Compositions and methods for treating neurocognitive disorders

Abstract

To provide methods for treating a subject having or at risk of developing a neurocognitive disorder, such as frontotemporal lobar degeneration or neuronal ceroid lipofuscinosis.SOLUTION: Provided is a method comprising administering, to a subject diagnosed as having a neurocognitive disorder (NCD), a composition comprising a population of cells comprising a transgene encoding a progranulin (PGRN) or a granulin (GRN).SELECTED DRAWING: Figure 1A

Description

[Technical field]

[0001] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. Said ASCII copy, created on January 29, 2020, is titled "51182-019WO2_Sequence_Listing_1.29.20_ST25" and is 22,275 bytes in size.

[0002] FIELD OF THEINVENTION The present disclosure relates to compositions and methods for treating neurocognitive disorders such as frontotemporal lobar degeneration and neuronal ceroid lipofuscinosis. [Background technology]

[0003] Neurodegeneration is a pathophysiological process observed in several diseases associated with progressive dementia, such as frontotemporal lobar degeneration and neuronal ceroid lipofuscinosis. A key feature of this process is the massive destruction of brain tissue and the accompanying neurodegeneration and neuronal death that causes a whole range of behavioral deficits, including, inter alia, cognitive decline, language impairment.

[0004] Frontotemporal lobar degeneration (FTLD) is a neurodegenerative disorder characterized by a complex clinical symptomatology that may include deficits in speech comprehension and production, poor motor planning and coordination, and / or loss of executive function characterized by lack of impulse control and a predominance of peremptory behavior. Clinical descriptions of FTLD are divided into three distinct variants: 1) behavioral-frontotemporal dementia characterized by prominent changes in behavior, personality, and prominent degeneration of the frontal lobe; 2) semantic dementia characterized by an insidious breakdown of language and prominent degeneration of the anterior temporal lobe; and 3) progressive incompetent aphasia characterized by speech and grammatical defects and corresponding degeneration of the left perisylvian cortex. Histological analysis of postmortem brain tissue from FTLD patients shows a complex and heterogeneous neuropathological profile with common indications of degeneration of neural tissue in the frontal and temporal lobes of the brain.

[0005] Neuronal ceroid lipofuscinosis (NCL) is a collective term for a clinically recognized collection of at least eight lysosomal storage disorders resulting from the accumulation of lipofuscin in cells of the body, such as neurons, liver, spleen, myocardium, and kidney cells. Lipofuscin is a lipopigment composed of fat and protein. Patients with NCL show significant neurodegeneration and progressive, irreversible loss of motor and cognitive abilities, although disease severity and clinical symptoms may depend on the specific NCL variant. Known variants of NCL include the juvenile variant also known as Santovuori-Haltia disease (SHD), the late-onset juvenile variant also known as Jansky-Bielschowsky disease (JBD), the Finnish late-onset juvenile variant (FLI), the variant with variant late (VLI), the CLN7 variant (CLN7), the CLN8 variant (CLN8), the Turkish late-onset juvenile variant (TLI), the juvenile variant also known as Batten disease (BD), and the adult variant also known as Koufs disease (KD). SHD is associated with early visual loss that progresses to complete retinal blindness by age 2, followed by a vegetative state at age 3, and brain death by age 4. This variant is also associated with spontaneous occurrence of epileptic seizures. The JBD variant occurs between 2 and 4 years of age and is associated with ataxia, seizures, progressive cognitive decline, and abnormal speech development, typically resulting in death by age 8. BD typically occurs between 4 and 10 years of age and includes symptoms such as vision loss, seizures, cognitive impairment, and early death. NCL patients with the KD variant generally have milder symptoms than the SHD and BD variants and have a life expectancy of approximately 40 years.

[0006] Existing treatments for FTLD and NCL seek to ameliorate disease symptoms, and there are no therapies that target the underlying neurodegeneration, thus highlighting the need for new therapeutic avenues. Summary of the Invention

[0007] The present disclosure provides a method for treating a neurocognitive disorder (NCD), such as frontotemporal lobar degeneration (FTLD) or neuronal ceroid lipofuscinosis (NCL), by administering cells, such as pluripotent cells (e.g., embryonic stem cells (ESCs) or induced pluripotent stem cells (ISPCs)), multipotent cells (e.g., CD34+ cells, such as hematopoietic stem cells (HSCs) or myeloid progenitor cells (MPCs)), blood lineage progenitor cells (BLPCS; e.g., monocytes), macrophages, microglial progenitor cells, or microglia, that comprise a transgene encoding progranulin (PGRN) or granulin (GRN). The cells can be administered to a subject with an NCD by one or more of a variety of routes, including directly (e.g., by intraventricular injection) or systemically (e.g., by intravenous administration) into the subject's central nervous system, among others. The disclosure also features compositions containing such cells, as well as kits containing these cells for treating NCDs.

[0008] In a first aspect, the present disclosure provides a method for treating a subject diagnosed with NCD (e.g., FTLD or NCL) by administering to the subject a composition comprising a population of cells that comprises a transgene encoding PGRN or GRN. In some embodiments, the transgene encoding PGRN or GRN can be expressed in macrophages or microglial cells. In some embodiments, the NCD is a severe NCD. In some embodiments, a severe NCD interferes with a subject's independence and / or normal daily functioning (e.g., social, occupational, or academic functioning, personal hygiene, grooming, dressing, toilet hygiene, functional mobility (e.g., ability to walk, get in and out of bed), and self-care. In some embodiments, a severe NCD is associated with a score obtained by the subject on a cognitive test that is at least 2 standard deviations away from the mean score of a reference population. In some embodiments, the NCD is a mild NCD. In some embodiments, a mild NCD does not interfere with a subject's independence and / or normal daily functioning. In some embodiments, a mild NCD is associated with a score obtained by the subject on a cognitive test that is at least 1-2 standard deviations away from the mean score of a reference population. In some embodiments, the cognitive test is an Eight-item Informant Interview to Differentiate Aging and Dementia (AD8), Annual Wellness Visit (AWV), General Practitioner Assessment of Cognition (GPCOG), Health Risk Assessment (HRA), Memory Impairment Screen (MIS), Mini Mental Status Exam (MMSE), Montreal Cognitive Assessment (MoCA), St. Louis University Mental Status Exam (SLUMS), and Short Informant Questionnaire on Cognitive Decline in the Elderly (Short IQCODE).In some embodiments, the NCD is associated with impairment in one or more of complex attention, executive function, learning and memory, language, sensorimotor function, and social cognition. In some embodiments, the NCD is not due to delirium or other psychiatric disorders (e.g., schizophrenia, bipolar disorder, or major depression). In some embodiments, the reference population is the general population. In some embodiments, the reference population is selected based on the subject's age, medical history, education, socioeconomic status, and lifestyle. In some embodiments, the NCD is a frontotemporal NCD. In some embodiments, the frontotemporal NCD is FTLD. In some embodiments, the NCD is due to a lysosomal disease. In some embodiments, the lysosomal disease is NCL.

[0009] In some embodiments, the PGRN is a full-length PGRN, e.g., a PGRN having the amino acid sequence of SEQ ID NO:1, or a variant thereof having at least 85% sequence identity thereto (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:1). In some embodiments, the PGRN comprises at least two (e.g., at least 2, 3, 4, 5, 6, 7, 8 or more) GRN domains having an amino acid sequence of any one of SEQ ID NOs: 2-9 or a variant thereof having at least 85% sequence identity thereto (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto). In some embodiments, the PGRN comprises at least two (e.g., 2, 3, 4, 5, 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least three (e.g., at least 3, 4, 5, 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least four (e.g., at least 4, 5, 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least five (e.g., at least 5, 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least six (e.g., at least 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least seven (e.g., at least 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least eight (e.g., at least 8 or more) GRN domains. In some embodiments, the PGRN comprises 2-16 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16) GRN domains. In some embodiments, the PGRN comprises 2-12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) GRN domains.In some embodiments, the PGRN comprises 2-8 (e.g., 2, 3, 4, 5, 6, 7, or 8) GRN domains. In some embodiments, the PGRN comprises 2-4 (e.g., 2, 3, or 4) GRN domains. In some embodiments, the PGRN comprises 2 GRN domains.

[0010] In some embodiments, the PGRN comprises a para-GRN domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the para-GRN domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the para-GRN domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the para-GRN domain has an amino acid sequence of SEQ ID NO:2.

[0011] In some embodiments, the PGRN comprises a GRN-1 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:3. In some embodiments, the GRN-1 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:3. In some embodiments, the GRN-1 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:3. In some embodiments, the GRN-1 domain has an amino acid sequence of SEQ ID NO:3.

[0012] In some embodiments, the PGRN comprises a GRN-2 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:4. In some embodiments, the GRN-2 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:4. In some embodiments, the GRN-2 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:4. In some embodiments, the GRN-2 domain has an amino acid sequence of SEQ ID NO:4.

[0013] In some embodiments, the PGRN comprises a GRN-3 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:5. In some embodiments, the GRN-3 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:5. In some embodiments, the GRN-3 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:5. In some embodiments, the GRN-3 domain has an amino acid sequence of SEQ ID NO:5.

[0014] In some embodiments, the PGRN comprises a GRN-4 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:6. In some embodiments, the GRN-4 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:6. In some embodiments, the GRN-4 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:6. In some embodiments, the GRN-4 domain has an amino acid sequence of SEQ ID NO:6.

[0015] In some embodiments, the PGRN comprises a GRN-5 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:7. In some embodiments, the GRN-5 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:7. In some embodiments, the GRN-5 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:7. In some embodiments, the GRN-5 domain has an amino acid sequence of SEQ ID NO:7.

[0016] In some embodiments, the PGRN comprises a GRN-6 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:8. In some embodiments, the GRN-6 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:8. In some embodiments, the GRN-6 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:8. In some embodiments, the GRN-6 domain has an amino acid sequence of SEQ ID NO:8.

[0017] In some embodiments, the PGRN comprises a GRN-7 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:9. In some embodiments, the GRN-7 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:9. In some embodiments, the GRN-7 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:9. In some embodiments, the GRN-7 domain has an amino acid sequence of SEQ ID NO:9.

[0018] In some embodiments, the GRN is a full-length GRN, such as a GRN having any one of the amino acid sequences of SEQ ID NOs: 2-9, or a variant thereof having at least 85% sequence identity thereto (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto). In some embodiments, the GRN is a para-GRN or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity thereto (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity thereto). In some embodiments, the para-GRN has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 2. In some embodiments, the para-GRN has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 2. In some embodiments, the para-GRN has the amino acid sequence of SEQ ID NO: 2.

[0019] In some embodiments, GRN is GRN-1 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:3. In some embodiments, GRN-1 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:3. In some embodiments, GRN-1 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:3. In some embodiments, GRN-1 has the amino acid sequence of SEQ ID NO:3.

[0020] In some embodiments, GRN is GRN-2 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:4. In some embodiments, GRN-2 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:4. In some embodiments, GRN-2 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:4. In some embodiments, GRN-2 has the amino acid sequence of SEQ ID NO:4.

[0021] In some embodiments, the GRN is GRN-3 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:5. In some embodiments, the GRN-3 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:5. In some embodiments, the GRN-3 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:5. In some embodiments, the GRN-3 has the amino acid sequence of SEQ ID NO:5.

[0022] In some embodiments, the GRN is GRN-4 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:6. In some embodiments, the GRN-4 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:6. In some embodiments, the GRN-4 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:6. In some embodiments, the GRN-4 has the amino acid sequence of SEQ ID NO:6.

[0023] In some embodiments, the GRN is GRN-5 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:7. In some embodiments, the GRN-5 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:7. In some embodiments, the GRN-5 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:7. In some embodiments, the GRN-5 has the amino acid sequence of SEQ ID NO:7.

[0024] In some embodiments, the GRN is GRN-6 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:8. In some embodiments, the GRN-6 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:8. In some embodiments, the GRN-6 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:8. In some embodiments, the GRN-6 has the amino acid sequence of SEQ ID NO:8.

[0025] In some embodiments, the GRN is GRN-7 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:9. In some embodiments, the GRN-7 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:9. In some embodiments, the GRN-7 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:9. In some embodiments, the GRN-7 has the amino acid sequence of SEQ ID NO:9.

[0026] In some embodiments, the order of the GRN domains in the PGRN polypeptide occurs in the same order as observed in wild-type human PGRN. In some embodiments, the order of the GRN domains in the PGRN polypeptide occurs in a different order than that found in wild-type human PGRN.

[0027] In some embodiments, the transgene encoding PGRN or GRN is codon-optimized. In some embodiments, the codon-optimized transgene encoding PGRN or GRN comprises a polynucleotide having at least 85% sequence identity (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 19. In some embodiments, the transgene encoding PGRN or GRN comprises a secretory signal peptide (e.g., a PGRN secretory signal peptide).

[0028] In some embodiments, PGRN or GRN is a PGRN or GRN fusion protein. In some embodiments, PGRN or GRN fusion protein comprises the low density lipoprotein receptor family (LDLRf) binding (Rb) domain of apolipoprotein E (ApoE), or a fragment, variant, or oligomer thereof. In some embodiments, the Rb domain of ApoE, or a fragment, variant, or oligomer thereof, is operably linked to the N-terminus of PGRN or GRN. In some embodiments, the Rb domain of ApoE, or a fragment, variant, or oligomer thereof, is operably linked to the C-terminus of PGRN or GRN. In some embodiments, PGRN or GRN fusion protein comprises at least one (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) oligomer of the Rb domain of ApoE. In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 25-185 of SEQ ID NO: 11. In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 50-180 of SEQ ID NO: 11. In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 75-175 of SEQ ID NO: 11. In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 100-170 of SEQ ID NO: 11.In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 125-160 of SEQ ID NO: 11. In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 130-150 of SEQ ID NO: 11. In some embodiments, the Rb domain comprises a region of ApoE having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 148-173, or a portion thereof including residues 159-167 of SEQ ID NO:11, or a variant thereof having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to residues 159-167 of SEQ ID NO:11. In some embodiments, the Rb domain comprises a region having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of residues 159-167 of SEQ ID NO:11.

[0029] In some embodiments, the PGRN or GRN fusion protein comprises PGRN or GRN and a glycosylation-independent lysosomal targeting (GILT) tag. In some embodiments, the GILT tag is operably linked to the N-terminus of PGRN or GRN. In some embodiments, the GILT tag is operably linked to the C-terminus of PGRN or GRN. In some embodiments, the GILT tag comprises a human IGF-II mutein having an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II (SEQ ID NO: 12). The mutein may have reduced binding affinity for the insulin receptor compared to the affinity of naturally occurring human IGF-II for the insulin receptor and / or may be resistant to furin cleavage. The mutein may bind to the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner. In some embodiments, the IGF-II mutein comprises a mutation within a region corresponding to amino acids 30-40 of SEQ ID NO: 12, wherein the mutation abolishes at least one furin protease cleavage site. In some embodiments, the mutation is an amino acid substitution, deletion, and / or insertion. In some embodiments, the mutation is a Lys or Ala amino acid substitution at a position corresponding to Arg37 or Arg40 of SEQ ID NO:12. In some embodiments, the mutation is a deletion or substitution of an amino acid residue corresponding to a position selected from the group consisting of 31-40, 32-40, 33-40, 34-40, 30-39, 31-39, 32-39, 34-37, 33-39, 35-39, 36-39, 37-40, 34-40, and combinations thereof of SEQ ID NO:12. In some embodiments, the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO:13. In some embodiments, the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO:14.In some embodiments, the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 15. In some embodiments, the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 16. In some embodiments, the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 17. In some embodiments, the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO:18.

[0030] In some embodiments, the transgene encoding PGRN or GRN further comprises a microRNA (miRNA) targeting sequence (e.g., a miR-126 targeting sequence). In some embodiments, the miRNA targeting sequence is located within the 3'-untranslated region (UTR) of the transgene.

[0031] In some embodiments, the PGRN or GRN penetrates the blood-brain barrier (BBB) ​​of a subject.

[0032] In some embodiments, the NCD is a PGRN-associated NCD. In some embodiments, the FTLD or NCL is a PGRN-associated FTLD or NCL.

[0033] In some embodiments, the subject suffering from PGRN-related FTLD or NCL has a mutation in PGRN gene.The mutation in PGRN gene can be a frameshift mutation.In some embodiments, the frameshift mutation is p.C31LfsX35 mutation, p.C31LfsX35 mutation, p.S82VfsX174 mutation, p.L271LfsX174 mutation, or p.T382NfsX32 mutation.

[0034] Additionally or alternatively, the mutation in the PGRN gene may be, for example, a missense mutation. For example, the subject may have a p.C521Y mutation, a p.A9D mutation, a p.P248L mutation, a p.R432C mutation, a p.C139R mutation, a p.C521Y mutation, or a p.C139R mutation.

[0035] In addition, or alternatively, the mutation in the PGRN gene may be, for example, a nonsense mutation. For example, the subject may have a p.Q125X mutation. In some embodiments, the subject may have a p.R493X mutation.

[0036] Additionally or alternatively, the mutation in the PGRN gene may be, for example, an insertion mutation. For example, the subject may have a c.1145insA mutation.

[0037] Additionally or alternatively, the mutation in the PGRN gene may be, for example, a transversion mutation. For example, the subject may have a p.0(IVS1+5G>C) mutation.

[0038] In some embodiments, the subject suffering from PGRN-associated FTLD or NCL may have any other pathogenic mutation in PGRN gene that is known to have a causative role in FTLD or NCL.For example, the subject having a pathogenic mutation in PGRN gene and can be treated with the compositions and methods described herein includes any one of the mutations discussed in Gijselinck et al., Human Mutation 29(12), 1373-1386, (2012), which is incorporated herein by reference as its disclosure relates to human PGRN mutations.

[0039] In some embodiments, the subject suffering from PGRN-related FTLD may have behavioral abnormal frontotemporal dementia (BVFTD) variant of FTLD.In some embodiments, the subject suffering from PGRN-related FTLD may have semantic dementia (SD) variant of FTLD.In some embodiments, the subject suffering from PGRN-related FTLD may have progressive non-verbal aphasia (PNA) variant of FTLD.

[0040] In some embodiments, a subject suffering from PGRN-associated NCL may have a Santavuori-Haltia disease variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a Batten disease variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a Kuhs disease variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a Jansky-Bielschowsky disease variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a Finnish late-onset childhood variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a late-onset childhood variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a CLN7 variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a CLN8 variant. In some embodiments, a subject suffering from PGRN-associated NCL may have a CLN10 variant. In some embodiments, the subject suffering from PGRN-associated NCL may have CLN11 variant.In some embodiments, the subject suffering from PGRN-associated NCL may have Turkish late-onset childhood variant.In some embodiments, the subject suffering from PGRN-associated NCL may have type 9 variant.

[0041] In some embodiments, the transgene encoding PGRN or GRN encodes wild-type human PGRN or GRN (e.g., any one of SEQ ID NOs: 1-9). In some embodiments, the transgene encoding PGRN comprises a polynucleotide encoding a polypeptide having at least two GRN domains (e.g., 2, 3, 4, 5, 6, 7, 8, or more GRN domains), e.g., a GRN domain having an amino acid sequence of any one of SEQ ID NOs: 2-9. In some embodiments, the transgene encoding PGRN comprises a polynucleotide encoding a polypeptide comprising at least two GRN domains (e.g., 2, 3, 4, 5, 6, 7, 8, or more GRN domains) arranged in the same order as observed in wild-type human PGRN. In some embodiments, the transgene encoding PGRN comprises a polynucleotide encoding a polypeptide comprising at least two GRN domains (e.g., 2, 3, 4, 5, 6, 7, 8, or more GRN domains) arranged in a different order than wild-type human PGRN. In some embodiments, the transgene encoding PGRN or GRN comprises a polynucleotide encoding a polypeptide including at least one amino acid substitution, e.g., one or more conservative amino acid substitutions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more amino acid substitutions, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more conservative amino acid substitutions), compared to any of the polypeptides having the sequence of any one of SEQ ID NOs: 1-8.

[0042] In some embodiments, the cell is a pluripotent cell. In some embodiments, the pluripotent cell is an ESC. In some embodiments, the pluripotent cell is an iPSC. In some embodiments, the cell is a CD34+ cell. In some embodiments, the cell is a pluripotent cell. In some embodiments, the pluripotent cell is a CD34+ cell. In some embodiments, the CD34+ cell is a hematopoietic stem cell. In some embodiments, the CD34+ cell is a myeloid progenitor cell. In some embodiments, the cell is a blood lineage progenitor cell (BLPC). In some embodiments, the BLPC is a monocyte. In some embodiments, the cell is a macrophage. In some embodiments, the cell is a microglial progenitor cell. In some embodiments, the cell is a microglia.

[0043] In some embodiments, the subject's endogenous population of microglia is ablated prior to administering the composition to the subject. In some embodiments, the method includes ablating the subject's endogenous population of microglia prior to administering the composition to the subject. In some embodiments, microglia are ablated using an agent selected from the group consisting of busulfan, PLX3397, PLX647, PLX5622, treosulfan, and clodronate liposomes, by radiation therapy, or a combination thereof.

[0044] In some embodiments, the composition is administered to the subject systemically. In some embodiments, the composition is administered to the subject by intravenous injection. In some embodiments, the composition is administered directly to the central nervous system of the subject. In some embodiments, the composition is administered to the subject directly into the cerebrospinal fluid. For example, the composition can be administered to the subject by intraventricular injection, intrathecal injection, stereotactic injection, or a combination thereof. In some embodiments, the composition can be administered to the subject by intraparenchymal injection.

[0045] In some embodiments, the composition is administered directly to the subject's bone marrow, for example, by intraosseous injection.

[0046] In some embodiments, the composition is administered to the subject by bone marrow transplant.

[0047] In some embodiments, the composition is administered to the subject by intraventricular injection. In some embodiments, the composition is administered to the subject by intravenous injection.

[0048] In some embodiments, the composition is administered to the subject by direct administration to the subject's central nervous system and by systemic administration. In some embodiments, the composition is administered to the subject by intraventricular and intravenous injection. In some embodiments, the composition is administered to the subject by intrathecal and intravenous injection. In some embodiments, the composition is administered to the subject by intraparenchymal and intravenous injection.

[0049] In some embodiments, the method includes administering a population of cells to a subject. In some embodiments, the population of cells is administered to a subject prior to administration of the composition. In some embodiments, the population of cells is administered to a subject after administration of the composition. In some embodiments, the cells are selected from the group consisting of embryonic stem cells, induced pluripotent stem cells, hematopoietic stem cells, and myeloid progenitor cells. In some embodiments, the cells are not modified to express a transgene encoding PGRN or GRN. In some embodiments, the cells are administered systemically to a subject. In some embodiments, the cells are administered to a subject by intravenous injection.

[0050] In some embodiments, endogenous PGRN or GRN is destroyed in cells prior to administration of the composition to a subject.

[0051] In some embodiments, the cell is contacted with a nuclease that catalyzes the cleavage of endogenous PGRN or GRN nucleic acid in the cell, thereby destroying endogenous PGRN or GRN.In some embodiments, the nuclease is a CRISPR-associated protein.In some embodiments, the CRISPR-associated protein is CRISPR-associated protein 9.In some embodiments, the CRISPR-associated protein is CRISPR-associated protein 12a.In some embodiments, the nuclease is a transcription activator-like effector nuclease, a meganuclease, or a zinc finger nuclease.

[0052] In some embodiments, endogenous PGRN or GRN is destroyed, for example, by contacting cells with an inhibitory RNA molecule for a time and in an amount sufficient to destroy the expression of endogenous PGRN or GRN. In some embodiments, the inhibitory RNA molecule is a short interfering RNA (siRNA), a short hairpin RNA (shRNA), or a miRNA.

[0053] In some embodiments, endogenous PGRN or GRN is destroyed in a subject before administering the composition to the subject. In some embodiments, endogenous PGRN or GRN is destroyed by administering an inhibitory RNA molecule to the subject. In some embodiments, the inhibitory RNA molecule is siRNA, shRNA, or miRNA. In some embodiments, endogenous PGRN or GRN is destroyed in a population of neurons of a subject before administering the composition to the subject. In some embodiments, endogenous PGRN or GRN is destroyed in a population of neurons, for example, by contacting the population of neurons with an inhibitory RNA molecule for a time and amount sufficient to destroy the expression of endogenous PGRN or GRN. In some embodiments, the inhibitory RNA molecule is siRNA, shRNA, or miRNA.

[0054] In some embodiments, the cells are autologous cells. In some embodiments, the cells are allogeneic cells.

[0055] In some embodiments, cells are transduced ex vivo to express PGRN or GRN.

[0056] In some embodiments, the cells are transduced with a viral vector selected from the group including adeno-associated virus (AAV), adenovirus, parvovirus, coronavirus, rhabdovirus, paramyxovirus, picornavirus, alphavirus, herpesvirus, poxvirus, and retroviridae viruses.

[0057] In some embodiments, the viral vector is a Retroviridae viral vector. In some embodiments, the Retroviridae viral vector is a lentiviral vector. In some embodiments, the Retroviridae viral vector is an alpharetroviral vector. In some embodiments, the Retroviridae viral vector is a gammaretroviral vector. In some embodiments, the Retroviridae viral vector comprises a central polypurine tract, a woodchuck hepatitis virus post-transcriptional regulatory element, a 5'-LTR, an HIV signal sequence, an HIV Psi signal 5'-splice site, a delta-GAG element, a 3'-splice site, and a 3'-self-inactivating LTR.

[0058] In some embodiments, the viral vector is an AAV selected from the group including AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAVS, AAV9, AAV10, and AAVrh74.

[0059] In some embodiments, the viral vector is a pseudotyped viral vector. In some embodiments, the viral vector is a pseudotyped AAV, pseudotyped adenovirus, pseudotyped parvovirus, pseudotyped coronavirus, pseudotyped rhabdovirus, pseudotyped paramyxovirus, pseudotyped picornavirus, pseudotyped alphavirus, pseudotyped herpesvirus, pseudotyped poxvirus, and pseudotyped retroviridae virus.

[0060] In some embodiments, cells are transfected ex vivo to express PGRN or GRN.

[0061] In some embodiments, the cells are transfected using an agent selected from the group including cationic polymers, diethylaminoethyl-dextran, polyethyleneimine, cationic lipids, liposomes, calcium phosphate, activated dendrimers, and magnetic beads; or a technique selected from the group including electroporation, Nucleofection, squeeze-poration, sonoporation, optical transfection, Magnetofection, and impalefection.

[0062] In some embodiments, the expression of PGRN or GRN in cells is mediated using a ubiquitous promoter. Exemplary ubiquitous promoters are elongation factor 1-alpha promoter and phosphoglycerate kinase 1 promoter. In some embodiments, the expression of PGRN in cells is mediated using a cell lineage-specific promoter. Exemplary cell lineage-specific promoters are PGRN promoter, CD11b promoter, CD68 promoter, C-X3-C motif chemokine receptor 1 promoter, allograft inflammatory factor 1 promoter, purinergic receptor P2Y12 promoter, transmembrane protein 119 promoter, and colony-stimulating factor 1 receptor promoter.

[0063] In some embodiments, the composition is administered to a subject in an amount sufficient to increase the amount of M2 microglia in the subject's brain relative to the amount of M1 microglia in the subject's brain, decrease the level of one or more pro-inflammatory cytokines in the subject's brain, increase the level of one or more anti-inflammatory cytokines in the subject's brain, improve cognitive processing ability in the subject, improve motor function in the subject, reduce neuronal loss in the subject, and / or decrease levels of alpha-synuclein protein, tau protein, TAR DNA binding protein 43 (TDP-43) positive inclusions, fused in sarcoma (FUS) positive inclusions, and / or ubiquitin positive inclusions and inclusions, or aggregates thereof, in the subject.

[0064] In some embodiments, the subject is a human.

[0065] In another aspect, the disclosure provides a pharmaceutical composition containing a population of cells comprising a transgene encoding PGRN or GRN.

[0066] In some embodiments of the preceding aspects, the cell is a pluripotent cell. In some embodiments, the pluripotent cell is an ESC. In some embodiments, the pluripotent cell is an iPSC. In some embodiments, the cell is a CD34+ cell. In some embodiments, the cell is a pluripotent cell. In some embodiments, the pluripotent cell is a CD34+ cell. In some embodiments, the CD34+ cell is a hematopoietic stem cell. In some embodiments, the CD34+ cell is a myeloid progenitor cell. In some embodiments, the cell is a blood lineage progenitor cell (BLPC). In some embodiments, the BLPC is a monocyte. In some embodiments, the cell is a macrophage. In some embodiments, the cell is a microglial progenitor cell. In some embodiments, the cell is a microglia.

[0067] In some embodiments, cells are transduced ex vivo to express PGRN or GRN. In some embodiments, cells are transfected ex vivo to express PGRN or GRN.

[0068] In some embodiments, the PGRN is a full-length PGRN, such as a PGRN having the amino acid sequence of SEQ ID NO:1, or a variant thereof having at least 85% sequence identity thereto (e.g., having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:1). In some embodiments, PGRN comprises at least two (e.g., at least 2, 3, 4, 5, 6, 7, 8 or more) GRN peptides having an amino acid sequence of any one of SEQ ID NOs: 2-9 or variants thereof having at least 85% sequence identity thereto (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto). In some embodiments, PGRN comprises at least two (e.g., at least 2, 3, 4, 5, 6, 7, 8 or more) GRN domains. In some embodiments, PGRN comprises at least three (e.g., at least 3, 4, 5, 6, 7, 8 or more) GRN domains. In some embodiments, PGRN comprises at least four (e.g., at least 4, 5, 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least five (e.g., at least 5, 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least six (e.g., at least 6, 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least seven (e.g., at least 7, 8 or more) GRN domains. In some embodiments, the PGRN comprises at least eight (e.g., at least 8 or more) GRN domains. In some embodiments, the PGRN comprises 2-16 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16) GRN domains. In some embodiments, the PGRN comprises 2-12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) GRN domains.In some embodiments, the PGRN comprises 2-8 (e.g., 2, 3, 4, 5, 6, 7, or 8) GRN domains. In some embodiments, the PGRN comprises 2-4 (e.g., 2, 3, or 4) GRN domains. In some embodiments, the PGRN comprises 2 GRN domains.

[0069] In some embodiments, the PGRN comprises a para-GRN domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the para-GRN domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the para-GRN domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:2. In some embodiments, the para-GRN domain has an amino acid sequence of SEQ ID NO:2.

[0070] In some embodiments, the PGRN comprises a GRN-1 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:3. In some embodiments, the GRN-1 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:3. In some embodiments, the GRN-1 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:3. In some embodiments, the GRN-1 domain has an amino acid sequence of SEQ ID NO:3.

[0071] In some embodiments, the PGRN comprises a GRN-2 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:4. In some embodiments, the GRN-2 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:4. In some embodiments, the GRN-2 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:4. In some embodiments, the GRN-2 domain has an amino acid sequence of SEQ ID NO:4.

[0072] In some embodiments, the PGRN comprises a GRN-3 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:5. In some embodiments, the GRN-3 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:5. In some embodiments, the GRN-3 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:5. In some embodiments, the GRN-3 domain has an amino acid sequence of SEQ ID NO:5.

[0073] In some embodiments, the PGRN comprises a GRN-4 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:6. In some embodiments, the GRN-4 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:6. In some embodiments, the GRN-4 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:6. In some embodiments, the GRN-4 domain has an amino acid sequence of SEQ ID NO:6.

