Methods and compositions for rejuvenating CNS glial populations through BCL11A transcription factor expression

Abstract

The present disclosure relates to methods for inducing rejuvenation in a population of glial progenitor cells and for treating a subject with a glial cell-related disorder, comprising administering to a population of adult glial progenitor cells an effective amount of an expression vector comprising a nucleotide sequence encoding B-cell lymphoma / leukemia 11 A (BCL11 A) and a regulatory element operably linked to the nucleotide sequence.

Description

[Technical Field]

[0001] This application claims priority from U.S. Provisional Patent Application No. 63 / 380,093, filed October 19, 2022, which is incorporated herein by reference.

[0002] This invention was made with government support under grants NS110776 and AG072298 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0003] The present application relates to methods and compositions for rejuvenating CNS glial populations using agents that express the B-cell lymphoma / leukemia 11A (BCL11A) transcription factor. [Background technology]

[0004] Glial progenitor cells (GPCs, also known as oligodendrocyte precursor cells and NG2 cells) colonize the human brain during development and remain abundant throughout adulthood. During development, human GPCs (hGPCs) are highly proliferative, bipotential cells that generate new oligodendrocytes and astrocytes. In rodents, this capacity declines during normal aging, with aging GPCs exhibiting diminished proliferative, migratory, and differentiated capacities. Similarly, adult human GPCs have been found to be less proliferative, less migratory, and more readily differentiated than their fetal counterparts when transplanted into congenital dysmyelinating mouse hosts. However, despite the distinct capabilities of fetal and adult hGPCs and the wealth of data regarding GPC transcription in rodent aging models, little data are available that address changes in GPC gene expression during human aging or provide a clear head-to-head comparison of transcription by fetal and adult human GPCs. The present disclosure aims to overcome these deficiencies in the art. Summary of the Invention

[0005] One aspect of the present disclosure relates to a method of inducing rejuvenation in a population of adult glial progenitor cells, comprising administering to the population of adult glial progenitor cells an effective amount of an expression vector comprising a nucleotide sequence encoding BCL11A and a regulatory element operably linked to the nucleotide sequence.

[0006] Another aspect of the present disclosure relates to a method of treating a subject having a glial cell-related disorder, comprising administering to a population of adult glial progenitor cells of the subject an effective amount of an expression vector comprising a nucleotide sequence encoding BCL11A and a regulatory element operably linked to the nucleotide sequence.

[0007] Another aspect of the present disclosure relates to an expression vector comprising a nucleic acid sequence encoding BCL11A and a regulatory element operably linked to the nucleic acid sequence.

[0008] Another aspect of the present disclosure relates to adult human glial progenitor cells harboring the expression vectors of the present application. [Brief explanation of the drawings]

[0009] [Figure 1]Figure 1 shows that BCL11A expression is associated with hGPC proliferation. Panel A) Expression of BCL11A by human glial progenitor cells (GPCs) relative to undifferentiated embryonic stem cells (ESCs) as a function of days in vitro (DIV) by RT-qPCR. BCL11A expression decreases during neural induction, increases during GPC specification, and declines again with cell age. Top, timeline defining differentiation stages. Panel B) Left, representative images of immunostaining for DAPI (blue) and MKI67 (green) in iPSC-derived hGPCs at 100 and 300 DIV. Right, quantification of MKI67+ cells relative to total DAPI+ cells. N=2. Panel C) Schematic showing the design of CBh-EGFP control virus (top) or CBh-BCL11A-T2A-EGFP. Panel D) Relative percentage of MKI67+ cells in the BCL11A+ (left) and GFP+ and GFP- fractions of LV-BCL11A-transduced hGPCs at DIV170, normalized to the LV-EGFP condition. Data are shown as mean ± SEM (N = 3). BCL11A was significantly overexpressed in LV-BCL11A-infected cultures, including by GFP+ cells (31.4 ± 2.8-fold, **P = 0.008, one-sample t-test), with a strikingly similar upward trend in GFP- cells (8.62 ± 2.66, one-sample t-test P = 0.103). In BCL11A-LV cultures, more MKI67+ cells were found among GFP+ cells (2.85 ± 0.61-fold greater than controls, paired t-test *P = 0.015) and GFP- cells (2.15 ± 0.42, *P = 0.017, paired t-test). Panel E) RT-qPCR results for BCL11A, MKI67, CDKN1A, CDKN2A, IL1A, and IL8 after treatment of hGPCs at DIV170 with etoposide and either LV-BCL11A or LV-EGFP, normalized to DMSO (N = 2-4 per target). BCL11A and MKI67 are downregulated, and CDKN1A is increased after etoposide treatment. Restoration of BCL11A expression by LV-BCL11A leads to a significant reduction in CDKN1A, along with a reduction in markers of inflammation and senescence (paired t-test, ***P<0.001). [Figure 2] Figure 1 shows that BCL11A overexpression induces a proliferative, migratory profile. Panel A) Volcano plot comparing differentially expressed genes in hGPCs 1 week after transduction with BCL11A or EGFP control. Panel B) Venn diagram showing the relative overlap between BCL11A-induced transcripts and transcripts enriched in primary fetal or adult hGPC isolates. Panel C) Heatmap of TPM for some of the top differentially expressed genes in BCL11A or control cultures. Highlighted in blue are down-regulated under BCL11A conditions and enriched in adult primary samples; highlighted in red are up-regulated by BCL11A overexpression and fetal enriched. Panel D) Upstream regulator analysis from Ingenuity Pathway Analysis (IPA), plotted from highest activation to highest repression. Predicted regulators include genes related to migration, proliferation, and chromatin regulation. Panel E) Significantly activated and repressed pathways as determined by IPA, showing a decrease in apoptotic and neural guidance pathways and an increase in proliferative and neuroglial pathways. [Figure 3] Single-cell RNA-seq of BCL11A-overexpressing cultures is shown. Panel A) UMAP plot showing cells from two sets of BCL11A (top) and GFP (bottom) hGPC cultures. Panel B) Dot plot showing markers of cell types expressed in vitro. Panel C) Violin plot showing genes significantly enriched in BCL11A-treated hGPCs (top) compared to their GFP-only controls (bottom). Panel D) UMAP plot colored by the ratio between fetal and adult AUC enrichment scores. Panel E) Fetal (left) or adult (right) AUC scores by Seurat cluster. Panel F) Comparison of overall AUC scores in BCL11A and GFP hGPCs. Panel G) Percentage of BCL11A- and GFP-treated cells in the most fetal-like and most adult-like clusters. [Figure 4]Figure 1 shows the in vivo migration of aging hGPCs upon BCL11A activation. Panel A) Schematic showing the design of the RFP tag used in hGPCs for mouse engraftment. Panel B) Experimental design. After 160 days in vitro, RFP+ hGPCs were engrafted into P1 mice. After 100 weeks, animals were injected with one virus in each hemisphere, LV-EGFP and LV-BCL11A-EGFP, to serve as an internal control. Three weeks later, mice were harvested for downstream processing. Panel C) Coronal images showing BCL11A overexpression in hemispheres infected with BCL11A (top) or GFP (bottom). The dashed line indicates the border of the corpus callosum (CC). Ctx - cortex, Str - striatum. Panel D) Coronal images showing MKI67+ cells in CC infected with BCL11A (left) and EGFP. Panel E) Quantification of BCL11A+ and MKI67+ cells per µm in the BCL11A or EGFP hemisphere (for BCL11A, LV-BCL11A cells / µm = 4.56e-5 ± 8.23e-6, LV-EGFP cells / µm = 5.06e-6 ± 1.45e-6, paired t-test *P = 0.031; for MKI67, LV-BCL11A cells / µm = 1.6e-5 ± 4.06e-6, LV-EGFP cells / µm = 6.63e-6 ± 1.26e-6, paired t-test P = 0.079, N = 3). Panel F) Coronal image of a chimerized CC. Human cells are marked in magenta with RFP and human nuclear antigen (hNA) and stained in green with OLIG2. Panel G) Coronal image of a chimerized CC. Human cells are marked with RFP and human nuclear antigen (hNA, magenta) and PDGFRa (green). [Figure 5]This shows that BCL11A enhances permissive chromatin states at genes for cell migration and survival. Panel A) UMAP plots of human cells isolated from three chimerized 2-year-old mouse brains, grouped by treatment condition (bottom). Dashed lines indicate clusters unique to the BCL11A condition. Panel B) Plot showing relative expression of markers of glial lineages, including astrocytes, GPCs, GPCs terminally differentiated into oligodendrocytes, and oligodendrocytes. Panel C) Violin plot showing transcripts enriched in cluster 8 (top) in BCL11A-treated hGPCs or their counterparts in the EGFP-treated condition. Panel D) UMAP plots of scATAC-seq of hGPC cultures expressing BCL11A or EGFP controls. Panel E) Differentially accessible loci in BCL11A (left) or EGFP (right) cells. Panel F) Relative abundance of K4me3, K27me3, and K9me3 signals detected genome-wide in hGPCs overexpressing BCL11A compared to controls (N=2). Panel G) Volcano plot highlighting loci differentially enriched for K4me3 (left), K27me3 (center), and K9me3 (right) in BCL11A or EGFP cells. Panel H) Representative tracks highlighting differences in chromatin mark enrichment in CTNNB1, TEAD2, and PIEZO1. DETAILED DESCRIPTION OF THE INVENTION

[0010] The sequence listing filed with the U.S. Patent Office as 1134-131 PCT.xml, created October 11, 2023, and having a size of 6,587 bytes, is incorporated herein by reference.

[0011] Reference will now be made in detail to certain aspects and exemplary embodiments of the present application, illustrating examples of the accompanying structures and drawings. Aspects of the present application will be described in conjunction with exemplary embodiments, including methods, materials, and examples; such description is non-limiting, and the scope of the present application is intended to encompass all equivalents, alternatives, and modifications, whether generally known or incorporated herein. The described aspects, features, advantages, and characteristics of the invention may be combined in any suitable manner in one or more additional embodiments. Those skilled in the relevant art will recognize that the invention may be practiced without one or more of the specific aspects or advantages of a particular embodiment. In other cases, additional aspects, features, and advantages may be recognized and claimed in certain embodiments that may not be present in all embodiments of the invention. Moreover, those skilled in the art will recognize many techniques and materials similar or equivalent to those described herein, which could be used in the practice of aspects and embodiments of the present application. The described aspects and embodiments of the present application are not limited to the methods and materials described.

[0012] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0013] I. Definition As used herein, the following terms or phrases (in parentheses) shall have the following meanings:

[0014] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the content clearly dictates otherwise. Thus, for example, reference to "a peptide" includes "one or more" peptides or "a plurality" of such peptides.

[0015] The term "about" or "approximately" includes being within a statistically meaningful range of a value. Such a range can be within an order of magnitude, preferably within 50%, more preferably within 20%, even more preferably within 10%, and even more preferably within 5% of a given value or range. The allowable variations encompassed by the term "about" or "approximately" will depend on the particular system under study and can be readily appreciated by one of ordinary skill in the art.

[0016] As used herein, the term "and / or" means that the listed items may be present or used individually or in any combination. In effect, the term means that "at least one of" or "one or more" of the listed items are used or present.

[0017] As will be understood by those skilled in the art, for any and all purposes, including providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations thereof. Any recited range can be readily recognized as fully descriptive and capable of being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc., of the same range. As a non-limiting example, each range discussed herein can be easily broken down into a lower third, middle third, upper third, etc. As will also be understood by those skilled in the art, all language such as "up to," "at least," etc., refers to a range that is inclusive of the recited numbers and can subsequently be broken down into subranges as described above. Finally, as will be understood by those skilled in the art, a range includes each individual member.

