Optimized GALC genes and expression cassettes and their use
Inventors
Gray, Steven James • Lykken, Erik • Vite, Charles H. • Bradbury, Allison
Assignees
University of North Carolina at Chapel Hill • University of Pennsylvania Penn
Publication Number
US-12359181-B2
Publication Date
2025-07-15
Expiration Date
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Abstract
This invention relates to polynucleotides comprising optimized GALC open reading frame (ORF) sequences, vectors comprising the same, and methods of using the same for delivery of the ORF to a cell or a subject and to treat disorders associated with aberrant expression of a GALC gene or aberrant activity of a GALC gene product in the subject, such as Krabbe disease (i.e., globoid cell leukodystrophy (GLD)).
Core Innovation
This invention relates to polynucleotides comprising optimized GALC open reading frame (ORF) sequences, vectors comprising the same, and methods of using the same for delivery of the ORF to a cell or a subject and to treat disorders associated with aberrant expression of a GALC gene or aberrant activity of a GALC gene product in the subject, such as Krabbe disease. One aspect of the invention relates to a polynucleotide comprising a canine or human GALC open reading frame, wherein the canine or human GALC open reading frame has been codon-optimized for expression in canine or human cells.
Krabbe disease (KD) is a rapidly progressive, terminal lysosomal storage disorder caused by mutations in the GALC gene that leads to accumulation of galactocerebroside and psychosine, demyelination, and progressive central nervous system dysfunction. There remains a need in the art for an effective treatment that targets the cause of the disease, i.e., GALC gene mutations, because current treatment options such as hematopoietic stem cell therapy (HSCT) have substantial limitations.
The present invention provides codon-optimized GALC genes, expression cassettes, and vectors capable of providing therapeutic levels of GALC expression for treating disorders associated with GALC expression such as Krabbe disease. Embodiments include expression cassettes comprising a codon-optimized canine or human GALC ORF operably linked to promoters and/or polyadenylation signals, AAV ITRs, and vectors (including AAV vectors), pharmaceutical formulations comprising the polynucleotide, expression cassette or vector, and methods of delivering and expressing the GALC ORF in cells and subjects.
Claims Coverage
Identified 3 inventive features from 2 independent claims.
Codon-optimized canine or human GALC open reading frame
A polynucleotide comprising a canine or human GALC open reading frame, wherein the GALC open reading frame is codon-optimized for expression in a canine or human cell.
Sequence comprising SEQ ID NO:1 or SEQ ID NO:2 or at least about 90% identity
Wherein said canine or human GALC open reading frame comprises the nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:2, or a nucleotide sequence having at least about 90% identity thereto.
Method of treating disorders associated with aberrant GALC expression or activity
A method of treating a disorder associated with aberrant expression of a GALC gene or aberrant activity of a GALC gene product in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of claim 18, such that the GALC open reading frame is expressed in the subject.
The independent claims disclose (1) a codon-optimized canine or human GALC open reading frame including specific sequences (SEQ ID NO:1 or SEQ ID NO:2 or ≥~90% identity) and (2) a method of treating disorders associated with aberrant GALC expression or activity by administering a pharmaceutical composition so that the GALC open reading frame is expressed in the subject.
Stated Advantages
Provides codon-optimized GALC genes, expression cassettes, and vectors capable of providing therapeutic levels of GALC expression for treating disorders such as Krabbe disease.
Targets the cause of the disease, i.e., GALC gene mutations.
Codon-optimized GALC sequence allows one to distinguish expression of the transduced sequence from expression of the endogenous GALC sequence in a subject.
As stated in the examples, gene therapy intervention is described as relatively safer, requires a shorter hospitalization, provides GALC enzyme to the tissue faster, yields sustained gene expression for a longer duration, provides better tissue distribution with potentially improved clinical benefits, and is expected to be more effective than HSCT in improving quality of life and event-free survival.
Documented Applications
Treating disorders associated with aberrant expression of a GALC gene or aberrant activity of a GALC gene product, including Krabbe disease (globoid cell leukodystrophy).
Expression of a GALC open reading frame in a cell by contacting the cell with the polynucleotide, expression cassette, and/or vector, thereby expressing the GALC open reading frame in the cell (in vitro, ex vivo, or in vivo).
Delivering a GALC open reading frame to a subject by administering the polynucleotide, expression cassette, vector, and/or transformed cell to the subject, thereby expressing the GALC open reading frame in the subject.
Use of the polynucleotide, expression cassette, vector, and/or transformed cell in pharmaceutical formulations in a pharmaceutically acceptable carrier for therapeutic administration.
Vectors comprising the polynucleotide or expression cassette, including AAV vectors (e.g., AAV9, AAVrh10, AAVOlig001) for delivery of codon-optimized GALC ORFs.
Production of virus vectors comprising providing an AAV template comprising the polynucleotide or expression cassette and ITR(s), Rep and Cap sequences, and helper functions under conditions sufficient for replication and packaging to produce recombinant AAV particles.
Transformed cells and transgenic animals comprising the polynucleotide, expression cassette, vector, and/or transformed cell of the invention, including use in laboratory animals (e.g., mouse, rat, rabbit, dog, monkey, or non-human primate).
Administration of the polynucleotide, expression cassette, vector, and/or transformed cell in combination with a bone marrow transplant (BMT/HSCT) as described in certain embodiments.
Veterinary and medical applications, including delivery to neonatal or prenatal subjects as described (e.g., administration as early as possible in the life of the subject, including newborn or in utero scenarios).
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