[0074] In some embodiments, the PGRN comprises a GRN-5 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:7. In some embodiments, the GRN-5 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:7. In some embodiments, the GRN-5 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:7. In some embodiments, the GRN-5 domain has an amino acid sequence of SEQ ID NO:7.

[0075] In some embodiments, the PGRN comprises a GRN-6 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:8. In some embodiments, the GRN-6 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:8. In some embodiments, the GRN-6 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:8. In some embodiments, the GRN-6 domain has an amino acid sequence of SEQ ID NO:8.

[0076] In some embodiments, the PGRN comprises a GRN-7 domain having an amino acid sequence that is at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:9. In some embodiments, the GRN-7 domain has an amino acid sequence that is at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:9. In some embodiments, the GRN-7 domain has an amino acid sequence that is at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) identical to the amino acid sequence of SEQ ID NO:9. In some embodiments, the GRN-7 domain has an amino acid sequence of SEQ ID NO:9.

[0077] In some embodiments, the GRN is a full-length GRN, such as a GRN having any one of the amino acid sequences of SEQ ID NOs: 2-9, or a variant thereof having at least 85% sequence identity thereto (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto). In some embodiments, the GRN is a para-GRN or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity thereto (SEQ ID NO: 2). In some embodiments, the para-GRN has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 2. In some embodiments, the para-GRN has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 2. In some embodiments, the para-GRN has the amino acid sequence of SEQ ID NO: 2.

[0078] In some embodiments, GRN is GRN-1 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:3. In some embodiments, GRN-1 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:3. In some embodiments, GRN-1 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:3. In some embodiments, GRN-1 has the amino acid sequence of SEQ ID NO:3.

[0079] In some embodiments, GRN is GRN-2 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:4. In some embodiments, GRN-2 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:4. In some embodiments, GRN-2 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:4. In some embodiments, GRN-2 has the amino acid sequence of SEQ ID NO:4.

[0080] In some embodiments, the GRN is GRN-3 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:5. In some embodiments, the GRN-3 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:5. In some embodiments, the GRN-3 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:5. In some embodiments, the GRN-3 has the amino acid sequence of SEQ ID NO:5.

[0081] In some embodiments, the GRN is GRN-4 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:6. In some embodiments, the GRN-4 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:6. In some embodiments, the GRN-4 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:6. In some embodiments, the GRN-4 has the amino acid sequence of SEQ ID NO:6.

[0082] In some embodiments, the GRN is GRN-5 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:7. In some embodiments, the GRN-5 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:7. In some embodiments, the GRN-5 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:7. In some embodiments, the GRN-5 has the amino acid sequence of SEQ ID NO:7.

[0083] In some embodiments, the GRN is GRN-6 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:8. In some embodiments, the GRN-6 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:8. In some embodiments, the GRN-6 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:8. In some embodiments, the GRN-6 has the amino acid sequence of SEQ ID NO:8.

[0084] In some embodiments, the GRN is GRN-7 or a variant thereof having at least 85% (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:9. In some embodiments, the GRN-7 has at least 90% (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:9. In some embodiments, the GRN-7 has at least 95% (e.g., at least 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO:9. In some embodiments, the GRN-7 has the amino acid sequence of SEQ ID NO:9.

[0085] In some embodiments, the order of the GRN domains in the PGRN polypeptide occurs in the same order as observed in wild-type human PGRN. In some embodiments, the order of the GRN domains in the PGRN polypeptide occurs in a different order than that found in wild-type human PGRN.

[0086] In some embodiments, the PGRN or GRN comprises a secretory signal peptide. In some embodiments, the secretory signal peptide is a PGRN secretory signal peptide.

[0087] In some embodiments, the PGRN or GRN is a PGRN or GRN fusion protein. In some embodiments, the PGRN or GRN fusion protein comprises the Rb domain of ApoE. In some embodiments, the Rb domain comprises a portion of ApoE having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO:11. In some embodiments, the Rb domain comprises a region having at least 70% sequence identity to the amino acid sequence of residues 159-167 of SEQ ID NO:11.

[0088] In some embodiments, the transgene encoding PGRN or GRN further comprises a miRNA targeting sequence in the 3'-UTR. In some embodiments, the miRNA targeting sequence is a miR-126 targeting sequence.

[0089] In some embodiments, endogenous PGRN or GRN is destroyed intracellularly.

[0090] In some embodiments, the pharmaceutical composition is formulated for systemic administration to a subject. In some embodiments, the pharmaceutical composition is formulated for administration to a subject by intravenous injection. In some embodiments, the pharmaceutical composition is formulated for administration to a subject's central nervous system. In some embodiments, the pharmaceutical composition is formulated for administration to a subject into the cerebrospinal fluid. In some embodiments, the pharmaceutical composition is formulated for administration to a subject by intraventricular injection, intrathecal injection, stereotactic injection, or a combination thereof. In some embodiments, the pharmaceutical composition is formulated for administration to a subject by intraparenchymal injection. In some embodiments, the pharmaceutical composition is formulated for administration directly to a subject's bone marrow. In some embodiments, the pharmaceutical composition is formulated for administration to a subject by intraosseous injection. In some embodiments, the pharmaceutical composition is administered to a subject by a bone marrow transplant comprising the pharmaceutical composition. In some embodiments, the pharmaceutical composition is formulated for administration to a subject by intraventricular and intravenous injection.

[0091] In some embodiments, the subject (e.g., human) has been diagnosed with an NCD. In some embodiments, the NCD is a severe NCD. In some embodiments, the severe NCD interferes with the subject's independence and / or normal daily functioning (e.g., social, occupational, or academic functioning, personal hygiene, grooming, dressing, toilet hygiene, functional mobility (e.g., ability to walk, get in and out of bed), and self-feeding). In some embodiments, the severe NCD is associated with a score obtained by the subject on a cognitive test that is at least 2 standard deviations away from the mean score of a reference population. In some embodiments, the NCD is a mild NCD. In some embodiments, the mild NCD does not interfere with the subject's independence and / or normal daily functioning. In some embodiments, the mild NCD is associated with a score obtained by the subject on a cognitive test that is at least 1-2 standard deviations away from the mean score of a reference population. In some embodiments, the cognitive test is selected from the group consisting of AD8, AWV, GPCOG, HRA, MIS, MMSE, MoCA, SLUMS, and Short IQCODE. In some embodiments, the NCD is associated with impairment in one or more of complex attention, executive function, learning and memory, language, sensorimotor function, and social cognition. In some embodiments, the NCD is not due to delirium or other psychiatric disorders (e.g., schizophrenia, bipolar disorder, or major depression). In some embodiments, the reference population is the general population. In some embodiments, the reference population is selected based on the subject's age, medical history, education, socioeconomic status, and lifestyle. In some embodiments, the NCD is a frontotemporal NCD. In some embodiments, the frontotemporal NCD is FTLD. In some embodiments, the NCD is due to a lysosomal disease. In some embodiments, the lysosomal disease is NCL.

[0092] In an additional aspect, the present disclosure provides a kit comprising a composition according to any of the above aspects and embodiments and a package insert, in some embodiments, the package insert instructs a user of the kit to carry out a method according to any of the above aspects and embodiments.

[0093] Additional embodiments of the present invention are provided in the paragraphs listed below.

[0094] E1. 1. A method of treating a subject diagnosed with a neurocognitive disorder (NCD), comprising administering to the subject a composition comprising a population of cells comprising a transgene encoding Progranulin (PGRN) or Granulin (GRN).

[0095] E2. The method according to E1, wherein said NCD is a severe NCD.

[0096] E3. The method of E2, wherein said severe NCD interferes with said subject's independence and / or normal daily functioning.

[0097] E4. The method of E2 or E3, wherein said severe NCD is associated with a score obtained by said subject on a cognitive test that is at least 2 standard deviations away from the mean score of a reference population.

[0098] E5. The method according to E1, wherein said NCD is a mild NCD.

[0099] E6. The method of E5, wherein said mild NCD does not interfere with said subject's independence and / or normal daily functioning.

[0100] E7. The method of E5 or E6, wherein said mild NCD is associated with a score obtained by said subject on a cognitive test that is 1 to 2 standard deviations away from the mean score of a reference population.

[0101] E8. The method of E4 or E7, wherein the reference population is the general population.

[0102] E9. The method of E4, E7, or E8, wherein the cognitive test is selected from the group consisting of Eight-item Informant Interview to Differentiate Aging and Dementia (AD8), Annual Wellness Visit (AWV), General Practitioner Assessment of Cognition (GPCOG), Health Risk Assessment (HRA), Memory Impairment Screen (MIS), Mini Mental Status Exam (MMSE), Montreal Cognitive Assessment (MoCA), St. Louis University Mental Status Exam (SLUMS), and Short Informant Questionnaire on Cognitive Decline in the Elderly (Short IQCODE).

[0103] E10. The method of any one of E1-E9, wherein the NCD is associated with impairment in one or more of complex attention, executive function, learning and memory, language, sensorimotor function, and social cognition.

[0104] E11. The method of any one of E1-E10, wherein said NCD is not due to delirium or other psychiatric disorder.

[0105] E12. The method according to any one of E1 to E11, wherein said NCD is a frontotemporal NCD.

[0106] E13. The method of claim 12, wherein said frontotemporal NCD is frontotemporal lobar degeneration (FTLD).

[0107] E14. The method according to any one of E1 to E11, wherein said NCD is due to a lysosomal disease.

[0108] E15. The method according to E14, wherein the lysosomal disease is neuronal ceroid lipofuscinosis (NCL).

[0109] E16. The method according to any one of E1 to E15, wherein the PGRN or the GRN comprises a secretory signal peptide.

[0110] E17. The method according to E16, wherein the secretory signal peptide is a PGRN secretory signal peptide.

[0111] E18. The method according to any one of E1 to E17, wherein the cell comprises a transgene encoding the PGRN.

[0112] E19. The method of E18, wherein the PGRN comprises at least two GRN domains.

[0113] E20. The method described in E19, wherein the PGRN comprises at least three GRN domains.

[0114] E21. The method of E20, wherein the PGRN comprises at least four GRN domains.

[0115] E22. The method of E21, wherein the PGRN comprises at least five GRN domains.

[0116] E23. The method of E22, wherein the PGRN comprises at least six GRN domains.

[0117] E24. The method of E23, wherein the PGRN comprises at least seven GRN domains.

[0118] E25. The method of E24, wherein the PGRN comprises at least eight GRN domains.

[0119] E26. The method according to any one of E1 to E25, wherein the PGRN comprises 2 to 16 GRN domains.

[0120] E27. The method of E26, wherein the PGRN comprises 2 to 12 GRN domains.

[0121] E28. The method of E27, wherein the PGRN comprises 2 to 8 GRN domains.

[0122] E29. The method of E28, wherein the PGRN comprises two to four GRN domains.

[0123] E30. The method according to E29, wherein the PGRN comprises two GRN domains.

[0124] E31. The method of any one of E1 to E30, wherein the PGRN comprises a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2.

[0125] E32. The method of E31, wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2.

[0126] E33. The method of E32, wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2.

[0127] E34. The method of E33, wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:2.

[0128] E35. The method according to any one of E1 to E34, wherein the PGRN comprises a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3.

[0129] E36. The method of E35, wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3.

[0130] E37. The method of E36, wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3.

[0131] E38. The method described in E37, wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:3.

[0132] E39. The method according to any one of E1 to E38, wherein the PGRN comprises a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4.

[0133] E40. The method of E39, wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4.

[0134] E41. The method of E40, wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4.

[0135] E42. The method described in E41, wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:4.

[0136] E43. The method according to any one of E1 to E42, wherein the PGRN comprises a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5.

[0137] E44. The method described in E43, wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5.

[0138] E45. The method described in E44, wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5.

[0139] E46. The method described in E45, wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:5.

[0140] E47. The method of any one of E1 to E46, wherein the PGRN comprises a GRN-4 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:6.

[0141] E48. The method described in E47, wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6.

[0142] E49. The method described in E48, wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:6.

[0143] E50. The method described in E49, wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:6.

[0144] E51. The method according to any one of E1 to E50, wherein the PGRN comprises a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7.

[0145] E52. The method of E51, wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7.

[0146] E53. The method of E52, wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7.

[0147] E54. The method described in E53, wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:7.

[0148] E55. The method according to any one of E1 to E54, wherein the PGRN comprises a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8.

[0149] E56. The method of E55, wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8.

[0150] E57. The method of E56, wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8.

[0151] E58. The method described in E49, wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:8.

[0152] E59. The method according to any one of E1 to E58, wherein the PGRN comprises a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9.

[0153] E60. The method described in E59, wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9.

[0154] E61. The method described in E60, wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9.

[0155] E62. The method described in E61, wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:9.

[0156] E63. The method according to any one of E1 to E62, wherein said PGRN has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:1.

[0157] E64. The method of E63, wherein said PGRN has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:1.

[0158] E65. The method of E64, wherein the PGRN has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:1.

[0159] E66. The method of E65, wherein the PGRN has the amino acid sequence of SEQ ID NO:1.

[0160] E67. The method according to any one of E1 to E66, wherein the PGRN is a full-length PGRN.

[0161] E68. The method of any one of E1 to E67, wherein said cell comprises a transgene encoding said GRN.

[0162] E69. The method according to any one of E1 to E68, wherein the GRN is a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2.

[0163] E70. The method of E69, wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2.

[0164] E71. The method of E70, wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2.

[0165] E72. The method of E71, wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:2.

[0166] E73. The method according to any one of E1 to E72, wherein the GRN is a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3.

[0167] E74. The method of E73, wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3.

[0168] E75. The method of E74, wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3.

[0169] E76. The method described in E75, wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:3.

[0170] E77. The method of any one of E1 to E76, wherein the GRN is a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4.

[0171] E78. The method of E77, wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4.

[0172] E79. The method of E78, wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4.

[0173] E80. The method described in E79, wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:4.

[0174] E81. The method according to any one of E1 to E80, wherein the GRN is a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5.

[0175] E82. The method of E81, wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5.

[0176] E83. The method of E82, wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5.

[0177] E84. The method described in E83, wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:5.

[0178] E85. The method according to any one of E1 to E84, wherein the GRN is a GRN-4 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:6.

[0179] E86. The method described in E85, wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6.

[0180] E87. The method described in E86, wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:6.

[0181] E88. The method described in E87, wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:6.

[0182] E89. The method according to any one of E1 to E88, wherein the GRN is a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7.

[0183] E90. The method described in E89, wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7.

[0184] E91. The method described in E90, wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7.

[0185] E92. The method of E91, wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:7.

[0186] E93. The method according to any one of E1 to E92, wherein the GRN is a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8.

[0187] E94. The method described in E93, wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8.

[0188] E95. The method described in E94, wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8.

[0189] E96. The method described in E95, wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:8.

[0190] E97. The method of any one of E1 to E96, wherein the GRN is a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9.

[0191] E98. The method of E97, wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9.

[0192] E99. The method of E98, wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9.

[0193] E100. The method described in E99, wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:9.

[0194] E101. The method according to any one of E1 to E100, wherein the GRN comprises a full-length GRN.

[0195] E102. The method of any one of E1 to E101, wherein said cell comprises a PGRN transgene having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO:10.

[0196] E103. The method of E102, wherein the cell comprises a PGRN transgene having at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO:10.

[0197] E104. The method of E103, wherein the cell comprises a PGRN transgene having at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO:10.

[0198] E105. The method according to E104, wherein the cell comprises a PGRN transgene having the nucleic acid sequence of SEQ ID NO:10.

[0199] E106. The method according to any one of E1 to E105, wherein the PGRN or the GRN is a PGRN or GRN fusion protein.

[0200] E107. The method of E106, wherein the PGRN or GRN fusion protein comprises the receptor binding (Rb) domain of apolipoprotein E (ApoE).

[0201] E108. The method of E107, wherein the Rb domain comprises a portion of ApoE having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO:11.

[0202] E109. The method of E107 or E108, wherein the Rb domain comprises a region having at least 70% sequence identity to the amino acid sequence of residues 159 to 167 of SEQ ID NO:11.

[0203] E110. The method according to any one of E1 to E109, wherein the transgene encoding the PGRN or the GRN further comprises a microRNA (miRNA) targeting sequence in the 3'-UTR.

[0204] E111. The method of E110, wherein the miRNA targeting sequence is a miR-126 targeting sequence.

[0205] E112. The method of any one of E1 to E111, wherein upon administration of the composition to the subject, the PGRN or the GRN penetrates the blood-brain barrier of the subject.

[0206] E113. The method according to any one of E13 to E112, wherein the FTLD or NCL is PGRN-associated FTLD or NCL.

[0207] E114. The method of E113, wherein the PGRN-associated FTLD is a behavioral abnormality frontotemporal variant of FTLD.

[0208] E115. The method of E113, wherein the PGRN-associated FTLD is a semantic dementia variant of FTLD.

[0209] E116. The method of E113, wherein said PGRN-associated FTLD is a progressive incompetent aphasia variant of FTLD.

[0210] E117. The method of E113, wherein the PGRN-associated NCL is Batten disease.

[0211] E118. The method according to any one of E1 to E117, wherein the cells are ESCs.

[0212] E119. The method according to any one of E1 to E117, wherein the cell is an iPSC.

[0213] E120. The method of any one of E1 to E117, wherein the cells are CD34+ cells.

[0214] E121. The method of E120, wherein the CD34+ cells are HSCs.

[0215] E122. The method of E120, wherein the CD34+ cells are MPCs.

[0216] E123. The method of any one of E1 to E122, wherein the endogenous microglia population in said subject is ablated prior to administration of said composition.

[0217] E124. The method of any one of E1 to E122, comprising removing a population of endogenous microglia in the subject prior to administering the composition to the subject.

[0218] E125. The method of E123 or E124, wherein the microglia are ablated using an agent selected from the group consisting of busulfan, PLX3397, PLX647, PLX5622, treosulfan, and clodronate liposomes, by radiation therapy, or a combination thereof.

[0219] E126. The method of any one of E1 to E125, wherein the composition is systemically administered to the subject.

[0220] E127. The method according to E126, wherein the composition is administered to the subject by intravenous injection.

[0221] E128. The method of any one of E1 to E125, wherein said composition is administered directly into the central nervous system of said subject.

[0222] E129. The method of E128, wherein the composition is administered to the subject by direct administration into cerebrospinal fluid.

[0223] E130. The method of E128 or E129, wherein the composition is administered to the subject by intraventricular injection, intrathecal injection, stereotactic injection, or a combination thereof.

[0224] E131. The method of E128, wherein the composition is administered to the subject by intraparenchymal injection.

[0225] E132. The method of any one of E1 to E125, wherein said composition is administered directly to the bone marrow of said subject.

[0226] E133. The method of E132, wherein the composition is administered to the subject by intraosseous injection.

[0227] E134. The method of any one of E1 to E125, wherein said composition is administered to said subject by bone marrow transplantation comprising said composition.

[0228] E135. The method of any one of E1 to E125, wherein said composition is administered to said subject by intraventricular injection.

[0229] E136. The method of any one of E1 to E125, wherein the composition is administered to the subject by intrathecal injection.

[0230] E137. The method of any one of E1 to E125, wherein the composition is administered to the subject by intraparenchymal injection.

[0231] E138. The method of any one of E1 to E125, wherein the composition is administered to the subject by intravenous injection.

[0232] E139. The method of any one of E1 to E125, wherein the composition is administered to the subject by direct administration to the central nervous system of the subject and by systemic administration.

[0233] E140. The method of E139, wherein the composition is administered to the subject by intraventricular and intravenous injection.

[0234] E141. The method of E139, wherein the composition is administered to the subject by intrathecal and intravenous injection.

[0235] E142. The method according to E139, wherein the composition is administered to the subject by intraparenchymal and intravenous injection.

[0236] E143. The method of any one of E1 to E142, further comprising administering to said subject the population of cells.

[0237] E144. The method of E143, wherein the population of cells is administered to the subject prior to administration of the composition.

[0238] E145. The method of E143, wherein after administration of the composition, the population of cells is administered to the subject.

[0239] E146. The method according to any one of E143 to E145, wherein the cells are selected from the group consisting of pluripotent cells, ESCs, IPSCs, multipotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, and microglia.

[0240] E147. The method of any one of E143 to E146, wherein the cell has not been modified to express a transgene encoding the PGRN or the GRN.

[0241] E148. The method according to any one of E143 to E147, wherein the cells are systemically administered to the subject.

[0242] E149. The method of E148, wherein the cells are administered to the subject by intravenous injection.

[0243] E150. The method of any one of E1 to E149, wherein endogenous PGRN or GRN is destroyed in the cells prior to administration of the composition to the subject.

[0244] E151. The method according to any one of E1 to E150, wherein the endogenous PGRN or GRN is destroyed in the subject prior to administration of the composition to the subject.

[0245] E152. The method of E151, wherein the endogenous PGRN or GRN is destroyed in a population of neurons in the subject prior to administration of the composition to the subject.

[0246] E153. The method of E150, wherein the endogenous PGRN or GRN is destroyed by contacting the cell with a nuclease that catalyzes cleavage of endogenous PGRN or GRN nucleic acid in the cell.

[0247] E154. The method of E153, wherein the nuclease is a clustered regularly interspaced short palindromic repeats (CRISPR) associated protein.

[0248] E155. The method of E154, wherein the CRISPR-associated protein is CRISPR-associated protein 9 (Cas9).

[0249] E156. The method of E154, wherein the CRISPR-associated protein is CRISPR-associated protein 12a (Cas12a).

[0250] E157. The method of E153, wherein the nuclease is a transcription activator-like effector nuclease, a meganuclease, or a zinc finger nuclease.

[0251] E158. The method according to any one of E150 to E152, wherein the endogenous PGRN or GRN is destroyed by administering an inhibitory RNA molecule to the cell, the subject, or the population of neurons.

[0252] E159. The method of E158, wherein the inhibitory RNA molecule is a short interfering RNA, a short hairpin RNA, or an miRNA.

[0253] E160. The method according to any one of E1 to E159, wherein the cells are autologous cells.

[0254] E161. The method according to any one of E1 to E159, wherein the cells are allogeneic cells.

[0255] E162. The method of any one of E1 to E61, wherein the cells are transduced ex vivo to express the PGRN or the GRN.

[0256] E163. The method of E162, wherein the cells are transduced with a viral vector selected from the group consisting of adeno-associated virus (AAV), adenovirus, parvovirus, coronavirus, rhabdovirus, paramyxovirus, picornavirus, alphavirus, herpesvirus, poxvirus, and retroviridae virus.

[0257] E164. The method according to E163, wherein said viral vector is a retroviridae viral vector.

[0258] E165. The method according to E164, wherein the Retroviridae viral vector is a lentiviral vector.

[0259] E166. The method according to E164, wherein said Retroviridae viral vector is an alpharetroviral vector.

[0260] E167. The method according to E164, wherein said Retroviridae viral vector is a gamma retroviral vector.

[0261] E168. The method according to any one of E164 to E167, wherein the Retroviridae viral vector comprises a central polypurine tract, a Woodchuck Hepatitis Virus post-transcriptional regulatory element, a 5'-LTR, an HIV signal sequence, an HIV Psi signal 5'-splice site, a delta-GAG element, a 3'-splice site, and a 3'-self-inactivating LTR.

[0262] E169. The method of E163, wherein the viral vector is an AAV selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, and AAVrh74.

[0263] E170. The method according to any one of E163 to E169, wherein the viral vector is a pseudotyped viral vector.

[0264] E171. The method of E170, wherein the pseudotyped viral vector is selected from the group consisting of pseudotyped AAV, pseudotyped adenovirus, pseudotyped parvovirus, pseudotyped coronavirus, pseudotyped rhabdovirus, pseudotyped paramyxovirus, pseudotyped picornavirus, pseudotyped alphavirus, pseudotyped herpesvirus, pseudotyped poxvirus, and pseudotyped retroviridae viruses.

[0265] E172. The method according to any one of E1 to E161, wherein the cells are transfected ex vivo to express the PGRN or the GRN.

[0266] E173. The method of E172, wherein the cells are transfected using: a) an agent selected from the group consisting of cationic polymers, diethylaminoethyl-dextran, polyethyleneimine, cationic lipids, liposomes, calcium phosphate, activated dendrimers, and magnetic beads; or b) a technique selected from the group consisting of electroporation, Nucleofection, squeeze-poration, sonoporation, optical transfection, Magnetofection, and impalefection.

[0267] E174. The method according to any one of E1 to E173, wherein expression of the PGRN or the GRN in the cell is mediated by a ubiquitous promoter.

[0268] E175. The method of E174, wherein the ubiquitous promoter is selected from the group consisting of elongation factor 1-alpha promoter and phosphoglycerate kinase 1 promoter.

[0269] E176. The method according to any one of E1 to E173, wherein expression of said PGRN or said GRN is mediated by a cell lineage specific promoter.

[0270] E177. The method of E176, wherein the cell lineage specific promoter is selected from the group consisting of PGRN promoter, CD11b promoter, CD68 promoter, C-X3-C motif chemokine receptor 1 promoter, allograft inflammatory factor 1 promoter, purinergic receptor P2Y12 promoter, transmembrane protein 119 promoter, and colony stimulating factor 1 receptor promoter.

[0271] E178. The method according to any one of E1 to E173, wherein expression of said PGRN or said GRN in said cell is mediated by a synthetic promoter.

[0272] E179. The method of any one of E1-E178, wherein the composition is administered to the subject in an amount sufficient to a) increase the amount of M2 microglia in the brain of the subject relative to the amount of M1 microglia in the brain of the subject; b) decrease the level of one or more pro-inflammatory cytokines in the brain of the subject; c) increase the level of one or more anti-inflammatory cytokines in the brain of the subject; d) improve the cognitive performance of the subject; e) improve the motor function of the subject; f) decrease neuronal loss in the subject; and / or g) decrease the level of alpha-synuclein protein, tau-positive neuronal inclusions, TAR DNA-binding protein 43 (TDP-43)-positive inclusions, fusion with sarcoma (FUS)-positive inclusions, and / or ubiquitin-positive inclusions, or aggregates thereof, in the subject.

[0273] E180. The method according to any one of E1 to E179, wherein the subject is a human.

[0274] E181. A pharmaceutical composition comprising a population of cells containing a transgene encoding PGRN or GRN, and further comprising one or more pharma- ceutically acceptable carriers, diluents, or excipients.

[0275] E182. The pharmaceutical composition of E181, wherein the PGRN or the GRN comprises a secretory signal peptide.

[0276] E183. The pharmaceutical composition of E182, wherein the secretory signal peptide is a PGRN secretory signal peptide.

[0277] E184. The pharmaceutical composition according to any one of E181 to E183, wherein the cell comprises a transgene encoding the PGRN.

[0278] E185. The pharmaceutical composition of E184, wherein the PGRN comprises at least two GRN domains.

[0279] E186. The pharmaceutical composition of E185, wherein the PGRN comprises at least three GRN domains.

[0280] E187. The pharmaceutical composition of E186, wherein the PGRN comprises at least four GRN domains.

[0281] E188. The pharmaceutical composition of E187, wherein the PGRN comprises at least five GRN domains.

[0282] E189. The pharmaceutical composition of E188, wherein the PGRN comprises at least six GRN domains.

[0283] E190. The pharmaceutical composition of E189, wherein the PGRN comprises at least seven GRN domains.

[0284] E191. The pharmaceutical composition of E190, wherein the PGRN comprises at least eight GRN domains.

[0285] E192. The pharmaceutical composition described in any one of E181 to E191, wherein the PGRN comprises 2 to 16 GRN domains.

[0286] E193. The pharmaceutical composition of E192, wherein the PGRN comprises 2 to 12 GRN domains.

[0287] E194. The pharmaceutical composition of E193, wherein the PGRN comprises 2 to 8 GRN domains.

[0288] E195. The pharmaceutical composition described in E194, wherein the PGRN contains 2 to 4 GRN domains.

[0289] E196. The pharmaceutical composition of E195, wherein the PGRN comprises two GRN domains.

[0290] E197. The pharmaceutical composition described in any one of E181 to E196, wherein the PGRN comprises a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2.

[0291] E198. The pharmaceutical composition described in E197, wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2.

[0292] E199. The pharmaceutical composition of E198, wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2.

[0293] E200. The pharmaceutical composition described in E199, wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:2.

[0294] E201. The pharmaceutical composition described in any one of E181 to E200, wherein the PGRN comprises a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3.

[0295] E202. The pharmaceutical composition of E201, wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3.

[0296] E203. The pharmaceutical composition of E202, wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3.

[0297] E204. The pharmaceutical composition described in E203, wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:3.

[0298] E205. A pharmaceutical composition described in any one of E181 to E204, wherein the PGRN comprises a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4.

[0299] E206. The pharmaceutical composition described in E205, wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4.

[0300] E207. The pharmaceutical composition described in E206, wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4.

[0301] E208. The pharmaceutical composition described in E207, wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:4.

[0302] E209. A pharmaceutical composition described in any one of E181 to E208, wherein the PGRN comprises a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5.

[0303] E210. The pharmaceutical composition described in E209, wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5.

[0304] E211. The pharmaceutical composition described in E210, wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5.

[0305] E212. The pharmaceutical composition described in E211, wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:5.

[0306] E213. A pharmaceutical composition described in any one of E181 to E212, wherein the PGRN comprises a GRN-4 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:6.

[0307] E214. The pharmaceutical composition described in E213, wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6.

[0308] E215. The pharmaceutical composition described in E214, wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:6.

[0309] E216. The pharmaceutical composition described in E215, wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:6.

[0310] E217. The pharmaceutical composition described in any one of E181 to E216, wherein the PGRN comprises a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7.

[0311] E218. The pharmaceutical composition described in E217, wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7.

[0312] E219. The pharmaceutical composition described in E218, wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7.

[0313] E220. The pharmaceutical composition described in E219, wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:7.

[0314] E221. A pharmaceutical composition described in any one of E181 to E220, wherein the PGRN comprises a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8.

[0315] E222. The pharmaceutical composition described in E221, wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8.

[0316] E223. The pharmaceutical composition described in E222, wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8.

[0317] E224. The pharmaceutical composition described in E223, wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:8.

[0318] E225. A pharmaceutical composition described in any one of E181 to E224, wherein the PGRN comprises a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9.

[0319] E226. The pharmaceutical composition described in E225, wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9.

[0320] E227. The pharmaceutical composition described in E226, wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9.

[0321] E228. The pharmaceutical composition described in E226, wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:9.

[0322] E229. The pharmaceutical composition according to any one of E181 to E228, wherein the PGRN is a full-length PGRN.

[0323] E230. The pharmaceutical composition described in E181 to E229, wherein the PGRN has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:1.

[0324] E231. The pharmaceutical composition described in E230, wherein the PGRN has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:1.

[0325] E232. The pharmaceutical composition described in E231, wherein the PGRN has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:1.

[0326] E233. The pharmaceutical composition described in E232, wherein the PGRN has the amino acid sequence of SEQ ID NO:1.

[0327] E234. The pharmaceutical composition described in any one of E181 to E233, wherein the cell comprises a transgene encoding the GRN.

[0328] E235. A pharmaceutical composition described in any one of E181 to E234, wherein the GRN is a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2.

[0329] E236. The pharmaceutical composition described in E235, wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2.

[0330] E237. The pharmaceutical composition described in E236, wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2.

[0331] E238. The pharmaceutical composition described in E237, wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:2.

[0332] E239. A pharmaceutical composition described in any one of E181 to E238, wherein the GRN is a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3.

[0333] E240. The pharmaceutical composition described in E239, wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3.