[0018] For purposes of understanding the scope of this application, the term "comprising," and its derivatives, as used herein, are intended to be open-ended terms specifying the presence of stated features, elements, components, groups, items, and / or steps, but not excluding the presence of other, unrecited features, elements, components, groups, items, and / or steps. The same also applies to words of similar meaning, such as "including," "involving," and "having," and their derivatives. The term "consisting of," and its derivatives, as used herein, are intended to be close-ended terms specifying the presence of stated features, elements, components, groups, items, and / or steps, but excluding the presence of other, unrecited features, elements, components, groups, items, and / or steps. The term "consisting essentially of" and its derivatives, as used herein, are intended to specify the presence of a described feature, element, component, group, item, and / or step and that do not materially affect the basic and novel characteristics of the feature, element, component, group, item, and / or step. In embodiments or claims in which the transitional term comprising (or the like) is used, such embodiments can also be envisioned by replacing the term "comprising" with the term "consisting of" or "consisting essentially of." The disclosed methods, kits, systems, and / or compositions can comprise, consist essentially of, or consist of the disclosed components.

[0019] In embodiments that include "additional" or "second" components, the second component, as used herein, is different from the other components or the first component. The "third" component is different from the other components, the first component, and the second component, and further listed or "additional" components are similarly different.

[0020] The term "complementary," when used in reference to nucleic acids, refers to the pairing of bases A with T or U, and G with C. The term "complementary" refers to nucleic acid molecules that are fully complementary, i.e., that form A with T or U pairs and G with C pairs throughout the reference sequence, as well as partially (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) complementary molecules.

[0021] The terms "nucleic acid," "nucleotide," and "polynucleotide" encompass both DNA and RNA, unless otherwise specified.

[0022] The terms "polypeptide," "peptide," or "protein" are used interchangeably herein to refer to a polymer of amino acid residues. This term encompasses all types of naturally occurring and synthetic proteins, including protein fragments of all lengths, fusion proteins, and modified proteins (including but not limited to glycoproteins), as well as other types of modified proteins (e.g., proteins resulting from phosphorylation, acetylation, myristoylation, palmitoylation, glycosylation, oxidation, formylation, amidation, polyglutamylation, ADP-ribosylation, pegylation, biotinylation, etc.).

[0023] The terms "disable," "disabled," or "eliminate" expression of a gene or gene product (e.g., RNA or protein) refer to the complete loss of transcription and / or translation of the gene, or the complete loss of the gene product (e.g., RNA or protein). Expression of the gene or gene product (e.g., RNA or protein), compared to a control, e.g., an unmodified cell, can be detected by standard methods known in the art, e.g., methods described herein.

[0024] The terms "express" and "expression" mean to enable or cause the information in a gene or DNA sequence to be produced, e.g., to produce an RNA or protein by activating cellular functions involved in transcription and / or translation of the corresponding gene or DNA sequence. A DNA sequence is expressed in or by a cell to form an "expression product," such as an RNA or protein. The expression product itself, e.g., the resulting protein, may also be said to be "expressed" by the cell. Expression products may be characterized as intracellular, extracellular, or transmembrane.

[0025] As used herein, the term "glial cells" refers to a population of non-neuronal cells that provide support and nutrition, maintain homeostasis, either form myelin or promote myelination, and participate in signal transduction in the nervous system. As used herein, "glial cells" encompass fully differentiated cells of the glial lineage, e.g., oligodendrocytes or astrocytes. For purposes of this application / disclosure, the term "glial cells" also refers to glial precursor cells at various stages of development or differentiation. Each of the glial cells defined in this paragraph may be referred to as an astroglial cell.

[0026] As used herein, the term "adult glial progenitor cells" refers to glial progenitor cells present in a mammal at any stage of postnatal development. In some embodiments, the term "adult glial progenitor cells" refers to glial progenitor cells present in a human subject who is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 years of age or older. In some embodiments, the term "adult glial progenitor cells" refers to glial progenitor cells present in a human subject who is 20 years of age or older, 25 years of age or older, 30 years of age or older, 35 years of age or older, 40 years of age or older, 45 years of age or older, or 50 years of age or older. In some embodiments, the term "adult glial progenitor cells" refers to glial progenitor cells present in an older human subject, e.g., an adult who is 55 years of age or older, 60 years of age or older, 65 years of age or older, 70 years of age or older, 75 years of age or older, or 80 years of age or older.

[0027] The term "functional variant" of a gene product (e.g., a protein) refers to a modified gene product (e.g., by deletion, substitution, insertion, glycosylation, etc.) that retains at least 50% of the biological activity of the unmodified (wild-type) gene product in a competition assay.

[0028] The term "effective amount" refers to the amount of an active compound or pharmaceutical agent that elicits the biological or pharmacological response in a tissue, system, animal, individual, or human that is being sought by a researcher, veterinarian, medical doctor, or other clinician.

[0029] The term "regulatory sequence" or "regulatory element" refers to a nucleic acid sequence or element that controls, regulates, causes, or enables the expression of a gene regulated by such regulatory sequence or element. Regulatory elements / sequences can be found 5' or 3' to the coding region of the regulated gene, or within the coding region, or within an intron. Examples of regulatory sequences / elements include, but are not limited to, promoters, enhancers, RNA polymerase initiation sites, ribosome binding sites, and other sequences that facilitate expression of an encoded polypeptide in a given expression system.

[0030] The term "promoter," as used herein, refers to a nucleotide sequence capable of controlling the expression of a coding sequence or functional RNA. Generally, a polynucleotide of interest is located 3' to the promoter sequence. In some embodiments, a promoter is derived entirely from a native gene. In some embodiments, a promoter is composed of different elements from different naturally occurring promoters. In some embodiments, a promoter comprises a synthetic nucleotide sequence. Those skilled in the art will understand that different promoters direct the expression of a gene in different tissues or cell types, or at different developmental stages, or in response to different environmental conditions, or in response to the presence or absence of drugs or transcriptional cofactors. Ubiquitous, cell type-specific, tissue-specific, developmental stage-specific, and conditional promoters, such as drug-responsive promoters (e.g., tetracycline-responsive promoters), are well known to those skilled in the art. Examples of promoters include, but are not limited to, the phophoglycerate kinase (PKG) promoter, CAG, NSE (neuron-specific enolase), synapsin, or NeuN promoter, the SV40 early promoter, the mouse mammary tumor virus long terminal repeat (LTR) promoter, the adenovirus major late promoter (AdMLP), the herpes simplex virus (HSV) promoter, the cytomegalovirus (CMV) promoter such as the CMV immediate early promoter region (CMVIE), the SFFV promoter, the Rous sarcoma virus (RSV) promoter, synthetic promoters, hybrid promoters, and the like. Promoters can be of human origin or from other species, including mice. Additionally, sequences derived from non-viral genes, such as the mouse metallothionein gene promoter, are also used herein. In some embodiments, the promoter is a heterologous promoter. In some embodiments, the promoter sequence consists of proximal and more distal upstream elements and may include enhancer elements.

[0031] The term "heterologous promoter," as used herein, refers to a promoter that is not found operably linked to a given coding sequence in nature.

[0032] The term "enhancer" refers to a nucleotide sequence capable of stimulating promoter activity and may be a native element of the promoter or a heterologous element inserted to enhance the level or tissue specificity of the promoter.

[0033] The terms "operably linked" or "operably linked" refer to the association of two or more nucleic acid fragments on a single nucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably linked to a coding sequence when it is capable of affecting the expression of that coding sequence (e.g., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory sequences in either sense or antisense orientation.

[0034] The term "transcription factor" refers to a DNA-binding protein that regulates the expression of a particular gene. Transcription factors can have a positive effect on gene transcription and therefore may be referred to as "activators" or "transcriptional activators." Transcription factors can also have a negative effect on gene expression and therefore may be referred to as "repressors" or "transcriptional repressors."

[0035] A rejuvenation process is observed when one or all of these markers of the fatigue phenotype are reduced or suppressed in the fatigued or senescent cell type due to the rejuvenation process.

[0036] Certain terms used in the specification, examples, and claims are collected here. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0037] The preferences and options for a given aspect, feature, embodiment, or parameter of the present disclosure should be considered to be disclosed in combination with any and all preferences and options for all other aspects, features, embodiments, and parameters of the present disclosure, unless the context indicates otherwise.

[0038] II. Methods Involving Promoting Expression and / or Activity of BCL11A Transcription Factor A. Methods for Inducing Rejuvenation in Glial Progenitor Cell Populations One aspect of the present disclosure relates to a method of inducing rejuvenation in a population of glial progenitor cells, hi some embodiments, the method comprises promoting the activity and / or enhancing the expression of B-cell lymphoma / leukemia 11A (BCL11A) in the population of glial progenitor cells at a level that induces rejuvenation of the glial progenitor cells.

[0039] BCL11A is a transcription factor expressed in the brain that forms a protein complex with CASK to regulate axon initiation and branching. Furthermore, BCL11A binds to the TBR1 regulatory region and inhibits the expression of TBR1, a neuron-specific protein implicated in intellectual disability and autism spectrum disorders.

[0040] Human BCL11A is located on chromosome 2p16.1 and is encoded by a gene highly conserved with mouse BCL11A (musBcl11a). Human BCL11A encodes a 125 kDa Kruppel-like zinc finger protein containing six C2H2 zinc fingers, a proline-rich region, and an acidic domain. BCL11A specifically binds to the 5'-GGCCGG-3' sequence and primarily functions as a transcriptional repressor. BCL11A is primarily expressed in the brain and most hematopoietic cells (including hematopoietic stem cells, general lymphoid precursors, B cells, and early T cell precursors), but is weakly expressed in T lymphocytes. As shown in Table 1, human BCL11A has multiple spliced ​​transcript variants. [Table 1] *Each of which is incorporated herein by reference in its entirety.

[0041] In some embodiments, the methods of the present application comprise expressing an effective amount of BCL11A protein in a population of glial progenitor cells. In some embodiments, the glial progenitor cells are adult glial progenitor cells.

[0042] In some embodiments, the expressing step comprises administering to the population of glial precursors or adult glial progenitor cells an effective amount of an expression vector capable of expressing BCL11A in the glial precursors or adult glial progenitor cells in an amount effective to induce rejuvenation of the glial precursors or adult glial progenitor cells. In some embodiments, the expression vector comprises a nucleotide sequence encoding BCL11A and a regulatory element operably linked to the nucleotide sequence. In some embodiments, the administering occurs ex vivo. In some embodiments, the administering occurs in vivo.

[0043] The terms "rejuvenating" or "rejuvenation", when used in the context of glial or adult glial cells or glial precursor or adult glial progenitor cells, refer to the reversal of the aging process in such cells and the return to a youthful cellular state, particularly with respect to their proliferative and / or differentiative capacity without loss of cellular identity.

[0044] The cellular aging phenotype in the glia or glial progenitor cells described above can be characterized by, among other things, the following markers: (1) increased expression of one or more markers of senescence, such as CDKN1A, CDKN2A, E2F6, ZNF274, IKZF3, and IL1A, or markers of glial differentiation, such as BCAS1, CLDN11, CNP, LPAR1, MAG, OGT, PLP1, PMP22, and MYRF, and (2) decreased expression of one or more youth markers associated with cellular youth, such as BCAN, CCND2, CDK1, CDK4, CDK5, CENPF, CHEK1, CHRDL1, FN1, HMGA2, LMNB1, MKI67, MYC, NFIB, TEAD1, TEAD2, and YAP1, and in particular PCDH15, PDGFRA, PTPRZ1, ST8SIA1, and CSPG4, of glial ontogeny.

[0045] As used herein, the rejuvenation process is observed when (1) the expression of one or more senescent markers is decreased, and / or (2) the expression of one or more young markers is increased. In some embodiments, the rejuvenation process is observed when an increase in the expression of one or more young markers is observed, consistent with regaining mitotic division and differentiation capacity. Examples of young markers consistent with regaining mitotic division and differentiation capacity include, but are not limited to, CCND2, CDK1, CDK4, CDK5, CENPF, CHEK1, FN1, HMGA2, LMNB1, MKI67, MYC, NFIB, PATZ1, TEAD1, TEAD2, TP53, and YAP1.

[0046] In some embodiments, the rejuvenation process is observed when increased expression of one or more juvenile markers consistent with a functional glial progenitor state is observed, including, but not limited to, BCAN, CA10, CHRDL1, CSPG4, NXPH1, PCDH15, PDGFRA, PTPRZ1, and ST8SIA1.

[0047] In some embodiments, an agent has a rejuvenating effect on (1) glial or adult glial cells or cell populations thereof, or (2) glial precursor or adult glial progenitor cells or cell populations thereof, if the expression of one or more juvenile markers is significantly increased in the aforementioned cells or cell populations after treatment with the agent.

