MicroRNAs for the generation of astrocytes
A method of generating a population of cells useful for treating a nerve disease or disorder in a subject, the method comprising up-regulating a level of at least one exogenous miRNA in mesenchymal stem cells (MSCs) and/or down-regulating a level of at least one miRNA using a polynucleotide agent that hybridizes to the miRNA, thereby generating the population of cells useful for treating the nerve disease or disorder. Isolated populations of cells with an astrocytic phenotype generated thereby and uses thereof are also provided.
1. An isolated population of genetically modified mesenchymal stem cells (MSCs) differentiated toward an astrocytic phenotype wherein each MSC comprises a combination of an exogenous microRNA (miR)-146 (SEQ ID NO:462) and an antagomir or RNA oligonucleotide that hybridizes to and inhibits an endogenous miR-302 (SEQ ID NO:369), wherein at least 50% of the MSCs express glial fibrillary acidic protein.
2. The isolated population of MSCs of claim 1 , wherein the at least 50% of the population of MSCs differentiated toward an astrocytic phenotype is further identified by expression of a marker selected from the group consisting of protein S100, glutamine synthetase, excitatory amino acid transporter 1 (EAAT1) and EAAT2.
3. A pharmaceutical composition comprising the isolated population of cells of claim 1 and a pharmaceutically acceptable carrier.
4. The isolated population of MSCs of claim 1 , wherein the at least 50% of the population of MSCs differentiated toward an astrocytic phenotype is further identified by astrocytic morphology.
5. A method of generating an isolated population of genetically modified mesenchymal stem cells (MSCs) differentiated toward an astrocytic phenotype, wherein at least 50% of the MSCs express glial fibrillary acidic protein, the method comprising introducing and expressing in MSCs a combination of an exogenous microRNA (miR)-146 (SEQ ID NO:462) and an antagomir or RNA oligonucleotide that hybridizes to and inhibits an endogenous miR-302 (SEQ ID NO:369), thereby generating an isolated population of genetically modified mesenchymal stem cells (MSCs) differentiated toward an astrocytic phenotype.
6. The method of claim 5 , wherein said MSCs are isolated from a tissue selected from the group consisting of bone marrow, adipose tissue, placenta, cord blood and umbilical cord.
7. The method of claim 5 , wherein said introducing comprises transfecting said MSCs with an expression vector which comprises a polynucleotide sequence which encodes a pre-miRNA of said miR-146 or a polynucleotide sequence which encodes said miR-146.
8. The method of claim 5 further comprising analyzing an expression of at least one marker selected from the group consisting of S100, glutamine sythetase, excitatory amino acid transporter 1 and EAAT2 following said generating.
9. The method of claim 5 , further comprising incubating said MSCs in a differentiation medium comprising at least one agent selected from the group consisting of platelet derived growth factor (PDGF), neuregulin, fibroblast growth factor 2 (FGF-b) and a c-AMP inducing agent following, prior to or concomitant with said expressing.