[0334] E241. The pharmaceutical composition described in E240, wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3.

[0335] E242. The pharmaceutical composition described in E241, wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:3.

[0336] E243. A pharmaceutical composition described in any one of E181 to E242, wherein the GRN is a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4.

[0337] E244. The pharmaceutical composition described in E243, wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4.

[0338] E245. The pharmaceutical composition described in E244, wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4.

[0339] E246. The pharmaceutical composition described in E245, wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:4.

[0340] E247. The pharmaceutical composition described in any one of E181 to E246, wherein the GRN is a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5.

[0341] E248. The pharmaceutical composition described in E247, wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5.

[0342] E249. The pharmaceutical composition described in E248, wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5.

[0343] E250. The pharmaceutical composition described in E249, wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:5.

[0344] E251. The pharmaceutical composition described in any one of E181 to E250, wherein the GRN-4 domain has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:6.

[0345] E252. The pharmaceutical composition described in E251, wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6.

[0346] E253. The pharmaceutical composition described in E252, wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:6.

[0347] E254. The pharmaceutical composition described in E253, wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:6.

[0348] E255. A pharmaceutical composition described in any one of E181 to E254, wherein the GRN is a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7.

[0349] E256. The pharmaceutical composition described in E255, wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7.

[0350] E257. The pharmaceutical composition described in E256, wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7.

[0351] E258. The pharmaceutical composition described in E257, wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:7.

[0352] E259. A pharmaceutical composition described in any one of E181 to E258, wherein the GRN is a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8.

[0353] E260. The pharmaceutical composition described in E259, wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8.

[0354] E261. The pharmaceutical composition described in E260, wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8.

[0355] E262. The pharmaceutical composition described in E261, wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:8.

[0356] E263. A pharmaceutical composition described in any one of E181 to E262, wherein the GRN is a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9.

[0357] E264. The pharmaceutical composition described in E263, wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9.

[0358] E265. The pharmaceutical composition described in E264, wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9.

[0359] E266. The pharmaceutical composition described in E265, wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:9.

[0360] E267. The pharmaceutical composition described in any one of E181 to E266, wherein the GRN is a full-length GRN.

[0361] E268. The pharmaceutical composition of any one of E181 to E267, wherein the cell comprises a PGRN transgene having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO:10.

[0362] E269. The pharmaceutical composition described in E268, wherein the PGRN transgene has at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO:10.

[0363] E270. The pharmaceutical composition described in E269, wherein the PGRN transgene has at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO:10.

[0364] E271. The pharmaceutical composition described in E270, wherein the PGRN transgene has the nucleic acid sequence of SEQ ID NO:10.

[0365] E272. The pharmaceutical composition described in any one of E181 to E271, wherein the PGRN or GRN is a PGRN or GRN fusion protein.

[0366] E273. The pharmaceutical composition of E272, wherein the PGRN or GRN fusion protein comprises the Rb domain of ApoE.

[0367] E274. The pharmaceutical composition of E273, wherein the Rb domain comprises a portion of ApoE having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO:11.

[0368] E275. The pharmaceutical composition of E273 or E274, wherein the Rb domain comprises a region having at least 70% sequence identity to the amino acid sequence of residues 159 to 167 of SEQ ID NO:11.

[0369] E276. The pharmaceutical composition described in any one of E181 to E275, wherein the introduced gene encoding PGRN or GRN further comprises an miRNA targeting sequence in the 3'-UTR.

[0370] E277. The pharmaceutical composition described in E276, wherein the miRNA targeting sequence is a miR-126 targeting sequence.

[0371] E278. The pharmaceutical composition according to any one of E181 to E277, wherein the cells are ESCs.

[0372] E279. The pharmaceutical composition according to any one of E181 to E277, wherein the cell is an iPSC.

[0373] E280. The pharmaceutical composition according to any one of E181 to E277, wherein the cells are CD34+ cells.

[0374] E281. The pharmaceutical composition of E280, wherein the CD34+ cells are HSCs.

[0375] E282. The pharmaceutical composition of E280, wherein the CD34+ cells are MPCs.

[0376] E283. The pharmaceutical composition described in any one of E181 to E282, wherein the cells are transfected ex vivo to express the PGRN or the GRN.

[0377] E284. The pharmaceutical composition described in any one of E181 to E282, wherein the cells are transduced ex vivo to express the PGRN or the GRN.

[0378] E285. The pharmaceutical composition of any one of E181 to E284, formulated for systemic administration to a human subject.

[0379] E286. The pharmaceutical composition of E285, formulated for administration to a human subject by intravenous injection.

[0380] E287. The pharmaceutical composition of any one of E181 to E284, formulated for administration to a human subject directly to the nervous system of said subject.

[0381] E288. The pharmaceutical composition of E287, formulated for administration to a human subject into cerebrospinal fluid.

[0382] E289. The pharmaceutical composition of E287 or E288, formulated for administration to a human subject by intraventricular injection, intrathecal injection, stereotactic injection, or a combination thereof.

[0383] E290. The pharmaceutical composition of E287, formulated for administration to a human subject by intraparenchymal injection.

[0384] E291. The pharmaceutical composition of any one of E181 to E284, formulated for administration directly to the bone marrow of a human subject.

[0385] E292. The pharmaceutical composition of E291, formulated for administration to a human subject by intraosseous injection.

[0386] E293. The pharmaceutical composition of any one of E181 to E284, formulated for administration to a human subject by bone marrow transplantation comprising said composition.

[0387] E294. The pharmaceutical composition of any one of E181 to E284, formulated for systemic administration to a human subject and for direct administration to the central nervous system of a human subject.

[0388] E295. The pharmaceutical composition of E294, which is formulated for administration by intraventricular and intravenous injection.

[0389] E296. The pharmaceutical composition of E294, which is formulated for administration by intrathecal and intravenous injection.

[0390] E297. The pharmaceutical composition according to E294, which is formulated for administration by intraparenchymal and intravenous injection.

[0391] E298. The pharmaceutical composition of any one of E285 to E297, wherein the human subject has been diagnosed with an NCD.

[0392] E299. The pharmaceutical composition according to E298, wherein said NCD is a severe NCD.

[0393] E300. The pharmaceutical composition of E299, wherein the severe NCD interferes with the subject's independence and / or normal daily functioning.

[0394] E301. The pharmaceutical composition of E299 or E300, wherein said severe NCD is associated with a score obtained by said subject on a cognitive test that is at least 2 standard deviations away from the mean score of a reference population.

[0395] E302. The pharmaceutical composition according to E298, wherein said NCD is a mild NCD.

[0396] E303. The pharmaceutical composition of E302, wherein said mild NCD does not interfere with the subject's independence and / or normal daily functioning.

[0397] E304. The pharmaceutical composition of E302 or E303, wherein said mild NCD is associated with a score obtained by said subject on a cognitive test that is at least 1-2 standard deviations away from the mean score of a reference population.

[0398] E305. The pharmaceutical composition according to E301 or E304, wherein said reference population is a general population.

[0399] E306. The pharmaceutical composition of E301, E304, or E305, wherein the cognitive test is selected from the group consisting of AD8, AWV, GPCOG, HRA, MIS, MMSE, MoCA, SLUMS, and Short IQCODE.

[0400] E307. The pharmaceutical composition of any one of E298 to E306, wherein the NCD is associated with impairment in one or more of complex attention, executive function, learning and memory, language, sensorimotor function, and social cognition.

[0401] E308. The pharmaceutical composition of any one of E298 to E307, wherein the NCD is not due to delirium or other psychiatric disorder.

[0402] E309. The pharmaceutical composition according to any one of E298 to E308, wherein the NCD is a frontotemporal NCD.

[0403] E310. The pharmaceutical composition described in E309, wherein the frontotemporal NCD is FTLD.

[0404] E311. The pharmaceutical composition according to any one of E298 to E308, wherein the NCD is due to a lysosomal disease.

[0405] E312. The pharmaceutical composition of E311, wherein the lysosomal disease is NCL.

[0406] E313. A kit comprising the pharmaceutical composition according to any one of E181 to E312 and an attached document.

[0407] E314. The kit described in E313, wherein the package insert instructs a user of the kit to perform a method according to any one of E1 to E180.

[0408] E315. The method according to any one of E1 to E180, wherein the PGRN or GRN fusion protein comprises PGRN or GRN and a GILT tag.

[0409] E316. The method of E315, wherein the GILT tag is operably linked to the N-terminus of the PGRN or GRN.

[0410] E317. The method of E315, wherein the GILT tag is operably linked to the C-terminus of the PGRN or GRN.

[0411] E318. The method according to any one of E315 to 317, wherein the GILT tag comprises a human IGF-II mutein having an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II (SEQ ID NO: 12).

[0412] E319. The method of any one of E315 to 318, wherein said IGF-II mutein comprises a mutation within a region corresponding to amino acids 30 to 40 of SEQ ID NO: 12, and said mutation destroys at least one Furin protease cleavage site.

[0413] E320. The method according to E319, wherein the mutation is an amino acid substitution, deletion, and / or insertion.

[0414] E321. The method according to E320, wherein the mutation is a Lys or Ala amino acid substitution at a position corresponding to Arg37 or Arg40 of SEQ ID NO:12.

[0415] E322. The method according to E320, wherein the mutation is a deletion or substitution of amino acid residues corresponding to positions selected from the group consisting of 31 to 40, 32 to 40, 33 to 40, 34 to 40, 30 to 39, 31 to 39, 32 to 39, 34 to 37, 33 to 39, 35 to 39, 36 to 39, 37 to 40, and 34 to 40 of SEQ ID NO: 12, or a combination thereof.

[0416] E323. The method described in any one of E315 to E322, wherein the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO:13.

[0417] E324. The method described in any one of E315 to E322, wherein the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO:14.

[0418] E325. The method described in any one of E315 to E322, wherein the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 15.

[0419] E326. The method described in any one of E315 to E322, wherein the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 16.

[0420] E327. The method described in any one of E315 to E322, wherein the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO:17.

[0421] E328. The method described in any one of E315 to E322, wherein the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO:18.

[0422] E329. The method according to any one of E1 to E180 or E315 to E328, wherein the cell is a pluripotent cell (e.g., ESC, iPSC), a multipotent cell (e.g., a CD34+ cell, such as an HSC or MPC), a BLPC, a monocyte, a macrophage, a microglial progenitor cell, or a microglia.

[0423] E330. The method according to any one of E1 to E180 or E315 to E329, wherein the transgene can be expressed in macrophages or microglial cells.

[0424] E331. The method of any one of E1 to E180 or E315 to E330, wherein the transgene is a codon optimized transgene.

[0425] E332. The method of E331, wherein the codon-optimized transgene comprises a polynucleotide having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO:19.

[0426] E333. The composition described in any one of E181 to E312, wherein the PGRN or GRN fusion protein comprises PGRN or GRN and a GILT tag.

[0427] E334. The composition described in E333, wherein the GILT tag is operably linked to the N-terminus of the PGRN or GRN.

[0428] E335. The composition described in E333, wherein the GILT tag is operably linked to the C-terminus of the PGRN or GRN.

[0429] E336. The composition according to any one of E333 to E335, wherein the GILT tag comprises a human IGF-II mutein having an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II (SEQ ID NO: 12).

[0430] E337. The composition of any one of E333 to E336, wherein said IGF-II mutein comprises a mutation within a region corresponding to amino acids 30 to 40 of SEQ ID NO: 12, and said mutation destroys at least one Furin protease cleavage site.

[0431] E338. The composition of E337, wherein the mutation is an amino acid substitution, deletion, and / or insertion.

[0432] E339. The composition of E338, wherein the mutation is a Lys or Ala amino acid substitution at a position corresponding to Arg37 or Arg40 of SEQ ID NO:12.

[0433] E340. The composition according to E338, wherein the mutation is a deletion or substitution of amino acid residues corresponding to positions selected from the group consisting of 31 to 40, 32 to 40, 33 to 40, 34 to 40, 30 to 39, 31 to 39, 32 to 39, 34 to 37, 33 to 39, 35 to 39, 36 to 39, 37 to 40, and 34 to 40 of SEQ ID NO: 12, or a combination thereof.

[0434] E341. The composition described in any one of E333 to E340, wherein the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 13.

[0435] E342. The composition described in any one of E333 to E340, wherein the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 14.

[0436] E343. The composition described in any one of E333 to E340, wherein the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 15.

[0437] E344. The composition described in any one of E333 to E340, wherein the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 16.

[0438] E345. The composition described in any one of E333 to E340, wherein the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO:17.

[0439] E346. The composition described in any one of E333 to E340, wherein the GILT tag has a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO:18.

[0440] E347. The composition of any one of E181 to E312 or E333 to E346, wherein the cell is a pluripotent cell (e.g., an ESC, an iPSC), a multipotent cell (e.g., a CD34+ cell, such as an HSC or MPC), a BLPC, a monocyte, a macrophage, a microglial progenitor cell, or a microglia.

[0441] E348. The composition of any one of E181 to E312 or E333 to E347, wherein the transgene is capable of being expressed in a macrophage or a microglial cell.

[0442] E349. The composition of any one of E1 to E180 or E333 to E348, wherein the transgene is a codon-optimized transgene.

[0443] E350. The composition of E349, wherein the codon-optimized transgene comprises a polynucleotide having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO:19. [Brief description of the drawings]

[0444] [Figure 1A] A series of plots showing transduction of human cells with lentiviral vectors containing a transgene encoding the human progranulin (PGRN) protein. Cell lysates were generated from human 239T cells transduced with lentiviral vectors encoding PGRN (MND.GRN) or green fluorescent protein (GFP; MND.GFP) at a multiplicity of infection (MOI) of 10, 50, 100, or 200. A separate set of control cells was not transduced (NTC). PGRN levels were quantified relative to actin using densitometry. [Figure 1B]Western blots using an antibody raised against human PGRN showed stable PGRN expression in 239T cells, with maximum expression observed at MOI 200. All groups showed statistically significant differences, except for NTC cells and MOI 10 GFP cells. Statistical analysis was performed using ANOVA. [Diagram 2] 1 is a Western blot showing expression of human PGRN in mouse lineage negative (Lin-) cells transduced with a lentiviral vector containing a transgene encoding human PGRN (i.e., the MND.GRN vector). Western blots using an antibody raised against human PGRN were used to analyze conditioned medium generated from Lin- mouse cells that were untransduced (-) or transduced (+) with the MND.GRN lentiviral vector, demonstrating release of human PGRN protein into the growth medium by the transduced cells. [Diagram 3] Western blot showing that immortalized cell lines transduced with lentiviral vectors containing a transgene encoding human PGRN are N-linked glycosylated. Cell lysates were generated from human 239T cell lines that were either untransduced (NT1, NT2, NT3, and NT4) or transduced with lentiviral vectors encoding human PGRN produced in four independent rounds of transduction (MND.GRN-1, MND.GRN-2, MND.GRN-3, and MND.GRN-4). Cell lysates were enzymatically digested with either EndoH (E.) or PNGase (P.) enzymes or heated (H.) and analyzed using Western blots with an antibody raised against human Progranulin. Enzymatic digestion with EndoH and PNGase indicates that human PGRN protein produced by transduced cells is N-linked glycosylated. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0445] definition As used herein, the terms "remove," "removing," "removal," and the like refer to the depletion of one or more cells in a population of cells in vivo or ex vivo. In some embodiments of the present disclosure, it may be desirable to remove endogenous cells in a subject (e.g., a subject undergoing treatment for a disease described herein, e.g., neurocognitive disorder (NCD; e.g., frontotemporal lobar degeneration (FTLD) or neuronal ceroid lipofuscinosis (NCL)) prior to administering a therapeutic population of cells to said subject. This may be advantageous, for example, to provide the newly administered cells with an environment in which they can engraft. Removal of the population of cells may be performed in a manner that selectively targets a specific cell type, for example, using an antibody-drug conjugate that binds to an antigen expressed on the target cell and subsequently causes death of the target cell. Additionally or alternatively, removal may be performed in a non-specific manner using cytotoxins that do not focus on a particular cell type, but instead may exert their cytotoxic effects on a variety of different cells. Exemplary agents that can be used to ablate a population of endogenous cells in a subject, such as a population of endogenous microglia or microglia precursor cells in a subject receiving therapy for the treatment of an NCD, are busulfan, PLX3397, PLX647, PLX5622, treosulfan, clodronate liposomes, and combinations thereof. Examples of ablation include depletion of at least 5% (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more) of cells in a population of cells in vivo or in vitro. Quantification of the number of cells in a sample of cells can be performed using various cell counting techniques, such as by using a counting chamber, a Coulter counter, flow cytometry, or other cell counting methods known in the art.

[0446] As used herein, "administration" refers to providing or giving to a subject a cell comprising a transgene (e.g., a transgene that can be expressed in macrophages or microglia) encoding a therapeutic agent (e.g., a progranulin (PGRN) or granulin (GRN)-encoding therapeutic agent, e.g., a pluripotent cell (e.g., an embryonic stem cell (ESC) or an induced pluripotent stem cell (ISPC)), a multipotent cell (e.g., a CD34+ cell, such as a hematopoietic stem cell (HSC) or a myeloid progenitor cell (MPC)), a blood lineage progenitor cell (BLPCS; e.g., a monocyte), a macrophage, a microglial progenitor cell, or a microglia) by any effective route. Exemplary routes of administration are described herein and below (e.g., intracerebroventricular (ICV) injection, intravenous (IV) injection, intrathecal (IT) injection, intraparenchymal (IP) injection, and stereotactic injection).

[0447] As used herein, "allogeneic" refers to cells, tissues, DNA, or factors taken or derived from a different subject of the same species. For example, in the situation where transduced PGRN-expressing or GRN-expressing cells are administered to a subject to treat NCD, the allogeneic cells may be cells obtained from a subject other than the subject and then transduced or transfected with a vector directing the expression of PGRN or GRN. The phrase "directing expression" refers to a polynucleotide that comprises a sequence that codes for a molecule to be expressed. The polynucleotide may comprise additional sequences that enhance the expression of the molecule of interest.

[0448] As used herein, "autologous" refers to cells, tissues, DNA, or factors that are taken or derived from an individual's own tissues, cells, or DNA.For example, in the situation where transduced PGRN-expressing or GRN-expressing cells are administered to a subject to treat NCD (e.g., FTLD or NCL), autologous cells can be cells that are obtained from the subject and then transduced or transfected with a vector that directs the expression of PGRN or GRN.

[0449] As used herein, the term "ApoE" refers to apolipoprotein E, a member of a class of proteins involved in lipid transport. Apolipoprotein E is a fat-binding protein (apolipoprotein) that is part of chylomicrons and intermediate density lipoproteins (IDL). They are essential for normal processing (catabolism) of triglyceride-rich lipoproteins. ApoE is encoded by the APOE gene. The term "ApoE" also refers to variants of wild-type ApoE protein, such as proteins having at least 85% identity (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.9% identity or more) to the amino acid sequence of wild-type ApoE shown in SEQ ID NO:11.

[0450] As used herein, the term "blood lineage progenitor cells" or "BLPCs" refers to any cell (e.g., mammalian cells) that can differentiate into one or more (e.g., 2, 3, 4, 5 or more) types of hematopoietic (i.e., blood) cells. BLPCs can differentiate into red blood cells, white blood cells (e.g., granulocytes (e.g., basophils, eosinophils, neutrophils, and mast cells) or agranulocytes (e.g., lymphocytes and monocytes), etc.), or platelets. BLPCs can also include differentiated blood cells (e.g., monocytes) that can further differentiate into another blood cell type (e.g., macrophages).

[0451] As used herein, the term "cell type" refers to a group of cells that share a statistically distinguishable phenotype based on gene expression data. For example, cells of a common cell type may share similar structural and / or functional characteristics, such as similar gene activation patterns and antigen presentation profiles. Cells of a common cell type may include those isolated from a common tissue (e.g., epithelial, nervous, connective, or muscle tissue) and / or those isolated from a common organ, tissue system, blood vessel, or other structure and / or region of the body.

[0452] As used herein, "codon optimization" refers to the process of modifying a nucleic acid sequence according to the principle that the frequency of occurrence of synonymous codons (e.g., codons that code for the same amino acid) in coding DNA is biased in different species. Such codon degeneracy allows the same polypeptide to be encoded by a variety of nucleotide sequences. Sequences modified in this manner are referred to herein as "codon optimization." This process may be performed on any of the sequences described herein to enhance expression or stability. Codon optimization may be performed, for example, in the manner described in U.S. Patent Nos. 7,561,972, 7,561,973, and 7,888,112, each of which is incorporated herein by reference in its entirety. The sequence surrounding the translation start site may be converted to a consensus Kozak sequence according to known methods. See, for example, Kozak et al, Nucleic Acids Res. 15: 8125-8148(1989), which is incorporated herein by reference in its entirety. Multiple stop codons may be incorporated.

[0453] As used herein, the term "cognitive test" refers to a test that can be performed by a skilled practitioner to evaluate the cognitive abilities of humans and other animals. Cognitive tests can be used to evaluate inductive reasoning skills, IQ, cognitive development, memory, knowledge organization, metacognition, thinking, and mental time measurement. Cognitive tests can be used to evaluate the processing abilities of subjects across multiple cognitive domains, including, but not limited to, executive function, learning and memory, language, sensory-motor function, and social cognition. Examples of cognitive tests include, but are not limited to, the Eight-item Informant Interview to Differentiate Aging and Dementia (AD8), Annual Wellness Visit (AWV), General Practitioner Assessment of Cognition (GPCOG), Health Risk Assessment (HRA), Memory Impairment Screen (MIS), Mini Mental Status Exam (MMSE), Montreal Cognitive Assessment (MoCA), St. Louis University Mental Status Exam (SLUMS), and Short Informant Questionnaire on Cognitive Decline in the Elderly (Short IQCODE). Those skilled in the art will recognize that other cognitive tests known in the art can also be used to assess cognitive function in a subject.

[0454] As used herein, the term "complex attention" refers to a cognitive function that describes a subject's (e.g., a human subject's) ability to hold information in memory for a short period of time and to manipulate that information (e.g., mental arithmetic). Impairments in complex attention can result in difficulties with focusing on conversations, difficulties with blocking out unwanted information, problems with prospective memory (e.g., memory for remembering something later), and inefficient memory for new information.

[0455] As used herein, the terms "pretreatment" and "pretreating" refer to a process in which a subject is prepared to receive a graft containing cells. Such procedures promote the engraftment of cell grafts, for example, by selectively depleting endogenous microglia or hematopoietic stem cells, thereby creating voids that are filled by exogenous cell grafts. According to the methods described herein, a subject can be pretreated for cell transplantation therapy by administering to the subject one or more agents capable of depleting endogenous microglia and / or hematopoietic stem or progenitor cells (e.g., busulfan, treosulfan, PLX3397, PLX647, PLX5622, and clodronate liposomes), radiation therapy, or a combination thereof. The pretreatment can be myeloablative or non-myeloablative. Other cell depleting agents and methods known in the art (e.g., antibody-drug conjugates) can also be used.

[0456] As used herein, the terms "conservative mutation," "conservative substitution," and "conservative amino acid substitution" refer to the replacement of one or more amino acids with one or more different amino acids that exhibit similar physicochemical properties, such as polarity, electrostatic charge, and steric bulk, which are summarized in Table 1 below for each of the 20 naturally occurring amino acids.

[0457] [Table 1]

[0458] From this table it can be seen that conservative amino acid families include: (i) G, A, V, L, and I; (ii) D and E; (iii) C, S, and T; (iv) H, K, and R; (v) N and Q; and (vi) F, Y, and W. Thus, a conservative variation or substitution is one that substitutes one amino acid for a member of the same amino acid family (e.g., Thr for Ser, Arg for Lys).

[0459] As used herein, the term "delirium or other psychiatric disorder" refers to a condition such as delirium (i.e., a syndrome involving impaired attention, consciousness, and cognition that develops over a short period of time (e.g., hours to days)) or another mental disorder (e.g., schizophrenia, bipolar disorder, and major depression) that is distinct from neurocognitive disorders and does not exhibit cognitive dysfunction as a core symptom. For example, a condition such as delirium or another psychiatric disorder may differ from an NCD in that cognitive dysfunction may be a symptom associated with the disease but is not a core feature of the disease. Delirium or another psychiatric disorder may differ from an NCD in terms of time to onset (e.g., hours to days for delirium versus months to years for NCD), etiology (e.g., substance-induced delirium), duration of symptoms (e.g., delirium may last days to hours, while NCDs may last for years), and resolution (e.g., delirium may resolve completely, while NCDs often do not).

[0460] As used herein, the term "disrupt" with respect to a gene refers to preventing the formation of a functional gene product. A gene product is functional if it fulfills its normal (wild-type) function. Disruption of a gene prevents the expression of a functional factor encoded by the gene and includes the insertion, deletion, or substitution of one or more bases in the sequence encoded by the gene and / or the promoter and / or operator required for the expression of the gene in the animal. The gene to be disrupted may be, for example, by removing at least a portion of the gene from the genome of the animal, modifying the gene to prevent the expression of the functional factor encoded by the gene, expressing a dominant negative factor by an interfering RNA, or an exogenous gene. Materials and methods for genetically modifying a cell to disrupt the expression of one or more genes are described in detail in US8,518,701; US9,499,808; and US2012 / 0222143, the disclosures of each of which are incorporated herein by reference in their entirety (in case of conflict, the present specification takes precedence).

[0461] As used herein, the terms "effective amount," "therapeutically effective amount," and "sufficient amount" of a composition, vector construct, viral vector, or cell described herein refer to an amount sufficient to produce a beneficial or desired result, including a clinical result, when administered to a subject, including a mammal, e.g., a human. Thus, an "effective amount" or its synonyms depend on the context in which it is applied. For example, in the context of treating an NCD (e.g., FTLD or NCL), it is the amount of the composition, vector construct, viral vector, or cell sufficient to achieve a treatment response compared to the response obtained when the composition, vector construct, viral vector, or cell is not administered. The amount of a given composition described herein that corresponds to such an amount will vary depending on various factors, such as a given drug, pharmaceutical formulation, route of administration, type of disease or disorder, subject identity (e.g., age, sex, weight) or recipient to be treated, but can nevertheless be routinely determined by one of skill in the art. Also, as used herein, a "therapeutically effective amount" of a composition, vector construct, viral vector, or cell of the present disclosure is an amount that produces a beneficial or desired result in a subject compared to a control. As defined herein, a therapeutically effective amount of a composition, vector construct, viral vector, or cell of the present disclosure can be readily determined by one of ordinary skill in the art by routine methods known in the art. Dosage regimens can be adjusted to obtain the optimal therapeutic response.

[0462] As used herein, the terms "embryonic stem cell" and "ES cell" refer to embryo-derived totipotent or pluripotent stem cells derived from the inner cell mass of a blastocyst that can be maintained in in vitro culture under appropriate conditions. ES cells can differentiate into cells of any of the three vertebrate germ layers, e.g., endoderm, ectoderm, and mesoderm. ES cells are also characterized by their ability to grow indefinitely under appropriate in vitro culture conditions. See, e.g., Thomson et al., Science 282:1145 (1998).

[0463] As used herein, the term "endogenous" describes a molecule (e.g., a polypeptide, nucleic acid, or cofactor) that is found naturally in a particular organism (e.g., a human) or in a particular location within an organism (e.g., an organ, tissue, or cell such as a human cell).

[0464] As used herein, the terms "engraft" and "engraftment" refer to the process of repopulating tissues with hematopoietic stem and progenitor cells (such cells either produced endogenously within the body or transplanted using any of the administration methods described herein (e.g., intravenous injection, intraventricular injection, intraosseous injection, and / or bone marrow transplantation). The terms encompass all events relating to or resulting in engraftment, such as tissue homing of cells and colonization of cells within a tissue of interest.

[0465] As used herein, the term "executive function" refers to a set of cognitive functions that facilitate cognitive control of behavior in a subject (e.g., human).Executive function includes, for example, goal-directed behavior selection and monitoring, attention control, cognitive inhibition, inhibitory control, working memory, and cognitive flexibility.Individuals normally acquire or perfect executive function throughout life, but this process may be derailed by the onset of NCD in a subject, which may adversely affect executive function.

[0466] As used herein, the term "express" refers to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, 5' capping, and / or 3' end processing); (3) translation of the RNA into a polypeptide or protein; and (4) post-translational modification of the polypeptide or protein. Expression of a gene of interest in a subject can be revealed, for example, by detecting an increase in the amount or concentration of mRNA encoding the corresponding protein (e.g., assessed using RNA detection procedures described herein or known in the art, such as quantitative polymerase chain reaction (qPCR) and RNA seq techniques), an increase in the amount or concentration of the corresponding protein (e.g., assessed using protein detection methods described herein or known in the art, such as enzyme-linked immunosorbent assay (ELISA), among others), and / or an increase in the activity of the corresponding protein (e.g., in the case of an enzyme, assessed using enzyme activity assays described herein or known in the art) in a sample obtained from the subject.

[0467] As used herein, the term "exogenous" describes a molecule (e.g., a polypeptide, nucleic acid, or cofactor) that is not naturally found in a particular organism (e.g., a human) or in a particular location within an organism (e.g., an organ, tissue, or cell such as a human cell). Exogenous materials include those provided to an organism from an outside source or extracted from an organism in culture.

[0468] As used herein, the term "functional potential" in relation to stem cells, such as hematopoietic stem cells, refers to functional properties of stem cells including: 1) multipotency (referring to the ability to differentiate into multiple different blood cell lineages, including, but not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes, red blood cells), platelets (e.g., megakaryoblasts, thrombocytic megakaryocytes, platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NK cells, B cells, and T cells)); 2) self-renewal (referring to the ability of stem cells to give rise to daughter cells that have the same potential as the mother cell and that may arise repeatedly throughout an individual's life without this potential being exhausted); and 3) the ability of the stem cell or its progeny, when reintroduced into a transplant recipient, to home to the stem cell niche and re-establish productive, sustained cell growth and differentiation.

[0469] As used herein, the term "furin-resistant IGF-II mutein" refers to an insulin-like growth factor II (IGF-II)-based peptide that contains an altered amino acid sequence compared to wild-type IGF-II (SEQ ID NO: 12) that either disables at least one native furin protease cleavage site or alters a sequence near or adjacent to a native furin protease cleavage site such that furin cleavage is blocked, inhibited, reduced, or slowed down compared to the wild-type human IGF-II peptide. As used herein, a furin-resistant IGF-II mutein is also referred to as an IGF-II mutein that is resistant to furin. An exemplary furin-resistant IGF-II mutein contains an amino acid substitution at a position corresponding to Arg37 and / or Arg40 of SEQ ID NO: 12.

[0470] As used herein, the term "Furin protease cleavage site" (also referred to as "Furin cleavage site" or "Furin cleavage sequence") refers to an amino acid sequence of a peptide or protein that serves as a recognition sequence for enzymatic protease cleavage by a furin or furin-like protease. Typically, a furin protease cleavage site has the consensus sequence Arg-XX-Arg, where X is any amino acid. The cleavage site is located after a carboxy-terminal arginine (Arg) residue in the sequence. In some embodiments, a furin cleavage site has the consensus sequence Lys / Arg-XXX-Lys / Arg-Arg, where X is any amino acid. The cleavage site is located after a carboxy-terminal arginine (Arg) residue in the sequence.

[0471] As used herein, the term "furin" refers to any protease capable of recognizing and cleaving the furin protease cleavage site as defined herein, including furin or furin-like proteases. Furin is also known as paired basic amino acid cleaving enzyme (PACE). Furin belongs to the subtilisin-like proprotein convertase family. The gene encoding furin is known as FUR (FES Upstream Region).