[0048] In some embodiments, an agent has a rejuvenating effect on (1) glial or adult glial cells or cell populations thereof, or (2) glial precursor or adult glial progenitor cells or cell populations thereof, if the expression of one or more senescence markers is significantly reduced in the cells or cell populations after treatment with the agent.

[0049] In some embodiments, an agent has a rejuvenating effect on (1) glial or adult glial cells or cell populations thereof, or (2) glial precursor or adult glial progenitor cells or cell populations thereof, if, after treatment with the agent, the expression of one or more juvenile markers is significantly increased and the expression of one or more senescent markers is significantly decreased in the cells or cell populations.

[0050] As used herein, expression of a gene is "significantly increased" if it is increased by 30% or more, 50% or more, 100% or more, 150% or more, 200% or more, 300% or more, 400% or more, 500% or more, 600% or more, 700% or more, 800% or more, 900% or more, or 1000% or more at the mRNA level, or by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100% or more at the protein level.

[0051] As used herein, expression of a gene is "significantly decreased" if it is decreased by 30% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more at the mRNA level, or by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more at the protein level.

[0052] In some embodiments, the methods of the present application comprise expressing in a population of glial progenitor cells an effective amount of BCL11A protein that results in increased expression of the MKI67 gene in the population of glial progenitor cells.

[0053] In some embodiments, the methods of the present application comprise expressing in a population of glial progenitor cells an effective amount of BCL11A protein, which results in activation of aged glial progenitor cells, hi some embodiments, the activation of aged glial progenitor cells is evidenced by mitotic expansion and / or migratory colony formation of aged glial progenitor cells.

[0054] In some embodiments, the methods of the present application comprise expressing in a population of glial progenitor cells an effective amount of BCL11A protein that results in migration of aged glial progenitor cells and thereby remyelination. One skilled in the art will recognize that remyelination by such already aged progenitor cells will provide therapeutic benefit in treating glial cell-related disorders or conditions.

[0055] Glial progenitor cells suitable for use in the methods disclosed herein include mammalian glial progenitor cells, such as human glial progenitor cells, rodent glial progenitor cells, non-human primate glial progenitor cells, ovine glial progenitor cells, bovine glial progenitor cells, butaglian progenitor cells, canine glial progenitor cells, and feline glial progenitor cells. In some embodiments, the adult glial progenitor cells are adult human glial progenitor cells.

[0056] In some embodiments, the expression vector comprises a nucleotide sequence encoding human BCL11A or a functional variant thereof. In some embodiments, the expression vector comprises a nucleotide sequence encoding human BCL11A. In some embodiments, the expression vector comprises a nucleotide sequence encoding SEQ ID NO:2.

[0057] In some embodiments, regulatory elements include ubiquitous promoters such as the chicken beta-actin (CBA) promoter, a hybrid form of the CBA promoter (CBh)CAG promoter, the cytomegalovirus (CMV) promoter, and the Rous sarcoma virus (RSV) promoter.

[0058] In some embodiments, the regulatory elements include promoters and / or enhancers of genes selectively expressed by glial progenitor cells, such as the promoters / enhancers of platelet-derived growth factor alpha (PDGFRA), zinc finger protein 488 (ZNF488), G protein-coupled receptor (GPR17), oligodendrocyte transcription factor 2 (OLIG2), chondroitin sulfate proteoglycan 4 (CSPG4), and SRY-box transcription factor 10 (SOX10).

[0059] In some embodiments, the regulatory element comprises an inducible promoter. An inducible promoter can directly or indirectly activate transcription of a nucleic acid molecule to which it is operably linked in response to a "modulator" (e.g., a chemical agent or a biomolecule, e.g., a metabolite, a small molecule) or stimulus. In the absence of a "modulator" or stimulus, the nucleotide sequence encoding BCL11A is not transcribed. The terms "not transcribed" or "not substantially expressed" mean that the level of transcription is at least 10-fold, 20-fold, 30-fold, 40-fold, or 50-fold lower than the level of transcription observed in the presence of the appropriate stimulus or modulator, and preferably at least 100-fold, 250-fold, or 500-fold or more lower than the level of transcription observed in the presence of the appropriate stimulus or modulator.

[0060] Inducible promoters suitable for use in the expression vectors of the present application include, but are not limited to, those regulated by hormones and hormone analogs, such as progesterone, ecdysone, and glucocorticoids, as well as promoters regulated by tetracycline, heat shock, heavy metal ions, interferon, and lactose operon-activating compounds. For a review of these systems, see Gingrich & Roder, "Inducible Gene Expression in the Nervous System of Transgenic Mice," Annu. Rev. Neurosci. 21:377-405 (1998), incorporated herein by reference in its entirety. Tissue-specific expression is well characterized in the field of gene expression, and tissue-specific and other inducible promoters are well known in the art.

[0061] In some embodiments, the regulatory element of the expression vector of the present application comprises an inducible promoter. If rejuvenation of glial progenitor cells is desired, this is preferably achieved by administering a suitable regulator (e.g., doxycycline, hormones) or other inducer to a subject using a route or other means that targets glial progenitor cells carrying the expression vector. Contacting glial progenitor cells carrying the expression vector with a regulator induces expression of the system designed to express BCL11A. However, it should be recognized by those skilled in the art that with other inducible vectors, the opposite is true: the regulator inhibits expression, and its removal allows expression. Suitable inducible promoters for inclusion in the systems of the present disclosure are well known in the art and include, but are not limited to, tetracycline-regulated operator systems, cumate-regulated operator systems, rapamycin-inducible systems, FKCsA-inducible systems, and ABA-inducible systems (see, e.g., Kallunki et al., "How to Choose the Right Inducible Gene Expression System for Mammalian Studies?" Cells 8(8):796 (2019); U.S. Pat. No. 8,728,759; and U.S. Pat. No. 7,745,592, which are incorporated by reference in their entireties).

[0062] In some embodiments, the expression vectors of the present application comprise a tetracycline-regulated operator system (tet-on promoter), and transcription of the gene of interest is activated in the presence of tetracycline. In some embodiments, the expression vectors of the present application comprise a tetracycline-regulated operator system (Tet-off promoter), and transcription of the gene of interest is activated in the absence of tetracycline.

[0063] In some embodiments, the expression vectors of the present application comprise a cumate-controlled operator system, and transcription of the gene of interest is activated in the presence of cumate. In some embodiments, the expression vectors of the present application comprise a cumate-controlled operator system, and transcription of the gene of interest is activated in the absence of cumate.

[0064] In some embodiments, the expression vectors of the present application comprise a rapamycin-regulated operator system, wherein transcription of the gene of interest is activated in the presence of rapamycin.

[0065] In some embodiments, the expression vector of the present application is a non-viral vector. In some embodiments, the non-viral vector is a plasmid.

[0066] In some embodiments, the expression vector of the present application is a viral vector. Examples of viral vectors include, but are not limited to, adenovirus vectors, adeno-associated virus ("AAV") vectors, retrovirus vectors, lentivirus vectors (i.e., lentiviral vectors), vaccinia virus vectors, herpes virus vectors, and any other viral vector suitable for introducing a gene of interest (e.g., BCL11A) described herein into a given organism or genetic background by any means that facilitates expression of the gene of interest.

[0067] In some embodiments, the expression vector of the vector of the present application is a lentiviral vector.

[0068] In some embodiments, the expression vector of the vector of the present application is an AAV vector.

[0069] In some embodiments, the expression vector of the vector of the present application is a retroviral vector.

[0070] Methods for generating and isolating viral expression vectors suitable for use as expression vectors are known in the art.

[0071] In some embodiments, the expressing step comprises administering to the population of glial progenitor cells an effective amount of an expression vector capable of expressing an agent that inhibits the activity of an endogenous repressor of BCL11A activity in adult glial progenitor cells. Examples of repressors of BCL11A activity include, but are not limited to, KLF1, POGZ, HRI, Mi2β, SOX2, and FOXQ1.

[0072] In some embodiments, the methods of the present application further comprise promoting expression of one or more pro-juvenation genes in the population of glial progenitor cells. As used herein, the term "pro-juvenation gene" refers to a gene that contributes to reversing and slowing aging, resulting in younger cells or tissues.

[0073] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of ARX, CEBPZ, DLX1, DLX2, ELK1, ETS1, ETV4, KLF16, MYBL2, MYC, NFYB, POU3F1, SMAD1, SOX3, SP5, TCF12, TFDP1, TP53, ZIC3, and ZNF195.

[0074] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of CEBPZ, MYBL2, MYC, NFYB, and ZNF195.

[0075] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of ACTB, AKR1C1, ANAPC11, AP2B1, APLP2, APOD, ARF5, ARL4A, ARPC3, ARPP19, ATOX1, ATP5F1E, ATP5MC1, ATP5MC3, ATP5MD, ATP5ME, ATP5MF, ATP5MG, ATP5MPL, ATP5PF, ATP6V0B, ATP6V0E1, ATXN7L3B, B2M, B3GAT2, BEX1, BEX3, BEX5, BLOC1S1, BMERB1, C18orf32, C1orf122, C1QBP, C4orf48, CADM4, CALM1, CALM3, CALR, CANX, CAV2, CC2D1A, CCND1, CCNI, CD63, CD82, CDC42, CDH2, CFL1, CHCHD2, CHGB, CIAO2B, CLCN3, CLTA, CLT C, CNN3, CNTN1, COTL1, COX4I1, COX6A1, COX6C, COX7A2, COX7C, COX8A, CPNE8, CPS1, CRNDE, CSPG4, CTHRC1, CUL4B, CYP51A1, DBI, DCX, DDAH1, DDX1, DENN D10, DMD, DMRT2, DNAJA2, DPYSL2, DRAP1, DSTN, DYNC1I2, EDF1, EDIL3, EEF1A1, EEF1B2, EEF2, EID1, EIF3J, ELOB, EMC10, EMP2, ESD, ETV1, FABP7, FAM17 1B, FAM177A1, FAU, FIS1, FXYD6, GADD45A, GAP43, GCSH, GNAS, GOLM1, GPM6B, GSTP1, H3-3A, H3-3B, HINT1, HNRNPA1, HNRNPA3, HNRNPAB, HNRNPC, HNRNPK, HNRNPM, HNRNPR, HSPA5, IGFBP2, ITGB8, ITM2A, ITM2B, JPT1, KDELR1, KLRK1-AS1, KRTCAP2, KTN1, LDHB, LHFPL3, LRRC4B, LY6H, MAP2, MARCKS, MARCKSL1 , MIA, MICOS10, MIF, MIR9-1HG, MMGT1, MPZL1, MT3, MTLN, MTRNR2L12, MTRNR2L8, MYL12A, MYL12B, NACA, NARS1, NCL, NDUFA1, NDUFA11, NDUFA13, NDUFA3,NDUFA4, NDUFB1, NDUFB11, NDUFB2, NDUFB6, NDUFB7, NDUFC2, NDUFS5, NEU4, NUCKS1, OAZ1, OLFM2, OSBPL8, OST4, OSTC, PABPC1 , PCBP2, PCDH10, PCDH11X, PCDH17, PCDHB2, PCDHGB6, PDGFRA, PDIA6, PEBP1, PEG10, PFN1, PGRMC1, PKIA, PLPP3, PLPPR1, PPIA , PRDX1, PRDX2, PRDX5, PSMB1, PSMB9, PTMS, PTN, PTPRA, RAB10, RAB14, RAB2A, RAB31, RAC1, RACK1, RMDN2, RAMP1, RO60, ROBO1 , RRAGB, RTN3, S100B, SARAF, SAT1, SBDS, SCARB2, SCP2, SCRG1, SEC62, SELENOK, SELENOT, SELENOW, SERF2, SERPINE2, SET, SH 3BGRL, SKP1, SLC25A6, SLIT2, SLITRK2, SMC3, SMDT1, SMOC1, SMS, SNCA, SNHG29, SNHG6, SNX3, SNX22, SOD1, SOX11, SOX2, SOX9 , SPCS2, SPCS3, SRP14, SSR4, STAG2, STMN1, SUPT16H, TALDO1, TBCB, TCEAL7, TCEAL8, TCEAL9, TIMP1, TLE5, TM4SF1, TM9SF3, T Selected from the group consisting of MA7, TMBIM6, TMCO1, TMEM147, TMEM258, TMEM50A, TMOD2, TMSB10, TMSB4X, TPT1, TRAF4, TRIO, TSC22D4, TSPAN6, TSPAN7, TTC3, TUBB, UBA52, UBL5, UQCR10, UQCR11, UQCRB, VIM, WSB2, WSCD1, YBX1, YWHAB, YWHAE, ZFAS1, ZNF428, and ZNF462.

