Methods and compositions for rejuvenating CNS glial populations by suppresion of transcription factors
Methods of inducing rejuvenation in a population of adult glial progenitor cells, and methods of treating a subject having a myelin deficiency are disclosed in this patent application. The method of inducing rejuvenation in a population of adult glial progenitor cells may comprise administering, to the population of adult glial progenitor cells, an effective amount of an agent that suppresses one or more transcription factors selected from the group consisting of (i) zinc finger protein 274 (ZNF274), (ii) Myc-associated factor X (MAX), (iii) E2F transcription factor 6 (E2F6), (iv) zinc finger protein Aiolos (IKZF3), and (v) signal transducer and activator of transcription 3 (STAT3).
1 . A method of repressing a senescence-associated gene expression signature in a population of adult glial progenitor cells, said method comprising:
administering, to the population of adult glial progenitor cells, an agent that suppresses one or more transcription factors selected from zinc finger protein 274 (ZNF274) and E2F transcription factor 6 (E2F6),
wherein the agent comprises one or more nucleic acid molecules that comprise a first nucleic acid sequence encoding a Cas protein and a second nucleic acid sequence encoding a guide RNA,
wherein the guide RNA is complementary to an E2F6 gene of Accession No: NM 001278277.2 or is complementary to a ZNF274 gene of Accession No: XR 001753588.2, wherein the guide RNA is 5 to 100 nucleic acid residues in length; and
wherein the senescence-associated gene expression signature comprises CDKN1A/p21.
2 . The method of claim 1 , wherein the first nucleic acid sequence and the second nucleic acid sequence are located on the same nucleic acid molecule.
3 . The method of claim 1 , wherein the Cas protein is a nuclease dead Cas protein.
4 . The method of claim 1 , wherein the one or more nucleic acid molecules are encoded by a non-viral expression vector.
5 . The method of claim 1 , wherein the one or more nucleic acid molecules are encoded by a viral expression vector.
6 . The method of claim 5 , wherein the viral expression vector is a lentiviral vector.
7 . The method of claim 5 , wherein the viral expression vector is an AAV vector.
8 . The method of claim 1 , wherein the first nucleic acid sequence encoding a Cas protein and the second nucleic acid sequence encoding a guide RNA are operably linked to a regulatory element.
9 . The method of claim 8 , wherein the regulatory element is a glial cell-specific promoter.
10 . The method of claim 8 , wherein the regulatory element is an inducible promoter.
11 . The method of claim 10 , wherein the inducible promoter is a tet-on or tet-off promoter.
12 . A method of suppressing expression of either CDKN1A/p21 or CDKN2A/p16, or both in a population of adult glial progenitor cells, said method comprising:
administering, to the population of adult glial progenitor cells, an agent that suppresses one or more transcription factors, wherein the one or more transcription factors are selected from zinc finger protein 274 (ZNF274) and E2F transcription factor 6 (E2F6),
wherein the agent comprises one or more nucleic acid molecules that comprise a first nucleic acid sequence encoding a Cas protein and a second nucleic acid sequence encoding a guide RNA comprising a nucleotide sequence that is complementary to a portion of a gene encoding one or more of the transcription factors; and
wherein the nucleotide sequence of the guide RNA is complementary to an E2F6 gene of Accession No: NM 001278277.2 or is complementary to a ZNF274 gene of Accession No: XR 001753588.2, wherein the guide RNA is 5 to 100 nucleic acid residues in length.