[0472] As used herein, the term "glycosylation-independent lysosomal targeting" or "GILT" refers to lysosomal targeting that is mannose-6-phosphate (M6P) independent. The GILT tag can be used to target a protein (e.g., GBA) expressed as a GILT-tagged fusion protein (e.g., GBA fusion protein linked to an IGF-II mutein) to lysosomes.

[0473] As used interchangeably herein, the terms "cation-independent mannose-6-phosphate receptor (CI-MPR)", "M6P / IGF-II receptor", "CI-MPR / IGF-II receptor", "IGF-II receptor" or "IGF2 receptor", or their abbreviations, refer to a cellular receptor that binds both M6P and IGF-II.

[0474] As used herein, the terms "frontotemporal lobar degeneration" and "FTLD" refer to a complex clinical syndrome characterized by the degeneration of brain tissue in the frontal and temporal lobes of the cerebral cortex. The terms "frontotemporal lobar degeneration" and "FTLD" can refer to any one of three clinically distinct variants of FTLD, including: 1) behavioral and personality changes, emotional blunting, social withdrawal, peremptory behavior, attention deficit, loss of inhibition, and prominent degeneration of the frontal lobe. In addition, BVFTD has a strong relationship with amyotrophic lateral sclerosis; 2) semantic dementia (SD) is characterized by verbal amnesic aphasia, progressive loss of semantic knowledge of words, objects, and concepts, and prominent degeneration of the anterior temporal lobe. Furthermore, the SD variant of FTLD exhibits flat affect, social deficits, perseverative behavior, and loss of inhibition; 3) progressive non-verbal aphasia (PNA) is characterized by motor impairments in speech production, reduced language expression, and significant degeneration of the perisylvian cortex. The histopathological profile of FTLD patients generally falls into one of three broad phenotypes, including those showing aggregation and deposition of (i) microtubule-associated tau protein inclusions; (ii) tau-negative, ubiquitin and TAR DNA-binding protein 43 (TDP-43)-positive protein inclusions, or (iii) ubiquitin and sarcoma fusion (FUS)-positive protein inclusions. For a comprehensive description of the clinical symptoms and histopathology of FTLD, see Rabinovici and Miller, CNS Drugs 24:375-398 (2010), the disclosure of which is incorporated herein by reference in its entirety.

[0475] As used herein, the term "general population" refers to the entire population of individuals with a particular characteristic of interest (e.g., age, medical history, education, socioeconomic status, or lifestyle, among others). Alternatively, the term "general population" may refer to a subset of the entire population of individuals with a particular characteristic of interest, such as, for example, a random sample with a defined sample size. In the methods disclosed herein, the general population can serve as a working standard (e.g., a reference population) against which a measured variable can be compared. For example, subjects who have been diagnosed with NCD can be evaluated for their cognition using a cognitive test disclosed herein, and the score obtained by the subject on the test can be compared to the performance of individuals in the general population (e.g., the entire general population or a random sample of the general population) on the same test. The size of the random sample of the general population can be determined by a skilled practitioner using methods well known in the art. For example, a skilled practitioner can perform a power analysis before collecting data (e.g., before administering a cognitive test to the subject) to determine the minimum sample required to detect a statistically significant effect with a desired level of confidence.

[0476] As used herein, the terms "granulin" and "GRN" refer to peptide products resulting from cleavage of the precursor protein PGRN. GRN peptides are involved in a variety of biological functions, including development, immunity, cell survival and proliferation, and tumorigenesis. Full-length wild-type human PGRN peptide has 7.5 GRN domains (e.g., 7 GRN domains, each about 60 amino acids long) and a 30 amino acid paragranulin (para-GRN) domain that can be individually cleaved by proteases. The terms "granulin" and "GRN" also refer to variants of wild-type human granulin peptide and nucleic acids encoding same, such as variant proteins having at least 85% sequence identity (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.9% identity or more) to any of the amino acid sequences of the wild-type GRN peptide (e.g., any one of SEQ ID NOs: 2-9), provided that the encoded GRN variant maintains the therapeutic function of the wild-type GRN. The terms "granulin" and "GRN" can also refer to GRN proteins in which the native secretory signal peptide is present. In addition, the terms "granulin" and "GRN" can refer to a "GRN fusion protein," which is a protein in which GRN is operably linked to another polypeptide, half-life modifying agent, or therapeutic agent, such as an ApoE receptor binding (Rb) domain (e.g., an Rb domain having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO: 11). As used herein, the term "GRN" can refer to the peptide or the gene encoding the protein, depending on the context, as will be appreciated by one of skill in the art.

[0477] As used herein, the terms "hematopoietic stem cells" and "HSCs" refer to immature blood cells that have the ability to self-renew and differentiate into mature blood cells of various cell lineages, including, but not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes, red blood cells), platelets (e.g., megakaryoblasts, thrombocytic megakaryocytes, platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NK cells, B cells, and T cells). It is known in the art that such cells may or may not include CD34+ cells. CD34+ cells are immature cells that express the CD34 cell surface marker. In humans, CD34+ cells are believed to include a subpopulation of cells with stem cell characteristics as defined above, while in mice, HSCs are CD34-. In addition, HSCs are also referred to as long-term repopulating HSCs (LT-HSCs) and short-term repopulating HSCs (ST-HSCs). LT-HSCs and ST-HSCs are differentiated based on functional potential and expression of cell surface markers. For example, human HSCs are CD34+, CD38-, CD45RA-, CD90+, CD49F+, and lin- (negative for mature cell lineage markers including CD2, CD3, CD4, CD7, CD8, CD10, CD11B, CD19, CD20, CD56, CD235A). In mice, bone marrow LT-HSCs are CD34-, SCA-1+, C-kit+, CD135-, Slamf1 / CD150+, CD48-, and lin- (negative for mature lineage markers including Ter119, CD11b, Gr1, CD3, CD4, CD8, B220, IL-7ra), whereas ST-HSCs are CD34+, SCA-1+, C-kit+, CD135-, Slamf1 / CD150+, and lin- (negative for mature lineage markers including Ter119, CD11b, Gr1, CD3, CD4, CD8, B220, IL-7ra). In addition, ST-HSCs are less quiescent (i.e., more active) and more proliferative than LT-HSCs under homeostatic conditions.However, LT-HSC has a higher self-renewal capacity (i.e., it can survive through adulthood and be continuously transplanted through successive recipients), whereas ST-HSC has a limited self-renewal capacity (i.e., it only survives for a limited period of time and does not have the ability to be continuously transplanted). Any of these HSCs can be used in any of the methods described herein. Optionally, ST-HSC is useful because it is highly proliferative and therefore can generate differentiated progeny more quickly.

[0478] As used herein, the term "HLA-matched" refers to a donor-recipient pair in which there is no mismatch of any of the HLA antigens between the donor and the recipient, such as a donor providing a hematopoietic stem cell graft to a recipient in need of hematopoietic stem cell transplantation therapy. HLA-matched (i.e., all six alleles matched) donor-recipient pairs have a lower risk of graft rejection because endogenous T and NK cells are less likely to recognize the foreign graft as foreign and therefore less likely to mount an immune response against the graft.

[0479] As used herein, the term "HLA-mismatched" refers to a donor-recipient pair in which at least one HLA antigen is mismatched between the donor and the recipient, particularly with respect to HLA-A, HLA-B, HLA-C, and HLA-DR, such as a donor providing a hematopoietic stem cell graft to a recipient in need of hematopoietic stem cell transplantation therapy. In some embodiments, one haplotype is matched and the other is mismatched. HLA-mismatched donor-recipient pairs may be at higher risk of graft rejection compared to HLA-matched donor-recipient pairs, since endogenous T cells and NK cells are more likely to recognize the foreign graft as foreign and therefore such T cells and NK cells are more likely to mount an immune response against the graft.

[0480] As used herein, the phrase "independence and / or normal daily functioning" refers to the ability of a subject (e.g., a human) to successfully perform daily activities without the assistance of a caregiver or social worker. Non-limiting examples of activities that allow an individual to independently perform daily functions include, for example, social, occupational, or academic functions, personal hygiene, grooming, dressing, toilet hygiene, functional mobility (e.g., walking ability, getting in and out of bed), and self-feeding. Subjects diagnosed with severe NCDs may have difficulty independently performing normal daily functions, while subjects diagnosed with mild NCDs may not have difficulty independently performing normal daily functions.

[0481] As used herein, the terms "induced pluripotent stem cell", "iPS cell" and "iPSC" refer to pluripotent stem cells that can be derived directly from differentiated somatic cells. Human iPS cells can be generated by introducing a specific set of reprogramming factors into non-pluripotent cells, which can include, for example, Oct3 / 4, Sox family transcription factors (e.g., Sox1, Sox2, Sox3, Sox15), Myc family transcription factors (e.g., c-Myc, 1-Myc, n-Myc), Krüppel-like family (KLF) transcription factors (e.g., KLF1, KLF2, KLF4, KLF5), and / or related transcription factors such as NANOG, LIN28 and / or Glis1. Human iPS cells can also be generated by the use of, for example, miRNAs, small molecules that mimic the action of transcription factors, or cell lineage specifiers. Human iPS cells are characterized by their ability to differentiate into any of the three vertebrate germ layers, such as endoderm, ectoderm, or mesoderm. Human iPS cells are also characterized by their ability to grow indefinitely under appropriate in vitro culture conditions. See, for example, Takahashi and Yamanaka, Cell 126:663 (2006).

[0482] As used herein, the term "IRES" refers to internal ribosome entry site. In general, IRES sequence is a function that allows eukaryotic ribosomes to bind to mRNA transcripts and start translation without binding to the 5' cap end. The mRNA that contains IRES sequence generates two translation products, one that starts from the 5' end of the mRNA and the other that starts from the internal translation mechanism mediated by IRES.

[0483] As used herein, the term "language" refers to the cognitive ability of a subject to learn and use complex systems of communication or to describe the rules governing these systems or the set of utterances that can be produced from such rules. Language abilities may be impaired in a subject with an NCD if the subject exhibits, for example, a limited vocabulary, an inability to produce complex grammar, frequent lexical errors, or aphasia, among other things.

[0484] As used herein, the phrase "learning and memory" refers to cognitive abilities that encompass the acquisition of skills or knowledge and the expression of acquired skills or knowledge (e.g., learning to say new words and speaking new words, respectively). "Learning and memory" can refer to two independent processes: 1) the acquisition of new skills or knowledge (i.e., learning); and 2) the processing, storage, and recall of learned skills or knowledge (i.e., remembering), which can differ by time scale (learning is generally slower and more effortful than memory recall or execution of learned skills) and neurobiological basis. Subjects diagnosed with NCDs may have impaired learning and memory compared to healthy subjects.

[0485] As used herein, the term "lysosomal disease" refers to a large set of about 50 genetic metabolic disorders resulting from abnormal lysosomal function. Abnormalities in genes involved in lipid (e.g., progranulin gene), glycoprotein, or mucopolysaccharide metabolism are common causes of lysosomal diseases. Accumulation of these molecules in cells ultimately leads to cell death. Common symptoms of lysosomal diseases are highly variable and depend on the specific disease, but may include developmental delay, movement disorders, seizures, dementia, hearing and / or visual dysfunction, liver or spleen enlargement, lung and heart problems, and abnormal bone development. Non-limiting examples of lysosomal diseases include neuronal ceroid lipofuscinosis, sphingolipidoses, galactosialidosis, gangliosidosis, Farber disease, Krabbe disease, Gaucher disease, lysosomal acid lipase deficiency, Niemann-Pick disease, sulfatidosis, mucopolysaccharidoses, mucopolysaccharidoses, lipodoses, alpha-mannosidosis, beta-mannosidosis, aspartylglucosaminuria, fucosidosis, lysosomal transport diseases, glycogen storage diseases, and cholesteryl ester storage diseases.

[0486] As used herein, the term "macrophage" refers to a type of white blood cell that engulfs and digests cell debris, foreign material, microorganisms, cancer cells, and anything else that does not have the types of proteins on its surface that are specific to healthy body cells, in a process called phagocytosis. Macrophages are found in essentially all tissues, where they patrol for potential pathogens by amoeboid-like movement. They take on various forms (with various names) throughout the body (e.g., histiocytes, Kupffer cells, alveolar macrophages, microglia, etc.), but are all part of the mononuclear phagocyte system. Besides phagocytosis, they play an important role in non-specific defense (innate immunity) and also help initiate specific defense mechanisms (adaptive immunity) by recruiting other immune cells, such as lymphocytes. For example, they are important as antigen presenters for T cells. Besides increasing inflammation and stimulating the immune system, macrophages also play an important anti-inflammatory role and can dampen immune responses through the release of cytokines.

[0487] As used herein, the term "microglia" or "microglial cells" refers to a type of resident macrophage cell and glial cell found in the brain and spinal cord that serves as the mainstay of immune defense in the central nervous system. The primary functions of microglial cells include immunosurveillance, phagocytosis, extracellular signaling (e.g., production and release of cytokines, chemokines, prostaglandins, and reactive oxygen species), antigen presentation, and promoting tissue repair and regeneration.

[0488] As used herein, the term "microglial progenitor cells" refers to progenitor cells that give rise to microglial cells. Microglial progenitor cells arise in the yolk sac during a limited period of embryonic development, infiltrate the brain mesenchyme, and perpetually renew themselves throughout life.

[0489] As used herein, the term "miRNA targeting sequence" refers to a nucleotide sequence located in the 3'-UTR of a target mRNA molecule that is hybridizable and complementary to a specific miRNA molecule (e.g., miR-126) and promotes RNA-induced silencing complex-dependent and Dicer-dependent mRNA destabilization and / or cleavage, thereby preventing expression of the mRNA transcript.

[0490] As used herein, the term "monocyte" refers to a type of white blood cell (i.e., leukocyte) that can differentiate into macrophages and myeloid lineage dendritic cells. Monocytes constitute an important component of the vertebrate adaptive immune response. Three distinct types of monocytes are known to exist, including classical monocytes (i.e., CD14++CD16-), characterized by strong expression of the CD14 cell surface receptor and no expression of CD16, non-classical monocytes (CD14+CD16++), which show low levels of CD14 expression and co-expression of C16, and intermediate monocytes (CD14++CD16+), which show high levels of CD14 expression and low levels of C16 expression. Monocytes perform a variety of functions that serve the immune system, including phagocytosis, antigen presentation, and cytokine secretion.

[0491] As used herein, the term "pluripotent cells" refers to cells that have the capacity to develop into multiple (e.g., 2, 3, 4, 5, or more) differentiated cell types, although not all of them. Non-limiting examples of pluripotent cells include cells of the hematopoietic lineage (e.g., granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythroid (e.g., reticulocytes, erythrocytes), platelets (e.g., megakaryoblasts, thrombocytic megakaryocytes, platelets), monocytic (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphoid (e.g., NK cells, B cells, and T cells). An example of a pluripotent cell is a CD34+ cell.

[0492] As used herein, the term "mutation" refers to a change in the nucleotide sequence of a gene. Mutations in genes can occur naturally, for example, as a result of DNA replication errors, DNA repair, radiation, and exposure to carcinogens, or mutations can be induced as a result of administration of a transgene that expresses a mutated gene. Examples of mutations can include frameshift, nonsense, missense, insertion, deletion, and transversion mutations. A frameshift mutation can refer to a change in a nucleotide sequence such that the position of the ribosomal reading frame on the mRNA shifts, resulting in improper translation of the RNA. A nonsense mutation can refer to a change in a single nucleotide of a gene that results in a premature stop codon in the transcript. A premature stop codon can result in the translation of a truncated protein product or nonsense-mediated decay of the transcript. A missense mutation can refer to a single nucleotide change in a gene that results in a codon that codes for a different amino acid, which can change the physicochemical properties of the protein product and / or render it nonfunctional. An insertion mutation may refer to the introduction of one or more nucleotides into the coding region of a gene, which may result in a frameshift and typically the production of a premature stop codon. A deletion mutation may refer to the removal of one or more nucleotides from the DNA sequence of a gene, which may result in a frameshift, typically a premature stop codon. A transversion mutation may refer to the change of a single purine nucleotide (e.g., adenine, guanine) to a single pyrimidine nucleotide (e.g., cytosine, thymine). A transversion mutation may not result in a change in the translated protein product (e.g., silent mutation) or may change the amino acid identity within a single codon, thereby altering the physicochemical properties and function of the translated protein product.Nomenclature for mutations and sequence variations uses a "reference sequence code" format, where the reference sequence can be "c" for coding DNA, "g" for genomic DNA, "m" for mitochondrial DNA, "r" for RNA, or "p" for protein, and the code can include symbols including ">" for substitution, "_" for range, ";" for additional changes in one allele, "," for additional transcripts / mosaicism, "()" for unknown changes, "[]" for allele, "del" for deletion, "dup" for duplication, "ins" for insertion, "inv" for inversion, "conv" for conversion, "ext" for extension, "X" for stop codon, "fsX" for frameshift resulting in a stop codon, "o" for reverse strand, and "t" for translocation. For example, the p.T382NfsX32 mutation in the PGRN gene corresponds to a change in the protein at amino acid 382 in which an asparagine has been replaced by a threonine as a result of a frameshift mutation, the length of the frameshift being 32 nucleotide base pairs including the stop codon.

[0493] As used herein, the term "myeloablative" or "myeloablation" refers to a conditioning regimen that substantially damages or destroys the hematopoietic system, typically by exposure to cytotoxic drugs (e.g., busulfan) or radiation. Myeloablation includes complete bone marrow destruction brought about by high doses of cytotoxic agents or total body irradiation that destroy the hematopoietic system.

[0494] As used herein, the term "neurocognitive disorder" or "NCD" refers to a set of clinical disorders or syndromes in which the primary clinical impairment is cognitive function, such as, for example, deficits in complex attention, executive function, learning and memory, language, sensorimotor function, and social cognition. NCDs are characterized as acquired conditions, not developmental conditions. For example, NCDs are conditions in which disrupted cognition has not been evident since birth or very early childhood, thus requiring that cognitive function in NCDs has declined from previously acquired levels. NCDs are distinguished from other disorders in which patients exhibit cognitive impairment, and NCDs include only disorders in which the core impairment is cognition. NCDs can be "severe NCDs" or "mild NCDs." "Severe NCDs" are characterized by significant cognitive decline that interferes with an individual's independence and / or normal daily functioning and is not due to delirium or other psychiatric disorders. Mild NCDs are characterized by moderate cognitive decline that does not interfere with an individual's independence and / or normal daily functioning and is not due to delirium or other psychiatric disorders. Severe and mild NCDs can also be distinguished based on quantitative cognitive testing of any one of the specific cognitive functions described above. For example, severe NCDs can be characterized by a score obtained on a cognitive test by a subject identified as having or at risk of developing an NCD being more than two standard deviations away from the mean score of a reference population (e.g., the mean score of the general population) or being in the third percentile of the distribution of scores of the reference population. Mild NCDs can be characterized by a score obtained on a cognitive test by a subject identified as having or at risk of developing an NCD being one to two standard deviations away from the mean score of a reference population (e.g., the mean score of the general population) or being between the third and sixteenth percentiles of the distribution of scores of the reference population. Non-limiting examples of cognitive tests that can be used to classify NCD patients as having either severe or mild NCD include AD8, AWV, GPCOG, HRA, MIS, MMSE, MoCA, SLUMS, and Short IQCODE.Additionally, NCD (e.g., severe or mild NCD) includes syndromic subtypes that represent a particular etiology of NCD, such as frontotemporal lobar degeneration (FTLD) or lysosomal disease (e.g., neuronal ceroid lipofuscinosis (NCL). As used herein, the terms "frontotemporal NCD" and "NCD due to lysosomal disease" correspond to NCDs resulting from frontotemporal lobar degeneration and lysosomal disease (e.g., NCL), respectively.

[0495] As used herein, the terms "neural ceroid lipofuscinosis" and "NCL" refer to a collection of at least eight clinically recognized lysosomal storage disorders resulting from the accumulation of lipofuscin in cells of the body, such as neurons, liver, spleen, myocardium, and kidney cells. NCL presents clinically with marked neurodegeneration and progressive and irreversible loss of motor and cognitive abilities, although disease severity and clinical symptoms may depend on the particular NCL variant. Known variants of NCL include the juvenile variant, also known as Santovuori-Haltia disease (SHD), the late-onset juvenile variant, also known as Jansky-Bielschowsky disease (JBD), the Finnish late-onset juvenile variant (FLI), the variant with variant late (VLI), the CLN7 variant (CLN7), the CLN8 variant (CLN8), the Turkish late-onset juvenile variant (TLI), the juvenile variant, also known as Batten disease (BD), and the adult variant, also known as Koufs disease (KD). SHD is associated with early visual loss that progresses to complete retinal blindness by age 2, followed by a vegetative state at age 3, and brain death by age 4. This variant is also associated with spontaneous occurrence of epileptic seizures. The JBD variant occurs between 2 and 4 years of age and is associated with ataxia, seizures, progressive cognitive decline, and abnormal speech development, typically resulting in death by age 8 years. BD typically occurs between 4 and 10 years of age and includes symptoms such as vision loss, seizures, cognitive impairment, and early death. NCL patients with the KD variant generally have milder symptoms than the SHD and BD variants and have a life expectancy of about 40 years. For a comprehensive description of NCL, see Mink et al., Journal of Child Neurology 28:1101-5 (2013), Nita et al., Epileptic Disorders 18:73-88 (2016), and Mole & Cotman, Biochimica et Biophysica Acta 1852:2237-41 (2015), the disclosures of which are incorporated herein by reference in their entireties.

[0496] As used herein, the terms "non-myeloablative" or "myelosuppressive" refer to conditioning regimens that do not eliminate substantially all hematopoietic cells of host origin.

[0497] As used herein, the term "pluripotent cell" refers to a cell that has the ability to develop into more than one differentiated cell type, such as hematopoietic (e.g., granulocytic (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythroid (e.g., reticulocytes, red blood cells), platelets (e.g., megakaryoblasts, thrombocytic megakaryocytes, platelets), monocyte (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytic (e.g., NK cells, B cells, and T cells) cell types. Exemplary cells are ESCs and iPSCs.

[0498] As used herein, the term "plasmid" refers to an extrachromosomal circular double-stranded DNA molecule into which additional DNA segments can be ligated. A plasmid is a type of vector, a nucleic acid molecule that can transport another nucleic acid to which it is linked. Certain plasmids can autonomously replicate in a host cell into which they are introduced (e.g., bacterial plasmids with a bacterial origin of replication and episomal mammalian plasmids). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and are thereby replicated along with the host genome. Certain plasmids can direct the expression of genes to which they are operably linked.

[0499] As used herein, the terms "Progranulin" and "PGRN" refer to a secreted trophic factor and precursor peptide for granulin. The gene is located on chromosome 17q21.31 and is also known as granulin precursor, proepithelin, PEPI, PC cell-derived growth factor, granulin-epithelin, CLN11, PCDFGF, GP88, GEP, granulin, and acrogranin. The terms "Progranulin" and "PGRN" refer to variants of wild-type human PGRN peptide and nucleic acids encoding same, e.g., variants having at least 85% sequence identity (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.9% identity or more) to the amino acid sequence of the wild-type PGRN peptide (e.g., SEQ ID NO: 1). The terms "progranulin" and "PGRN" also refer to a protein or a polynucleotide having at least 85% sequence identity (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.9% identity or more) to the nucleic acid sequence of a wild-type PGRN gene (e.g., SEQ ID NO: 2), provided that the encoded PGRN variant maintains the therapeutic function of wild-type PGRN. The terms "progranulin" and "PGRN" can also refer to a variant of PGRN having two or more (e.g., 2, 3, 4, 5, 6, 7, 8, or more) granulin (GRN) domains having the amino acid sequence of any one of SEQ ID NOs: 2-9. The terms "Progranulin" and "PGRN" may also refer to variants of PGRN having 2 to 16 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16) GRN domains having any one of the amino acid sequences of SEQ ID NOs: 2 to 9. The terms "Progranulin" and "PGRN" may also refer to PGRN protein in which a native secretory signal peptide is present.In addition, the terms "Progranulin" and "PGRN" can refer to a "PGRN fusion protein," which is a protein in which PGRN is operably linked to another polypeptide, half-life modifying agent, or therapeutic agent, such as an ApoE receptor binding (Rb) domain (e.g., an Rb domain having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO: 11). As used herein, the term "PGRN" can refer to the peptide or the gene encoding the protein, depending on the context, as will be understood by one of skill in the art.

[0500] As used herein, a patient suffering from "Progranulin-related FTLD or NCL" and "PGRN-related FTLD or NCL" is a patient who has been diagnosed with FTLD or NCL and also contains a deleterious mutation in the PGRN gene. More than 70 pathogenic mutations have been reported in the PGRN gene, the majority of which result in a premature stop codon and nonsense-mediated decay of truncated PGRN mRNA. PGRN mutations are described in Gijselinck et al., Human Mutation 29:1373-1386 (2012) and Pottier et al., Journal of Neurochemistry. 138:32-53 (2016), the disclosure of which is incorporated herein by reference, as it relates to human PGRN mutations.

[0501] As used herein, the term "promoter" refers to a recognition site on DNA to which RNA polymerase binds. The polymerase drives the transcription of the transgene. Exemplary promoters suitable for use in the compositions and methods described herein are described, for example, in Sandelin et al., Nature Reviews Genetics 8:424 (2007), which is incorporated herein by reference as its disclosure relates to nucleic acid regulatory elements. In addition, the term "promoter" can refer to synthetic promoters, which are regulatory DNA sequences that do not naturally occur in biological systems. Synthetic promoters include parts of naturally occurring promoters in combination with non-naturally occurring polynucleotide sequences and can be optimized to express recombinant DNA using a variety of transgenes, vectors, and target cell types.

[0502] "Percent sequence identity" with respect to a reference polynucleotide or polypeptide sequence is defined as the percentage of nucleic acids or amino acids in a candidate sequence that are identical to the nucleic acids or amino acids in the reference polynucleotide or polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent nucleic acid and amino acid sequence identity can be accomplished in a variety of ways that are within the ability of one of skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, or Megalign software. One of skill in the art can determine appropriate parameters for aligning sequences, including any algorithms required to obtain maximum alignment over the entire length of the sequences being compared. For example, percent sequence identity values ​​can be generated using the sequence comparison computer program BLAST. As an example, the percent sequence identity of a given nucleic acid or amino acid sequence A to, with, or against a given nucleic acid or amino acid sequence B (which may alternatively be expressed as a given nucleic acid or amino acid sequence A having a certain percent sequence identity to, with, or against a given nucleic acid or amino acid sequence B) is calculated as follows: Multiply by 100(fraction X / Y) where X is the number of nucleotides or amino acids in a programmatic alignment of A and B that are scored as identical matches by a sequence alignment program (e.g., BLAST), and Y is the total number of nucleic acids in B. It will be appreciated that if the length of nucleic acid or amino acid sequence A is not equal to the length of nucleic acid or amino acid sequence B, then the percent sequence identity of A to B will not equal the percent sequence identity of B to A.

[0503] As used herein, the term "pharmacologically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable for contact with the tissues of a subject, such as a mammal (e.g., a human), without undue toxicity, irritation, allergic response, or other problematic complications, commensurate with a reasonable benefit / risk ratio.

[0504] As used herein, the potent "ApoE-derived receptor binding peptide (Rb)" has the ability to translocate proteins across the BBB to the brain when engineered as a fusion protein. Thus, this method can function to selectively open the BBB for therapeutic agents (e.g., soluble PGRN or GRN) when engineered as a fusion protein. Because this peptide utilizes the Rb domain of ApoE, rather than the entire ApoE protein, it can be easily conjugated to diagnostic or therapeutic agents without compromising or interfering with the important biological functions of ApoE. This pathway is also an alternative uptake route that can facilitate further / secondary brain distribution of agents after they reach the CNS due to the widespread expression of LDLRf members in the brain parenchyma. An exemplary Rb domain can be found in the N-terminus of ApoE. For example, an Rb domain useful in conjunction with the compositions and methods described herein is a polypeptide having the amino acid sequence of residues 1-191 of SEQ ID NO:11, residues 25-185 of SEQ ID NO:11, residues 50-180 of SEQ ID NO:11, residues 75-175 of SEQ ID NO:11, residues 100-170 of SEQ ID NO:11, or residues 125-165 of SEQ ID NO:11, as well as variants thereof, such as polypeptides having at least 85% sequence identity (e.g., at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity) with any of these sequences. An exemplary Rb domain is a region of ApoE having the amino acid sequence of residues 159-167 of SEQ ID NO:11.

[0505] As used herein, the term "regulatory sequence" includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals) that control the transcription or translation of antibody chain genes. Such regulatory sequences are described, for example, in Perdew et al., Regulation of Gene Expression (Humana Press, New York, NY, (2014)), which is incorporated herein by reference.

[0506] As used herein, the term "sample" refers to a specimen isolated from a subject (e.g., blood, blood components (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissue (e.g., placenta or skin), pancreatic juice, chorionic villus samples, and cells).

[0507] As used herein, the term "secretory signal peptide" refers to a short (usually 16-60 amino acids) peptide region within a precursor protein that directs secretion of the precursor protein from the cytoplasm to the periplasmic or extracellular space of the host. Such secretory signal peptides are generally positioned at the amino terminus of the precursor protein. In some embodiments, the secretory signal peptide is linked to the amino terminus. Typically, the secretory signal peptide is cleaved during passage through the cellular secretory pathway. Cleavage is not essential as long as the secreted protein retains its desired activity. Exemplary secretory signal peptides include the PGRN secretory signal peptide.

[0508] As used herein, the term "social cognition" refers to cognitive function encompassing a set of skills that determine how a subject (e.g., a human) processes, remembers, and adapts to information about other conspecific subjects (e.g., other humans) and social situations. Non-limiting examples of social cognition include, for example, emotional responses to social stimuli, processing abilities for theory of mind tasks, the ability to recognize faces, impulse control in social situations, and joint attention. Subjects who have been diagnosed with an NCD may exhibit impaired social cognition compared to healthy subjects.

[0509] As used herein, the terms "stem cell" and "undifferentiated cell" refer to cells in an undifferentiated or partially differentiated state that have the developmental potential to differentiate into multiple cell types. Stem cells can proliferate while maintaining their functional potential to give rise to more such stem cells. Stem cells can divide asymmetrically, known as obligate asymmetric differentiation, with one daughter cell retaining the functional potential of the parent stem cell and the other daughter cell expressing another specific function, phenotype and / or developmental potential that is somewhat different from the parent cell. The daughter cells themselves can be induced to proliferate while also retaining one or more cells with the developmental potential of the parent, and to give rise to progeny that later differentiate into one or more mature cell types. Differentiated cells may themselves be derived from pluripotent cells, etc., that are themselves derived from pluripotent cells. Alternatively, some of the stem cells in a population can divide symmetrically into two stem cells. Thus, the term "stem cell" refers to any subset of cells that, under certain circumstances, have the developmental potential to differentiate into a more specialized or differentiated phenotype, and, under certain circumstances, retain the ability to proliferate without substantial differentiation. In some embodiments, the term stem cell generally refers to a naturally occurring parent cell whose descendants (progeny cells) specialize by differentiation, often in different directions, by acquiring completely different characteristics, as occurs in the gradual diversification of embryonic cells and tissues. Some differentiated cells also have the ability to give rise to cells of greater developmental potential. Such ability may be natural or may be artificially induced by treatment with various factors. Cells that begin as stem cells can progress to a differentiated phenotype, but can also be "reverted" and then induced to re-express the stem cell phenotype, a term often referred to by those skilled in the art as "dedifferentiation" or "reprogramming" or "reverse differentiation."