[0076] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of APOD, B2M, BEX3, BEX5, CCND1, CTHRC1, EDIL3, EMC10, FABP7, GADD45A, ITM2A, LRRC4B, LY6H, MIA, MT3, NEU4, OLFM2, PTMS, RAMP1, SNX3, TRAF4, TRIO, UBA52, and YWHAB.

[0077] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of ANAPC11, APOD, ATP5MC3, B2M, CALM1, MT3, NEU4, PEBP1, RAMP1, SOD1, and TBCB.

[0078] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of APOD, BEX3, BEX5, CCND1, CTHRC1, EDIL3, EMC10, GADD45A, ITM2A, MIA, TRAF4, and TRIO.

[0079] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of B2M, FABP7, LRRC4B, LY6H, MT3, NEU4, OLFM2, PTMS, RAMP1, SNX3, UBA52, and YWHAB.

[0080] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of LY6H, MIA, GADD45A, ITM2A, and ITM2B.

[0081] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of HDAC2, EZH2, MYC, HMGA2, NFIB, and TEAD2.

[0082] In some embodiments, the methods of the present application further comprise inhibiting expression of one or more anti-rejuvenation genes in the population of glial progenitor cells. As used herein, the term "anti-rejuvenation gene" refers to a gene that has the opposite function of a pro-rejuvenation gene.

[0083] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ABCG1, ADGRB1, ADGRG1, AKAP9, AL360181.3, ANKRD10, ARGLU1, ARL4C, ARL16, ARMCX6, ATP1A2, ATP1B3, ATP10B, B3GNT7, BHLHE41, BPTF, BRI3, BX664615.2, BX890604.1, C1QL2, CAMK2N1, CCDC85B, CCNL1, CHCHD10, CHORDC1, CIRBP, CLDN10, COL9A1, COL9A2, CXADR, DAN CR, DCXR, DHX36, DLL3, DNAJA1, DNM3, ECH1, EGR1, EIF1AX, ELAVL3, EMID1, ETFB, FABP5, FAM133A, FAM133B, FBXO2, FERMT1, FIBIN, FOS, FOSB, FSCN1, F SIP2, GABPB1-AS1, GALR1, GNG8, GNPTAB, GOLGA8A, GOLGA8B, GPR155, GRID2, GRM7, HAPLN1, HMX1, HSPA1A, HSPA1B, HSPH1, HTRA1, IGFBP2, JAG1, JUN, JU NB, KCNIP4, KCNQ1OT1, KLF3-AS1, LAMP2, LINC01116, LINC01301, LINC01896, LRP4, LRRC7, MACF1, MALAT1, MAP3K13, MASP1, MDH1, MT1E, MT2A, MYT1, N ASP, NKTR, NUTM2A-AS1, OFD1, PCDHB5, PCDHGA3, PCDHGB6, PEPD, PHGDH, PLCG2, PMP2, PNISR, PPP1R14A, PTGDS, RAB3IP, RAF1, RAP1GAP, RARRES2, RBM25 , RBMX, REV3L, RHOBTB3, RIMS2, RIT2, RRBP1, RSRP1, S100A1, S100A16, SAT1, SCG2, SEMA3E, SERTAD1, SEZ6L, SEZ6L2, SH3GLB2, SNHG15, SNRNP70, SPARCL1, SRSF5, STAT3, STXBP6, SYNRG, THBS4, TLE4, TMEM176B, TPI1, TSC22D3, USP11, VCAN, WFDC1, WSB1, ZFYVE16, ZNF528, and ZNF528-AS1.

[0084] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ADGRG1 ARL4C, ARMCX6, ATP1A2, ATP1B3, B3GNT7, CXADR, DLL3, FABP5, FIBIN, IGFBP2, LRRC7, MAP3K13, MT1E, MT2A, PCDHGA3, PCDHGB6, PLCG2, PTGDS, SAT1, SEZ6L, SPARCL1, THBS4, and TLE4.

[0085] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ARGLU1, EGR1, FSIP2, HSPH1, MACF1, NKTR, RBMX, STAT3, TLE4, and WSB1.

[0086] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ADGRG1, ATP1A2, ATP1B3, B3GNT7, CXADR, DLL3, FABP5, MT1E, MT2A, PTGDS, SEZ6L, and THBS4.

[0087] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ARL4C, ARMCX6, FIBIN, IGFBP2, LRRC7, MAP3K13, PCDHGA3, PCDHGB6, PLCG2, SAT1, SPARCL1, and TLE4.

[0088] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ZNF274, MAX, E2F6, IKZF3, and STAT3.

[0089] In some embodiments, inhibiting the expression of one or more anti-rejuvenation genes comprises administering to the population of glial progenitor cells an expression vector encoding one or more microRNAs. As used herein, the term "microRNA" or "miRNA" refers to a class of small RNA molecules that can negatively regulate gene expression. In some embodiments, the one or more microRNAs are selected from the group consisting of miR-193a-5P, miR-23b-3p, miR-4687-3p, miR-4651, miR-4270, and miR-24-3p. It may be necessary to expand this group.

[0090] In some embodiments, inhibiting the expression of one or more rejuvenation-inhibiting genes comprises administering to the population of glial progenitor cells an expression vector encoding one or more shRNAs. Short hairpin RNA (shRNA) molecules contain sense and antisense sequences from a target gene connected by a loop. Once transcribed, shRNA molecules are transported from the nucleus to the cytoplasm, where the enzyme Dicer processes them into small / short interfering RNAs (siRNAs). As used herein, the term "short hairpin RNA interference" or "shRNAi" refers to a process mediated by a type of small RNA molecule that negatively regulates gene expression.

[0091] In some embodiments, inhibiting the expression of one or more rejuvenation-inhibiting genes comprises administering to the population of glial progenitor cells one or more antisense oligonucleotides or an expression vector encoding one or more antisense oligonucleotides. As used herein, the term "antisense oligonucleotide" or "ASO" refers to small (approximately 18-30 nucleotides), synthetic, single-stranded nucleic acid polymers of diverse chemistries that can be used to modulate gene expression through various mechanisms. ASOs can be subdivided into two major categories: RNase H-competent and steric-blocked. The endogenous RNase H enzyme RNASEH1 recognizes the RNA-DNA heteroduplex substrate formed when DNA-based oligonucleotides bind to their cognate mRNA transcripts and catalyzes RNA degradation. Cleavage at the ASO binding site causes destruction of the target RNA, thereby silencing target gene expression. Stereoblocked oligonucleotides are ASOs designed to bind to target transcripts with high affinity but lack RNase H capability and therefore do not induce target transcript degradation. Thus, such oligonucleotides contain either nucleotides that do not form RNase H substrates when paired with RNA, or a mixture of nucleotide chemistries (i.e., "mixmers") such that runs of consecutive DNA-like bases are avoided.

[0092] In some embodiments, inhibiting the expression of one or more rejuvenation-inhibiting genes comprises administering to the population of glial progenitor cells an expression vector encoding a nuclease-based gene editing system. As used herein, the term "nuclease-based gene editing system" refers to a system comprising a nuclease or a derivative thereof that can be recruited to a target sequence within a genome. Examples of nuclease-based gene editing systems include, but are not limited to, clustered regularly interspaced short palindromic repeats-associated ("Cas") proteins (e.g., Cas9, Cas12a, and Cas12b)-associated systems (CRISPR-CAS systems), zinc finger nuclease-associated systems ("ZFN systems"), and transcription activator-like effector nuclease-associated systems ("TALEN systems").

[0093] B. Methods of treating glial cell-associated disorders. Another aspect of the present disclosure relates to a method of treating a subject having a glial cell-related disorder, comprising administering to a population of glial progenitor cells of the subject an effective amount of an agent that promotes the activity and / or enhances the expression of BCL11A in the population of glial progenitor cells.

[0094] In some embodiments, the glial cell-associated disorder is selected from the group consisting of multiple sclerosis, neuromyelitis optica, transverse myelitis, optic neuritis, subcortical infarction, diabetic leukoencephalopathy, hypertensive leukoencephalopathy, age-related white matter disease, spinal cord injury, radiation or chemotherapy-induced demyelination, post-infectious and post-vaccination leukoencephalitis, periventricular leukomalacia, childhood leukodystrophies (e.g., Pelizaeus-Merzbacher disease, Tay-Sachs disease, Sandhoff gangliosidosis, Krabbe disease, metachromatic leukodystrophy, mucopolysaccharidoses, Niemann-Pick A disease, adrenoleukodystrophy, Canavan disease, vanishing white matter disease, and Alexander disease), lysosomal storage diseases, congenital dysmyelination, inflammatory demyelination, vascular demyelination, and cerebral palsy.

[0095] In some embodiments, the glial cell-associated disorder is a neurodegenerative disease selected from the group consisting of Huntington's disease, frontotemporal dementia, Parkinson's disease, multiple system atrophy, and amyotrophic lateral sclerosis.

[0096] In some embodiments, the glial cell-associated disorder is a neuropsychiatric disorder selected from the group consisting of schizophrenia, autism spectrum disorder, and bipolar disorder.

[0097] In some embodiments, the glial cell-associated disorder is a myelin disease, and the myelin disease is a leukodystrophy or a white matter disease.

[0098] In some embodiments, "treating" a subject with a glial cell-related disorder includes (1) preventing, delaying, or reducing the likelihood of the onset and / or appearance of at least one clinical or subclinical symptom of a glial cell-related disorder occurring in a subject who may be affected by or is predisposed to a glial cell-related disorder but who has not yet experienced or is not displaying a clinical or subclinical symptom of the glial cell-related disorder, or (2) inhibiting the glial cell-related disorder, i.e., preventing, reducing, or delaying the onset of dysmyelination or its recurrence, or at least one clinical or subclinical symptom thereof, or (3) alleviating the glial cell-related disorder, i.e., causing regression of the glial cell-related disorder or at least one clinical or subclinical symptom thereof. The benefit to the treated subject is statistically significant or at least perceptible to the patient or physician.

[0099] As used herein, the term "subject" refers to an individual organism, e.g., an individual mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a non-human primate. In some embodiments, the subject is a rodent. In some embodiments, the subject is a sheep, goat, cat, or dog. In some embodiments, the subject is a research animal. In some embodiments, the subject has been genetically engineered, e.g., is a genetically engineered non-human subject.

[0100] The subject can be an adult subject, hi some embodiments, the subject is at least 1 year old, at least 2 years old, at least 4 years old, at least 6 years old, at least 8 years old, at least 10 years old, at least 12 years old, at least 15 years old, at least 18 years old, at least 20 years old, at least 25 years old, at least 30 years old, at least 35 years old, at least 40 years old, at least 45 years old, at least 50 years old, at least 55 years old, at least 60 years old, at least 65 years old, at least 70 years old, at least 75 years old, at least 80 years old, at least 85 years old, at least 90 years old, at least 95 years old, at least 100 years old, or older.

[0101] In some embodiments, the subject is an adult subject between 18 and 100 years old, 20 and 100 years old, 30 and 100 years old, 40 and 100 years old, 50 and 100 years old, 50 and 100 years old, 60 and 100 years old, 70 and 100 years old, 80 and 100 years old, or 90 and 100 years old.

[0102] In some embodiments, the agent that promotes the activity and / or enhances the expression of BCL11A is an agent that expresses an effective amount of BCL11A protein, which results in increased expression of the MKI67 gene in glial progenitor cells.

