Gene therapy for cns degeneration
AssigneeSpacecraft Seven, LLC
InventorsAnnahita KERAVALA, Paven BATTIPROLU, Raj PRABHAKAR, Roderick WONG, Piratip PRATUMSUWAN, Naveen YALAMANCHI
Life of this patent
Estimating the term…
Abstract
The present disclosure relates generally to compositions and methods for treating, preventing, inhibiting, or delaying central nervous system degeneration. The disclosure relates to a recombinant gene therapy vector comprising a PARK2, PINK1, DJ-1, LRRK2, SCNA, c-Rel, ATG7, VMAT2, or GBA gene, or functional fragment or variant thereof. The disclosure also relates to CRISPR/Cas-based gene editing systems for treating, preventing, inhibiting, or delaying central nervous system degeneration.
Claims131
A recombinant gene therapy vector comprising an expression cassette comprising a transgene encoding an E3 ubiquitin protein ligase (PARK2), a PTEN-induced putative kinase 1 (PINK1), a protein deglycase DJ-1 (DJ-1), a Leucine Rich Repeat Kinase 2 (LRRK2), an alpha-synuclein (SCNA), a Proto-oncogene c-Rel (c-Rel), a Ubiquitin-like modifier-activating enzyme (ATG7), a Synaptic vesicular amine transporter (VMAT2), or a glucocerebrosidase (GBA), wherein the transgene polynucleotide is operably linked to a eukaryotically active promoter sequence.
The vector of any one of claims 1 to 6, wherein the expression cassette further comprises a 3′ untranslated region selected from Table 7.
The method of any of claims 28 to 59, wherein the UPDRS score of a subject treated by the method is improved compared to the UPDRS score of a subject not so treated.
A method of inhibiting degeneration or death of a dopaminergic neuron comprising a mutation in a gene associated with a Parkinson's Disease (PD), wherein the mutated gene is:
- a Parkinson protein 2, E3 ubiquitin protein ligase (PARK2) gene, a PTEN-induced putative kinase 1 (PINK1) gene, a protein deglycase DJ-1 (DJ-1) gene, a Leucine Rich Repeat Kinase 2 (LRRK2) gene, an alpha-synuclein (SCNA) gene, a Proto-oncogene c-Rel (c-Rel) gene, a Ubiquitin-like modifier-activating enzyme (ATG7) gene, Synaptic vesicular amine transporter (VMAT2) gene, or glucocerebrosidase (GBA) gene, comprising:
- contacting the neuron with a recombinant gene therapy vector comprising a polynucleotide encoding a wild-type protein expressed by a wild-type version of the mutated gene, or a functional variant or fragment thereof;
- wherein following contact with the recombinant gene therapy vector, the neuron expresses the wild-type protein, or functional variant or fragment thereof.
A recombinant gene therapy vector comprising a polynucleotide encoding a wild-type Parkinson protein 2, E3 ubiquitin protein ligase (PARK2) gene, a PTEN-induced putative kinase 1 (PINK1) gene, a protein deglycase DJ-1 (DJ-1) gene, a Leucine Rich Repeat Kinase 2 (LRRK2) gene, an alpha-synuclein (SCNA) gene, a Proto-oncogene c-Rel (c-Rel) gene, a Ubiquitin-like modifier-activating enzyme (ATG7) gene, Synaptic vesicular amine transporter (VMAT2) gene, or glucocerebrosidase (GBA) gene, or a functional variant or fragment thereof;
- wherein the polynucleotide is operatively linked to a eukaryotically active promoter;
- and wherein a neuron transduced with said recombinant gene therapy vector expresses the wild-type protein, or functional variant or fragment thereof.
A method of treating or inhibiting onset of a Parkinson's Disease (PD) in a subject suffering from or at risk of the PD, comprising:
- administering a recombinant gene therapy vector comprising a polynucleotide sequence encoding a wild-type Parkinson protein 2, E3 ubiquitin protein ligase (PARK2), a PTEN-induced putative kinase 1 (PINK1), a protein deglycase DJ-1 (DJ-1), a Leucine Rich Repeat Kinase 2 (LRRK2), an alpha-synuclein (SCNA), a Proto-oncogene c-Rel (c-Rel), a Ubiquitin-like modifier-activating enzyme (ATG7), Synaptic vesicular amine transporter (VMAT2), or glucocerebrosidase (GBA) protein, or a functional variant or fragment thereof, to the subject;
- wherein administration of the recombinant gene therapy vector treats or inhibits onset of the Parkinson's Disease in the subject.
The method of claim 101, wherein the recombinant gene therapy vector further comprises one or more of:
- a neuron-specific promoter, optionally selected from the group consisting of hSYN1 (human synapsin), INA (alpha-internexin), NES (nestin), TH (tyrosine hydroxylase), FOXA2 (Forkhead box A2), CaMKII (calmodulin-dependent protein kinase II), and NSE (neuron-specific enolase) promoters;
- a ubiquitous promotor selected from the group consisting of CMV, CAG, UBC, PGK, EF1-alpha, GAPDH, SV40, HBV, and chicken beta-actin, human beta-actin promoters;
- an enhancer;
- an intron;
- a poly-A signal;
- a sequence encoding an 2A peptide;
- and
- a WPRE (Woodchuck hepatitis virus posttranscriptional regulatory element).
A method of inhibiting degeneration or death of a dopaminergic neuron having a mutated Parkin (PRKN) gene, comprising contacting the neuron with a gene editing system comprising:
- a. a Cas protein or a polynucleotide encoding a Cas protein;
- b. a guide-RNA (gRNA);
- and
- c. a repair template comprising a functional Parkinson protein 2, E3 ubiquitin protein ligase (PARK2) gene, a PTEN-induced putative kinase 1 (PINK1) gene, a protein deglycase DJ-1 (DJ-1) gene, a Leucine Rich Repeat Kinase 2 (LRRK2) gene, an alpha-synuclein (SCNA) gene, a Proto-oncogene c-Rel (c-Rel) gene, a Ubiquitin-like modifier-activating enzyme (ATG7) gene, Synaptic vesicular amine transporter (VMAT2) gene, or glucocerebrosidase (GBA) gene, or a functional variant or fragment thereof;
- wherein the gene editing system is capable of repairing an endogenous gene in the neuron or inserting a functional gene into the genome of the neuron.
A gene editing system for a cell comprising:
- a. a Cas protein or a polynucleotide encoding a Cas protein;
- b. a guide-RNA (gRNA);
- and
- c. a repair template comprising a functional Parkinson protein 2, E3 ubiquitin protein ligase (PARK2) gene, a PTEN-induced putative kinase 1 (PINK1) gene, a protein deglycase DJ-1 (DJ-1) gene, a Leucine Rich Repeat Kinase 2 (LRRK2) gene, an alpha-synuclein (SCNA) gene, a Proto-oncogene c-Rel (c-Rel) gene, a Ubiquitin-like modifier-activating enzyme (ATG7) gene, Synaptic vesicular amine transporter (VMAT2) gene, or glucocerebrosidase (GBA) gene, or a functional variant or fragment thereof;
- wherein the gene editing system is capable of repairing an endogenous gene in the cell or inserting a functional gene into the genome of the cell.
Dependencies are inferred from the claim text.