[0510] As used herein, the term "transfection" refers to any of a wide variety of techniques commonly used to introduce exogenous DNA into prokaryotic or eukaryotic host cells, such as electroporation, lipofection, calcium phosphate precipitation, DEAE-dextran transfection, Nucleofection, squeeze-poration, sonoporation, optical transfection, Magnetofection, impalefection, and the like.

[0511] As used herein, the term "transgene" refers to a recombinant nucleic acid (e.g., DNA or cDNA) that encodes a gene product (e.g., PGRN or GRN). The gene product may be an RNA, a peptide, or a protein. In addition to the coding region for the gene product, the transgene may include or be operably linked to one or more elements that facilitate or enhance expression, such as a promoter, enhancer(s), destabilization domain(s), response element(s), reporter element(s), insulator element(s), polyadenylation signal(s), and / or other functional elements. Embodiments of the present disclosure may utilize any known suitable promoter, enhancer(s), destabilization domain(s), response element(s), reporter element(s), insulator element(s), polyadenylation signal(s), and / or other functional elements.

[0512] As used herein, the term "subject" or "patient" refers to an animal (e.g., a mammal, such as a human). The subject treated according to the methods described herein may be a subject who has been diagnosed with an NCD or who is at risk for developing these conditions. Diagnosis may be performed by any method or technique known in the art. One of skill in the art will understand that the subject treated according to the present disclosure may have undergone standard testing or may have been identified as at risk by the presence of one or more risk factors associated with a disease or condition without testing.

[0513] As used herein, the terms "transduction" and "transducing" refer to the method of introducing a viral vector construct, or a portion thereof, into a cell and the subsequent expression in the cell of a transgene encoded by the vector construct, or a portion thereof.

[0514] As used herein, "treatment" or "treating" refers to an approach to obtain a beneficial or desired result, e.g., a clinical result. Beneficial or desired results include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable; reduction in the extent of the disease or condition; stabilization (i.e., not worsening) of the disease, disorder, or condition; prevention of the progression of the disease or condition; delay or slowing of the progression of the disease or condition; amelioration or palliation of the disease or condition; and remission (whether partial or complete). "Ameliorating" or "alleviating" a disease or condition means that the extent and / or undesirable clinical signs of the disease, disorder, or condition are reduced and / or the time course of progression is slowed or prolonged, compared to the extent or time course in the absence of treatment. "Treatment" can also mean prolonging survival, compared to the expected survival in the absence of treatment. Those in need of treatment include those already with the condition or disorder, as well as those susceptible to having the condition or disorder, or those in need of prevention of the condition or disorder.

[0515] As used herein, the term "vector" includes nucleic acid vectors, e.g., DNA vectors such as plasmids, RNA vectors, viruses, or other suitable replicons (e.g., viral vectors). Various vectors have been developed for delivering polynucleotides encoding exogenous proteins to prokaryotic or eukaryotic cells. Examples of such expression vectors are disclosed, for example, in WO1994 / 011026, which is incorporated herein by reference as it relates to vectors suitable for expressing genes of interest. Expression vectors suitable for use in the compositions and methods described herein include polynucleotide sequences as well as additional sequence elements used, for example, for protein expression and / or integration of these polynucleotide sequences into the genome of mammalian cells. Certain vectors that can be used for expression of PGRN or GRN as described herein include plasmids that include regulatory sequences that direct gene transcription, such as promoter and enhancer regions. Other useful vectors for expression of PGRN or GRN include polynucleotide sequences that increase the translation rate of these genes or improve the stability or nuclear export of mRNA resulting from gene transcription. These sequence elements include, for example, 5' and 3' untranslated regions, internal ribosome entry sites (IRES), and polyadenylation signal sites to direct efficient transcription of genes carried in the expression vector. Expression vectors suitable for use in the compositions and methods described herein may also contain a polynucleotide encoding a marker for selecting cells containing such a vector. Examples of suitable markers are genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, kanamycin, nourseothricin, or zeocin.

[0516] Detailed Description Described herein are compositions and methods for treating neurocognitive disorders (NCDs), such as frontotemporal lobar degeneration (FTLD) or neuronal ceroid lipofuscinosis (NCL), in a subject (such as a mammalian subject, e.g., a human). The compositions and methods described herein can be used to treat NCDs (e.g., FTLD or NCLs (e.g., progranulin (PGRN)-associated FTLD or NCL)) in a subject (e.g., a human subject) by administering a cell, e.g., a pluripotent cell, an embryonic stem cell (ESC), an induced pluripotent stem cell (iPSC), a pluripotent cell, a CD34+ cell, a hematopoietic stem cell (HSC), a myeloid progenitor cell (MPC), a blood lineage progenitor cell (BLPC), a monocyte, a macrophage, a microglial progenitor cell, or a microglia, that includes a transgene (e.g., a transgene that can be expressed in macrophages or microglial cells) that encodes PGRN or granulin (GRN). For example, compositions containing cells that have been modified ex vivo to express PGRN or GRN are described herein. In the next section, compositions and methods useful for treating NCD are described in more detail.

[0517] Neurocognitive disorders Neurocognitive disorders (NCDs) are defined as a set of disorders characterized by cognitive impairment as a core symptom, and which show cognitive decline (e.g., acquired impairment) compared to a previous higher level of cognition, rather than developmental impairment. NCDs are broadly divided into severe or mild syndromes (e.g., severe NCD and mild NCD) based on the degree of impairment diagnosed in subjects. Furthermore, NCDs can be classified based on their etiology. For example, non-limiting examples of NCDs can include frontotemporal NCD, NCD due to lysosomal disease (e.g., NCL), NCD due to AD, vascular NCD, NCD with Lewy bodies, NCD due to Parkinson's disease, frontotemporal NCD, NCD due to traumatic brain injury, NCD due to HIV infection, substance / drug-induced NCD, NCD due to Huntington's disease, NCD due to prion disease, NCD due to another medical condition, NCD due to multiple etiologies, and unspecified NCD. The compositions and methods disclosed herein are useful for treating an NCD (e.g., FTLD or NCL).

[0518] Frontotemporal lobar degeneration FTLD is a clinical syndrome characterized by progressive neurodegeneration in the frontal and temporal lobes of the cerebral cortex. The manifestations of FTLD are complex and heterogeneous, and can be expressed as any one of three clinically distinct variants of FTLD, including: 1) behavioral and personality changes, blunted affect, social withdrawal, perseverative behavior, attention deficit, loss of inhibition, and prominent degeneration of the frontal lobe; 2) semantic dementia (SD), characterized by verbal amnesic aphasia, progressive loss of semantic knowledge of words, objects, and concepts, and prominent degeneration of the anterior temporal lobe. In addition, the SD variant of FTLD shows flat affect, social deficits, perseverative behavior, and loss of inhibition; or 3) progressive non-verbal aphasia (PNA), characterized by motor impairment in speech production, reduced language expression, and prominent degeneration of the perisylvian cortex. Neuronal loss in the brains of FTLD patients is associated with one of three distinct neuropathologies: 1) the presence of tau-positive neurons and glial inclusions; 2) ubiquitin (ub)-positive and TAR DNA-binding protein 43 (TDP43)-positive, but tau-negative inclusions; or 3) ub- and fusion with sarcoma (FUS)-positive, but tau- and TDP-43-negative inclusions. These neuropathologies are deemed important in the pathogenesis of FTLD.

[0519] Nearly half of FTLD patients have a first-degree family member with dementia, ALS, or Parkinson's disease, suggesting a strong genetic link to the etiology of the disease.Several mutations on chromosome 17q21 have been associated with FTLD presentation.

[0520] Neuronal ceroid lipofuscinosis NCL is a collective term referring to a collection of at least eight clinically recognized lysosomal storage disorders resulting from the accumulation of lipofuscin in cells of the body, such as neurons, liver, spleen, myocardium, and kidney cells. Clinically, NCL presents with marked neurodegeneration and progressive and irreversible loss of motor and cognitive abilities, although disease severity and clinical manifestations may depend on the particular NCL variant.

[0521] Known variants of NCL include the juvenile variant, also known as Santovuori-Haltia disease (SHD), the late-onset juvenile variant, also known as Jansky-Bielschowsky disease (JBD), the Finnish late-onset juvenile variant (FLI), the variant with variant late (VLI), the CLN7 variant (CLN7), the CLN8 variant (CLN8), the Turkish late-onset juvenile variant (TLI), the juvenile variant, also known as Batten disease (BD), and the adult variant, also known as Koufs disease (KD). SHD is associated with early visual loss that progresses to complete retinal blindness by age 2, followed by a vegetative state at age 3, and brain death by age 4. This variant is also associated with spontaneous occurrence of epileptic seizures. The JBD variant occurs between 2 and 4 years of age and is associated with ataxia, seizures, progressive cognitive decline, and abnormal speech development, typically resulting in death by age 8. BD typically occurs between 4 and 10 years of age and includes symptoms such as vision loss, seizures, cognitive impairment, and early death. NCL patients with the KD variant generally have milder symptoms than the SHD and BD variants and have a life expectancy of approximately 40 years.

[0522] Progranulin-related frontotemporal dementia and neuronal ceroid lipofuscinosis Studies investigating the association between chromosome 17q21 and FTLD have found several FTLD-associated mutations in the PGRN gene. These mutations often result in the aggregation and accumulation of ub-positive, TDP43-positive, tau-negative neuropathological inclusions in the brains of FTLD patients. PGRN is a secreted precursor peptide to several mature GRN proteins and is thought to function primarily as a neurotrophic growth factor, promoting neuronal differentiation and survival. PGRN has also been demonstrated to serve anti-inflammatory and neuroprotective functions. Although PGRN is ubiquitously expressed, as a result of its association with FTLD, much attention has been focused on the central nervous system (CNS), where PGRN is expressed in multiple cell types, including neurons, glia, and endothelial cells. In FTLD, more than 70 loss-of-function mutations in the PGRN gene have been identified, the vast majority of which result in haploinsufficiency and a reduction in serum PGRN levels by more than 50%. PGRN mutations are described in Gijselinck et al., Human Mutation 29:1373-86(2008), the disclosure of which is incorporated herein by reference as it relates to human PGRN mutations. The effect of PGRN mutations is dose-dependent, as homozygous patients who completely lack functional PGRN protein develop NCL, suggesting an additional role for this protein in normal lysosomal function. For a comprehensive description of NCL and related gene mutations, see Mink et al. Journal of Child Neurology 28:1101-5(2013), Nita et al. Epileptic Disorders 18:73-88(2016), and Mole & Cotman, Biochimica et Biophysica Acta 1852:2237-41(2015), and Ward et al., Science Translational Medicine 9(385):eaah5642, the disclosures of which are incorporated herein by reference in their entirety.

[0523] Clinical management of FTLD mainly uses selective serotonin reuptake inhibitors (SSRIs) and antipsychotics to manage the emotional and behavioral changes associated with FTLD. Similarly, most interventional treatments targeting NCL pathology aim to reduce the severity of disease symptoms, such as epileptic seizures, movement disorders, enhance immune response, and pain management. However, this strategy targets ameliorating disease symptoms without addressing its onset and progression. Unlike these treatments, the compositions and methods described herein offer the advantage of treating another biochemical phenomenon that may underlie the onset of NCD, such as FTLD or NCL. Thus, the compositions and methods described herein target the physiological cause of the disease and represent a potential curative treatment.

[0524] The compositions and methods described herein can be used to treat NCDs by administering cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) that contain a transgene encoding PGRN or GRN (such as a transgene that can be expressed in macrophages or microglial cells). These compositions and methods can be used to treat NCDs with any etiology, e.g., genetic mutation, environmental toxins, or sporadic. These compositions and methods can also be used to treat subjects with PGRN-associated FTLD or NCL. The compositions and methods described herein can be used to treat subjects with normal PGRN or GRN activity, reduced PGRN or GRN activity, and subjects whose PGRN mutation status and / or PGRN or GRN activity levels are unknown. The compositions and methods described herein can also be administered as a preventative treatment to subjects at risk of developing an NCD, e.g., subjects with PGRN mutations, subjects with reduced PGRN or GRN activity, and subjects with mutations in one or more of the genes associated with an NCD (e.g., FTLD or NCL).

[0525] Progranulin and Granulin Constructs A transgene-containing construct that can be used in conjunction with the compositions and methods described herein comprises a polynucleotide encoding wild-type human PGRN (the amino acid sequence of which is shown below as SEQ ID NO: 1), or a variant thereof, e.g., a protein having at least 85% sequence identity (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 1. In some embodiments, the PGRN comprises at least two GRN domains (e.g., at least 2, 3, 4, 5, 6, 7, 8, or more GRN domains) having the amino acid sequence of any one of SEQ ID NOs: 2-9. In some embodiments, the PGRN comprises 2-16 GRN domains (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 GRN domains).

[0526] In some embodiments, a polynucleotide encoding wild-type PGRN or GRN may be a polynucleotide that has been codon-optimized to confer resistance to degradation by endogenous PGRN- or GRN-directed nucleases and inhibitory RNAs as described in detail below. In some embodiments, a codon-optimized transgene encoding PGRN or GRN comprises a polynucleotide having at least 85% sequence identity (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 19.

[0527] The wild-type human PGRN protein has the following amino acid sequence (GenBank Accession No: NP_002078.1): MWTLVSWVALTAGLVAGTRCPDGQFCPVACCLDPGGASYSCCRPLLDKWPTTLSRHLGGPCQVDAHCSAGHSCIFTVSGTSSCCPFPEAVACGDGHHCCPRGFHCSADGRSCFQRSGNNSVGAIQCPDSQFECPDFSTCCVMVDGSWG CCPMPQASCCEDRVHCCPHGAFCDLVHTRCITPTGTHPLAKKLPAQRTNRAVALSSSVMCPDARSRCPDGSTCCELPSGKYGCCPMPNATCSDHLHCCPQDTVCDLIQSKCLSKENATTDLLTKLPAHTVGDVKCDMEVSCPDGYTC CRLQSGAWGCCPFTQAVCCEDHIHCCPAGFTCDTQKGTCEQGPHQVPWMEKAPAHLSLPDPQALKRDVPCDNVSSCPSSDTCCQLTSGEWGCCPIPEAVCCSDHQHCCPQGYTCVAEGQCQRGSEIVAGLEKMPARRASLSHPRDIGC DQHTSCPVGQTCCPSLGGSWACCQLPHAVCCEDRQHCCPAGYTCNVKARSCEKEVVSAQPATFLARSPHVGVKDVECGEGHFCHDNQTCCRDNRQGWACCPYRQGVCCADRRHCCPAGFRCAARGTKCLRREAPRWDAPLRDPALRQLL (SEQ ID NO:1)

[0528] Wild-type human paragranulin (para-GRN) has the following amino acid sequence: TRCPDGQFCPVACCLDPGGASYSCCRPLLD (SEQ ID NO:2)

[0529] The wild-type human granulin-1 (GRN-1) peptide has the following amino acid sequence: GGPCQVDAHCSAGHSCIFTVSGTSSCCPFPEAVACGDGHHCCPRGFHCSADGRSCF (SEQ ID NO:3)

[0530] The wild-type human granulin-2 (GRN-2) peptide has the following amino acid sequence: AIQCPDSQFECPDFSTCCVMVDGSWGCCPMPQASCCEDRVHCCPHGAFCDLVHTRCI (SEQ ID NO:4)

[0531] The wild-type human granulin-3 (GRN-3) peptide has the following amino acid sequence: VMCPDARSRCPDGSTCCELPSGKYGCCPMPNATCCSDHLHCCPQDTVCDLIQSKCL (SEQ ID NO:5)

[0532] The wild-type human granulin-4 (GRN-4) peptide has the following amino acid sequence: DVKCDMEVSCPDGYTCCRLQSGAWGCCPFTQAVCCEDHIHCCPAGFTCDTQKGTCE (SEQ ID NO:6)

[0533] The wild-type human granulin-5 (GRN-5) peptide has the following amino acid sequence: VPCDNVSSCPSSDTCCQLTSGEWGCCPIPEAVCCSDHQHCCPQGYTCVAEGQCQ (SEQ ID NO:7)

[0534] The wild-type human granulin-6 (GRN-6) peptide has the following amino acid sequence: IGCDQHTSCPVGQTCCPSLGGSWACCQLPHAVCCEDRQHCCPAGYTCNVKARSCE (SEQ ID NO:8)

[0535] The wild-type human granulin-7 (GRN-7) peptide has the following amino acid sequence: DVECGEGHFCHDNQTCCRDNRQGWACCPYRQGVCCADRRHCCPAGFRCAARGTKCL (SEQ ID NO:9)

[0536] Wild-type human PGRN (CCDS identifier: 11483.1) has the following nucleic acid sequence: (SEQ ID NO:10)

[0537] The codon-optimized human PGRN has the following nucleic acid sequence: (SEQ ID NO:19)

[0538] According to the methods described herein, a subject is administered a polynucleotide encoding the amino acid sequence of SEQ ID NO:1, or a polynucleotide encoding a polypeptide having at least 85% sequence identity (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO:1, or a polynucleotide encoding a polypeptide including one or more conservative amino acid substitutions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more conservative amino acid substitutions) relative to SEQ ID NO:1, or an amino acid sequence of any one of SEQ ID NOs:2-9 or SEQ ID NOs:2-9. Cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) containing a polynucleotide encoding a polypeptide comprising one or more GRN domains with variants thereof having at least 85% sequence identity (e.g., at least 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to any one of the following may be administered, provided that the encoded PGRN or GRN variant maintains the therapeutic function of wild-type PGRN or GRN. The neurotrophic activity of wild-type PGRN is important for neurotrophic support and maintenance of lysosomal function. Loss of PGRN leads to neurodegeneration and lysosomal storage disease in a dose-dependent manner.

[0539] host cell Cells that can be used in conjunction with the compositions and methods described herein include cells (e.g., pluripotent cells, ESCs, iPSCs, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, or microglial progenitor cells) or differentiated cells (e.g., macrophages or microglia). For example, one type of cell that can be used in conjunction with the compositions and methods described herein is a pluripotent cell. A pluripotent cell is a cell that has the ability to develop into more than one differentiated cell type. Examples of pluripotent cells are ESCs and iPSCs. ESCs and iPSCs have the ability to differentiate into cells of the ectoderm, which gives rise to the skin and nervous system, the endoderm, which forms the digestive tract and airway, the endocrine glands, the liver, and the pancreas, and the mesoderm, which forms bone, cartilage, muscle, connective tissue, and most of the circulatory system. Another type of cell that can be used in conjunction with the compositions and methods described herein is a pluripotent cell. A pluripotent cell is a cell that has the ability to differentiate into multiple cell types, but not all of them. A non-limiting example of a pluripotent cell is a CD34+ cell (eg, HSC or MPC).

[0540] The cells that can be used in conjunction with the compositions and methods described herein include HSCs and MPCs. HSCs are immature blood cells that have the ability to self-renew and differentiate into mature blood cells, including a variety of cell lineages, including, but not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes, erythrocytes), platelets (e.g., megakaryoblasts, platelet-producing megakaryocytes, platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NK cells, B cells, and T cells). Human HSCs are CD34+. In addition, HSCs also refer to long-term repopulating HSCs (LT-HSCs) and short-term repopulating HSCs (ST-HSCs). Any of these HSCs can be used in conjunction with the compositions and methods described herein.

[0541] HSCs can differentiate into myeloid progenitor cells that are also CD34+. Myeloid progenitor cells can further differentiate into granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes, erythrocytes), platelets (e.g., megakaryoblasts, platelet-producing megakaryocytes, and platelets), monocytes (e.g., monocytes and macrophages), dendritic cells, and microglia. Common myeloid progenitor cells can be characterized by cell surface molecules, including lin-, SCA1-, c-kit+, CD34+, and CD16 / 32. mid It is known that.

[0542] HSCs and myeloid progenitor cells can be obtained from blood products. Blood products are products obtained from the body or body organs that contain cells of hematopoietic origin. Such sources include unfractionated bone marrow, umbilical cord, placenta, peripheral blood, or mobilized peripheral blood. All of the aforementioned crude or unfractionated blood products can be enriched for cells with characteristics of HSCs or myeloid progenitor cells in several ways. For example, more mature differentiated cells can be selected based on the cell surface molecules they express. Blood products can be fractionated by positively selecting for CD34+ cells, which include a subpopulation of hematopoietic stem cells that are capable of self-renewal, multipotency, and that can home into the hematopoietic stem cell niche when reintroduced into the transplant recipient to re-establish productive and sustained hematopoiesis. Such selection is accomplished, for example, using commercially available magnetic anti-CD34 beads (Dynal, Lake Success, NY). Myeloid progenitor cells can also be isolated based on the markers they express. Unfractionated blood products can be obtained directly from donors or retrieved from cryopreservation stores. HSCs and myeloid progenitor cells can also be obtained by differentiation of ES cells, iPS cells, or other reprogrammed mature cell types.

[0543] Cells that can be used in conjunction with the compositions and methods described herein include allogeneic and autologous cells. All of the aforementioned cell types can be differentiated into microglia. The cells described herein can also be differentiated into microglial progenitor cells or microglial stem cells. Differentiation can occur ex vivo or in vivo. Methods for ex vivo differentiation of human ESCs and iPSCs are known to those of skill in the art and are described in Muffat et al., Nature Medicine 22:1358-1367 (2016) and Pandya et al., Nature Neuroscience (2017) (epub ahead of print), which are incorporated herein by reference for their disclosures regarding methods for differentiating cells into microglia.

[0544] Microglia Cells that can be used in conjunction with the compositions and methods described herein include those that can differentiate into microglia or are differentiated microglia. Microglia are bone marrow-derived cells that function as immune cells or resident macrophages of the central nervous system. Microglia are genetically and functionally very similar to macrophages and share the ability to dynamically shift between pro-inflammatory and anti-inflammatory states. The pro-inflammatory state is known as classical activation, or M1, and the anti-inflammatory state is called alternative activation, or M2. Microglia can be shifted between the two states by extracellular signals, such as signals from neighboring neurons or astrocytes, cellular debris, toxins, infection, ischemia, and trauma, among others. M1 microglia are often observed in diseased brains, especially in diseases involving neuroinflammation, such as FTLD or NCL. The classically activated M1 phenotype is observed in progranulin null mice, CLN3(Δex7 / 8) mice, and CLN6 mice. nclfM1 microglia are also observed in mouse models of FTLD and NCL, such as in mice. Although it is unclear whether M1 microglia are a cause or a consequence of neuroinflammation, classical activation of microglia can secrete proinflammatory cytokines, such as TNF-α, IL-1β, and IL-6, chemokines, and nitric oxide, which can lead to persistent inflammation, neuronal damage, and further activation of M1 microglia. This positive feedback loop can be detrimental to brain tissue, and therefore methods to reduce M1 activation and / or increase M2 activation may help patients with diseases characterized by neuroinflammation, such as NCDs.

[0545] Expression of Progranulin or Granulin in Mammalian Cells PGRN activity is reduced in FTLD and NCL patients, and FTLD brains contain classically activated M1 microglia. The compositions and methods described herein target these dysfunctions by administering cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitors, or microglia) that contain transgenes (e.g., transgenes that can be expressed in macrophages or microglial cells) that code for PGRN or GRN. To utilize these agents in therapeutic applications in the treatment of NCD (e.g., FTLD or NCL), these agents can be directed to the interior of cells, and in certain instances, to specific organelles. A wide range of methods have been established for delivering such proteins to mammalian cells and for stably expressing genes encoding such proteins in mammalian cells.

[0546] Polynucleotides encoding progranulin or granulin One of the platforms that can be used to achieve therapeutically effective intracellular concentrations of PGRN or GRN in mammalian cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) is by stable expression of genes encoding these agents (e.g., by integration into the nuclear or mitochondrial genome of mammalian cells). These genes are polynucleotides that code for the primary amino acid sequence of the corresponding protein. To introduce such foreign genes into mammalian cells, these genes can be incorporated into vectors. Vectors can be introduced into cells by various methods, including transformation, transfection, direct uptake, projectile bombardment, and by encapsulation of the vector in liposomes. Examples of suitable methods for transfecting or transforming cells are calcium phosphate precipitation, electroporation, microinjection, infection, lipofection, and direct uptake. Such methods are described in further detail in, for example, Green et al., Molecular Cloning: A Laboratory Manual, Fourth Edition (Cold Spring Harbor University Press, New York (2014)); and Ausubel et al., Current Protocols in Molecular Biology (John Wiley & Sons, New York (2015)), the disclosures of each of which are incorporated herein by reference.

[0547] PGRN or GRN can also be introduced into mammalian cells by targeting a vector containing a gene encoding such an agent to cell membrane phospholipids. For example, the vector molecule can be linked to the VSV-G protein, a viral protein that has affinity for all cell membrane phospholipids, to target the vector to phospholipids on the extracellular surface of the cell membrane. Such constructs can be produced using methods well known to those skilled in the art.

[0548] The recognition and binding of the polynucleotide encoding PGRN or GRN by mammalian RNA polymerase is important for gene expression. It can therefore contain sequence elements within the polynucleotide that exhibit high affinity for the transcription factors that recruit RNA polymerase and promote the assembly of the transcription complex at the transcription initiation site. Such sequence elements include, for example, mammalian promoters whose sequences can be recognized and bound by specific transcription initiation factors and ultimately RNA polymerase. Examples of mammalian promoters are described in Smith et al., Mol. Sys. Biol., 3:73(2007) online publication, the disclosure of which is incorporated herein by reference.

[0549] Suitable polynucleotides for use in the compositions and methods described herein also include those encoding PGRN or GRN downstream of a mammalian promoter. Useful promoters for expressing PGRN or GRN in mammalian cells include, for example, elongation factor 1-alpha (EF1α) promoter, phosphoglycerate kinase 1 (PGK) promoter, CD68 molecule (CD68) promoter (see Dahl et al., Molecular Therapy 23:835 (2015), which is incorporated herein by reference for its relevance to the use of PGK and CD68 promoters to express PGRN), CD11b promoter, PGRN promoter, C-X3-C motif chemokine receptor 1 (CX3CR1) promoter, allograft inflammatory factor 1 (AIF1) promoter, purinergic receptor P2Y12 (P2Y12) promoter, transmembrane protein 119 (TMEM119) promoter, and colony-stimulating factor 1 receptor (CSF1R) promoter. Alternatively, promoters derived from viral genomes can also be used for stable expression of these agents in mammalian cells. Examples of functional viral promoters that can be used to promote mammalian expression of these agents are the adenovirus late promoter, the vaccinia virus 7.5K promoter, the simian virus 40 (SV40) promoter, the cytomegalovirus promoter, the tk promoter of herpes simplex virus (HSV), the mouse mammary tumor virus (MMTV) promoter, the long terminal repeat (LTR) promoter of human immunodeficiency virus (HIV), the promoter of Moloney virus, the Epstein-Barr virus (EBV), the Rous sarcoma virus (RSV), and the cytomegalovirus (CMV) promoter. Alternatively, synthetic promoters optimized for use in mammalian cells can be used for stable expression of PGRN or GRN transgenes.

[0550] Once a polynucleotide encoding PGRN or GRN has been integrated into the nuclear DNA of a mammalian cell, transcription of the polynucleotide can be induced by methods known in the art. For example, expression can be induced by exposing the mammalian cell to an external chemical reagent, such as an agent that modulates the binding of transcription factors and / or RNA polymerase to the mammalian promoter to regulate gene expression. The chemical reagent serves to promote the binding of RNA polymerase and / or transcription factors to the mammalian promoter, for example by removing repressor proteins bound to the promoter. Alternatively, the chemical reagent can serve to increase the affinity of the mammalian promoter for RNA polymerase and / or transcription factors, such that the transcription rate of genes placed downstream of the promoter increases in the presence of the chemical reagent. Examples of chemical reagents that enhance polynucleotide transcription by the aforementioned mechanisms are tetracycline and doxycycline. These reagents are commercially available (Life Technologies, Carlsbad, CA) and can be administered to mammalian cells to promote gene expression according to established protocols.

[0551] Other DNA sequence elements that can be included in polynucleotides for use in the compositions and methods described herein are enhancer sequences. Enhancers represent another class of regulatory elements that induce conformational changes in polynucleotides, including genes of interest, such that the DNA adopts a three-dimensional orientation favorable for the binding of transcription factors and RNA polymerase at the transcription start site. Thus, polynucleotides for use in the compositions and methods described herein include those that encode PGRN or GRN and additionally include mammalian enhancer sequences. Many enhancer sequences are currently known from mammalian genes, examples being enhancers derived from genes encoding mammalian globin, elastase, albumin, alpha-fetoprotein, and insulin. Enhancers for use in the compositions and methods described herein also include those derived from the genetic material of viruses capable of infecting eukaryotic cells. Examples are the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers. Additional enhancer sequences that induce activation of eukaryotic gene transcription are disclosed in Yaniv et al., Nature 297:17 (1982). The enhancer can be spliced, for example, at the 5' or 3' position of the gene into the vector containing the polynucleotide encoding the water-forming NADH oxidase. In a preferred orientation, the enhancer is located 5' to the promoter, which is then placed 5' to the polynucleotide encoding PGRN or GRN.

[0552] Cell-specific gene expression Interfering RNA (RNAi) has been widely used to knock down the expression of endogenous genes by delivering small interfering RNA (siRNA) to cells to cause the degradation of complementary mRNA. An additional application is to exploit the diversity of endogenous microRNAs (miRNAs) to negatively regulate the expression of exogenously introduced transgenes tagged with artificial miRNA target sequences. These miRNA target tagged transgenes can be negatively regulated depending on the activity of a given miRNA, which can be specific to tissue, cell lineage, activation, or differentiation stage. These artificial miRNA target sequences (miRTs) can be recognized as targets by specific miRNAs to induce post-transcriptional gene silencing. While strong transgene expression in targeted cells can result in beneficial therapeutic outcomes, off-target expression, such as ectopic or unregulated transgene expression in HSPCs or other progenitor cells, can have cytotoxic effects and result in counter-selection of transgene-containing cells leading to altered cell behavior and reduced therapeutic efficacy. Incorporation of miRT for miRNAs that are broadly expressed in HSPCs and progenitor cells, but not in cells of myeloid lineage, allows for the repression of transgene expression in HSPCs and other progenitor cells, allowing for silent, long-term storage of transgene-containing hematopoietic progeny, while allowing robust transgene expression in mature differentiated target cells. miR-126 is highly expressed in HSPCs, other progenitor cells, and cells of the erythroid lineage, but is absent in cells of the myeloid lineage (e.g., macrophages and microglia) (Gentner et al., Science Translational Medicine. 2:58ra34 (2010)). For example, miR-126 targeting sequences incorporated within a transgene allow for targeted expression of the transgene in cells of the myeloid lineage and repression of expression in HSPCs and other progenitor cells, thus minimizing off-target cytotoxic effects. In some embodiments, a transgene encoding a PGRN or GRN agent may include a miR-126 targeting sequence.