[0103] In some embodiments, the agent that promotes BCL11A activity and / or enhances its expression is an agent that expresses an effective amount of BCL11A protein, which results in activation of aged glial progenitor cells, hi some embodiments, activation of aged glial progenitor cells is evidenced by mitotic expansion and / or migratory colony formation of aged glial progenitor cells.

[0104] In some embodiments, the agent that promotes the activity and / or enhances the expression of BCL11A is an agent that expresses an effective amount of BCL11A protein that results in remyelination of aged glial cells.

[0105] In some embodiments, the agent is an expression vector comprising a nucleotide sequence encoding BCL11A and a regulatory element operably linked to the nucleotide sequence.

[0106] In some embodiments, the expression vector comprises a nucleotide sequence encoding human BCL11A or a functional variant thereof. In some embodiments, the expression vector comprises a nucleotide sequence encoding human BCL11A. In some embodiments, the expression vector comprises a nucleotide sequence encoding SEQ ID NO:2.

[0107] In some embodiments, regulatory elements include ubiquitous promoters such as the chicken beta-actin (CBA) promoter, a hybrid form of the CBA promoter (CBh)CAG promoter, the cytomegalovirus (CMV) promoter, and the Rous sarcoma virus (RSV) promoter.

[0108] In some embodiments, the regulatory elements include promoters and / or enhancers of genes selectively expressed by glial progenitor cells, such as the promoters / enhancers of platelet-derived growth factor alpha (PDGFRA), zinc finger protein 488 (ZNF488), G protein-coupled receptor (GPR17), oligodendrocyte transcription factor 2 (OLIG2), chondroitin sulfate proteoglycan 4 (CSPG4), and SRY-box transcription factor 10 (SOX10).

[0109] In some embodiments, the regulatory element comprises an inducible promoter. In some embodiments, the inducible promoter is a tet-on or tetp-off promoter. In some embodiments, the inducible promoter comprises a cumate-controlled operator system, where transcription of the gene of interest is activated in the presence of cumate. In some embodiments, the expression vector of the present application comprises a cumate-controlled operator system, where transcription of the gene of interest is activated in the absence of cumate. In some embodiments, the inducible promoter comprises a rapamycin-controlled operator system, where transcription of the gene of interest is activated in the presence of rapamycin.

[0110] In some embodiments, the expression vector is a non-viral vector. In some embodiments, the non-viral vector is a plasmid.

[0111] In some embodiments, the expression vector is a viral vector, hi some embodiments, the viral vector is a lentiviral vector, an AAV vector, or a retroviral vector.

[0112] In some embodiments, the agent comprises an expression vector capable of expressing an agent that inhibits the activity of an endogenous repressor of BCL11A activity in adult glial progenitor cells. Examples of repressors of BCL11A activity include, but are not limited to, KLF1, POGZ, HRI, Mi2β, SOX2, and FOXQ1.

[0113] In some embodiments, the method further comprises promoting expression of one or more pro-juvenation genes in the population of glial progenitor cells.

[0114] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of ARX, CEBPZ, DLX1, DLX2, ELK1, ETS1, ETV4, KLF16, MYBL2, MYC, NFYB, POU3F1, SMAD1, SOX3, SP5, TCF12, TFDP1, TP53, ZIC3, and ZNF195.

[0115] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of CEBPZ, MYBL2, MYC, NFYB, and ZNF195.

[0116] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of ACTB, AKR1C1, ANAPC11, AP2B1, APLP2, APOD, ARF5, ARL4A, ARPC3, ARPP19, ATOX1, ATP5F1E, ATP5MC1, ATP5MC3, ATP5MD, ATP5ME, ATP5MF, ATP5MG, ATP5MPL, ATP5PF, ATP6V0B, ATP6V0E1, ATXN7L3B, B2M, B3GAT2, BEX1, BEX3, BEX5, BLOC1S1, BMERB1, C18orf32, C1orf122, C1QBP, C4orf48, CADM4, CALM1, CALM3, CALR, CANX, CAV2, CC2D1A, CCND1, CCNI, CD63, CD82, CDC42, CDH2, CFL1, CHCHD2, CHGB, CIAO2B, CLCN3, CLTA, CLT C, CNN3, CNTN1, COTL1, COX4I1, COX6A1, COX6C, COX7A2, COX7C, COX8A, CPNE8, CPS1, CRNDE, CSPG4, CTHRC1, CUL4B, CYP51A1, DBI, DCX, DDAH1, DDX1, DENN D10, DMD, DMRT2, DNAJA2, DPYSL2, DRAP1, DSTN, DYNC1I2, EDF1, EDIL3, EEF1A1, EEF1B2, EEF2, EID1, EIF3J, ELOB, EMC10, EMP2, ESD, ETV1, FABP7, FAM17 1B, FAM177A1, FAU, FIS1, FXYD6, GADD45A, GAP43, GCSH, GNAS, GOLM1, GPM6B, GSTP1, H3-3A, H3-3B, HINT1, HNRNPA1, HNRNPA3, HNRNPAB, HNRNPC, HNRNPK, HNRNPM, HNRNPR, HSPA5, IGFBP2, ITGB8, ITM2A, ITM2B, JPT1, KDELR1, KLRK1-AS1, KRTCAP2, KTN1, LDHB, LHFPL3, LRRC4B, LY6H, MAP2, MARCKS, MARCKSL1 , MIA, MICOS10, MIF, MIR9-1HG, MMGT1, MPZL1, MT3, MTLN, MTRNR2L12, MTRNR2L8, MYL12A, MYL12B, NACA, NARS1, NCL, NDUFA1, NDUFA11, NDUFA13, NDUFA3,NDUFA4, NDUFB1, NDUFB11, NDUFB2, NDUFB6, NDUFB7, NDUFC2, NDUFS5, NEU4, NUCKS1, OAZ1, OLFM2, OSBPL8, OST4, OSTC, PABPC1 , PCBP2, PCDH10, PCDH11X, PCDH17, PCDHB2, PCDHGB6, PDGFRA, PDIA6, PEBP1, PEG10, PFN1, PGRMC1, PKIA, PLPP3, PLPPR1, PPIA , PRDX1, PRDX2, PRDX5, PSMB1, PSMB9, PTMS, PTN, PTPRA, RAB10, RAB14, RAB2A, RAB31, RAC1, RACK1, RMDN2, RAMP1, RO60, ROBO1 , RRAGB, RTN3, S100B, SARAF, SAT1, SBDS, SCARB2, SCP2, SCRG1, SEC62, SELENOK, SELENOT, SELENOW, SERF2, SERPINE2, SET, SH 3BGRL, SKP1, SLC25A6, SLIT2, SLITRK2, SMC3, SMDT1, SMOC1, SMS, SNCA, SNHG29, SNHG6, SNX3, SNX22, SOD1, SOX11, SOX2, SOX9 , SPCS2, SPCS3, SRP14, SSR4, STAG2, STMN1, SUPT16H, TALDO1, TBCB, TCEAL7, TCEAL8, TCEAL9, TIMP1, TLE5, TM4SF1, TM9SF3, T Selected from the group consisting of MA7, TMBIM6, TMCO1, TMEM147, TMEM258, TMEM50A, TMOD2, TMSB10, TMSB4X, TPT1, TRAF4, TRIO, TSC22D4, TSPAN6, TSPAN7, TTC3, TUBB, UBA52, UBL5, UQCR10, UQCR11, UQCRB, VIM, WSB2, WSCD1, YBX1, YWHAB, YWHAE, ZFAS1, ZNF428, and ZNF462.

[0117] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of APOD, B2M, BEX3, BEX5, CCND1, CTHRC1, EDIL3, EMC10, FABP7, GADD45A, ITM2A, LRRC4B, LY6H, MIA, MT3, NEU4, OLFM2, PTMS, RAMP1, SNX3, TRAF4, TRIO, UBA52, and YWHAB.

[0118] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of ANAPC11, APOD, ATP5MC3, B2M, CALM1, MT3, NEU4, PEBP1, RAMP1, SOD1, and TBCB.

[0119] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of APOD, BEX3, BEX5, CCND1, CTHRC1, EDIL3, EMC10, GADD45A, ITM2A, MIA, TRAF4, and TRIO.

[0120] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of B2M, FABP7, LRRC4B, LY6H, MT3, NEU4, OLFM2, PTMS, RAMP1, SNX3, UBA52, and YWHAB.

[0121] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of LY6H, MIA, GADD45A, ITM2A, and ITM2B.

[0122] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of HDAC2, EZH2, MYC, HMGA2, NFIB, and TEAD2.

[0123] In some embodiments, the method further comprises inhibiting expression of one or more anti-rejuvenation genes in the population of glial progenitor cells.

[0124] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ABCG1, ADGRB1, ADGRG1, AKAP9, AL360181.3, ANKRD10, ARGLU1, ARL4C, ARL16, ARMCX6, ATP1A2, ATP1B3, ATP10B, B3GNT7, BHLHE41, BPTF, BRI3, BX664615.2, BX890604.1, C1QL2, CAMK2N1, CCDC85B, CCNL1, CHCHD10, CHORDC1, CIRBP, CLDN10, COL9A1, COL9A2, CXADR, DAN CR, DCXR, DHX36, DLL3, DNAJA1, DNM3, ECH1, EGR1, EIF1AX, ELAVL3, EMID1, ETFB, FABP5, FAM133A, FAM133B, FBXO2, FERMT1, FIBIN, FOS, FOSB, FSCN1, F SIP2, GABPB1-AS1, GALR1, GNG8, GNPTAB, GOLGA8A, GOLGA8B, GPR155, GRID2, GRM7, HAPLN1, HMX1, HSPA1A, HSPA1B, HSPH1, HTRA1, IGFBP2, JAG1, JUN, JU NB, KCNIP4, KCNQ1OT1, KLF3-AS1, LAMP2, LINC01116, LINC01301, LINC01896, LRP4, LRRC7, MACF1, MALAT1, MAP3K13, MASP1, MDH1, MT1E, MT2A, MYT1, N ASP, NKTR, NUTM2A-AS1, OFD1, PCDHB5, PCDHGA3, PCDHGB6, PEPD, PHGDH, PLCG2, PMP2, PNISR, PPP1R14A, PTGDS, RAB3IP, RAF1, RAP1GAP, RARRES2, RBM25 , RBMX, REV3L, RHOBTB3, RIMS2, RIT2, RRBP1, RSRP1, S100A1, S100A16, SAT1, SCG2, SEMA3E, SERTAD1, SEZ6L, SEZ6L2, SH3GLB2, SNHG15, SNRNP70, SPARCL1, SRSF5, STAT3, STXBP6, SYNRG, THBS4, TLE4, TMEM176B, TPI1, TSC22D3, USP11, VCAN, WFDC1, WSB1, ZFYVE16, ZNF528, and ZNF528-AS1.

[0125] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ADGRG1 ARL4C, ARMCX6, ATP1A2, ATP1B3, B3GNT7, CXADR, DLL3, FABP5, FIBIN, IGFBP2, LRRC7, MAP3K13, MT1E, MT2A, PCDHGA3, PCDHGB6, PLCG2, PTGDS, SAT1, SEZ6L, SPARCL1, THBS4, and TLE4.

[0126] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ARGLU1, EGR1, FSIP2, HSPH1, MACF1, NKTR, RBMX, STAT3, TLE4, and WSB1.

[0127] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ADGRG1, ATP1A2, ATP1B3, B3GNT7, CXADR, DLL3, FABP5, MT1E, MT2A, PTGDS, SEZ6L, and THBS4.

[0128] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ARL4C, ARMCX6, FIBIN, IGFBP2, LRRC7, MAP3K13, PCDHGA3, PCDHGB6, PLCG2, SAT1, SPARCL1, and TLE4.

[0129] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ZNF274, MAX, E2F6, IKZF3, and STAT3.

[0130] In some embodiments, inhibiting expression of one or more rejuvenation inhibitor genes comprises administering to the population of glial progenitor cells a small molecule inhibitor of a rejuvenation inhibitor gene, hi some embodiments, the small molecule inhibitor is an AHR inhibitor selected from the group consisting of BAY-218, perillaldehyde StemRegenin 1 (SR1), KYN-101, CH-223191, BAY 2416964, PDM2, and GNF351.

