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EP-3411059-B1 - METHOD TO ENHANCE THE EFFICIENCY OF SYSTEMIC AAV GENE DELIVERY TO THE CENTRAL NERVOUS SYSTEM

EP3411059B1EP 3411059 B1EP3411059 B1EP 3411059B1EP-3411059-B1

Inventors

  • ESTEVES, Miguel Sena
  • BATISTA, Ana Rita
  • CHOUDHURY, Sourav Roy

Dates

Publication Date
20260506
Application Date
20170202

Claims (11)

  1. A targeting composition for use in a method for delivering a transgene to CNS tissue in a subject, the method comprising administering to the subject: (i) an effective amount of a targeting composition, wherein the targeting composition comprises a hydrophilic peptide consisting of the amino acid sequence of SEQ ID NO: 17; and, (ii) an effective amount of a recombinant AAV (rAAV) that is capable of targeting CNS tissue, wherein the rAAV comprises at least one inverted terminal repeat, at least one capsid protein, and a nucleic acid comprising a promoter operably linked to a transgene.
  2. The targeting composition for use of claim 1, wherein the at least one capsid protein has a serotype selected from the group consisting of AAV1, AAV2, AAV2i8, AAV2.5, AAV6, AAV8, AAVrh8, AAV9, AAVrh10, AAV-B1, AA V9.45A-String (e.g., AAV9.45-AS), AAV9.45Angiopep, AAV9.47-Angiopep, and AAV9.47-AS.
  3. The targeting composition for use of claim 1 or 2, wherein the at least one capsid protein comprises or consists of a sequence selected from the group consisting of SEQ ID NO: 1-14.
  4. The targeting composition for use of any one of claims 1-3, wherein the at least one capsid protein comprises an N-terminally grafted heterologous targeting peptide.
  5. The targeting composition for use of claim 4, wherein the N-terminally grafted heterologous targeting peptide comprises or consists of a sequence selected from the group consisting of SEQ ID NOs: 15-16.
  6. The targeting composition for use of any one of claims 1-5, wherein the at least one capsid protein comprises or consists of a sequence selected from the group consisting of SEQ ID NO: 10-14.
  7. The targeting composition for use of any one of claims 1-6, wherein the transgene is a CNS-associated transgene selected from the group consisting of ALS2, ANG, ATXN2, C9orf72, DCTN1, FIG4, FUS, NEFH, OPTN, PFN1, PRPH, SETX, SIGMAR1, SMN1, SOD1, SPG11, TARDBP, UBQLN2, VAPB, VCP, Htt, Xbp1s, CRAG, MAN2B1, MAN2B2, MAN2C1, AGA, CLN1, CLN2, CLN3, CLN5, CLN6, MFSD8, CLN8, CTSD, MANBA, CTNS, LAMP2, GLA, ASAH1, FUCA1, CTSA, GBA, GALC, GLB1, HEXA, HEXB, ARSA, IDUA, IDS, SGSH, NAGLU, HGSNAT, GNS, GALNS, ARSB, GUSB, HYAL1, SMPD1, NPC1, NPC2, GAA, NAGA, SLCA17A5, and LAL (LIPA).
  8. The targeting composition for use of any one of claims 1-7, wherein the administration of the nucleic acid and the targeting composition is each independently selected from the group consisting of: intravenous administration, intravascular administration, intra-arterial administration, intracerebral administration, intraventricular administration, and intrathecal administration.
  9. The targeting composition for use according to any one of claims 1-8, wherein the administration to the CNS further comprises administering an effective amount of an inhibitor of endocytosis to the subject, optionally wherein the inhibitor of endocytosis is a small molecule selected from the group consisting of: chloroquine, hypertonic sucrose, chlorpromazine, monodansylcadaverine, phenylarsine oxide, monensin, a phenolthiazine compound, methyl-β-cyclodextrin, filipin, cytochalasin D, latrunculin, amiloride, dynasore, dynole, and dyngoe.
  10. The targeting composition for use of any one of claims 1-9, wherein the targeting composition of (i) is administered to the subject prior to the nucleic acid of (ii), optionally wherein the targeting composition of (i) is administered to the subject between 60 minutes and 1 second before the nucleic acid of (ii).
  11. The targeting composition for use of any one of claims 1-9, wherein the nucleic acid of (ii) is administered prior to the targeting composition of (i), optionally wherein the nucleic acid of (ii) is administered to the subject between 60 minutes and 1 second before the targeting composition of (i).