[0553] Secretory signal peptide The polynucleotide encoding PGRN or GRN may include one or more polynucleotides encoding secretory signal peptides. The secretory signal peptides may have an amino acid sequence of 5-30 residues in length and may be located upstream (i.e., 5') of the polynucleotide encoding PGRN or GRN. These secretory signal peptides allow recognition of the nascent polypeptide during synthesis by the signal recognition particle, resulting in translocation to the ER, packaging into transport vesicles, and finally secretion. Exemplary secretory signal peptides for protein secretion are those derived from PGRN, IGF-II, alpha-1 antitrypsin, IL-2, IL-6, CD5, immunoglobulins, trypsinogen, serum albumin, prolactin, elastin, tissue plasminogen activator signal peptide (tPA-SP), and insulin. In some embodiments, cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) containing a transgene encoding a secreted form of PGRN or GRN can be utilized as a therapeutic strategy to correct a protein deficiency (e.g., PGRN or GRN) by injecting the missing protein into the bloodstream. As blood perfuses the patient's tissues, PGRN or GRN is taken up by the cells and transported to their site of action.

[0554] ApoE tagging of secreted progranulin or granulin for blood-brain barrier penetration In some embodiments, PGRN or GRN (e.g., PGRN or GRN fusion protein) is modified to penetrate the blood-brain barrier (BBB). Modifications to mediate BBB penetration are well known in the art. An exemplary modification is the use of a tag containing the Rb domain of ApoE (amino acid residues 148-173 of SEQ ID NO: 11). The complete ApoE peptide sequence is shown below. MKVLWAALLVTLAGCQAKVEQAVETEPEPELRQQTEWQSGQRWELALGRFWDYLRWVQTLSEQVQEELLSSQVTQELRALMDETMKELKAYKSELEEQLTPVAEETRARLSKELQAAQARLGADMEDVCGRLVQYRGEVQAMLGQSTEELRVRLASH LRKLRKRLLRDADDLQKRLAVYQAGAREGAERGLSAIRERLGPLVEQGRVRAATVGSLAGQPLQERAQAWGERLRARMEEMGSRTRDRLDEVKEQVAEVRAKLEEQAQQIRLQAEAFQARLKSWFEPLVEDMQRQWAGLVEKVQAAVGTSAAPVPSDNH (SEQ ID NO:11)

[0555] ApoE is a key protein involved in lipid transport, and its cellular internalization is mediated by several members of the low-density lipoprotein (LDL) receptor gene family, including the LDL receptor, the very low-density lipoprotein receptor (VLDLR), and the LDL receptor-related proteins (LRPs, including LRP1, LRP2, and LRP8). LDL receptors have been found to be highly expressed in brain capillary endothelial cells (BCECs), with down-regulation of expression observed in peripheral blood vessels. Restricted expression of LRPs and VLDLRs has also been noted in the liver and brain, when detected in BCECs, neurons, and glial cells. Except for LDLR, several members of the low-density lipoprotein receptor family (LDLRf) proteins, including LRP1 and VLDLR, are highly expressed in BCECs forming the BBB. These proteins may bind to ApoE and promote their transcytosis to the abluminal side of the BBB.

[0556] In addition, receptor-associated proteins (RAPs), antagonists and ligands of both LRP1 and VLDLR, have been shown to be more permeable across the BBB than transferrin in vivo and in vitro (Pan et al., J. Cell Sci. 117:5071-8 (2004)), indicating that these lipoprotein receptors (LDLRfs) may be efficient BBB delivery targets, despite their lower expression than the transferrin receptor. As described herein, a potent receptor-binding peptide (Rb) derived from ApoE, when engineered as a fusion protein, has the ability to translocate proteins across the BBB to the brain. Thus, this method, when engineered as a fusion protein, may function to selectively open the BBB for therapeutic agents. Because this peptide utilizes the Rb domain of ApoE, rather than the entire ApoE protein, it can be easily conjugated to diagnostic or therapeutic agents without compromising their biological functions or interfering with the important biological functions of ApoE. This route is also an alternative uptake route that can facilitate further / secondary brain distribution after the drug reaches the CNS due to the widespread expression of LDLRf members in the brain parenchyma. Regardless of the administration strategy, e.g., enzyme replacement therapy or cell-based gene-based therapy, both the amount and distribution of therapeutic agent in the brain parenchyma should have a significant impact on the clinical outcome of disease treatment. A detailed description of the development and use of the Rb domain of ApoE in targeted delivery of proteins across the BBB can be found in U.S. Patent Application Publication No. 20140219974, the entirety of which is incorporated herein by reference.

[0557] In some embodiments, the PGRN or GRN fusion protein has a peptide sequence comprising the LDLRf Rb domain of SEQ ID NO: 11, or a fragment, variant, or oligomer thereof. Exemplary receptor binding domains can be found at the N-terminus of ApoE, e.g., between amino acid residues 1-191 of SEQ ID NO: 11, between amino acid residues 25-185 of SEQ ID NO: 11, between amino acid residues 50-180 of SEQ ID NO: 11, between amino acid residues 75-175 of SEQ ID NO: 11, between amino acid residues 100-170 of SEQ ID NO: 11, or between amino acid residues 125-165 of SEQ ID NO: 11. Exemplary receptor binding domains have the amino acid sequence of residues 159-167 of SEQ ID NO: 11.

[0558] In some embodiments, the peptide sequence comprising the receptor binding domain of ApoE may comprise at least one amino acid mutation, deletion, addition, or substitution. In some embodiments, the amino acid substitution may be a combination of two or more mutations, deletions, additions, or substitutions. In some embodiments, at least one substitution is a conservative substitution. In some embodiments, at least one amino acid addition comprises the addition of a selected sequence already found in the Rb domain of ApoE. Those skilled in the art will recognize the appropriate modifications that can be made to the sequence while retaining some degree of biochemical activity for transport across the BBB.

[0559] Glycosylation-independent lysosomal targeting Glycosylation-independent lysosomal targeting (GILT) technology can be utilized to target therapeutic enzymes (e.g., PGRN or GRN) to lysosomes. Specifically, GILT technology uses a peptide tag instead of M6P to engage CI-MPR for lysosomal targeting. Typically, a GILT tag is a protein, peptide, or other moiety that binds to CI-MPR in a mannose-6-phosphate-independent manner. Advantageously, this technology mimics the normal biological mechanism for importing lysosomal enzymes, but in a mannose-6-phosphate-independent manner. In some embodiments, PGRN or GRN is secreted as a PGRN or GRN fusion protein containing PGRN or GRN and a GILT tag. In some embodiments, the GILT tag is fused to the N-terminus of the PGRN or GRN protein. In some embodiments, the GILT tag is fused to the C-terminus of the PGRN or GRN protein. In some embodiments, the GILT tag is derived from human insulin-like growth factor II (IGFII). Human IGF-II is a high affinity ligand of CI-MPR, also called IGF-II receptor. Binding of GILT-tagged therapeutic enzymes to M6P / IGF-II receptor targets the protein to lysosomes via the endocytic pathway. A detailed description of GILT technology and GILT tags can be found in U.S. Publication Nos. 20030082176, 20040006008, 20040005309, 20050281805, and 2009043207, all teachings of which are incorporated herein by reference in their entirety.

[0560] Furin-resistant GILT tag The GILT tag from IGF-II may undergo proteolytic cleavage by Furin during production in mammalian cells. The Furin protease typically recognizes and cleaves a cleavage site having the consensus sequence Arg-XX-Arg, where X is any amino acid. The cleavage site is located after the carboxy-terminal arginine (Arg) residue of the sequence. In some embodiments, the Furin cleavage site has the consensus sequence Lys / Arg-XXX-Lys / Arg-Arg, where X is any amino acid. The cleavage site is located after the carboxy-terminal arginine (Arg) residue of the sequence. The mature human IGF-II peptide sequence is shown below. AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSRGIVEECCFRSCDLALLETYCATPAKSE (SEQ ID NO:12)

[0561] Mature human IGF-II contains two potential overlapping furin cleavage sites between residues 34-40 (bold). A modified GILT tag that is resistant to cleavage by furin still retains the ability to bind to the CI-MPR in a mannose-6-phosphate-independent manner. Specifically, a furin-resistant GILT tag can be designed by mutating the amino acid sequence at one or more furin cleavage sites such that the mutation disables at least one furin cleavage site. Thus, in some embodiments, a furin-resistant GILT tag is a furin-resistant IGF-II mutein that includes a mutation that disables at least one furin protease cleavage site or alters the sequence adjacent to a furin protease cleavage site such that furin cleavage is blocked, inhibited, reduced, or slowed down compared to a wild-type IGF-II peptide (e.g., wild-type human mature IGF-II). The appropriate mutation does not affect the ability of the furin-resistant GILT tag to bind to the human cation-independent mannose-6-phosphate receptor. In some embodiments, the furin-resistant IGF-II muteins suitable for use with the compositions and methods described herein are capable of inhibiting IGF-II levels in a mannose-6-phosphate-independent manner at pH 7.4 of 10-15%. -7Less than or equal to (e.g., 10 -8 , 10 -9 , 10 -10 , 10 -11 The furin-resistant IGF-II mutein binds to the human cation-independent mannose-6-phosphate receptor with a dissociation constant of 30-40, or less. In some embodiments, the furin-resistant IGF-II mutein comprises a mutation within a region corresponding to amino acids 30-40 of SEQ ID NO: 12 (e.g., 31-40, 32-40, 33-40, 34-40, 30-39, 31-39, 32-39, 34-37, 32-39, 33-39, 34-39, 35-39, 36-39, 37-40, 34-40). In some embodiments, a suitable mutation disables at least one furin protease cleavage site. The mutation may be an amino acid substitution, deletion, or insertion. For example, any one amino acid in the region corresponding to residues 30-40 (e.g., 31-40, 32-40, 33-40, 34-40, 30-39, 31-39, 32-39, 34-37, 32-39, 33-39, 34-39, 35-39, 36-39, 37-40, 34-40) of SEQ ID NO: 12 can be substituted with any other amino acid or deleted. For example, a substitution at position 34 can affect furin recognition of the first cleavage site. Insertion of one or more additional amino acids within each recognition site can disable one or both furin cleavage sites. One or more of the residues at the degenerate positions can also be deleted to disable both furin cleavage sites.

[0562] In some embodiments, the furin-resistant IGF-II mutein comprises an amino acid substitution at a position corresponding to Arg37 or Arg40 of SEQ ID NO: 12. In some embodiments, the furin-resistant IGF-II mutein comprises a Lys or Ala substitution at position Arg37 or Arg40. Other substitutions are possible, including combinations of Lys and / or Ala mutations at both positions 37 and 40, or substitutions of amino acids other than Lys or Ala. In some embodiments, furin-resistant IGF-II muteins suitable for use in conjunction with the compositions and methods described herein may comprise additional mutations. For example, up to 30% or more of the residues in SEQ ID NO: 12 may be altered (e.g., up to 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30% or more of the residues may be altered). Thus, a furin-resistant IGF-II mutein suitable for use in conjunction with the compositions and methods described herein may have an amino acid sequence that is at least 70%, including at least 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, identical to SEQ ID NO: 12. In some embodiments, a furin-resistant IGF-II mutein suitable for use in conjunction with the compositions and methods described herein is specifically targeted to the CI-MPR. The furin-resistant IGF-II mutein binds to the CI-MPR with high affinity (e.g., 10 at pH 7.4). -7Mutations of IGF-II polypeptides that result in proteins that bind to IGF-II (with a dissociation constant of M or less) but with lower affinity compared to native IGF-II to other receptors to which IGF-II is known to bind are particularly useful. For example, furin-resistant IGF-II muteins suitable for use in conjunction with the compositions and methods described herein can be engineered to have lower binding affinity for the IGF-I receptor compared to the affinity of naturally occurring human IGF-II for the IGF-I receptor. Additional mutation strategies have been utilized and are discussed in detail in U.S. Patent Application Publication No. 2009043207, which is incorporated herein by reference. For example, substitution of IGF-II residues Tyr27 with Leu, Leu43 with Val, or Ser26 with Phe reduces the affinity of IGF-II for the IGF-I receptor by 94-fold, 56-fold, and 4-fold, respectively (Torres et al., J. Mol. Biol. 248(2):385-401 (1995)). Deletion of residues 1-7 of human IGF-II resulted in a 30-fold decrease in affinity for the human IGF-I receptor, accompanied by a 12-fold increase in affinity for the rat IGF-II receptor (Hashimoto et al., J. Biol. Chem. 270(30):18013-8 (1995)). The NMR structure of IGF-II shows that Thr7 is located close to residues 48Phe and 50Ser, as well as close to the 9Cys-47Cys disulfide bridge. It is thought that interactions of Thr7 with these residues may stabilize the flexible N-terminal hexapeptide required for IGF-I receptor binding (Terasawa et al., EMBO J. 13(23)5590-7 (1994)). Together, this interaction can modulate binding to the IGF-II receptor. Truncating the C-terminus of IGF-II (residues 62-67) also appears to reduce the affinity of IGF-II for the IGF-I receptor by 5-fold (Roth et al., Biochem. Biophys.Res. Commun. 181(2):907-14 (1991)). The binding faces of IGF-I and the cation-independent M6P receptor are on separate faces of IGF-II.Based on the structural and mutational data, a functional cation-independent M6P binding domain can be constructed that is significantly smaller than human IGF-II. For example, the amino-terminal amino acids (e.g., 1-7 or 2-7) and / or the carboxy-terminal residues 62-67 can be deleted or replaced. In addition, amino acids 29-40 can possibly be deleted or replaced without altering the folding of the rest of the polypeptide or binding to the cation-independent M6P receptor. Thus, a targeting moiety can be constructed that includes amino acids 8-28 and 41-61. These stretches of amino acids could possibly be directly linked or separated by a linker. Alternatively, amino acids 8-28 and 41-61 can be provided on separate polypeptide chains. The comparable domain of insulin, which is homologous to IGF-II and has a tertiary structure closely related to that of IGF-II, has sufficient structural information to permit proper refolding into the appropriate tertiary structure even when present in separate polypeptide chains (Wang et al., Trends Biochem. Sci. 16(8):279-281 (1991)). Thus, for example, amino acids 8-28, or conservatively substituted variants thereof, could be fused to a lysosomal enzyme; the resulting fusion protein could be mixed with amino acids 41-61, or conservatively substituted variants thereof, and administered to a patient. To avoid sequestration of IGF-II / GILT constructs, IGF-II can also be modified to minimize binding to serum IGF binding proteins (Baxter, Am. J. Physiol Endocrinol Metab. 278(6):967-76(2000)). Many studies have located the residues in IGF-II required for binding to IGF binding proteins. Constructs with mutations in these residues can be screened for retention of high affinity binding to the M6P / IGF-II receptor and low affinity for the IGF binding protein.For example, replacement of Phe 26 of IGF-II with Ser has been reported to reduce the affinity of IGF-II for IGFBP-1 and -6 but have no effect on binding to the M6P / IGF-II receptor (Bach et al., J. Biol. Chem. 268(13):9246-54 (1993)). Other substitutions, such as Lys at Glu9, may also be advantageous. Similar mutations in regions of IGF-I that are highly conserved with IGF-II, either separately or in combination, result in greatly reduced IGF-BP binding (Magee et al., Biochemistry 38(48):15863-70 (1999)).

[0563] An alternative approach is to identify the minimal region of IGF-II that can bind with high affinity to the M6P / IGF-II receptor. Most of the residues involved in binding of IGF-II to the M6P / IGF-II receptor are clustered on one side of IGF-II (Terasawa et al., EMBO J. 13(23):5590-7 (1994)). Although the IGF-II tertiary structure is usually maintained by three intramolecular disulfide bonds, peptides incorporating amino acid sequences on the M6P / IGF-II receptor binding surface of IGF-II can be designed to fold properly and have binding activity. Such minimal binding peptides are highly preferred lysosomal targeting domains. For example, a preferred lysosomal targeting domain is amino acids 8-67 of human IGF-II. Peptides designed based on the region around amino acids 48-55, which bind to the M6P / IGF-II receptor, are also desirable lysosomal targeting domains. Alternatively, random libraries of peptides can be screened for the ability to bind to the M6P / IGF-II receptor by either yeast two-hybrid or phage display type assays.

[0564] Many of the furin-resistant IGF-II muteins described herein have reduced or decreased binding affinity for the insulin receptor. Thus, in some embodiments, peptide tags suitable for use with the compositions and methods described herein have reduced or decreased binding affinity for the insulin receptor compared to the affinity of naturally occurring human IGF-II for the insulin receptor. In some embodiments, peptide tags suitable for use with the compositions and methods described herein that have reduced or decreased binding affinity for the insulin receptor include peptide tags that have a binding affinity for the insulin receptor that is 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 12-fold, 14-fold, 16-fold, 18-fold, 20-fold, 50-fold, or more than 100-fold lower than that of wild-type mature human IGF-II. Binding affinity for the insulin receptor can be measured using a variety of in vitro and in vivo assays known in the art.

[0565] In some embodiments, the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 13, as shown below. GGGGAGGGGAGGGGAGGGGAGGGPSLCGGELVDTLQFVCGDRGFYFSRPASRVSARSRGIVEECCFRSCDLALLETYCATPAKSE (SEQ ID NO: 13)

[0566] In some embodiments, the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 14, as shown below. GAPGGGSPAPAPTPAPTPAPAGGGPSGAPLCGGELVDTLQFVCGDRGFYFSRPASRVSARSRGIVEECCFRSCDLALLETYCATPAKSE (SEQ ID NO: 14)

[0567] In some embodiments, the GILT tag has an amino acid sequence having at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the amino acid sequence of SEQ ID NO: 15, as shown below. GAPGGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTGPSGAPLCGGELVDTLQFVCGDRGFYFSRPASRVSARSRGIVEECCFRSCDLALLETYCATPAKSE (SEQ ID NO: 15)

[0568] In some embodiments, the GILT tag is encoded by a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 16, as shown below. GGCGGAGGCGGAGCTGGTGGCGGCGGAGCAGGCGGTGGTGGTGCAGGCGGCGGAGGTGCTGGCGGAGGACCATCTCTTTGTGGCGGAGAACTGGTGGACACCCTGCAGTTCGTGTGTGGCGACAGAGGCT TCTACTTTAGCAGACCCGCCAGCAGAGTGTCCGCCAGATCTAGAGGAATCGTGGAAGAGTGCTGCTTCAGAAGCTGCGACCTGGCACTGCTGGAAACCTACTGTGCCACACCAGCCAAGAGCGAGTGATG (SEQ ID NO:16)

[0569] In some embodiments, the GILT tag is encoded by a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 17, as shown below. GGAGCACCAGGCGGAGGATCTCCAGCTCCTGCTCCTACACCAGCTCCAGCACCGACGCCTGCTCCAGCTGGCGGAGGACCTTCTGGTGCACCTCTTTGTGGCGGAGAGCTGGTGGATACCCTGCAGTTCGTGTGTGGCG ACCGGGGCTTCTACTTTAGCAGACCTGCCAGCAGAGTGTCCGCCAGATCTAGAGGCATCGTGGAAGAGTGCTGCTTCAGAAGCTGCGACCTGGCACTGCTGGAAACCTACTGTGCCACACCAGCCAAGAGCGAGTGATGA (SEQ ID NO:17)

[0570] In some embodiments, the GILT tag is encoded by a nucleic acid sequence having at least 85% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity) to the nucleic acid sequence of SEQ ID NO: 18, as shown below. GGAGCACCAGGCGGATCTCCAGCAGGATCTCCAACCTCTACCGAGGAAGGCACAAGCGAGTCTGCCACACCTGAGTCTGGACCTGGCACAAGCACAGAGCCTAGCGAAGGATCTGCCCCAGGTTCTCCTGCCGGCTCTCCTACAAGTACAGGACCTTCTGGCGCTCCACTGTGTGGC GGAGAACTGGTGGATACCCTGCAGTTCGTGTGCGGCGACAGAGGCTTCTACTTTAGCAGACCCGCCAGCAGAGTGTCCGCCAGATCTAGAGGAATCGTGGAAGAGTGCTGCTTCAGAAGCTGCGATCTGGCACTGCTGGAAACCTACTGTGCCACACCAGCCAAGAGCGAGTGATGA (SEQ ID NO:18)

[0571] Vectors for expressing progranulin or granulin In addition to achieving high transcription and translation rates, stable expression of exogenous genes in mammalian cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglial cells) can be achieved by integrating a polynucleotide containing the gene into the nuclear genome of the mammalian cells. Various vectors have been developed for delivering and integrating polynucleotides encoding exogenous proteins into the nuclear DNA of mammalian cells. Examples of expression vectors are disclosed, for example, in WO1994 / 011026, which is incorporated herein by reference. Expression vectors for use in the compositions and methods described herein include polynucleotide sequences encoding PGRN or GRN, as well as additional sequence elements used, for example, for the expression of these agents and / or the integration of these polynucleotide sequences into the genome of mammalian cells. Certain vectors that can be used for the expression of PGRN or GRN include plasmids that include regulatory sequences that direct gene transcription, such as promoter and enhancer regions. Other vectors useful for expressing PGRN or GRN include polynucleotide sequences that enhance the translation rate of these genes or improve the stability or nuclear export of the mRNA resulting from gene transcription. These sequence elements include, for example, 5' and 3' untranslated regions, IRES, and polyadenylation signal sites to direct efficient transcription of the genes carried on the expression vector. Expression vectors suitable for use in the compositions and methods described herein may also include polynucleotides that code for markers for selecting cells containing such vectors. Examples of suitable markers are genes that code for resistance to antibiotics such as ampicillin, chloramphenicol, kanamycin, and nourseothricin.

[0572] Viral vector for expressing progranulin or granulin Viral genomes provide a rich source of vectors that can be used for efficient delivery of exogenous genes into mammalian cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglial cells). Viral genomes are particularly useful vectors for gene delivery because polynucleotides contained within such genomes are typically integrated into the nuclear genome of mammalian cells by generalized or specialized transduction. These processes occur as part of the natural viral replication cycle and do not require additional proteins or reagents to induce gene integration. Examples of viral vectors are retroviruses (e.g., Retroviridae viral vectors), adenoviruses (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvoviruses (e.g., adeno-associated viruses), coronaviruses, negative strand RNA viruses such as orthomyxoviruses (e.g., influenza viruses), rhabdoviruses (e.g., rabies and vesicular stomatitis viruses), paramyxoviruses (e.g., measles and Sendai), positive strand RNA viruses such as picornaviruses and alphaviruses, as well as double-stranded DNA viruses including adenoviruses, herpesviruses (e.g., herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxviruses (vaccinia, modified vaccinia Ankara (MVA), fowlpox, canarypox). Other viruses include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, human papillomavirus, human foamy virus, and hepatitis virus.Examples of retroviruses are avian leukemia sarcoma, avian C virus, mammalian C, B, D virus, oncoretrovirus, HTLV-BLV group, lentivirus, alpharetrovirus, gammaretrovirus, spumavirus (Coffin, JM, Retroviridae: The viruses and their replication, Virology, Third Edition (Lippincott-Raven, Philadelphia, (1996))). Other examples are murine leukemia virus, murine sarcoma virus, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, gibbon monkey leukemia virus, Mason-Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentivirus. Other examples of vectors are described, for example, in McVey et al., (US 5,801,030), the teachings of which are incorporated herein by reference.

[0573] Retroviral Vectors The delivery vector used in the methods and compositions described herein may be a retroviral vector. One type of retroviral vector that can be used in the methods and compositions described herein is a lentiviral vector. A subset of retroviruses, lentiviral vectors (LVs), transduce a wide range of dividing and non-dividing cell types with high efficiency, resulting in stable, long-term expression of transgenes. An overview of the optimization strategy for packaging and transducing LVs is provided in Delenda, The Journal of Gene Medicine 6: S125 (2004), the disclosure of which is incorporated herein by reference.

[0574] The use of lentivirus-based gene transfer techniques relies on the in vitro generation of recombinant lentiviral particles carrying a highly deleted viral genome in which the transgene of interest is housed. In particular, recombinant lentiviruses are recovered by in trans co-expression in permissive cell lines of (1) a packaging construct, i.e., a vector expressing the Gag-Pol precursor together with Rev (alternatively expressed in trans); (2) a vector expressing an envelope receptor, generally of heterologous nature; and (3) a transfer vector, a viral cDNA in which all open reading frames have been removed but which maintains sequences necessary for replication, encapsidation, and expression, and into which the sequences to be expressed have been inserted.

[0575] The LV used in the methods and compositions described herein may include one or more of the following: 5'-long terminal repeat (LTR), HIV signal sequence, HIV Psi signal 5'-splice site (SD), delta-GAG element, Rev responsive element (RRE), 3'-splice site (SA), elongation factor (EF) 1-alpha promoter, and 3'-self-inactivating LTR (SIN-LTR). The lentiviral vector optionally includes a central polypurine tract (cPPT) and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), as described in US 6,136,597, the disclosure of which is incorporated herein by reference as it relates to the WPRE. The lentiviral vector may further include a pHR' backbone, which may include, for example, as shown below.

[0576] The lentigene LV described in Lu et al., Journal of Gene Medicine 6:963 (2004) can also be used to express DNA molecules and / or transduce cells. The LV used in the methods and compositions described herein may include a 5'-long terminal repeat (LTR), an HIV signal sequence, an HIV Psi signal 5'-splice site (SD), a delta-GAG element, a Rev responsive element (RRE), a 3'-splice site (SA), an elongation factor (EF) 1-alpha promoter, and a 3'-self-inactivating LTR (SIN-LTR). Optionally, it will be readily apparent to one of skill in the art that one or more of these regions may be replaced with another region that performs a similar function.

[0577] PGRN or GRN should be expressed at a sufficiently high level. The expression of the transgene is driven by a promoter sequence. Optionally, the LV comprises a CMV promoter. The promoter may be an EF1α or PGK promoter. In another embodiment, the promoter is a microglia-specific promoter, such as a CD68 promoter, a CX3CR1 promoter, a CD11b promoter, an AIF1 promoter, a P2Y12 promoter, a TMEM119 promoter, or a CSF1R promoter. Optionally, the LV comprises a synthetic promoter optimized for use in mammalian cells. Those skilled in the art will be familiar with many promoters that are suitable in the vector constructs described herein.

[0578] Enhancer elements can be used to increase expression of modified DNA molecules or to enhance lentiviral integration efficiency. LVs used in the methods and compositions described herein may include a nef sequence. LVs used in the methods and compositions described herein may include a cPPT sequence that enhances vector integration. The cPPT acts as a second origin of (+)-strand DNA synthesis and introduces a partial strand overlap in the middle of the native HIV genome. Introducing a cPPT sequence into the transfer vector backbone significantly increased nuclear transport and the total amount of genome integrated into the DNA of the target cell. LVs used in the methods and compositions described herein may include a woodchuck posttranscriptional regulatory element (WPRE). The WPRE functions at the transcriptional level by promoting nuclear export of transcripts and / or by increasing the efficiency of polyadenylation of nascent transcripts, thus increasing the total amount of mRNA in the cell. Addition of the WPRE to the LV significantly improves transgene expression levels from several different promoters both in vitro and in vivo. LVs used in the methods and compositions described herein may include both a cPPT sequence and a WPRE sequence. The vector may also contain an IRES sequence, which allows expression of multiple polypeptides from a single promoter.

[0579] In addition to IRES sequences, other elements that allow for the expression of multiple polypeptides are useful. The vectors used in the methods and compositions described herein may contain multiple promoters that allow for the expression of more than one polypeptide. The vectors used in the methods and compositions described herein may contain protein cleavage sites that allow for the expression of more than one polypeptide. Examples of protein cleavage sites that allow for the expression of more than one polypeptide are described in Klump et al., Gene Ther.;8:811 (2001), Osborn et al., Molecular Therapy 12:569 (2005), Szymczak and Vignali Expert Opin Biol Ther. 5:627 (2005), and Szymczak et al. Nat Biotechnol. 22:589 (2004), which are incorporated herein by reference as their disclosures relate to protein cleavage sites that allow for the expression of more than one polypeptide. It will be readily apparent to one of skill in the art that other elements that allow for the expression of multiple polypeptides identified in the future will be useful and can be utilized in vectors suitable for use in the compositions and methods described herein.

[0580] The vectors used in the methods and compositions described herein may be clinical grade vectors.

[0581] Viral Regulatory Elements The viral regulatory element is a component of the delivery vehicle used to introduce the nucleic acid molecule into the host cell (e.g., pluripotent cell, ESC, iPSC, pluripotent cell, CD34+ cell, HSC, MPC, BLPC, monocyte, macrophage, microglial progenitor cell, or microglial cell). The viral regulatory element is optionally a retroviral regulatory element. For example, the viral regulatory element may be the LTR and gag sequence from HSC1 or MSCV. The retroviral regulatory element may be derived from a lentivirus or may be a heterologous sequence identified from other genomic regions. Those skilled in the art will also understand that other viral regulatory elements may be used with the nucleic acid molecules described herein as they are identified.

[0582] Adeno-associated virus vectors for nucleic acid delivery The nucleic acids of the compositions and methods described herein can be incorporated into rAAV vectors and / or virions to facilitate their introduction into cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglial cells). AAV vectors can be used in the central nervous system, and suitable promoters and serotypes are discussed in Pignataro et al., J Neural Transm (2017) (epub ahead of print), which is incorporated herein by reference as its disclosure relates to promoters and AAV serotypes useful in CNS gene therapy. The rAAV vectors useful in the compositions and methods described herein are recombinant nucleic acid constructs that include (1) a heterologous sequence to be expressed (e.g., a polynucleotide encoding PGRN or GRN) and (2) viral sequences that facilitate integration and expression of the heterologous gene. The viral sequences may include sequences of AAV required in cis for DNA replication and packaging into virions (e.g., functional ITRs). Such rAAV vectors may also contain marker or reporter genes. Useful rAAV vectors are deleted in whole or in part from one or more of AAV WT genes, but retain functional adjacent ITR sequences. AAV ITRs may be of any serotype suitable for a particular application. Methods for using rAAV vectors are described, for example, in Tai et al., J. Biomed. Sci. 7:279 (2000), and Monahan and Samulski, Gene Delivery 7:24 (2000), the disclosures of which are incorporated herein by reference as they relate to AAV vectors for gene delivery.

[0583] The nucleic acids and vectors described herein can be incorporated into rAAV virions to facilitate the introduction of the nucleic acid or vector into cells. The capsid protein of AAV constitutes the outer non-nucleic acid portion of the virion and is encoded by the AAV cap gene. The cap gene encodes three viral coat proteins, VP1, VP2, and VP3, required for the assembly of the virion. The construction of rAAV virions is described, for example, in US5,173,414; US5,139,941; US5,863,541; US5,869,305; US6,057,152; and US6,376,237, the disclosures of which are incorporated herein by reference as they relate to AAV vectors for gene delivery, as well as in Rabinowitz et al., J. Virol. 76:791 (2002) and Bowles et al., J. Virol. 77:423 (2003).

[0584] rAAV virions useful in conjunction with the compositions and methods described herein include those derived from various AAV serotypes, including AAV 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and rh74. For targeting cells located in or delivered to the central nervous system, AAV2, AAV9, and AAV10 may be particularly useful. The construction and use of AAV vectors of various serotypes and AAV proteins are described, for example, in Chao et al., Mol. Ther. 2:619 (2000); Davidson et al., Proc. Natl. Acad. Sci. USA 97:3428 (2000); Xiao et al., J. Virol. 72:2224 (1998); Halbert et al., J. Virol. 74:1524 (2000); Halbert et al., J. Virol. 75:6615 (2001); and Auricchio et al., Hum. Molec. Genet. 10:3075 (2001), the disclosures of each of which are incorporated by reference herein as they relate to AAV vectors for gene delivery.