[0131] In some embodiments, inhibiting expression of one or more anti-rejuvenation genes comprises administering to the population of glial progenitor cells an expression vector encoding one or more microRNAs. In some embodiments, the one or more microRNAs are selected from the group consisting of miR-193a-5P, miR-23b-3p, miR-4687-3p, miR-4651, miR-4270, and miR-24-3p - this group needs to be expanded.

[0132] In some embodiments, the step of inhibiting expression of one or more anti-rejuvenation genes comprises administering to the population of glial progenitor cells an expression vector encoding one or more shRNAs.

[0133] In some embodiments, the step of inhibiting expression of one or more anti-rejuvenation genes comprises administering to the population of glial progenitor cells one or more antisense oligonucleotides or an expression vector encoding one or more antisense oligonucleotides.

[0134] In some embodiments, inhibiting expression of one or more rejuvenation-inhibiting genes comprises administering to the population of glial progenitor cells an expression vector encoding a nuclease-based gene editing system. Examples of nuclease-based gene editing systems include, but are not limited to, a CRISPR-CAS system, a ZFN system, and a TALEN system.

[0135] III. Expression Vectors Another aspect of the present application relates to an expression vector as described herein.

[0136] In some embodiments, the expression vector comprises a nucleotide sequence encoding human BCL11A or a functional variant thereof. In some embodiments, the expression vector comprises a nucleotide sequence encoding human BCL11A. In some embodiments, the expression vector comprises a nucleotide sequence encoding SEQ ID NO:2.

[0137] In some embodiments, regulatory elements include ubiquitous promoters such as the chicken beta-actin (CBA) promoter, a hybrid form of the CBA promoter (CBh)CAG promoter, the cytomegalovirus (CMV) promoter, and the Rous sarcoma virus (RSV) promoter.

[0138] In some embodiments, the regulatory elements include promoters and / or enhancers of genes selectively expressed by glial progenitor cells, such as the promoters / enhancers of platelet-derived growth factor alpha (PDGFRA), zinc finger protein 488 (ZNF488), G protein-coupled receptor (GPR17), oligodendrocyte transcription factor 2 (OLIG2), chondroitin sulfate proteoglycan 4 (CSPG4), and SRY-box transcription factor 10 (SOX10).

[0139] In some embodiments, the regulatory element comprises an inducible promoter. In some embodiments, the inducible promoter is a tet-on or tetp-off promoter. In some embodiments, the inducible promoter comprises a cumate-controlled operator system, where transcription of the gene of interest is activated in the presence of cumate. In some embodiments, the expression vector of the present application comprises a cumate-controlled operator system, where transcription of the gene of interest is activated in the absence of cumate. In some embodiments, the inducible promoter comprises a rapamycin-controlled operator system, where transcription of the gene of interest is activated in the presence of rapamycin.

[0140] In some embodiments, the expression vector is capable of expressing an agent that inhibits the activity of an endogenous repressor of BCL11A activity in adult glial progenitor cells, including, but not limited to, KLF1, POGZ, HRI, Mi2β, SOX2, and FOXQ1.

[0141] In some embodiments, the expression vector is capable of expressing one or more rejuvenation-promoting genes in a population of glial progenitor cells.

[0142] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of ARX, CEBPZ, DLX1, DLX2, ELK1, ETS1, ETV4, KLF16, MYBL2, MYC, NFYB, POU3F1, SMAD1, SOX3, SP5, TCF12, TFDP1, TP53, ZIC3, and ZNF195.

[0143] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of CEBPZ, MYBL2, MYC, NFYB, and ZNF195.

[0144] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of ACTB, AKR1C1, ANAPC11, AP2B1, APLP2, APOD, ARF5, ARL4A, ARPC3, ARPP19, ATOX1, ATP5F1E, ATP5MC1, ATP5MC3, ATP5MD, ATP5ME, ATP5MF, ATP5MG, ATP5MPL, ATP5PF, ATP6V0B, ATP6V0E1, ATXN7L3B, B2M, B3GAT2, BEX1, BEX3, BEX5, BLOC1S1, BMERB1, C18orf32, C1orf122, C1QBP, C4orf48, CADM4, CALM1, CALM3, CALR, CANX, CAV2, CC2D1A, CCND1, CCNI, CD63, CD82, CDC42, CDH2, CFL1, CHCHD2, CHGB, CIAO2B, CLCN3, CLTA, CLT C, CNN3, CNTN1, COTL1, COX4I1, COX6A1, COX6C, COX7A2, COX7C, COX8A, CPNE8, CPS1, CRNDE, CSPG4, CTHRC1, CUL4B, CYP51A1, DBI, DCX, DDAH1, DDX1, DENN D10, DMD, DMRT2, DNAJA2, DPYSL2, DRAP1, DSTN, DYNC1I2, EDF1, EDIL3, EEF1A1, EEF1B2, EEF2, EID1, EIF3J, ELOB, EMC10, EMP2, ESD, ETV1, FABP7, FAM17 1B, FAM177A1, FAU, FIS1, FXYD6, GADD45A, GAP43, GCSH, GNAS, GOLM1, GPM6B, GSTP1, H3-3A, H3-3B, HINT1, HNRNPA1, HNRNPA3, HNRNPAB, HNRNPC, HNRNPK, HNRNPM, HNRNPR, HSPA5, IGFBP2, ITGB8, ITM2A, ITM2B, JPT1, KDELR1, KLRK1-AS1, KRTCAP2, KTN1, LDHB, LHFPL3, LRRC4B, LY6H, MAP2, MARCKS, MARCKSL1 , MIA, MICOS10, MIF, MIR9-1HG, MMGT1, MPZL1, MT3, MTLN, MTRNR2L12, MTRNR2L8, MYL12A, MYL12B, NACA, NARS1, NCL, NDUFA1, NDUFA11, NDUFA13, NDUFA3,NDUFA4, NDUFB1, NDUFB11, NDUFB2, NDUFB6, NDUFB7, NDUFC2, NDUFS5, NEU4, NUCKS1, OAZ1, OLFM2, OSBPL8, OST4, OSTC, PABPC1 , PCBP2, PCDH10, PCDH11X, PCDH17, PCDHB2, PCDHGB6, PDGFRA, PDIA6, PEBP1, PEG10, PFN1, PGRMC1, PKIA, PLPP3, PLPPR1, PPIA , PRDX1, PRDX2, PRDX5, PSMB1, PSMB9, PTMS, PTN, PTPRA, RAB10, RAB14, RAB2A, RAB31, RAC1, RACK1, RMDN2, RAMP1, RO60, ROBO1 , RRAGB, RTN3, S100B, SARAF, SAT1, SBDS, SCARB2, SCP2, SCRG1, SEC62, SELENOK, SELENOT, SELENOW, SERF2, SERPINE2, SET, SH 3BGRL, SKP1, SLC25A6, SLIT2, SLITRK2, SMC3, SMDT1, SMOC1, SMS, SNCA, SNHG29, SNHG6, SNX3, SNX22, SOD1, SOX11, SOX2, SOX9 , SPCS2, SPCS3, SRP14, SSR4, STAG2, STMN1, SUPT16H, TALDO1, TBCB, TCEAL7, TCEAL8, TCEAL9, TIMP1, TLE5, TM4SF1, TM9SF3, T Selected from the group consisting of MA7, TMBIM6, TMCO1, TMEM147, TMEM258, TMEM50A, TMOD2, TMSB10, TMSB4X, TPT1, TRAF4, TRIO, TSC22D4, TSPAN6, TSPAN7, TTC3, TUBB, UBA52, UBL5, UQCR10, UQCR11, UQCRB, VIM, WSB2, WSCD1, YBX1, YWHAB, YWHAE, ZFAS1, ZNF428, and ZNF462.

[0145] In some embodiments, the one or more additional rejuvenation-promoting genes are selected from the group consisting of APOD, B2M, BEX3, BEX5, CCND1, CTHRC1, EDIL3, EMC10, FABP7, GADD45A, ITM2A, LRRC4B, LY6H, MIA, MT3, NEU4, OLFM2, PTMS, RAMP1, SNX3, TRAF4, TRIO, UBA52, and YWHAB.

[0146] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of ANAPC11, APOD, ATP5MC3, B2M, CALM1, MT3, NEU4, PEBP1, RAMP1, SOD1, and TBCB.

[0147] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of APOD, BEX3, BEX5, CCND1, CTHRC1, EDIL3, EMC10, GADD45A, ITM2A, MIA, TRAF4, and TRIO.

[0148] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of B2M, FABP7, LRRC4B, LY6H, MT3, NEU4, OLFM2, PTMS, RAMP1, SNX3, UBA52, and YWHAB.

[0149] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of LY6H, MIA, GADD45A, ITM2A, and ITM2B.

[0150] In some embodiments, the one or more additional pro-rejuvenating genes are selected from the group consisting of HDAC2, EZH2, MYC, HMGA2, NFIB, and TEAD2.

[0151] In some embodiments, the expression vector is capable of expressing one or more repressors of rejuvenation suppressor genes in a population of glial progenitor cells.

[0152] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ABCG1, ADGRB1, ADGRG1, AKAP9, AL360181.3, ANKRD10, ARGLU1, ARL4C, ARL16, ARMCX6, ATP1A2, ATP1B3, ATP10B, B3GNT7, BHLHE41, BPTF, BRI3, BX664615.2, BX890604.1, C1QL2, CAMK2N1, CCDC85B, CCNL1, CHCHD10, CHORDC1, CIRBP, CLDN10, COL9A1, COL9A2, CXADR, DAN CR, DCXR, DHX36, DLL3, DNAJA1, DNM3, ECH1, EGR1, EIF1AX, ELAVL3, EMID1, ETFB, FABP5, FAM133A, FAM133B, FBXO2, FERMT1, FIBIN, FOS, FOSB, FSCN1, F SIP2, GABPB1-AS1, GALR1, GNG8, GNPTAB, GOLGA8A, GOLGA8B, GPR155, GRID2, GRM7, HAPLN1, HMX1, HSPA1A, HSPA1B, HSPH1, HTRA1, IGFBP2, JAG1, JUN, JU NB, KCNIP4, KCNQ1OT1, KLF3-AS1, LAMP2, LINC01116, LINC01301, LINC01896, LRP4, LRRC7, MACF1, MALAT1, MAP3K13, MASP1, MDH1, MT1E, MT2A, MYT1, N ASP, NKTR, NUTM2A-AS1, OFD1, PCDHB5, PCDHGA3, PCDHGB6, PEPD, PHGDH, PLCG2, PMP2, PNISR, PPP1R14A, PTGDS, RAB3IP, RAF1, RAP1GAP, RARRES2, RBM25 , RBMX, REV3L, RHOBTB3, RIMS2, RIT2, RRBP1, RSRP1, S100A1, S100A16, SAT1, SCG2, SEMA3E, SERTAD1, SEZ6L, SEZ6L2, SH3GLB2, SNHG15, SNRNP70, SPARCL1, SRSF5, STAT3, STXBP6, SYNRG, THBS4, TLE4, TMEM176B, TPI1, TSC22D3, USP11, VCAN, WFDC1, WSB1, ZFYVE16, ZNF528, and ZNF528-AS1.

[0153] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ADGRG1 ARL4C, ARMCX6, ATP1A2, ATP1B3, B3GNT7, CXADR, DLL3, FABP5, FIBIN, IGFBP2, LRRC7, MAP3K13, MT1E, MT2A, PCDHGA3, PCDHGB6, PLCG2, PTGDS, SAT1, SEZ6L, SPARCL1, THBS4, and TLE4. In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ARGLU1, EGR1, FSIP2, HSPH1, MACF1, NKTR, RBMX, STAT3, TLE4, and WSB1.

[0154] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ADGRG1, ATP1A2, ATP1B3, B3GNT7, CXADR, DLL3, FABP5, MT1E, MT2A, PTGDS, SEZ6L, and THBS4.

[0155] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ARL4C, ARMCX6, FIBIN, IGFBP2, LRRC7, MAP3K13, PCDHGA3, PCDHGB6, PLCG2, SAT1, SPARCL1, and TLE4.

[0156] In some embodiments, the one or more rejuvenation suppressor genes are selected from the group consisting of ZNF274, MAX, E2F6, IKZF3, and STAT3.

