Search

US-20260125456-A1 - CHIMERIC ANTIGEN RECEPTORS FOR TREATMENT OF NEURODEGENERATIVE DISEASES AND DISORDERS

US20260125456A1US 20260125456 A1US20260125456 A1US 20260125456A1US-20260125456-A1

Abstract

The present disclosure generally relates to novel chimeric antigen receptors (“CARs”), modified regulatory T cells (“Tregs”) expressing such CARs and/or Tregs which are engineered to express neurodegenerative disease modifying molecules, e.g., which express molecules which prevent oxidative/inflammatory activity, or which promote neuronal growth/survival such as nerve growth factors or non-classical neurotrophic factors. The present disclosure also generally relates to compositions containing such modified Tregs, and methods of use thereof as therapeutics, in particular for treating and preventing neurodegenerative diseases and symptoms associated with therewith, and/or for slowing the onset of such neurodegenerative diseases, particularly in persons at risk because of genetic factors or in persons exhibiting early signs of developing such a neurodegenerative disease.

Inventors

  • Charles SENTMAN
  • David GRABER
  • W. James Cook

Assignees

  • THE TRUSTEES OF DARTMOUTH COLLEGE

Dates

Publication Date
20260507
Application Date
20250610

Claims (20)

  1. 1 - 157 . (canceled)
  2. 158 . A modified Regulatory T cell (Treg) comprising a chimeric antigen receptor (CAR), the CAR comprising an antigen binding domain that specifically binds to an epitope on a protein(s) associated with the pathology of a neurodegenerative disease or condition.
  3. 159 . The modified Treg of claim 158 , wherein the protein(s) is expressed or overexpressed at a site of neurodegeneration.
  4. 160 . The modified Treg of claim 158 , wherein the protein(s) is selected from the group consisting of human amyloid beta, amyloid-beta 1-42, alpha-synuclein, superoxide dismutase-1 (SOD-1), hyperphosphorylated tau protein; TAR DNA-binding protein 43 (TDP-43): chromosome 9 open reading frame 72 (c9orf72), β-Synuclein, y-Synuclein, RNA-binding protein fused in sarcoma (FUS), ubiquitin, ubiquilin-2, p62, optineurin, ataxin-2, parkin, Serine/threonine-protein kinase PINK1, and Leucine-rich repeat serine/threonine-protein kinase 2 (LRRK2), Huntingtin with tandem glutamine repeats, prion proteins, transthyretin, dentatorubral pallidoluysian atrophy (DRPLA) protein, androgen receptor, an ataxin, P/Q-type calcium channel al A subunit, TATAbox-binding protein, glial fibrillary acidic protein, DNA excision repair protein ERCC-6, survival motor neuron protein, and cystatin C.
  5. 161 . The modified Treg of claim 158 , wherein the CAR specifically binds to superoxide dismutase-1 (SOD-1).
  6. 162 . The modified Treg of claim 158 , wherein the antigen binding domain comprises the complementarity determining regions (CDR) of the scFv: DG01 (SEQ ID NO:1), DG02 (SEQ ID NO:2), DG03 (SEQ ID NO:3), DG04 (SEQ ID NO:4), DG05 (SEQ ID NO:5), DG06 (SEQ ID NO:6), DG07 (SEQ ID NO:7), DG08 (SEQ ID NO:8), DG09 (SEQ ID NO:9), DG10 (SEQ ID NO:10), and DG11 (SEQ ID NO:11).
  7. 163 . The modified Treg of claim 158 , wherein the antigen binding domain comprises the amino acid sequence of the scFv according to DG01 (SEQ ID NO:1), DG02 (SEQ ID NO:2), DG03 (SEQ ID NO:3), DG04 (SEQ ID NO:4), DG05 (SEQ ID NO:5), DG06 (SEQ ID NO:6), DG07 (SEQ ID NO:7), DG08 (SEQ ID NO:8), DG09 (SEQ ID NO:9), DG10 (SEQ ID NO:10), and DG11 (SEQ ID NO:11), or amino acid sequence comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereof.
  8. 164 . The modified Treg of claim 158 , wherein the CAR comprises a hinge domain, optionally a CD28 hinge domain.
  9. 165 . The modified Treg of claim 158 , wherein the CAR comprises a transmembrane domain, optionally the transmembrane domain of CD28.
  10. 166 . The modified Treg of claim 158 , wherein the CAR comprises an intracellular signaling domain, optionally comprising a cytoplasmic signaling domain from a lymphocyte receptor chain, a TCR/CD3 complex protein, an Fc receptor subunit, an IL-2 receptor subunit, CD3z, FcRg, FcRb, CD3g, CD3d, CD3e, CD5, CD22, CD66d, CD79a, CD79b, CD278 (ICOS), Fc e RI, DAP 10, or DAP12.