[0585] Pseudotyped rAAV vectors are also useful in conjunction with the compositions and methods described herein. Pseudotyped vectors include AAV vectors of a given serotype pseudotyped with a capsid gene from a serotype other than the given serotype (e.g., AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, and AAV10, among others). Techniques for the construction and use of pseudotyped rAAV virions are known in the art and are described, for example, in Duan et al., J. Virol. 75:7662 (2001); Halbert et al., J. Virol. 74:1524 (2000); Zolotukhin et al., Methods, 28:158 (2002); and Auricchio et al., Hum. Molec. Genet. 10:3075 (2001).

[0586] AAV virions with mutations in the virion capsid can be used to infect specific cell types more effectively than non-mutated capsid virions. For example, suitable AAV mutants may have ligand insertion mutations to facilitate targeting of AAV to specific cell types. The construction and characterization of AAV capsid mutants, including insertion mutants, alanine screening mutants, and epitope tag mutants, are described in Wu et al., J. Virol. 74:8635 (2000). Other rAAV virions that can be used in the methods described herein include capsid hybrids generated by molecular breeding of viruses and even by exon shuffling. See, for example, Soong et al., Nat. Genet., 25:436 (2000) and Kolman and Stemmer, Nat. Biotechnol. 19:423 (2001).

[0587] Methods for delivering exogenous nucleic acids to target cells Techniques that can be used to introduce polynucleotides, such as codon-optimized DNA or RNA (e.g., mRNA, tRNA, siRNA, miRNA, shRNA, chemically modified RNA), into mammalian cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglial cells) are well known in the art. For example, electroporation can be used to permeabilize mammalian cells (e.g., human target cells) by applying an electrostatic potential to the cells of interest. Mammalian cells, such as human cells, subjected to an external electric field in this manner are then susceptible to uptake of exogenous nucleic acids. Electroporation of mammalian cells is described in detail, for example, in Chu et al., Nucleic Acids Research 15:1311 (1987), the disclosure of which is incorporated herein by reference. A similar technique, Nucleofection™, utilizes an applied electric field to stimulate uptake of exogenous polynucleotides into the nucleus of eukaryotic cells. Nucleofection™ and protocols useful for carrying out this technique are described in detail, for example, in Distler et al., Experimental Dermatology 14:315 (2005), as well as US2010 / 0317114, the disclosures of each of which are incorporated herein by reference.

[0588] Another useful technique for transfection of target cells is squeeze-poration. This technique induces rapid mechanical deformation of cells to stimulate the uptake of exogenous DNA through membrane pores that form in response to applied stress. This technique is advantageous in that no vector is required to deliver nucleic acid to cells, such as human target cells. Squeeze-poration is described in detail, for example, in Sharei et al., Journal of Visualized Experiments 81:e50980 (2013), the disclosure of which is incorporated herein by reference.

[0589] Lipofection is another technique useful for transfection of target cells. This method involves loading nucleic acid into liposomes that often present cationic functional groups, such as quaternary or protonated amines, toward the exterior of the liposome. This promotes electrostatic interactions between the liposome and the cell due to the anionic nature of the cell membrane, ultimately leading to the uptake of exogenous nucleic acid, for example, by direct fusion of the liposome with the cell membrane or by endocytosis of the complex. Lipofection is described in detail, for example, in U.S. Pat. No. 7,442,386, the disclosure of which is incorporated herein by reference. A similar technique that utilizes ionic interactions with the cell membrane to induce the uptake of exogenous nucleic acid is to contact cells with cationic polymer-nucleic acid complexes. Exemplary cationic molecules that associate with polynucleotides to impart a positive charge favorable for interaction with cell membranes are activated dendrimers (e.g., as described in Dennig, Topics in Current Chemistry 228:227 (2003), the disclosure of which is incorporated herein by reference), polyethyleneimine, and diethylaminoethyl (DEAE)-dextran, the use of which as transfection agents is described in detail, for example, in Gulick et al., Current Protocols in Molecular Biology 40:1:9.2:9.2.1 (1997), the disclosure of which is incorporated herein by reference. Magnetic beads are another tool that can be used to transfect target cells in a gentle and efficient manner, since this method utilizes an applied magnetic field to direct the uptake of nucleic acids. This technology is described in detail, for example, in US2010 / 0227406, the disclosure of which is incorporated herein by reference.

[0590] Another useful tool for inducing the uptake of exogenous nucleic acids by target cells is laser transfection, also called phototransfection, a technique that involves exposing cells to electromagnetic radiation of specific wavelengths in order to gently permeabilize the cells and allow polynucleotides to penetrate the cell membrane. The biological activity of this technique appears to be similar to, and in some cases superior to, electroporation.

[0591] Impalefection is another technique that can be used to deliver genetic material to target cells. It relies on the use of nanomaterials such as carbon nanofibers, carbon nanotubes, and nanowires. Needle-like nanostructures are synthesized perpendicular to the surface of a substrate. DNA containing the genes intended for intracellular delivery is attached to the nanostructure surface. A tip with an array of these needles is then pressed against a cell or tissue. Cells impaled with the nanostructures can express the delivered gene(s). An example of this technique is described in Shalek et al., PNAS 107:25 1870 (2010), the disclosure of which is incorporated herein by reference.

[0592] Magnetofection can also be used to deliver nucleic acids to target cells. The principle of magnetofection is to associate nucleic acids with cationic magnetic nanoparticles. The magnetic nanoparticles are made of iron oxide, which is completely biodegradable, and are coated with specific cationic proprietary molecules that vary depending on the application. Their association with gene vectors (DNA, siRNA, viral vectors, etc.) is achieved by salt-induced colloidal aggregation and electrostatic interactions. The magnetic particles are then concentrated at the target cells under the influence of an external magnetic field generated by a magnet. This technology is described in detail in Scherer et al., Gene Therapy 9:102 (2002), the disclosure of which is incorporated herein by reference.

[0593] Another useful tool for inducing the uptake of exogenous nucleic acid by target cells is sonoporation, which is a technique that uses sound (typically ultrasonic frequencies) to change the permeability of cell plasma membrane, making cells permeable and allowing polynucleotides to penetrate the cell membrane.This technique is described in detail, for example, in Rhodes et al., Methods in Cell Biology 82:309 (2007), the disclosure of which is incorporated herein by reference.

[0594] Microvesicles represent another potential carrier that can be used to modify the genome of target cells according to the methods described herein.For example, microvesicles induced by the simultaneous overexpression of glycoprotein VSV-G and genome modification proteins, such as nucleases, can be used to efficiently deliver proteins that later catalyze the site-specific cleavage of endogenous polynucleotide sequences to cells, preparing the genome of cells for the covalent integration of a polynucleotide of interest, such as a gene or regulatory sequence.The use of such vesicles, also called gesicles, in the genetic modification of eukaryotic cells is described in detail, for example, in Quinn et al., Genetic Modification of Target Cells by Direct Delivery of Active Protein [abstract]: Methylation changes in early embryonic genes in cancer [abstract] : Proceedings of the 18th Annual Meeting of the American Society of Gene and Cell Therapy;2015 May 13, Abstract No. 122.

[0595] Modulating gene expression using gene editing techniques Disruption of endogenous progranulin or granulin In some embodiments, endogenous PGRN or GRN is destroyed (e.g., in a subject undergoing treatment, such as in a population of neurons in a subject undergoing treatment, or in cells administered to the subject (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia)). An exemplary method for destroying endogenous PGRN or GRN expression is to administer an inhibitory RNA molecule to a subject or contact the population of neurons of the subject or the population of cells administered to the subject. The inhibitory RNA molecule may function to destroy endogenous PGRN or GRN, for example, acting via an RNA interference (RNAi) pathway. The inhibitory RNA molecule may reduce the expression level (e.g., protein level or mRNA level) of endogenous PGRN or GRN. For example, the inhibitory RNA molecule includes short interfering RNA, short hairpin RNA, and / or microRNA that target full-length endogenous PGRN or GRN. siRNAs are double-stranded RNA molecules typically with a length of about 19-25 base pairs. shRNAs are RNA molecules containing hairpin turns that reduce expression of target genes via RNAi. shRNAs can be delivered to cells, for example, by transfection, electroporation, or transduction, in the form of plasmids, e.g., viral or bacterial vectors. MicroRNAs are non-coding RNA molecules typically with a length of about 22 nucleotides. miRNAs bind to target sites on mRNA molecules and silence the mRNA, for example, by causing cleavage of the mRNA, destabilization of the mRNA, or inhibition of translation of the mRNA. Inhibitory RNA molecules can be modified to include modified nucleotides, e.g., 2'-fluoro, 2'-o-methyl, 2'-deoxy, unlocked nucleic acid, 2'-hydroxy, phosphorothioate, 2'-thiouridine, 4'-thiouridine, 2'-deoxyuridine. Without wishing to be bound by theory, it is believed that certain modifications may increase nuclease resistance and / or serum stability, or may reduce immunogenicity.

[0596] In some embodiments, the inhibitory RNA molecule reduces the level and / or activity or function of endogenous PGRN or GRN. In embodiments, the inhibitory RNA molecule inhibits the expression of endogenous PGRN or GRN. In other embodiments, the inhibitory RNA molecule increases the degradation of endogenous PGRN or GRN and / or reduces the stability of endogenous PGRN or GRN. The inhibitory RNA molecule can be chemically synthesized or transcribed in vitro.

[0597] In some embodiments, endogenous PGRN or GRN is destroyed in cells that contain PGRN or GRN transgenes, for example, using gene editing techniques described herein. In some embodiments, endogenous PGRN or GRN is destroyed globally in a subject, for example, using gene editing techniques described herein. In some embodiments, endogenous PGRN or GRN is destroyed in a population of neurons in a subject, for example, using gene editing techniques described herein. In some embodiments, destruction of endogenous PGRN or GRN in a subject, neuron, and / or cell that contains PGRN or GRN transgenes is performed before administering the cells to a subject.

[0598] The generation and use of inhibitory therapeutics based on non-coding RNA, such as ribozymes, RNAse P, siRNA, miRNA, etc., are also known in the art, as described, for example, in Sioud, RNA Therapeutics: Function, Design, and Delivery (Methods in Molecular Biology), Humana Press (2010).

[0599] Nuclease-Mediated Gene Regulation Another useful tool for disruption and / or integration of target genes into the genome of cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglial cells) is the clustered regularly interspaced short palindromic repeats (CRISPR) / Cas system, which originally evolved as an adaptive defense mechanism in bacteria and archaea against viral infection. The CRISPR / Cas system contains palindromic repeat sequences within plasmid DNA and CRISPR-associated proteins (Cas; e.g., Cas9 or Cas12a). This DNA and protein assembly directs site-specific DNA cleavage of the target sequence by first integrating foreign DNA into the CRISPR locus. Polynucleotides containing these foreign sequences and the repeat-spacer elements of the CRISPR locus can then be transcribed into host cells to generate guide RNAs, which can then be annealed to the target sequence and localize Cas nuclease to this site. In this way, highly site-specific Cas-mediated DNA cleavage can be engineered in foreign polynucleotides, since the interaction that brings Cas into close proximity with the target DNA molecule is governed by RNA:DNA hybridization. As a result, the CRISPR / Cas system can theoretically be designed to cleave any target DNA molecule of interest (e.g., endogenous PGRN or GRN). This technology has been utilized to edit eukaryotic genomes (Hwang et al. Nature Biotechnology 31:227 (2013), the disclosure of which is incorporated herein by reference) and can be used as an efficient means to site-specifically edit cellular genomes to perform DNA cleavage prior to the incorporation of genes encoding target genes. The use of CRISPR / Cas to regulate gene expression is described, for example, in US 8,697,359, the disclosure of which is incorporated herein by reference.Alternative methods for disrupting target DNA by site-specific cleavage of genomic DNA prior to integration of a gene of interest into a cell include the use of zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Unlike the CRISPR / Cas system, these enzymes do not contain a guided polynucleotide to localize to a specific target sequence. Instead, target specificity is controlled by a DNA-binding domain within these enzymes. The use of ZFNs and TALENs in genome editing applications is described, for example, in Urnov et al. Nature Reviews Genetics 11:636 (2010); and Joung et al. Nature Reviews Molecular Cell Biology 14:49 (2013), the disclosures of both of which are incorporated herein by reference. In some embodiments, endogenous PGRN or GRN can be disrupted in cells containing PGRN or GRN transgenes using these gene editing techniques described herein.

[0600] Transposon-mediated gene regulation In addition to viral vectors, various additional tools have been developed that can be used to incorporate exogenous genes into cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia). One such method that can be used to incorporate a polynucleotide encoding a target gene into a cell involves the use of a transposon. A transposon is a polynucleotide that encodes a transposase enzyme and contains a polynucleotide sequence or gene of interest flanked by 5' and 3' excision sites. Once the transposon is delivered to a cell, expression of the transposase gene begins, resulting in an active enzyme that cleaves the gene of interest from the transposon. This activity is mediated by site-specific recognition of the transposon excision site by the transposase. In certain cases, these excision sites may be terminal repeats or inverted terminal repeats. Once excised from the transposon, the gene of interest can be integrated into the genome of a mammalian cell by transposase-catalyzed cleavage of a similar excision site present in the nuclear genome of the cell. This allows the gene of interest to be inserted into the nuclear DNA cleaved at the complementary excision site, after which the integration process is completed with covalent ligation of a phosphodiester bond that connects the gene of interest to the DNA of the mammalian cell genome. In certain cases, the transposon may be a retrotransposon, such that the gene encoding the target gene is first transcribed into an RNA product and then reverse transcribed into DNA before integration into the mammalian cell genome. Transposon systems include piggyback transposons (described in detail, for example, in WO2010 / 085699) and Sleeping Beauty transposons (described in detail, for example, in US2005 / 0112764), the disclosures of each of which are incorporated herein by reference.

[0601] Diagnostic methods Methods well known in the art, such as those described in The Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition and the International Classification of Diseases, 11 thUsing the method described in Revision, etc., a subject can be diagnosed as having an NCD (e.g., FTLD or NCL). For example, the diagnosis of an NCD in a subject can be guided by a neuropsychological test that evaluates the degree of cognitive impairment in the subject. The cognitive function of a subject can be evaluated by performing a cognitive test that evaluates processing ability across one or more cognitive domains, including but not limited to complex attention, executive function, learning and memory, language, sensory-motor function, and social cognition. The cognitive function in a subject can be compared to a standard (e.g., a reference population, such as the general population) appropriate for the subject's age, medical history, education, socio-economic status, and lifestyle to determine a diagnosis of an NCD in the subject. A subject can be diagnosed as having a severe NCD or a mild NCD. A severe NCD is characterized by a significant cognitive decline that interferes with an individual's independence and / or normal daily functioning and is not due to delirium or other mental disorders. A mild NCD is characterized by a moderate cognitive decline that does not interfere with a subject's independence and / or normal daily functioning and is not due to delirium or other mental disorders. Severe NCDs may be characterized by a score obtained by a subject on a cognitive test that is more than two standard deviations away from the mean score of a reference population (e.g., the mean score of the general population) or that is in the third percentile of the distribution of scores in the reference population. Mild NCDs may be characterized by a score obtained by a subject on a cognitive test that is one to two standard deviations away from the mean score of a reference population (e.g., the mean score of the general population) or that is in the third to sixteenth percentile of the distribution of scores in the reference population.Non-limiting examples of cognitive tests include the Eight-item Informant Interview to Differentiate Aging and Dementia (AD8), Annual Wellness Visit (AWV), General Practitioner Assessment of Cognition (GPCOG), Health Risk Assessment (HRA), Memory Impairment Screen (MIS), Mini Mental Status Exam (MMSE), Montreal Cognitive Assessment (MoCA), St. Louis University Mental Status Exam (SLUMS), and Short Informant Questionnaire on Cognitive Decline in the Elderly (Short IQCODE).In addition, or alternatively, the use of F18-fluorodeoxyglucose PET scans or MRI scans can also be used to determine the presence of neurodegeneration in subjects with NCD.

[0602] Furthermore, the subject can be examined for the presence of biomarkers specific to the particular NCD of interest.For example, the subject can be examined for the presence of biomarkers that indicate that the subject has FTLD, such as tau-positive neuronal and glial inclusions, ub-positive and TDP43-positive but tau-negative inclusions, ub- and FUS-positive but tau-negative inclusions, mutations in the PGRN gene disclosed herein and / or mutations at chromosome 17q21 disclosed herein, etc. To determine whether the subject has NCL, the subject can also be examined for the presence of lipofuscin inclusions in body cells, such as neurons, liver, spleen, myocardium, and kidney cells, as well as mutations in one or more CLN genes or PGRN genes.

[0603] Treatment Method Selecting a target The subject that can be treated as described herein is the subject that has NCD (e.g., FTLD or NCL) or is at risk of developing.The type of FTLD can be PGRN-related FTLD, including but not limited to behavioral abnormal frontotemporal dementia, semantic dementia, and progressive non-verbal aphasia variant of FTLD.The type of NCL can be PGRN-related NCL, including but not limited to Santavuori-Haltia disease, Jansky-Bielschowsky disease, Batten disease, Kuhs disease, Finnish late-onset childhood NCL, atypical late-onset childhood NCL, CLN7 NCL, CLN8 NCL, Turkish late-onset childhood NCL, type 9 NCL, CLN10 NCL, and CLN11 NCL.

[0604] In addition, the type of NCD may be sporadic NCD caused by environmental toxins, such as herbicides or pesticides, or NCD associated with non-PGRN mutations, such as mutations in one or more genes associated with NCDs. The compositions and methods described herein can be used to treat subjects with normal PGRN or GRN activity, reduced PGRN or GRN activity, and subjects with unknown PGRN mutation status and / or PGRN or GRN activity levels. The compositions and methods described herein can also be administered as preventive treatment to subjects at risk of developing NCDs, such as subjects with PGRN mutations, subjects with reduced PGRN or GRN activity, subjects with one or more mutations in genes associated with NCDs, or subjects exposed to environmental toxins associated with NCDs. Subjects at risk of NCD may show early symptoms, but may not yet show symptoms when treatment is administered.

[0605] In some embodiments, the methods and compositions described herein are used to detect and treat a variety of inflammatory bowel disorders, including, for example, frameshift mutations (e.g., p.C31LfsX35, p.C31LfsX35, p.S82VfsX174, p.L271LfsX174, and / or p.T382NfsX32 mutations), missense mutations (p.C521Y, p.A9D, p.P248L, p.R432C, p.C139R, p.C521Y, and / or p.C139R mutations), nonsense mutations (e.g., p.Q125X or p.R493 X mutation), insertion mutation (e.g., c.1145insA mutation), and / or transversion mutation (e.g., p.0(IVS1+5G>C mutation). In some embodiments, the methods and compositions described herein can be administered to subjects with any other pathogenic mutation in the PGRN gene. For example, the pathogenic mutation in the PGRN gene can be any of the mutations discussed in Gijselinck et al., Human Mutation 29:1373-1386, (2012), which is incorporated by reference herein as the disclosure relates to human PGRN mutations.

[0606] Route of administration The cells and compositions described herein can be administered to a subject with an NCD (e.g., FTLD or NCL) by a variety of routes, such as intraventricular, intrathecal, intraparenchymal, stereotactic, intravenous, intraosseous, or by bone marrow transplantation. In some embodiments, the cells and compositions described herein can be administered systemically (e.g., intravenously), directly to the central nervous system (CNS) (e.g., intraventricular, intrathecal, intraparenchymal, or stereotactic), or directly to bone marrow (e.g., intraosseous). In some embodiments, the cells and compositions described herein are administered to a subject intraventricularly into the lateral cerebral ventricles (a description of this method can be found in Capotondo et al., Science Advances 3:e1701211 (2017), which is incorporated herein by reference, as it relates to intraventricular infusion of hematopoietic stem and progenitor cells into the lateral cerebral ventricles in a mouse model). The optimal route of administration in any given case depends on the particular cells or compositions administered, the subject, the pharmaceutical formulation method, the method of administration (e.g., time of administration and route of administration), the age, weight, sex of the subject, the severity of the disease being treated, the diet of the subject, and the excretion rate of the subject. Multiple routes of administration can be used to treat a single subject, for example, intraventricular or stereotactic injection and intravenous injection, intraventricular or stereotactic injection and intraosseous injection, intraventricular or stereotactic injection and bone marrow transplant, intraventricular or stereotactic injection and intraparenchymal injection, intrathecal injection and intravenous injection, intrathecal injection and intraosseous injection, intrathecal injection and bone marrow transplant, intrathecal injection and intraparenchymal injection, intraparenchymal injection and intravenous injection, intraparenchymal injection and intraosseous injection, or intraparenchymal injection and bone marrow transplant. Multiple routes of administration can be used to treat a single subject at one time, or the subject can be treated via one route of administration initially and then via another route of administration at a second visit, for example, one week, two weeks, one month, six months, or one year later. For treatment of an NCD, the cells may be administered to a subject once, or the cells may be administered to a subject one or more times (eg, 2-10 times) weekly, monthly, or yearly.

[0607] Pretreatment It may be advantageous to deplete or remove endogenous microglia and / or hematopoietic stem and progenitor cells prior to administration of cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) or compositions. Microglia and / or hematopoietic stem and progenitor cells can be removed using chemical agents (e.g., busulfan, treosulfan, PLX3397, PLX647, PLX5622, or clodronate liposomes), irradiation, or combinations thereof. Agents used for cell removal may be BBB-permeable (e.g., busulfan) or may lack the ability to cross the BBB (e.g., treosulfan). Exemplary microglia and / or hematopoietic stem and progenitor cell ablative agents are busulfan (Capotondo et al., PNAS 109:15018 (2012), which is incorporated by reference as the disclosure thereof pertains to the use of busulfan to ablate microglia), treosulfan, PLX3397, PLX647, PLX5622, or clodronate liposomes. Other agents for depleting endogenous microglia and / or hematopoietic stem and progenitor cells include cytotoxins that can be covalently attached to an antibody or antigen-binding fragment thereof capable of binding to an antigen expressed by hematopoietic stem cells to form an antibody-drug conjugate. Cytotoxins suitable for antibody drug conjugates include DNA intercalating agents (e.g., anthracyclines), agents capable of disrupting the mitotic spindle (e.g., vinca alkaloids, maytansine, maytansinoids, and their derivatives), RNA polymerase inhibitors (e.g., amatoxins such as a-amanitin and its derivatives), agents capable of interfering with protein biosynthesis (e.g., agents exhibiting rRNA N-glycosidase activity, such as saporin and ricin A chain), among others known in the art.The depletion may eliminate all microglia and / or hematopoietic stem and progenitor cells or may reduce the number of microglia and / or hematopoietic stem and progenitor cells by at least 5% (e.g., at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more). The agent or agents that deplete microglia and / or hematopoietic stem and progenitor cells can be administered at least one week (e.g., 1, 2, 3, 4, 5, or 6 weeks or more) prior to administration of the cells or compositions described herein. Cells administered in the manner described herein can replace the depleted microglia and / or hematopoietic stem and progenitor cells and can repopulate the brain after intraventricular, stereotactic, intravenous, or intraosseous injection, or after bone marrow transplantation. Cells administered intravenously, intraosseously, or by bone marrow transplantation can cross the blood-brain barrier, enter the brain, and differentiate into microglia. Cells administered to the brain, for example intracerebroventricularly or stereotactically, may differentiate into microglia in vivo or may differentiate into microglia ex vivo.

[0608] Stem Cell Recapture The methods described herein may include administering a population of cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) to a subject. The cells may be cells that have not been modified to express a transgene encoding PGRN or GRN. The cells may destroy endogenous PGRN or GRN. After conditioning as described herein, the cells may be administered systemically (e.g., intravenously) or by bone marrow transplantation to reconstitute the bone marrow compartment. For example, the cells may migrate into a stem cell niche and increase the amount of cells of the hematopoietic lineage at such sites by, for example, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, 310%, 320%, 330%, 340%, 350%, 360%, 370%, 380%, 390%, 400%, 410%, 420%, 430%, 440%, 450%, 460%, 470%, 480%, 490%, 500%, 510%, 520%, 53 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61 %, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or more. Administration can be performed before or after administration of a composition described herein.

[0609] Donor cell selection In some embodiments, the subject is a donor. In such a case, the removed cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) can be reinjected into the subject (after modification (e.g., integration of transgenes encoding PGRN or GRN and / or disruption of endogenous PGRN or GRN), and the cells will then home to hematopoietic tissue and establish productive hematopoiesis, thereby expanding or repopulating the defective or missing cell line (e.g., population of microglia) in the subject. In this scenario, the transplanted cells are least likely to undergo graft rejection, since the injected cells are derived from the subject and express the same HLA class me and class II antigens as the subject expresses. Alternatively, the subject and the donor can be different. In some embodiments, the subject and the donor can be related, e.g., HLA-matched. As described herein, HLA-matched donor-recipient pairs have a lower risk of graft rejection because endogenous T cells and NK cells in the transplant recipient are less likely to recognize foreign hematopoietic stem or progenitor cells as foreign and therefore less likely to mount an immune response to the transplant. An exemplary HLA-matched donor-recipient pair is a genetically related donor and recipient, such as a familial donor-recipient pair (e.g., a sibling donor-recipient pair). In some embodiments, the subject and donor are HLA-mismatched, which occurs when at least one HLA antigen is mismatched between the donor and the recipient, particularly with respect to HLA-A, HLA-B, and HLA-DR. For example, one haplotype may be matched between the donor and the recipient, and the other may be mismatched, to reduce the likelihood of graft rejection.

[0610] Pharmaceutical Compositions and Dosages The number of cells administered to a subject to treat an NCD (e.g., FTLD or NCL, e.g., PGRN-associated FTLD or NCL) as described herein can depend, for example, on the expression level of PGRN or GRN, the subject, the pharmaceutical formulation method, the method of administration (e.g., time and route of administration), the age, weight, and sex of the subject, the severity of the disease being treated, and whether the subject has been treated with an agent that eliminates endogenous microglia. The number of cells administered can be, for example, 1×10 6 cells / kg~1×10 12 cells / kg, or more (e.g., 1 x 10 7 cells / kg, 1×10 8 cells / kg, 1×10 9 cells / kg, 1×10 10 cells / kg, 1×10 11 cells / kg, 1×10 12 The dosage may be 1×10 cells / kg or more). The cells may be administered in an undifferentiated state or after partial or complete differentiation into microglia. The number of cells may be administered at any suitable dose after pretreatment. A non-limiting example of a dose is about 1×10 5 Approximately 1 x 10 cells / kg of recipient 7 Cells / kg (e.g., approximately 2 x 10 5 cells / kg ~ approx. 9×10 6 cells / kg, approximately 3×10 5 cells / kg ~ approx. 8×10 6 cells / kg, approximately 4×10 5 cells / kg~about 7×10 6 cells / kg, approximately 5×10 5 cells / kg~about 6×10 6 cells / kg, approximately 5×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 6×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 7×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 8×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 9×10 5 cells / kg ~ approx. 1×10 7cells / kg, and approximately 1 x 10 6 cells / kg ~ approx. 1×10 7 Additional exemplary doses are, inter alia, about 1×10 10 Approximately 1 x 10 cells / kg of recipient 12 Cells / kg (e.g., approximately 2 x 10 10 cells / kg ~ approx. 9×10 11 cells / kg, approximately 3×10 10 cells / kg ~ approx. 8×10 11 cells / kg, approximately 4×10 10 cells / kg~about 7×10 11 cells / kg, approximately 5×10 10 cells / kg~about 6×10 11 cells / kg, approximately 5×10 10 cells / kg ~ approx. 1×10 12 cells / kg, approximately 6×10 10 cells / kg ~ approx. 1×10 12 cells / kg, approximately 7×10 10 cells / kg ~ approx. 1×10 12 cells / kg, approximately 8×10 10 cells / kg ~ approx. 1×10 12 cells / kg, approximately 9×10 10 cells / kg ~ approx. 1×10 12 cells / kg, and approximately 1 x 10 11 cells / kg ~ approx. 1×10 12 cells / kg).

[0611] The cells and compositions described herein can be administered in an amount sufficient to ameliorate one or more pathological features in NCD. Administration of the cells or compositions described herein can increase the amount of M2 microglia in the subject's brain compared to the amount of M1 microglia in the subject's brain, decrease the level of proinflammatory cytokines in the subject's brain, increase the level of anti-inflammatory cytokines in the subject's brain, improve the cognitive performance of the subject, improve the motor function of the subject, reduce alpha-synuclein protein levels, tau positive neuronal inclusion levels, TAR DNA binding protein 43 (TDP-43) positive inclusion levels, sarcoma fusion (FUS) positive inclusion levels, and / or ubiquitin positive inclusion levels or aggregation thereof in the subject, reduce brain tissue loss in the subject, improve vision, improve language skills, and / or reduce the severity or frequency of epileptic seizure levels in the subject. The numbers of M1 and M2 microglia were assessed using ELISA to compare the levels of cytokines, chemokines, and other pro- and anti-inflammatory mediators in the cerebrospinal fluid (CSF) of subjects before and after treatment, and translocator protein-activating protein (TSPO), a protein highly expressed in classically activated M1 microglia, was assayed using, for example, TSPO radioligand. 11 The effects of C-(R)-PK11195 can be assessed by using PET imaging to observe or by analyzing the levels of M1 and M2 associated genes and proteins in tissue samples using standard techniques, such as Western blot analysis, immunohistochemistry, or quantitative RT-PCR. Cognitive and motor function can be assessed using standard neurological tests before and after treatment, and monomeric and oligomeric α-synuclein can be detected in plasma and CSF using ELISA. Neurodegeneration can be assessed using F18-fluorodeoxyglucose PET scans or MRI scans. Subjects can be evaluated 1, 2, 3, 4, 5, 6 or more months after administration of the population of cells, depending on the route of administration used for the treatment. Depending on the results of the evaluation, the subject can receive additional treatment.

[0612] kit The compositions described herein can be provided in a kit for use in treating NCD (e.g., FTLD or NCL). The compositions can include host cells described herein (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) that include a transgene encoding PGRN or GRN (e.g., a transgene that can be expressed in macrophages or microglia), and can optionally have endogenous PGRN or GRN disrupted. The cells can be cryopreserved, for example, in dimethylsulfoxide (DMSO), glycerol, or another cryoprotectant. The kit can include a package insert that instructs the user of the kit, such as a physician, to carry out the methods described herein. The kit can optionally include a syringe or other device for administering the composition. EXAMPLES

[0613] The following examples are set forth to provide one of ordinary skill in the art with an illustration of how the compositions and methods described herein can be used, made, and evaluated, and are intended to be merely illustrative of the disclosure and are not intended to limit the scope of what the inventors regard as the disclosure.

[0614] Example 1. Generation of cells containing a transgene encoding progranulin or granulin An exemplary method for generating cells (e.g., pluripotent cells, embryonic stem cells (ESCs), induced pluripotent stem cells (ISPCs), pluripotent cells, CD34+ cells, hematopoietic stem cells (HSCs), myeloid progenitor cells (MPCs), blood lineage progenitor cells, monocytes, macrophages, microglial progenitor cells, or microglia) containing a transgene encoding progranulin (PGRN) or granulin (GRN) for use in the compositions and methods described herein is by transduction. Retroviral vectors (e.g., lentiviral, alpharetroviral, or gammaretroviral vectors) containing a microglia-specific promoter, such as the CD68 promoter, and a polynucleotide encoding PGRN or GRN can be engineered using standard techniques known in the art. After engineering the retroviral vector, the retrovirus can be used to transduce cells to generate a population of cells expressing PGRN or GRN.