[0157] In some embodiments, the repressor comprises one or more microRNAs. In some embodiments, the one or more microRNAs are selected from the group consisting of miR-193a-5P, miR-23b-3p, miR-4687-3p, miR-4651, miR-4270, and miR-24-3p. - This group needs to be expanded.

[0158] In some embodiments, the repressor comprises one or more shRNAs.

[0159] In some embodiments, the repressor comprises one or more antisense oligonucleotides.

[0160] In some embodiments, the repressor comprises a nuclease-based gene editing system. Examples of nuclease-based gene editing systems include, but are not limited to, a CRISPR-CAS system, a ZFN system, and a TALEN system.

[0161] In some embodiments, the expression vector of the present application is a non-viral vector. In some embodiments, the non-viral vector is a plasmid. In some embodiments, the non-viral vector is a bacterial vector.

[0162] In some embodiments, the expression vector of the present application is a viral vector. In some embodiments, the viral vector is a lentiviral vector, an AAV vector, or a retroviral vector.

[0163] In some embodiments, the expression vector of the present application is a lentiviral vector.

[0164] In some embodiments, the expression vector of the present application is an AAV vector.

[0165] Another aspect of the present application relates to cells harboring the expression vectors of the present application. As used herein, the phrase "cells harboring an expression vector" refers to cells that contain the expression vector in either an integrated or non-integrated (epichromosomal) form.

[0166] A further aspect of the present disclosure is directed to a preparation of glial progenitor cells expressing a genetic construct according to the present disclosure.

[0167] The following examples are intended to illustrate the practice of embodiments of the present disclosure, but are not intended to limit its scope in any way. [Example]

[0168] Example 1: Materials and Methods hGPC production and aging Human embryonic stem cells (hESCs, line WA09 / H9) and induced pluripotent stem cells (iPSCs, line C27) were expanded in feeder-free conditions in mTeSR1 medium and then differentiated into hGPCs. To assess the effect of age on hGPCs in vitro, hGPCs maintained on mouse laminin were grown until 160 DIV and passaged every 3 weeks by manual dissection or until 90% confluency at 360 DIV. Cells were harvested for analysis at 0, 30, 60, 90, 180, and 360 DIV.

[0169] In vitro lentiviral production and infection This study identified BCL11A isoform 2 as the most abundant gene in isolated hESC-derived GPCs. Based on this finding, BCL11A isoform 2 (NCBI Reference Sequence: NM_018014.4) was cloned into a pTANK backbone and then cloned into T2A-EGFP, driven by the ubiquitously expressed CBh promoter. To generate an EGFP-expressing control virus, EGFP alone was cloned downstream of the CBh promoter. Viral particles were produced by transfection of 293HEK cells, concentrated, and titrated (LV-BCL11A: 3.83 x 10^9 PFU / mL, LV-EGFP: 4.50 x 10^9 PFU / mL). GPCs were transduced in vitro by adding either LV-BCL11A or LV-EGFP to the culture medium at an MOI of 2 for 24 hours. One week after infection, cells were fixed with 4% paraformaldehyde (PFA) or dissociated for downstream analysis and processing.

[0170] Etoposide treatment in hGPC At 160 DIV, hGPCs were treated with either 5 μM etoposide or dimethyl sulfoxide (DMSO) as a control, added to standard glial culture medium. Five days after treatment, etoposide and DMSO were removed by replacement with fresh medium, and cells in both conditions were treated with LV-BCL11A or LV-EGFP. One week after infection, cells were dissociated and processed for RT-QPCR as described herein.

[0171] Generation and analysis of glial chimeric mice To establish human glial chimeric mice, hGPCs were derived from iPSCs (line C27) tagged with a cassette expressing membrane-bound tRFP and intracellular mScarlet integrated into the AAVS1 locus, allowing visualization of human cells by fluorescence-associated cell sorting and histology. On postnatal day 1, Rag1 immunodeficient mice were engrafted into the corpus callosum (CC) with 300k cells. After engraftment, chimerized mice were aged for 2 years and then received stereotactic injections of LV-BCL11A virus in the left hemisphere and LV-EGFP control virus in the right hemisphere. Virus was deposited at 1µL per stop in the striatum, CC, and cortex. Three weeks after injection, mice were euthanized and either microdissected for cell sorting or perfused with 4% PFA for cryosectioning and immunohistochemistry.

[0172] Immunostaining and quantification After in vitro fixation, cell cultures were washed with PBS and incubated in staining buffer (0.3% Triton X-100 and 1% BSA in PBS). Primary antibodies against BCL11A, GFP, or MKI67 were added for 1 hour, and then the cells were washed again with PBS. Secondary antibodies and DAPI were added for 30 minutes, and then the cells were washed and stored in PBS containing thimerosal. Immunostained cultures were imaged with an ImageXpress High-Content Imaging System and quantified using MetaXpress software.

[0173] In vivo cryosections were mounted and stained for GFP, human nuclear antigen, MKI67, BCL11A, PDGFRA, or OLIG2 by overnight incubation at 4°C in permeabilization / blocking buffer (0.1% Triton X-100 and 10% normal goat serum in PBS). After rinsing with PBS, sections were incubated with secondary antibodies, rinsed, and mounted for imaging. Images were processed using ImageJ. For each mouse, MKI67+ or BCL11A+ cells were counted in CCs in two sections around the injection site, and the area of ​​the CCs was measured.

[0174] Cell sorting For RNA and chromatin analysis in bulk or single cells, cells were isolated from the cultures by fluorescence-linked cell sorting. Briefly, hGPC cultures were dissociated into single cells using 50% Accutase in DPBS, then filtered and stained with DAPI to distinguish cells that were no longer intact. EGFP+ / DAPI- cells were collected in either a minimal volume of PBS for single-cell RNA-seq, lysis buffer for bulk RNA extraction, or nuclear isolation buffer for CUT&Tag (Cleavage Under Targets and Tagmentation).

[0175] Glial chimeric mice. Cells were isolated from chimerized mice by microdissection of the injection tract and dissociation in papain for 45 minutes at 37°C, followed by perfusion with cold HBSS and gentle trituration midway through the incubation period. Papain was quenched with ovomucoid, and the dissociated tissue was then filtered to select for RFP+ / DAPI-infected or GFP+ / DAPI-LV-infected human cells. Viability was confirmed by trypan blue in a hemocytometer before proceeding to single-cell RNA sequencing.

[0176] Quantitative real-time PCR and RNA sequencing RNA was extracted from either untreated hGPC cultures or GFP+ cells using the Qiagen RNeasy Micro kit. For Q-RT PCR, RNA was reverse transcribed and then mixed with SYBR Green master mix and primers targeting BCL11A, MKI67, CDKN1A, CDKN2A, IL1A, or IL8. For RNA-seq, total RNA was prepared using the Illumina TruSeq kit.

[0177] Single-cell RNA sequencing and ATAC-seq GFP+ cells sorted from GPC cultures or dissociated tissues were counted and then processed using the 10X Genomics Chromium v3.1 Single Cell 3' workflow according to the manufacturer's instructions. Single-cell ATAC sequencing was completed using the 10X Genomics Chromium 3'v1.1 Single Cell ATAC kit.

[0178] CUT&Tag To further characterize the effect of BCL11A overexpression on chromatin state, we performed CUT&Tag on LV-transduced hGPCs using antibodies targeting H3K4me3, H3K27me3, and H3K9me3 according to the EpiCypher CUTANA CUT&Tag protocol. Nuclei were isolated, adsorbed to concanavalin A magnetic beads, and chromatin was tagged with pAG-Tn5.

[0179] Bioinformatics analysis Bulk RNA-seq data were aligned using STAR and imported into R for analysis using DESeq2. Network analysis was performed using Qiagen Ingenuity Pathway Analysis. Single-cell RNA-seq and ATAC-seq data were processed with a custom STAR-based aligner to distinguish human from mouse cells and analyzed using Seurat and Signac in R. Fetal and adult gene signature enrichment scores were calculated using AUCell. CUT&Tag data were aligned using bowtie2 and processed with samtool and bedtool. Peaks were called using SEACR and annotated with ChIPseeker. Differential peak enrichment was modeled using DESeq2, and tracks were visualized using the Interactive Genome Viewer (IGV).

[0180] Example 2: BCL11A expression corresponds to hGPC proliferation In this study, we first investigated whether BCL11A is related to hGPC mitotic capacity and self-renewal. To that end, we evaluated BCL11A expression in hGPCs by RT-qPCR during both hGPC differentiation and subsequent in vitro aging. BCL11A expression was low during neural induction, increased during glial specification, peaked in juvenile hGPCs at 100 DIV, and declined to much lower levels at 360 DIV (Figure 1, panel A). The change in BCL11A expression was accompanied by a decrease in cell cycling between 100 and 300 DIV, as assessed by MKI67 ICC (Figure 1, panel B).

[0181] Next, we generated lentiviruses co-expressing BCL11A and EGFP (LV-BCL11A) along with an EGFP-only control virus (LV-EGFP) to identify infected cells (Figure 1, panel C). One week after infection, hGPCs transduced with LV-BCL11A showed a significant increase in MKI67+ / GFP+ cells compared to the LV-EGFP control (fold change = 2.86 ± 0.61, paired t-test *P = 0.015, N = 3). This was accompanied by a significant increase in cell number in BCL11A / GFP-treated cultures compared to their LV-EGFP-treated counterparts (fold change relative to LV-EGFP = 31.4 ± 2.8, **P = 0.008 by t-test, N = 3). Interestingly, untransduced GFP- cells in these cultures that received LV-BCL11A also showed a significant increase in MKI67+ cells (log2 fold change relative to LV-EGFP = 2.15 ± 0.42, paired t-test *P = 0.017), suggesting that BCL11A overexpression may promote proliferation through paracrine as well as intracellular mechanisms (Figure 1, panel D).

[0182] To further investigate the relationship between self-renewal and BCL11A expression, we treated hGPCs with etoposide, a DNA-damaging agent that halts cell division and induces senescence marks. Following etoposide treatment, we observed a decrease in BCL11A expression (fold change = 0.30 ± 0.14) compared to DMSO-treated controls. Concomitant with the decrease in BCL11A expression, MKI67 significantly decreased (fold change = 0.35 ± 0.04, *P = 0.04 by one-sample t-test, N = 2), and p21 significantly increased (fold change = 2.14 ± 0.05, *P = 0.026, t-test), while other markers of senescence, including p16, IL1A, and IL8, showed an upward trend. Treatment with LV-BCL11A restored BCL11A expression to control levels but was not sufficient to restore MKI67 expression. However, there was a significant decrease in p21 expression relative to LV-EGFP (paired t-test, ***P<0.001), along with a downward trend in all of p16, IL1A, and IL8, suggesting a protective effect against etoposide-induced senescence (Figure 1, Panel E).

[0183] Example 3: BCL11A overexpression induces a youth-associated gene expression signature RNA sequencing from isolated LV-BCL11A- or LV-EGFP-treated hGPCs revealed 985 differentially expressed genes in BCL11A-overexpressing samples and 690 genes that were lower in BCL11A-infected hGPCs compared with GFP-treated controls. Upregulated genes included markers of GPC identity, such as PDGFRA and ASCL1, regulators of cell-cell communication, such as NOTCH2, and genes with roles in proliferation and migration, including TOP2A and TEAD2 (Figure 2, panels A and C). In contrast, genes higher in control cultures included markers of mature cell identity, such as MYT1L and BIN1 (Figure 2, panels A and C). We next investigated whether genes upregulated after BCL11A overexpression comprised a juvenile-associated expression signature. This study utilized a generated dataset comparing the transcriptomes of hGPCs isolated from primary fetal or adult tissues to assess the overlap between genes induced by BCL11A and genes differentially expressed in fetal samples. Of the 985 genes upregulated in hGPCs overexpressing BCL11A, 270 (27.4%) were upregulated in fetal samples, as opposed to 46 (4.7%) in adult samples (Figure 2, Panel B).