  11. 167 . The modified Treg of claim 158 , wherein the CAR comprises a co-stimulatory domain, optionally wherein the co-stimulatory domain comprises the co-stimulatory domain of an MHC class I molecule, a TNF receptor protein, an immunoglobulin-like protein, a cytokine receptor, an integrin, a signaling lymphocytic activation molecule (SLAM protein), an activating NK cell receptor, or a Toll ligand receptor.
  12. 168 . The modified Treg of claim 158 , wherein the CAR comprises a co-stimulatory domain, optionally wherein the co-stimulatory domain comprises the co-stimulatory domain of B7-H3, BAFFR, BTLA, BLAME (SLAMF8), CD2, CD4, CD5, CD7, CD 8 a, CD8 p, CD11a, LFA-1 (CD11a/CD18), CD11b, CD11c, CD11d, CD18, CD19, CD19a, CD27, CD28, CD29, CD30, CD40, CD49a, CD49D, CD49f, CD69, CD84, CD96 (Tactile), CD 100 (SEMA4D), CD103, CRTAM, 0×40 (CD134), 4-1BB (CD137), SLAM (SLAMF1, CD 150, IPO-3), CD 160 (BY55), SELPLG (CD162), DNAM1 (CD226), Ly9 (CD229), SLAMF4 (CD244, 2B4), ICOS (CD278), CEACAM1, CDS, CRTAM, DAP 10, GADS, GITR, HVEM (LIGHTR), IA4, ICAM-1, IL2R b, IL2R g, IL7R a, ITGA4, ITGA6, ITGAD, ITGAE, ITGAL, ITGAM, ITGAX, ITGB1, ITGB2, ITGB7, KIRDS2, LAT, LFA-1, LIGHT, LTBR, NKG2C, NKG2D, NKp30, NKp44, NKp46, NKp80 (KLRF1), PAG/Cbp, PD-1, PSGL1, SLAMF6 (NTB-A, Ly108), SLAMF7, SLP-76, TNFR2, TRAN CE/RANKL, VLA1, VLA-6, or a ligand that specifically binds with CD83.
  13. 169 . The modified Treg of claim 158 , wherein the CAR comprises an intracellular signaling domain and/or co-stimulatory domain selected from CD28-CD3, 4-1BB-CD3, DAP10-CD3, CD44-CD3, CTLA4-CD3, CD28, DAP10, 4-1BB and CD3γ.
  14. 170 . The modified Treg of claim 158 , wherein the Treg does not express a functional TCR.
  15. 171 . The modified Treg of claim 158 , comprising a heterologous construct expressing a neurodegenerative disease modifying molecule (NDMM), optionally wherein the NDMM is a pro-neuronal factor, neurotrophic factor, or nerve growth factor.
  16. 172 . The modified Treg of claim 158 , comprising a heterologous construct expressing a NDMM selected from the group consisting of brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), glial cell-line derived neurotrophic factor (GDNF), interleukin-1 receptor antagonist (IL-1ra); interleukin-6 (IL-6), activated protein C (APC), thrombomodulin, tissue plasminogen activator (tPA), Protein deglycase DJ-1, a tissue inhibitor of metalloproteinases (TIMP), insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), bone morphogenetic protein (BMP), erythropoietin (EPO), thrombopoietin (TPO), and granulocyte-colony stimulating factor (G-CSF).
  17. 173 . The modified Treg of claim 171 , wherein the NDMM is (i) Nrf2 (Keap1 inhibitor peptide) (SEQ ID NO: 51), and/or a construct comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereof; (ii) human catalase construct (SEQ ID NO:52), and/or a construct comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereof; (iii) BDNF (SEQ ID NO: 53), and/or a construct comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereof; or (iv) IGF-1 (SEQ ID NO: 54), and/or a construct comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereof.
  18. 174 . The modified Treg of claim 158 , wherein the CAR comprises the amino acid sequence of DG03-CD28-CD37 (DG03-28-3) (SEQ ID NO:22); DG03-CD28tm-DAP10-CD3 (DG03-28tm-10-3ζ) (SEQ ID NO:45); DG03-CD28tm-CD44-CD3 (DG03-28tm-44-3) (SEQ ID NO:46); DG03-CD28tm-4-1-BB-CD37 (DG03-28tm-BB-3) (SEQ ID NO:47); DG03-CD28tm-CD3 (DG03-28tm-3) (SEQ ID NO:48); DG03-CD28 (DG03-28) (SEQ ID NO:49); and DG03-CD28tm (SEQ ID NO:50), or a CAR comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs:22, or 45-50.
  19. 175 . The modified Treg of claim 158 , wherein the CAR comprises the amino acid sequence of DG05-CD28-CD3ζ (DG05-28-3z) (SEQ ID NO:24); DG05-CD28tm-DAP10-CD3ζ (DG05-28tm-10-3ζ) (SEQ ID NO:40); DG05-0028tm-0044-CD3ζ (DG05-28tm-44-34) (SEQ ID NO:41); DG05-CD28tm-CD3ζ (DG05-28tm-3ζ) (SEQ ID NO:42); DG05-CD28 (DG05-28) (SEQ ID NO:43); and DG05-CD28tm (DG05-28tm) (SEQ ID NO:44), or a CAR comprising at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs:24, or 40-44.
  20. 176 . The modified Treg of claim 158 , wherein the Treg comprises a suicide gene, optionally wherein the suicide gene is expressed under the control of an inducible promoter.

