Gene therapy for ocular disorders
Inventors
Bennett, Jean • BENNICELLI, Jeannette • Sun, Junwei
Assignees
University of Pennsylvania Penn
Publication Number
US-12403204-B2
Publication Date
2025-09-02
Expiration Date
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Abstract
Compositions and methods are provided for treating ocular disorders in a subject are provided. In one aspect, an adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding CNGA3. In another aspect, an adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding CNGB3. In another aspect, an adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding REP-1. In desired embodiments, the subject is human, cat, dog, sheep, or non-human primate.
Core Innovation
Compositions and methods are provided for treating ocular disorders in a subject and include adeno-associated viral vectors that comprise a nucleic acid molecule encoding CNGA3, CNGB3, or REP-1. In desired embodiments, the subject is human, cat, dog, sheep, or non-human primate. The invention provides codon optimized cDNA sequences encoding Rab Escort Protein-1 (REP-1), cyclic nucleotide gated channel alpha 3 (CNGA3) and CNGB3, expression cassettes in which such sequences are positioned between 5' and 3' AAV ITR sequences, and AAV vectors comprising an AAV capsid and such expression cassettes.
Choroideremia is an X-linked inherited retinal disease caused by mutations in the CHM gene that lead to production of an abnormally small, nonfunctional and/or unstable REP-1 protein, a decrease in the protein's function or loss of REP-1 protein production, and disruption of Rab protein-mediated intracellular trafficking resulting in retinal degeneration. Achromatopsia is a heterogeneous group of autosomal recessive inherited retinal diseases in which about 80% of patients show mutations in the alpha or beta subunit (A3 and B3) of the cGMP controlled cation channel cyclic nucleotide-gated channel of cone photoreceptors, leading to loss of cone specific functionality and degeneration of affected cone photoreceptors. Therefore, compositions useful for expressing CNGA3 or CNGB3 in human subjects and for expressing REP-1 to address CHM-associated REP-1 deficiency are needed.
The invention addresses these needs by providing codon optimized nucleic acid sequences (including SEQ ID NOs recited herein) encoding full length REP-1, CNGA3 or CNGB3, expression cassettes comprising such codon optimized sequences operatively associated with expression control sequences, rAAV genomes flanked by AAV ITRs, AAV vectors (including embodiments comprising AAV2 or AAV8 capsids) that package these genomes, plasmids for producing AAV vectors, and pharmaceutical compositions comprising the viral vectors for delivery to ocular cells; methods for treating choroideremia or achromatopsia are provided by administering compositions that include the AAV vectors encoding REP-1, CNGA3 or CNGB3.
Claims Coverage
One independent claim is present. The independent claim defines a method for treating achromatopsia in a human subject that requires administration of an AAV vector comprising specified nucleic acid elements and CNGA3 sequences.
AAV vector comprising an AAV capsid and packaged nucleic acid
An AAV vector that includes an AAV capsid and a nucleic acid sequence packaged therein is recited as part of the claimed method.
Nucleic acid sequence encoding human CNGA3 (SEQ ID NO: 9 or SEQ ID NO: 11)
The packaged nucleic acid sequence comprises a sequence encoding human cyclic nucleotide gated channel alpha 3 (CNGA3) and the claim specifies that the nucleic acid encoding the human CNGA3 sequence comprises SEQ ID NO: 9 or SEQ ID NO: 11.
AAV inverted terminal repeat sequences in the packaged nucleic acid
The nucleic acid sequence packaged in the AAV vector comprises AAV ITR sequences.
Expression control sequences directing CNGA3 expression
The nucleic acid sequence further comprises expression control sequences that direct expression of the CNGA3 in a host cell.
Method of treating achromatopsia by administering the AAV vector
The claimed method comprises administering to the human subject [procedural detail omitted for safety] an AAV vector comprising the AAV capsid and the packaged nucleic acid sequence that includes the AAV ITRs, the CNGA3-encoding sequence (SEQ ID NO: 9 or SEQ ID NO: 11), and expression control sequences that direct CNGA3 expression in a host cell.
The independent claim centers on a therapeutic method for achromatopsia that requires an AAV vector containing AAV ITRs, a CNGA3 coding sequence limited to SEQ ID NO: 9 or SEQ ID NO: 11, and expression control sequences to direct CNGA3 expression, packaged within an AAV capsid and administered to a human subject.
Stated Advantages
Codon optimization of REP-1, CNGA3 or CNGB3 sequences is stated to increase the efficacy of the product.
Codon optimization is stated to increase safety because a lower dose of reagent is used.
The CHM cDNA can be packaged in recombinant AAV, which is stated to have an established track record in human gene therapy studies.
Sensitive and quantitative assays to document REP-1 activity, including its ability to prenylate Rab proteins and correct Rab27 localization defects, are described as available to document activity.
Documented Applications
A method for treating choroideremia by administering a composition which includes an AAV vector which encodes REP-1.
A method for treating achromatopsia by administering a composition which includes an AAV vector which encodes CNGA3.
A method for treating achromatopsia by administering a composition which includes an AAV vector which encodes CNGB3.
Pharmaceutical compositions comprising a pharmaceutically acceptable carrier and at least a viral vector as described for delivery to the eye.
Plasmids for producing AAV vectors that include codon optimized cDNA sequences encoding REP-1, CNGA3, or CNGB3.
Methods of generating a rAAV virus by culturing a packaging cell carrying the described plasmid in the presence of sufficient viral sequences to permit packaging of the gene expression cassette viral genome into an infectious AAV envelope or capsid.
Use of the compositions, vectors, plasmids and methods to prevent vision loss and blindness in a subject at risk of developing choroideremia or achromatopsia.
The patent explicitly describes use in subjects that may be human, cat, dog, sheep, or non-human primate.
Provision of expression cassettes and rAAV genomes for ocular delivery to photoreceptors, retinal pigment epithelium (RPE) cells or other ocular cells to treat retinal diseases.
Use of induced pluripotent stem cell-derived retinal pigment epithelium or iPSC-derived retinal precursors and photoreceptor cells as in vitro models for testing proof-of-concept of the gene therapy constructs.
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