[0184] To define the network and upstream effectors responsible for inducing a juvenile-like transcriptional state, this study used Ingenuity Pathway Analysis. Regulators predicted to have significant upstream activity in BCL11A-overexpressing cells included YAP1, MYC, EZH2, CTNNB1, and TEAD2. Senescence and apoptosis markers CDKN1A and TP53, along with E2F6 and IKZF3, genes identified as drivers of the adult hGPC phenotype, were among those identified as repressed after BCL11A activation (Figure 2, Panel D). The most highly activated pathways were those related to migration, invasion, and oligodendrocyte identity, whereas pathways related to apoptosis, neuronal identity, and senescence were repressed (Figure 2, Panel E).

[0185] Example 4: Single-cell RNA sequencing reveals a shift to younger cell identity after BCL11A activation Next, using single-cell RNA sequencing, we investigated the effect of BCL11A overexpression on hGPC identity in vitro. After filtering out low-quality cells, we recovered 11,512 LV-BCL11A cells and 12,700 LV-GFP cells in 19 clusters (Figure 3, Panel A). The cultures contained multiple GPC subtypes: Clusters 3, 9, and 10 were PDGFRA / ASCL1-expressing GPCs; clusters 5 and 15 were proliferative GPCs; cluster 16 was radial glia-like GPCs marked with VIM and HOPX; cluster 6 was pro-astrocytic GPCs; cluster 16 was radial glia-like cells; clusters 0, 1, 4, 7, 8, 10, 11, 12, and 17 were pro-neuronal GPCs; clusters 2, 13, and 14 were TAC1-positive neural precursors; and cluster 18 was OLIG2-high cells (Figure 3, Panel B). Glial markers, specifically markers of radial glial identity, were among the most enriched genes in BCL11A-treated cells, whereas genes associated with neuronal fate were relatively reduced (Figure 3, Panel C). Next, we compared BCL11A overexpression with fetal and adult hGPC identity by generating fetal and adult enrichment scores using AUCell based on differentially expressed genes between fetal and adult hGPCs (Figure 3, Panel D). Clusters 5 and 15 were most enriched for fetal signatures, while cluster 13 was most enriched for adult identity (Figure 3, Panel E). Overall, BCL11A overexpression appeared to induce a shift toward fetal and away from adult identity (Figure 3, Panel F). LV-BCL11A-treated cells comprised the majority (63%) of the most fetal-enriched cluster and a minority (16%) of the most adult-enriched cluster (Figure 3, Panel G).

[0186] Example 5: In vivo migration of aged hGPCs in gliotic chimeric mice Based on these data, we next investigated whether BCL11A could reinitiate or accelerate self-renewal in aging GPCs. To this end, mice were engrafted with RFP+ iPSC-derived hGPCs on postnatal day 1 and then aged for 2 years (Figure 4, Panel A). We then treated these 2-year-old chimeric mice with LV-BCL11A or LV-EGFP by injection into the CC. Three weeks later, mice were harvested for single-cell transcriptomics or histological analysis (Figure 4, Panel B). Immunostaining confirmed significant overexpression of BCL11A in the LV-BCL11A-treated hemisphere compared with the LV-EGFP-treated controls (LV-BCL11A cells / µm² = 4.56e-5 ± 8.23e-6, LV-EGFP cells / µm² = 5.06e-6 ± 1.45e-6, *P = 0.031 by paired t-test, N = 3). This was associated with a trend toward more MKI67+ cells in the CC of the BCL11A-injected hemisphere (LV-BCL11A cells / µm² = 1.6e-5 ± 4.06e-6, LV-EGFP cells / µm² = 6.63e-6 ± 1.26e-6, P = 0.079 by paired t-test, N = 3) (Figure 4, Panel C). Donor cell distribution was widespread and relatively uniform, with no tumors or heterotopia. This study also observed a relative increase in RFP, OLIG2+, and PDGFRa+ cells in the LV-BCL11A-treated hemisphere, suggesting that BCL11A transduction activates aging GPCs, resuming mitotic expansion and migratory colonization of the host brain. Furthermore, membrane tagging of human donor cells allowed us to define the majority of their morphology within the white matter as myelinating oligodendrocytes, suggesting at least a partial reversal of the typical age-related loss of myelin in these aging mice (Figure 4, panels D and E). Human cells co-colonized with MBP in the LV-BCL11A-treated CC and striatum, indicating that the migrating cells were capable of producing myelin.This effect was still present at 6 weeks postinfection (Fig. 4 , panels F and G), with more RFP + and OLIG2 + cells detected in the LV-BCL11A-infected hemisphere than in the LV-EGFP-treated control hemisphere.

[0187] Example 6: BCL11A upregulates genes associated with migration and proliferation in vivo This study isolated RFP+ / GFP+ cells from 2-year-old chimerized mice treated with LV-BCL11A or LV-EGFP, and then performed scRNA-seq to identify BCL11A-associated changes in gene expression and phenotype in vivo. In total, this study captured 2,260 human cells from the BCL11A-treated hemisphere and 2,696 human cells from the GFP-treated control (Figure 5, Panel A). These included AQP4+ astrocytes, PDGFRA+ GPCs, late GPR17+ GPCs maturing along the oligodendrocyte lineage, and a cluster of more mature MBP-defined oligodendrocytes (Figure 5, Panel B). Notably, this study noticed a cluster of early-matured GPCs differentially present in the BCL11A condition (Figure 5, Panel A, bottom). This cluster was further investigated and compared with the most similar group of cells from the GFP control condition. Compared to their GFP-treated counterparts, BCL11A-treated clusters were enriched for glial progenitor genes (PDGFRA, ASCL1) and genes involved in maintaining stemness, migration, and cell-cell signaling, including HES5 and CTNNB1. The most significantly upregulated genes were markers of astrocytes and radial glia (FABP7, VIM, HOPX), suggesting a less differentiated hGPC identity. In contrast, control cells were enriched for markers of more mature glial (MOG, MYRF) and occasional neuronal (DSCAML1) differentiation (Figure 5, Panel C).

[0188] Example 7: BCL11A expression generates a permissive chromatin state in juvenile-associated genes We next sought to investigate how BCL11A expression regulates chromatin states to induce a migratory, proliferative state in hGPCs. We performed scATAC-seq on 15,801 cells recovered from LV-GFP-infected cultures and 11,465 cells recovered from LV-BCL11A-transduced cultures (Figure 5, panel D). Following BCL11A overexpression, we observed increased accessibility at loci including HMGA2 and NFIB, two targets identified as drivers of fetal hGPC identity. GFP cultures also showed increased accessibility at genes associated with pro-neuronal pathways, including ADGRL2 and GAD1 (Figure 5, panel E). To understand the genome-wide effects of BCL11A activation, we also assayed the activating chromatin mark H3K4me3 and the repressive marks K27me3 and K9me3. BCL11A-treated cells showed a relative enrichment in K4me3 and a relative decrease in K9me3, suggesting a genome-wide net enhancement of transcription (Figure 5, panel F). Interestingly, OLIG1, a key determinant of glial lineage progression, was significantly enriched in both K4me3 and K27me3 after BCL11A overexpression, indicating possible bivalent regulation of its expression (Figure 5, panel G). Genes identified as differentially upregulated after BCL11A overexpression also showed remodeling in their chromatin state, with a loss of repressive marks around CTNNB1, TEAD2, and PIEZO1, accompanied by a concomitant increase in K4me3 signal.

[0189] Taken together, these data indicate that BCL11A overexpression results in a permissive chromatin state around essential genes for cell-cell communication, migration, and stem cell maintenance, including drivers of Wnt / β-catenin and YAP / TAZ signaling. Given the transcriptional upregulation of components and downstream targets of these pathways in vivo and in vitro, their activation underlies the observed migration of hGPCs after BCL11A expression, shifting the cells toward a fetal-like state. [Table 2] TIFF2025536331000004.tif249170

[0190] While various embodiments have been described above, it should be understood that such disclosure is presented by way of example only, and not limitation. Thus, the breadth and scope of the subject compositions and methods should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0191] The foregoing description is intended to teach those skilled in the art how to carry out the present invention and does not intend to describe in detail all obvious modifications and variations thereof that will become apparent to those skilled in the art upon reading the description. However, all such obvious modifications and variations are intended to be included within the scope of the present invention as defined by the following claims. The claims are intended to cover the elements and steps in any order that is effective to fulfill the objectives intended therein, unless the context specifically dictates otherwise.

Claims

1. 1. A method of inducing rejuvenation in a population of adult glial progenitor cells, said method comprising: The method comprises administering to the population of adult glial progenitor cells an effective amount of an expression vector comprising a nucleotide sequence encoding B-cell lymphoma / leukemia 11A (BCL11A) and a regulatory element operably linked to the nucleotide sequence.

2. 1. A method of treating a subject having a glial cell-associated disorder, comprising: The method comprises administering to a population of adult glial progenitor cells of the subject an effective amount of an expression vector comprising a nucleotide sequence encoding B-cell lymphoma / leukemia 11A (BCL11A) and a regulatory element operably linked to the nucleotide sequence.

3. 3. The method of claim 2, wherein the glial cell-related disorder is a dysmyelination selected from the group consisting of multiple sclerosis, neuromyelitis optica, transverse myelitis, optic neuritis, subcortical infarction, diabetic leukoencephalopathy, hypertensive leukoencephalopathy, age-related white matter disease, spinal cord injury, radiation or chemotherapy-induced demyelination, post-infectious and post-vaccination leukoencephalitis, periventricular leukomalacia, childhood leukodystrophy, lysosomal storage disease, congenital dysmyelination, inflammatory demyelination, vascular demyelination, and cerebral palsy.

4. 3. The method of claim 2, wherein the glial cell-associated disorder is a neurodegenerative disease selected from the group consisting of Huntington's disease, frontotemporal dementia, Parkinson's disease, multiple system atrophy, and amyotrophic lateral sclerosis.

5. 5. The method of claim 4, wherein the glial cell-associated disorder is Huntington's disease.

6. 3. The method of claim 2, wherein the subject is a human and the glial cell-associated disorder is a neuropsychiatric disorder selected from the group consisting of schizophrenia, autism spectrum disorder, and bipolar disorder.

7. The method of any one of claims 1 to 6, wherein the nucleotide sequence encoding BCL11A comprises the nucleotide sequence encoding human BCL11A of SEQ ID NO: 2, or a functional variant thereof.

8. The method according to any one of claims 1 to 7, wherein the expression vector is a non-viral expression vector.

9. The method of any one of claims 1 to 7, wherein the agent is a viral expression vector.

10. 10. The method of claim 9, wherein the viral expression vector is a lentiviral vector.

11. 10. The method of claim 9, wherein the viral expression vector is an AAV vector.

12. The method of any one of claims 1 to 11, wherein the regulatory element is a glial cell-specific promoter.

13. The method of any one of claims 1 to 11, wherein the regulatory element is an inducible promoter.

14. The method of claim 13, wherein the inducible promoter is a tet-on or tet-off promoter.

15. 15. The method of any one of claims 1 to 14, further comprising expressing in the population of adult glial progenitor cells one or more genes selected from the group consisting of histone deacetylase 2 (HDAC2), histone-lysine N-methyltransferase EZH2 (EZH2), myc proto-oncogene protein (MYC), high mobility group protein HMGI-C (HMGA2), nuclear factor type 1B (NFIB), and transcriptional enhancer factor TEF-410 (TEAD2).

16. 16. The method of any one of claims 1 to 15, further comprising expressing in the population of adult glial progenitor cells one or more glial cell-specific genes selected from the group consisting of PDGFRA, ZNF488, GPR17, OLIG2, CSPG4, and SOX10.

17. An expression vector comprising: a nucleic acid sequence encoding B-cell lymphoma / leukemia 11A (BCL11A); a regulatory element operably linked to the nucleic acid sequence, The expression vector is a lentiviral vector or an AAV vector.

18. 18. The expression vector of claim 17, wherein the regulatory element comprises a glial progenitor cell-specific promoter.

19. 18. The expression vector of claim 17, wherein the regulatory element comprises an inducible promoter.

20. The expression vector of claim 19, wherein the nucleotide sequence encoding BCL11A comprises a nucleotide sequence encoding human BCL11A of SEQ ID NO: 2, or a functional variant thereof.

21. An adult human glial precursor cell carrying the expression vector according to any one of claims 17 to 20.

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