Splice variants of GDNF and uses thereof
View Patent ↗The present invention relates to Glial Cell Line-Derived Neurotrophic Factor (GDNF) protein and gene and is, in particular, directed to a novel splice variant of GDNF protein, which is encoded by a novel splice variant pre-(γ)pro-GDNF, and secreted under biological regulation.
1. A method of treating Parkinson's disease in a human subject, said method comprising:
administering by injection into the striatum of the brain of the subject suffering from Parkinson's disease a therapeutically effective amount of a polynucleotide encoding pre-(β)pro-GDNF protein,
wherein the polynucleotide encoding the pre-(β)pro-GDNF protein is a polynucleotide encoding a polypeptide comprising at least one of the amino acid sequences selected from the group consisting of SEQ ID NO: 31, SEQ ID NO: 35, and SEQ ID NO: 52,
wherein the polynucleotide is in a recombinant adeno-associated virus suitable to introduce the polynucleotide into brain cells, and
wherein the polynucleotide is operatively linked to a cytomegalovirus promoter expression regulatory element,
wherein the encoded pre-(β)pro-GDNF protein promotes the survival of dopaminergic neurons in Parkinson's Disease.
2. The method according to claim 1 , wherein the polynucleotide encoding the pre-(β)pro-GDNF protein is selected from the group consisting of the polynucleotides as set forth in SEQ ID NO: 32, SEQ ID NO: 36 and SEQ ID NO: 51.
3. The method according to claim 1 , wherein the polynucleotide encoding the pre-(β)pro-GDNF protein is a polynucleotide encoding a polypeptide selected from the group consisting of the proteins set forth in SEQ ID NO: 35 and SEQ ID NO: 52.
4. A method for promoting survival of dopamine neurons in a brain comprising:
administering a therapeutically effective amount to promote survival of dopamine neurons of a polynucleotide encoding pre-(β)pro-GDNF protein by injection into the striatum of the brain,
wherein the polynucleotide encoding the pre-(β)pro-GDNF is a polynucleotide encoding a polypeptide comprising at least one of the amino acid sequences selected from the group consisting of SEQ ID NO: 31, SEQ ID NO: 35, and SEQ ID NO: 52,
wherein the polynucleotide is operatively linked to an expression regulatory element in a recombinant adeno-associated virus suitable for introducing the polynucleotide into a neuron, and
wherein the expression regulatory element is cytomegalovirus promoter.
5. The method according to claim 4 , wherein administration is to a patient afflicted with Parkinson's disease.
6. The method according to claim 4 , wherein the polynucleotide encoding the pre-(β)pro-GDNF protein is selected from the group consisting of the polynucleotides as set forth in SEQ ID NO: 32, SEQ ID NO: 36 and SEQ ID NO: 51.
7. The method according to claim 4 , wherein the polynucleotide encoding the pre-(β)pro-GDNF protein is a polynucleotide encoding a polypeptide selected from the group consisting of the proteins set forth in SEQ ID NO: 35 and SEQ ID NO: 52.