Description

RELATED APPLICATIONS This application is a continuation of U.S. application Ser. No. 16/968,451 filed Aug. 7, 2020, which is a National Phase application under U.S.C. § 371 of PCT/US2019/017489, filed Feb. 11, 2019, which claims the benefit of and priority to U.S. Provisional Application No. 62/628,632, filed on Feb. 9, 2018, the contents of which are incorporated by reference in their entireties. GRANTS The instant application was made with government support under grant no. R21 NS102556 awarded by the National Institutes of Health (“NIH”). The government has certain rights in the invention. SEQUENCE DISCLOSURE The Sequence Listing submitted as an XML file named “DTM_2018-117_CON_US_ST26.xml,” created on Oct. 9, 2025, and having a size of 328,762 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.834(c)(1). FIELD OF THE ART The present disclosure generally relates to novel chimeric antigen receptors (“CARs”), nucleic acids encoding such CARs and constructs containing same, modified regulatory T cells (“Tregs”) expressing such CARs and/or Tregs which are engineered to express neurodegenerative disease modifying molecules, e.g., which prevent oxidative/inflammatory activity, or which promote neuronal growth/survival such as nerve growth factors or non-classical neurotrophic factors. The present disclosure also generally relates to compositions containing such modified Tregs, and methods of use thereof as therapeutics, in particular for treating and preventing neurodegenerative diseases and symptoms associated therewith, and/or for slowing the onset of such neurodegenerative diseases, particularly in persons at risk because of genetic factors or in persons exhibiting early signs of developing such neurodegenerative diseases. BACKGROUND Neurodegenerative diseases can generally be characterized by a slow progressive loss of neurons in the central nervous system (CNS), which often leads to deficits in specific brain functions (e.g. memory, movement, cognition) performed by the affected CNS region. These neurodegenerative diseases include, for example, Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis, Huntington's disease, and multiple system atrophy. Neurodegenerative diseases usually extend over a decade, and the actual onset of neurodegeneration may precede clinical manifestations by many years. Alzheimer's disease (AD) is a progressive neurodegenerative disease that is one of the primary reasons for memory dysfunction and dementia after 60 years of age. Neuronal dysfunction and death in the frontal cortex and hippocampus, along with microglia-mediated neuroinflammation and formation of aberrant protein aggregates and fibrils are hallmarks of AD. Advancing age increases its prevalence, with an estimated 4.5 million individuals over the age of 65 living with clinical AD in the United States. This number is projected to rise to over 13 million, and to over 130 million worldwide, by 2050 (Hebert et al., Alzheimer disease in the United States (2010-2050) estimated using the 2010 census. Neurology 2013, 80:1778-83) Sporadic and familial forms of AD have an overproduction and/or decreased clearance of extracellular amyloid-beta (Aβ) peptides and intraneuronal tangles of twisted tau protein fibers. The genetic basis for inheritable autosomal dominant, early-onset AD involves mutations in genes that alter Aβ production, aggregation, or clearance: these genes include amyloid precursor protein (APP), presenilin-1 (PS1), and presenilin-2 (PS2). Aβ peptides self-oligomerize into small aggregates that can develop into diffuse plaques. Multiple antibodies that bind Aβ in its monomeric, oligomeric, and plaque forms have been created (Montoliu-Gaya L, Villegas S: Aβ-Immunotherapeutic strategies: a wide range of approaches for Alzheimer's disease treatment. Expert reviews in molecular medicine 2016, 18:e13). Neuroinflammation is known to occur in AD, and when associated near Aβ plaques there is a greater neurodegeneration (Heneka M T, et al.: Neuroinflammation in Alzheimer's disease. The Lancet Neurology 2015, 14:388-405; Kreisl W C, et al.: Distinct patterns of increased translocator protein in posterior cortical atrophy and amnestic Alzheimer's disease. Neurobiol. Aging 2017, 51:132-40). Data suggest that inflammatory microglia—the resident macrophages of the central nervous system—have a role in neurodegeneration and cognitive decline (Kreisl W C et al.: Distinct patterns of increased translocator protein in posterior cortical atrophy and amnestic Alzheimer's disease. Neurobiol Aging 2017, 51:132-40; Paolicelli R C, et al.: TDP-43 Depletion in Microglia Promotes Amyloid Clearance but Also Induces Synapse Loss. Neuron 2017, 95:297,308. e6). There are approximately 7.5 million people with Parkinson's disease worldwide and the disease prevalence increases progressively as age increases (Hebert L E et al.: Alzheimer disease in the United States (2010-20