Description

BACKGROUND The discovery that certain viral vectors (e.g., AAV vectors, such as AAV9) cross the blood brain barrier (BBB) after intravascular infusion and transduce cells in the CNS started a revolution in the field of gene therapy for neurological diseases. However, the efficiency of viral vector-mediated neuronal transduction is generally very low in most CNS regions, both in the neonatal period and in adult animals, where transgene expression is limited mostly to astrocytes and endothelial cells. Attempts have been made to enhance AAV-mediated CNS gene transfer after systemic administration, for example through the development of more potent capsids, and the combination of rAAV with agents known to disrupt the BBB, such as mannitol. Generally, the combination of AAV with BBB disrupting drugs has shown only modest to no effect on the efficiency of CNS gene transfer. (EP 2 453 923 discloses ApoE-based carrier peptides including K16ApoE (SEQ ID NO: 42) for use in delivering biologically active molecules such as plasmids and oligonucleotides across the blood-brain barrier (BBB). The carrier peptides form noncovalent complexes with the active molecules. (Mousazadeh M. et al., J. Drug Target. 2007 January; 15(3): 226-230) discloses an ApoE-derived peptide conjugated to polylysine (apoEdp-PLL) used to carry genetic material, i.e., plasmid PCDNA3-1 containing the β-galactosidase gene, across the blood brain barrier. (Zou L.-L. et al., Curr. Neuropharmacol. 2013 March; 11(2): 197-208) is a review about cell-penetrating peptides and their use in delivering therapeutic molecules such as plasmids and nucleic acids to the CNS. Several hydrophilic peptides between 4 and 50 amino acids in length linked to a BBB receptor targeting agent are disclosed which are used to deliver plasmid DNA comprising a transgene to the CNS. New approaches are needed to achieve high efficiency gene transfer to the CNS. SUMMARY The invention is as defined in the appended claims. Aspects of the disclosure relate to compositions and methods for increased efficiency of gene transfer to the CNS. The disclosure is based, in part, on the surprising discovery that viral vector-mediated delivery of nucleic acids to the CNS of a subject can be enhanced by administering the viral vector with a composition comprising a highly hydrophilic molecule conjugated to a blood brain barrier (BBB)-receptor ligand (e.g., K16ApoE). Accordingly, provided but not claimed is a method for delivering a transgene to CNS tissue in a subject, the method comprising administering to the subject: (i) an effective amount of a targeting composition, wherein the targeting composition comprises a hydrophilic peptide between 4 and 50 amino acids in length linked to a blood brain barrier (BBB) receptor targeting agent; and, an effective amount of a nucleic acid comprising a promoter operably linked to a transgene. Also provided but not claimed is a method for treating central nervous system (CNS) disease in a subject the method comprising administering to the subject ((i) an effective amount of a targeting composition, wherein the targeting composition comprises a hydrophilic peptide between 4 and 50 amino acids in length linked to a blood brain barrier (BBB) receptor targeting agent; and, an effective amount of a nucleic acid comprising a promoter operably linked to a CNS-associated transgene. The CNS disease may be selected from the group consisting of: Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), and lysosomal storage disorder (LSD). The lysosomal storage disorder may be selected from the group consisting of: GM1 gangliosidosis, Tay-Sachs disease, and Sandhoff disease. The nucleic acid may be delivered via a retrovirus vector, an adenovirus vector, a Herpes simplex virus (HSV) vector, or an adeno-associated virus (AAV) vector. Also provided but not claimed is a composition comprising (i) a nucleic acid encoding a transgene; and, (ii) a targeting composition, wherein the targeting composition comprises a hydrophilic peptide between 4 and 50 amino acids in length linked to a blood brain barrier (BBB) receptor targeting agent. The compositions described herein may be contained within a host cell (e.g., a bacterial cell, an insect cell, or a eukaryotic cell, such as a mammalian cell). The composition may further comprise an effective amount of an inhibitor of endocytosis (e.g., chloroquine). The composition may further comprise a pharmaceutically acceptable excipient. The nucleic acid may be contained in a retrovirus vector, an adenovirus vector, a Herpes simplex virus (HSV) vector, or an adeno-associated virus (AAV) vector. The viral vector may be a recombinant AAV (rAAV) comprising (i) at least one inverted terminal repeat; (ii) at least one capsid protein; and, (iii) a nucleic acid comprising a promoter operably linked to a transgene. The at least one capsid protein of a viral vector described herein can be of AAV serotype. The at least one capsid protein may