof, wherein the neurological condition is characterized by loss of neurons and presence of reactive NG2 cells, comprising:
 administering a therapeutically effective dose of transcription factor NeuroD1 to a reactive NG2 cell of the subject,   wherein administering the therapeutically effective dose of NeuroD1 comprises administering an expression vector comprising a human NG2 promoter, and a nucleic acid sequence encoding a NeuroD1 protein, thereby producing a neuronal phenotype in the reactive NG2 cell to ameliorate the neurological condition of the subject.   
     
     
         64 . The method of  claim 63 ,
 wherein the nucleic acid sequence comprises a nucleic acid having a sequence that has at least 70% identity to a DNA sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, a complement thereof, and a fragment thereof, or   wherein the nucleic acid sequence encodes a protein comprising a polypeptide having a sequence that has at least 70% identity to a polypeptide sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, and a functional fragment thereof.