[0615] An additional exemplary method for generating cells containing a transgene encoding PGRN or GRN for use in the compositions and methods described herein is transfection. Using molecular biology techniques known in the art, plasmid DNA can be generated that contains a promoter, such as a microglia-specific promoter (e.g., CD68 promoter), and a polynucleotide encoding PGRN or GRN. For example, the PGRN gene can be amplified from a human cell line using PCR-based techniques known in the art, or the PGRN or GRN gene can be synthesized, for example, using solid-phase polynucleotide synthesis procedures. The PGRN or GRN gene and promoter can then be ligated into a plasmid of interest, for example, using an appropriate restriction endonuclease-mediated cleavage and ligation protocol. After the plasmid DNA has been manipulated, the plasmid can be used to transfect cells, for example, using electroporation or another transfection technique described herein, to generate a population of cells expressing PGRN or GRN. In both exemplary methods described herein, PGRN or GRN can be expressed as a PGRN or GRN fusion protein. The PGRN or GRN fusion protein may contain a peptide sequence that includes the LDLRf Rb domain of ApoE to allow the penetration of the PGRN or GRN fusion protein through the blood-brain barrier. Alternatively, the fusion protein may contain PGRN or GRN and a glycosylation-independent lysosomal targeting (GILT) tag. An exemplary GILT tag is a mutein derived from human insulin-like growth factor II (IGF-II) that has an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II. These IGF-II muteins have reduced binding activity for the insulin receptor compared to the affinity of naturally occurring human IGF-II for the insulin receptor, are resistant to furin cleavage, and bind to the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner. The GILT tag facilitates the delivery of the secreted GBA fusion protein to lysosomes.

[0616] Example 2. Administration of a population of cells containing a transgene encoding progranulin or granulin to a subject suffering from a neurocognitive disorder According to the method disclosed herein, a practitioner can treat a subject, such as a human subject, to reduce or alleviate the symptoms of neurocognitive disorder (NCD), such as frontotemporal lobar degeneration (FTLD) or neuronal ceroid lipofuscinosis (NCL).To this end, a practitioner can administer a population of cells (e.g., pluripotent cells, ESCs, iPSCs, pluripotent cells, CD34+ cells, HSCs, MPCs, BLPCs, monocytes, macrophages, microglial progenitor cells, or microglia) that contain a transgene that codes for PGRN or GRN (e.g., a transgene that can be expressed in macrophages or microglia) to a human subject.Using the techniques described herein or known in the art, cells can be transduced or transfected ex vivo to express PGRN or GRN. A population of cells containing a transgene encoding PGRN or GRN can be administered to a subject, for example, systemically (e.g., intravenously), directly to the central nervous system (CNS) (e.g., intraventricularly or stereotactically), or directly to the bone marrow (e.g., intraosseous) to treat an NCD. Cells can also be administered to a patient by multiple routes of administration, for example, intravenously and intraventricularly. Cells can be administered at a concentration of 1×10 6 cells / kg~1×10 12 cells / kg or more (e.g., 1 x 10 7 cells / kg, 1×10 8 cells / kg, 1×10 9 cells / kg, 1×10 10 cells / kg, 1×10 11 cells / kg, 1×10 12 The cells are administered in a therapeutically effective amount, such as 100,000 cells / kg, or more.

[0617] Prior to administering the population of cells to the subject, one or more agents, e.g., busulfan, treosulfan, PLX3397, PLX647, PLX5622, and / or clodronate liposomes, may also be administered to the subject to ablate the subject's endogenous microglia and / or hematopoietic stem and progenitor cells. Other cell ablation methods known in the art, such as irradiation, may be used alone or in combination with one or more of the aforementioned agents to ablate the subject's microglia and / or hematopoietic stem and progenitor cells. These agents and / or treatments may ablate at least 5% (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 99%, or more) of the endogenous microglia and / or hematopoietic stem and progenitor cells as assessed by PET imaging techniques known in the art. After removal of the microglia, the population of cells is administered to the subject, whereby the cells repopulate the brain and differentiate into microglia. The population of cells can be administered to the subject, for example, one week to one month (e.g., one week, two weeks, three weeks, four weeks) or more after removal of the microglia.

[0618] Following removal of the subject's endogenous microglia and / or hematopoietic stem and progenitor cells, a population of cells can be administered to the subject systemically (e.g., intravenously) or by bone marrow transplantation to reconstitute the bone marrow compartment. The number of cells can be administered at any suitable dose after conditioning. A non-limiting example of a dose is about 1×10 5 Approximately 1 x 10 cells / kg of recipient 7 cells / kg (e.g., about 2×10 5 cells / kg ~ approx. 9×10 6 cells / kg, approximately 3×10 5 cells / kg ~ approx. 8×10 6 cells / kg, approximately 4×10 5 cells / kg~about 7×10 6 cells / kg, approximately 5×10 5 cells / kg~about 6×10 6 cells / kg, approximately 5×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 6×105 cells / kg ~ approx. 1×10 7 cells / kg, approximately 7×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 8×10 5 cells / kg ~ approx. 1×10 7 cells / kg, approximately 9×10 5 cells / kg ~ approx. 1×10 7 cells / kg, or app...

Claims

1. 1. A method of treating a subject diagnosed with a neurocognitive disorder (NCD), comprising administering to the subject a composition comprising a population of cells comprising a transgene encoding progranulin (PGRN) or granulin (GRN).

2. 2. The method of claim 1, wherein the NCD is a severe NCD.

3. 3. The method of claim 2, wherein the severe NCD interferes with the subject's independence and / or normal daily functioning.

4. 4. The method of claim 2 or 3, wherein said severe NCD is associated with a score obtained by said subject on a cognitive test that is at least 2 standard deviations away from the mean score of a reference population.

5. 2. The method of claim 1, wherein the NCD is a mild NCD.

6. 6. The method of claim 5, wherein the mild NCD does not interfere with the subject's independence and / or normal daily functioning.

7. 7. The method of claim 5 or 6, wherein said mild NCD is associated with a score obtained by said subject on a cognitive test that is 1-2 standard deviations away from the mean score of a reference population.

8. The method of claim 4 or 7, wherein the reference population is the general population.

9. The cognitive tests include Eight-item Informant Interview to Differentiate Aging and Dementia (AD8), Annual Wellness Visit (AWV), General Practitioner Assessment of Cognition (GPCOG), Health Risk Assessment (HRA), Memory Impairment Screen (MIS), Mini Mental Status Exam (MMSE), Montreal Cognitive Assessment (MoCA), St.

9. The method of claim 4, 7, or 8, wherein the said test is selected from the group consisting of: Louis University Mental Status Exam (SLUMS), and Short Informant Questionnaire on Cognitive Decline in the Elderly (Short IQCODE).

10. 10. The method of any one of claims 1-9, wherein the NCD is associated with impairment in one or more of complex attention, executive function, learning and memory, language, sensorimotor function, and social cognition.

11. 11. The method of any one of claims 1 to 10, wherein the NCD is not due to delirium or other psychiatric disorder.

12. The method of any one of claims 1 to 11, wherein the NCD is a frontotemporal NCD.

13. 13. The method of claim 12, wherein the frontotemporal NCD is frontotemporal lobar degeneration (FTLD).

14. The method of any one of claims 1 to 11, wherein the NCD is due to a lysosomal disorder.

15. 15. The method of claim 14, wherein the lysosomal disease is neuronal ceroid lipofuscinosis (NCL).

16. The method of any one of claims 1 to 15, wherein the PGRN or the GRN comprises a secretory signal peptide.

17. The method of claim 16 , wherein the secretory signal peptide is a PGRN secretory signal peptide.

18. The method of any one of claims 1 to 17, wherein the cell comprises a transgene encoding the PGRN.

19. 20. The method of claim 18, wherein the PGRN comprises at least two GRN domains, optionally wherein the PGRN comprises at least three GRN domains, optionally wherein the PGRN comprises at least four GRN domains, optionally wherein the PGRN comprises at least five GRN domains, optionally wherein the PGRN comprises at least six GRN domains, optionally wherein the PGRN comprises at least seven GRN domains, or optionally wherein the PGRN comprises at least eight GRN domains.

20. 20. The method of any one of claims 1 to 19, wherein the PGRN comprises 2-16 GRN domains, optionally wherein the PGRN comprises 2-12 GRN domains, optionally wherein the PGRN comprises 2-8 GRN domains, optionally wherein the PGRN comprises 2-4 GRN domains, or optionally wherein the PGRN comprises 2 GRN domains.

21. 21. The method of any one of claims 1 to 20, wherein the PGRN comprises a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2, optionally wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2, optionally wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2, or optionally wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:

2.

22. 22. The method of any one of claims 1 to 21, wherein the PGRN comprises a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3, or optionally wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:

3.

23. 23. The method of any one of claims 1 to 22, wherein the PGRN comprises a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4, or optionally wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:

4.

24. 24. The method of any one of claims 1 to 23, wherein the PGRN comprises a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5, or optionally wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:

5.

25. 25. The method of any one of claims 1 to 24, wherein the PGRN comprises a GRN-4 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:6, or optionally wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:

6.

26. 26. The method of any one of claims 1 to 25, wherein the PGRN comprises a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7, or optionally wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:

7.

27. 27. The method of any one of claims 1 to 26, wherein the PGRN comprises a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8, or optionally wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:

8.

28. 28. The method of any one of claims 1 to 27, wherein the PGRN comprises a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9, or optionally wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:

9.

29. 29. The method of any one of claims 1-28, wherein the PGRN has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:1, optionally wherein the PGRN has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:1, optionally wherein the PGRN has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:1, or optionally wherein the PGRN has the amino acid sequence of SEQ ID NO:

1.

30. The method of any one of claims 1 to 29, wherein the PGRN is a full-length PGRN.

31. The method of any one of claims 1 to 30, wherein the cell comprises a transgene encoding the GRN.

32. 32. The method of any one of claims 1 to 31, wherein the GRN is a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2, optionally wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2, optionally wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2, or optionally wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:

2.

33. 33. The method of any one of claims 1 to 32, wherein the GRN is a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3, or optionally wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:

3.

34. 34. The method of any one of claims 1 to 33, wherein the GRN is a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4, or optionally wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:

4.

35. 35. The method of any one of claims 1 to 34, wherein the GRN is a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5, or optionally wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:

5.

36. 36. The method of any one of claims 1 to 35, wherein the GRN is a GRN-4 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 6, or optionally wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:

6.

37. 37. The method of any one of claims 1 to 36, wherein the GRN is a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7, or optionally wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:

7.

38. 38. The method of any one of claims 1 to 37, wherein the GRN is a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8, or optionally wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:

8.

39. 39. The method of any one of claims 1 to 38, wherein the GRN is a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9, or optionally wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:

9.

40. The method of any one of claims 1 to 39, wherein the GRN comprises a full-length GRN.

41. 41. The method of any one of claims 1-40, wherein the cell comprises a PGRN transgene having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO: 10, optionally wherein the cell comprises a PGRN transgene having at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO: 10, optionally wherein the cell comprises a PGRN transgene having at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO: 10, optionally wherein the cell comprises a PGRN transgene having the nucleic acid sequence of SEQ ID NO:

10.

42. The method of any one of claims 1 to 40, wherein the cell comprises a codon-optimized PGRN transgene.

43. 43. The method of claim 42, wherein the codon optimized transgene has at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO: 19, optionally wherein the codon optimized transgene has at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO: 19, optionally wherein the codon optimized transgene has at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO: 19, or optionally wherein the codon optimized transgene has the nucleic acid sequence of SEQ ID NO:

19.

44. The method of any one of claims 1 to 43, wherein the PGRN or the GRN is a PGRN or GRN fusion protein.

45. 45. The method of claim 44, wherein the PGRN or the GRN fusion protein comprises the receptor binding (Rb) domain of apolipoprotein E (ApoE).

46. 46. ​​The method of claim 45, wherein the Rb domain comprises a portion of ApoE having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO:

11.

47. The method of claim 45 or 46, wherein the Rb domain comprises a region having at least 70% sequence identity to the amino acid sequence of residues 159-167 of SEQ ID NO:

11.

48. 45. The method of claim 44, wherein the PGRN or GRN fusion protein comprises PGRN or GRN and a glycosylation-independent lysosomal targeting (GILT) tag.

49. 49. The method of claim 48, wherein the GILT tag comprises a human IGF-II mutein having an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II (SEQ ID NO: 12) and having a reduced binding affinity for the insulin receptor compared to the affinity of naturally occurring human IGF-II for the insulin receptor, wherein the IGF-II mutein is resistant to furin cleavage and binds to the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate dependent manner.

50. 50. The method of claim 49, wherein said IGF-II mutein comprises a mutation within a region corresponding to amino acids 30-40 of SEQ ID NO:12, and said mutation destroys at least one Furin protease cleavage site.

51. 51. The method of claim 50, wherein the mutation is an amino acid substitution, deletion, and / or insertion.

52. 52. The method of claim 51 , wherein the mutation is a Lys or Ala amino acid substitution at a position corresponding to Arg37 or Arg40 of SEQ ID NO:

12.

53. 52. The method of claim 51, wherein the mutation is a deletion or substitution of amino acid residues corresponding to positions selected from the group consisting of 31-40, 32-40, 33-40, 34-40, 30-39, 31-39, 32-39, 34-37, 33-39, 35-39, 36-39, 37-40, 34-40 of SEQ ID NO: 12, and combinations thereof.

54. 54. The method of any one of claims 48 to 53, wherein the GILT tag has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 13; optionally, the GILT tag has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 13; optionally, the GILT tag has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 13; optionally, the GILT tag has the amino acid sequence of SEQ ID NO:

13.

55. 54. The method of any one of claims 48 to 53, wherein the GILT tag has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 14; optionally, the GILT tag has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 14; optionally, the GILT tag has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14; optionally, the GILT tag has the amino acid sequence of SEQ ID NO:

14.

56. 54. The method of any one of claims 48 to 53, wherein the GILT tag has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 15; optionally, the GILT tag has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 15; optionally, the GILT tag has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 15; optionally, the GILT tag has the amino acid sequence of SEQ ID NO:

15.

57. 54. The method of any one of claims 48 to 53, wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 16; optionally, the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 16; optionally, the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 16; optionally, the GILT tag is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO:

16.

58. 54. The method of any one of claims 48 to 53, wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 17; optionally, the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 17; optionally, the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 17; optionally, the GILT tag is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO:

17.

59. 54. The method of any one of claims 48 to 53, wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 18; optionally, the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 18; optionally, the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 18; optionally, the GILT tag is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO:

18.

60. 60. The method of any one of claims 1 to 59, wherein the transgene encoding the PGRN or the GRN further comprises a microRNA (miRNA)-126 (miR-126) targeting sequence in the 3'-UTR.

61. 61. The method of any one of claims 1 to 60, wherein the PGRN or GRN penetrates the blood-brain barrier of the subject upon administration of the composition to the subject.

62. The method of any one of claims 13 to 61, wherein the FTLD or NCL is PGRN-associated FTLD or NCL.

63. 63. The method of claim 62, wherein the PGRN-related FTLD is a behavioral abnormality frontotemporal variant of FTLD.

64. 63. The method of claim 62, wherein the PGRN-related FTLD is the semantic dementia variant of FTLD.

65. 63. The method of claim 62, wherein the PGRN-related FTLD is the progressive non-verbal aphasia variant of FTLD.

66. 63. The method of claim 62, wherein the PGRN-associated NCL is Batten disease.

67. The method of any one of claims 1 to 54, wherein the cell is a pluripotent or multipotent cell.

68. 68. The method of claim 67, wherein the pluripotent cells are CD34+ cells.

69. 69. The method of claim 68, wherein the CD34+ cells are hematopoietic stem cells (HSCs) or myeloid progenitor cells (MPCs).

70. 68. The method of claim 67, wherein the pluripotent cell is an embryonic stem cell (ESC) or an induced pluripotent stem cell (iPSC).

71. 67. The method of any one of claims 1 to 66, wherein the cell is a blood lineage progenitor cell (BLPC), a microglial progenitor cell, a monocyte, a macrophage, or a microglia.

72. 72. The method of claim 71, wherein the BLPC is a monocyte.

73. 73. The method of any one of claims 1 to 72, wherein the subject has a population of endogenous microglia depleted prior to administration of the composition.

74. 73. The method of any one of claims 1 to 72, comprising removing a population of endogenous microglia in the subject prior to administering the composition to the subject.

75. 74. The method of claim 72 or 73, wherein the microglia are ablated using an agent selected from the group consisting of busulfan, PLX3397, PLX647, PLX5622, treosulfan, and clodronate liposomes, by radiation therapy, or a combination thereof.

76. 76. The method of any one of claims 1-75, wherein the composition is administered to the subject by systemic administration, by direct administration to the subject's central nervous system, by direct administration to the subject's bone marrow, or by bone marrow transplantation comprising the composition.

77. 77. The method of any one of claims 1 to 76, further comprising administering to the subject a population of cells.

78. 78. The method of claim 77, wherein the population of cells is administered to the subject prior to administration of the composition or after administration of the composition.

79. 79. The method of claim 77 or 78, wherein the cell is a pluripotent or multipotent cell.

80. 80. The method of claim 79, wherein the pluripotent cells are CD34+ cells.

81. 81. The method of claim 80, wherein the CD34+ cells are hematopoietic stem cells (HSCs) or myeloid progenitor cells (MPCs).

82. 80. The method of claim 79, wherein the pluripotent cell is an embryonic stem cell (ESC) or an induced pluripotent stem cell (iPSC).

83. 80. The method of any one of claims 77 to 79, wherein the cell is a blood lineage progenitor cell (BLPC), a microglial progenitor cell, a monocyte, a macrophage, or a microglia.

84. 84. The method of claim 83, wherein the BLPC is a monocyte.

85. The method of any one of claims 77-84, wherein the cell has not been modified to express a transgene encoding the PGRN or the GRN.

86. 86. The method of any one of claims 1-85, wherein endogenous PGRN or GRN is disrupted in the cell, in the subject, or in a population of neurons in the subject prior to administration of the composition to the subject.

87. 87. The method of claim 86, wherein the endogenous PGRN or GRN is destroyed by contacting the cell with a nuclease that catalyzes cleavage of endogenous PGRN or GRN nucleic acid in the cell.

88. 88. The method of claim 87, wherein the nuclease is clustered regularly interspaced short palindromic repeats (CRISPR) associated protein 9 (Cas9) and the CRISPR-associated protein is CRISPR-associated protein 12a (Cas12a), a transcription activator-like effector nuclease, a meganuclease, or a zinc finger nuclease.

89. 89. The method of any one of claims 86-88, wherein the endogenous PGRN or GRN is disrupted by administering an inhibitory RNA molecule to the cell, the subject, or the population of neurons.

90. 90. The method of claim 89, wherein the inhibitory RNA molecule is a short interfering RNA, a short hairpin RNA, or a miRNA.

91. The method of any one of claims 1 to 90, wherein the cells are autologous or allogeneic cells.

92. The method of any one of claims 1 to 91, wherein the cells are transfected or transduced ex vivo to express the PGRN or the GRN.

93. 93. The method of claim 92, wherein the cells are transduced with a viral vector selected from the group consisting of adeno-associated virus (AAV), adenovirus, parvovirus, coronavirus, rhabdovirus, paramyxovirus, picornavirus, alphavirus, herpesvirus, poxvirus, and retroviridae viruses.

94. 94. The method of claim 93, wherein the viral vector is a retroviridae viral vector.

95. 95. The method of claim 94, wherein the Retroviridae viral vector is a lentiviral vector, an alpharetroviral vector, or a gammaretroviral vector.

96. The method of claim 94 or 95, wherein the Retroviridae viral vector comprises a central polypurine tract, a woodchuck hepatitis virus post-transcriptional regulatory element, a 5'-LTR, an HIV signal sequence, an HIV Psi signal 5'-splice site, a delta-GAG element, a 3'-splice site, and a 3'-self-inactivating LTR.

97. 94. The method of claim 93, wherein the viral vector is an AAV selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, and AAVrh74.

98. The method of any one of claims 93 to 97, wherein the viral vector is a pseudotyped viral vector.

99. 99. The method of claim 98, wherein the pseudotyped viral vector is selected from the group consisting of pseudotyped AAV, pseudotyped adenovirus, pseudotyped parvovirus, pseudotyped coronavirus, pseudotyped rhabdovirus, pseudotyped paramyxovirus, pseudotyped picornavirus, pseudotyped alphavirus, pseudotyped herpesvirus, pseudotyped poxvirus, and pseudotyped Retroviridae viruses.

100. 93. The method of claim 92, wherein the cells are transfected using: a) an agent selected from the group consisting of cationic polymers, diethylaminoethyl-dextran, polyethyleneimine, cationic lipids, liposomes, calcium phosphate, activated dendrimers, and magnetic beads; or b) a technique selected from the group consisting of electroporation, Nucleofection, squeeze-poration, sonoporation, optical transfection, Magnetofection, and impalefection.

101. The method of any one of claims 1 to 100, wherein expression of the PGRN or the GRN in the cell is mediated by a ubiquitous promoter, a cell lineage-specific promoter, or a synthetic promoter.

102. 102. The method of claim 101, wherein the ubiquitous promoter is selected from the group consisting of elongation factor 1-alpha promoter and phosphoglycerate kinase 1 promoter.

103. 102. The method of claim 101, wherein the cell lineage specific promoter is selected from the group consisting of PGRN promoter, CD11b promoter, CD68 promoter, C-X3-C motif chemokine receptor 1 promoter, allograft inflammatory factor 1 promoter, purinergic receptor P2Y12 promoter, transmembrane protein 119 promoter, and colony stimulating factor 1 receptor promoter.

104. A pharmaceutical composition comprising a population of cells containing a transgene encoding PGRN or GRN, and further comprising one or more pharma- ceutically acceptable carriers, diluents, or excipients.

105. The pharmaceutical composition of claim 104, wherein the PGRN or the GRN comprises a PGRN secretory signal peptide.

106. The pharmaceutical composition of claim 104 or 105, wherein the cell comprises a transgene encoding the PGRN.

107. The pharmaceutical composition of claim 106, wherein the PGRN comprises at least two GRN domains, optionally, the PGRN comprises at least three GRN domains, optionally, the PGRN comprises at least four GRN domains, optionally, the PGRN comprises at least five GRN domains, optionally, the PGRN comprises at least six GRN domains, optionally, the PGRN comprises at least seven GRN domains, or optionally, the PGRN comprises at least eight GRN domains.

108. The pharmaceutical composition of any one of claims 104-107, wherein the PGRN comprises 2-16 GRN domains, optionally wherein the PGRN comprises 2-12 GRN domains, optionally wherein the PGRN comprises 2-8 GRN domains, optionally wherein the PGRN comprises 2-4 GRN domains, or optionally wherein the PGRN comprises 2 GRN domains.

109. The pharmaceutical composition of any one of claims 104-108, wherein the PGRN comprises a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2, optionally the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2, optionally the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2, or optionally the para-GRN domain has the amino acid sequence of SEQ ID NO:

2.

110. The pharmaceutical composition of any one of claims 104 to 109, wherein the PGRN comprises a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3, or optionally wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:

3.

111. The pharmaceutical composition of any one of claims 104-110, wherein the PGRN comprises a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:4, or optionally wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:

4.

112. The pharmaceutical composition of any one of claims 104-111, wherein the PGRN comprises a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5, or optionally wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:

5.

113. The pharmaceutical composition of any one of claims 104-112, wherein the PGRN comprises a GRN-4 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:6, or optionally wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:

6.

114. The pharmaceutical composition of any one of claims 104 to 113, wherein the PGRN comprises a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:7, or optionally wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:

7.

115. The pharmaceutical composition of any one of claims 104-114, wherein the PGRN comprises a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8, or optionally wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:

8.

116. The pharmaceutical composition of any one of claims 104-115, wherein the PGRN comprises a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9, or optionally wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:

9.

117. The pharmaceutical composition of any one of claims 104 to 116, wherein the PGRN is a full-length PGRN.

118. The pharmaceutical composition of claim 117, wherein the PGRN has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:1, optionally, the PGRN has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:1, optionally, the PGRN has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:1, optionally, the PGRN has the amino acid sequence of SEQ ID NO:1, or optionally, the cell comprises a transgene encoding the GRN.

119. 119. The pharmaceutical composition of any one of claims 104-118, wherein the GRN is a para-GRN domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:2, optionally wherein the para-GRN domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2, optionally wherein the para-GRN domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:2, or optionally wherein the para-GRN domain has the amino acid sequence of SEQ ID NO:

2.

120. The pharmaceutical composition of any one of claims 104 to 119, wherein the GRN is a GRN-1 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:3, optionally wherein the GRN-1 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:3, or optionally wherein the GRN-1 domain has the amino acid sequence of SEQ ID NO:

3.

121. The pharmaceutical composition of any one of claims 104 to 120, wherein the GRN is a GRN-2 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 4, optionally wherein the GRN-2 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 4, or optionally wherein the GRN-2 domain has the amino acid sequence of SEQ ID NO:

4.

122. The pharmaceutical composition of any one of claims 104 to 121, wherein the GRN is a GRN-3 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:5, optionally wherein the GRN-3 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5, or optionally wherein the GRN-3 domain has the amino acid sequence of SEQ ID NO:

5.

123. The pharmaceutical composition of any one of claims 104 to 122, wherein the GRN-4 domain has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 6, optionally wherein the GRN-4 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 6, optionally wherein the GRN-4 domain has the amino acid sequence of SEQ ID NO:

6.

124. The pharmaceutical composition of any one of claims 104 to 123, wherein the GRN is a GRN-5 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 7, optionally wherein the GRN-5 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 7, or optionally wherein the GRN-5 domain has the amino acid sequence of SEQ ID NO:

7.

125. The pharmaceutical composition of any one of claims 104 to 124, wherein the GRN is a GRN-6 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:8, optionally wherein the GRN-6 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:8, or optionally wherein the GRN-6 domain has the amino acid sequence of SEQ ID NO:

8.

126. The pharmaceutical composition of any one of claims 104 to 125, wherein the GRN is a GRN-7 domain having an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:9, optionally wherein the GRN-7 domain has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:9, or optionally wherein the GRN-7 domain has the amino acid sequence of SEQ ID NO:

9.

127. The pharmaceutical composition of any one of claims 104 to 126, wherein the GRN is a full-length GRN.

128. The pharmaceutical composition of any one of claims 104-126, wherein the cell comprises a PGRN transgene having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO: 10, optionally wherein the PGRN transgene has at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO: 10, optionally wherein the PGRN transgene has at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO: 10, or optionally wherein the PGRN transgene has the nucleic acid sequence of SEQ ID NO:

10.

129. The pharmaceutical composition of any one of claims 104-128, wherein the cell comprises a codon-optimized PGRN transgene.

130. The pharmaceutical composition of claim 129, wherein the codon-optimized transgene has at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO: 19, optionally, the codon-optimized transgene has at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO: 19, optionally, the codon-optimized transgene has at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO: 19, or optionally, the codon-optimized transgene has the nucleic acid sequence of SEQ ID NO:

19.

131. The pharmaceutical composition of any one of claims 104 to 130, wherein the PGRN or the GRN is a PGRN or GRN fusion protein.

132. The pharmaceutical composition of claim 131, wherein the PGRN or the GRN fusion protein comprises the Rb domain of ApoE.

133. 133. The pharmaceutical composition of claim 132, wherein the Rb domain comprises a portion of ApoE having an amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO:

11.

134. The pharmaceutical composition of claim 132 or 133, wherein the Rb domain comprises a region having at least 70% sequence identity to the amino acid sequence of residues 159-167 of SEQ ID NO:

11.

135. The pharmaceutical composition of claim 131, wherein the PGRN or GRN fusion protein comprises PGRN or GRN and a glycosylation-independent lysosomal targeting (GILT) tag.

136. 136. The pharmaceutical composition of claim 135, wherein the GILT tag comprises a human IGF-II mutein having an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II (SEQ ID NO: 12) and having a reduced binding affinity for the insulin receptor compared to the affinity of naturally occurring human IGF-II for the insulin receptor, wherein the IGF-II mutein is resistant to furin cleavage and binds to the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate dependent manner.

137. 137. The pharmaceutical composition of claim 136, wherein said IGF-II mutein comprises a mutation within a region corresponding to amino acids 30-40 of SEQ ID NO: 12, and said mutation destroys at least one Furin protease cleavage site.

138. The pharmaceutical composition of claim 137, wherein the mutation is an amino acid substitution, deletion, and / or insertion.

139. The pharmaceutical composition of claim 138, wherein the mutation is a Lys or Ala amino acid substitution at a position corresponding to Arg37 or Arg40 of SEQ ID NO:

12.

140. The pharmaceutical composition of claim 138, wherein the mutation is a deletion or substitution of an amino acid residue corresponding to a position selected from the group consisting of 31-40, 32-40, 33-40, 34-40, 30-39, 31-39, 32-39, 34-37, 33-39, 35-39, 36-39, 37-40, 34-40 of SEQ ID NO: 12, and combinations thereof.

141. The pharmaceutical composition of any one of claims 135-140, wherein the GILT tag has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 13, optionally wherein the GILT tag has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 13, optionally wherein the GILT tag has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 13, optionally wherein the GILT tag has the amino acid sequence of SEQ ID NO:

13.

142. The pharmaceutical composition of any one of claims 135 to 140, wherein the GILT tag has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 14, optionally wherein the GILT tag has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 14, optionally wherein the GILT tag has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14, optionally wherein the GILT tag has the amino acid sequence of SEQ ID NO:

14.

143. The pharmaceutical composition of any one of claims 135 to 140, wherein the GILT tag has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 15, optionally wherein the GILT tag has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 15, optionally wherein the GILT tag has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 15, optionally wherein the GILT tag has the amino acid sequence of SEQ ID NO:

15.

144. The pharmaceutical composition of any one of claims 135 to 140, wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 16, optionally wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 16, optionally wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 16, optionally wherein the GILT tag is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO:

16.

145. The pharmaceutical composition of any one of claims 135 to 140, wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 17, optionally wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 17, optionally wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 17, optionally wherein the GILT tag is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO:

17.

146. The pharmaceutical composition of any one of claims 135 to 140, wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 18, optionally wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 18, optionally wherein the GILT tag is encoded by a polynucleotide having a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 18, optionally wherein the GILT tag is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO:

18.

147. The pharmaceutical composition of any one of claims 104 to 146, wherein the transgene encoding PGRN or GRN further comprises a miR-126 targeting sequence in the 3'-UTR.

148. The composition of any one of claims 104 to 147, wherein the cell is a pluripotent or multipotent cell.

149. The composition of claim 148, wherein the pluripotent cells are CD34+ cells.

150. The composition of claim 149, wherein the CD34+ cells are HSCs or MPCs.

151. The composition of claim 148, wherein the pluripotent cells are ESCs or iPSCs.

152. The composition of any one of claims 104 to 147, wherein the cell is a BLPC, a microglial progenitor cell, a macrophage, or a microglia.

153. The composition of claim 152, wherein the BLPC is a monocyte.

154. The pharmaceutical composition of any one of claims 104-153, wherein the cells are transfected or transduced ex vivo to express the PGRN or the GRN.

155. A kit comprising the pharmaceutical composition according to any one of claims 104 to 154 and an accompanying instruction.

156. The kit of claim 155, wherein the package insert instructs a user of the kit to carry out the method of any one of claims 1 to 103.

Images