Gene therapy for combined methylmalonic acidemia/aciduria and hyperhomocysteinemia/homocystinuria, cobalamin C type, and deficiency of MMACHC
Inventors
Venditti, Charles P. • Sloan, Jennifer L.
Assignees
US Department of Health and Human Services
Publication Number
US-11510998-B2
Publication Date
2022-11-29
Expiration Date
2036-04-27
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Abstract
The present invention provides a synthetic MMACHC polynucleotide comprising a polynucleotide encoding MMACHC that is codon-optimized for expression in a human. Also provided is a polypeptide encoded by a synthetic MMACHC polynucleotide, an expression vector comprising a MMACHC gene sequence under the control of a chicken beta actin (CBA) promoter, and an expression vector comprising a synthetic MMACHC polynucleotide. Methods of treating cobalamin C deficiency and for detecting or tracking exogenous MMACHC are also provided.
Core Innovation
The invention provides a synthetic MMACHC polynucleotide encoding the MMACHC protein that is codon-optimized for expression in humans. The invention also includes polypeptides encoded by the synthetic MMACHC, expression vectors comprising the synthetic MMACHC nucleotide sequence under control of a chicken beta actin (CBA) promoter, and compositions including these polynucleotides, polypeptides, or vectors with pharmaceutically acceptable carriers. Methods for treating cobalamin C deficiency by administering these compositions and methods for detecting or tracking exogenous MMACHC are also provided.
Patients with combined methylmalonic acidemia/aciduria and hyperhomocysteinemia/homocystinuria cobalamin C type (cblC) suffer from metabolic decompensation, thromboembolic events, multisystemic disease, and require lifelong intramuscular injections of cobalamin plus other medications. Despite treatment, they experience progressive neurological and ocular complications, including legal blindness by age ten and cognitive impairments. The burden of existing treatments and the lack of effective therapies for neurological and ocular manifestations represent a significant medical need.
To address this, the invention introduces codon-optimized synthetic MMACHC nucleotide sequences, vectors for expression of MMACHC, and methods of administering these to treat conditions arising from MMACHC deficiency. The invention further develops viable mouse models exhibiting characteristic phenotypes of cblC for use in research and therapeutic testing. The use of codon optimization enhances expression levels, and gene therapies using adeno-associated viral vectors expressing codon-optimized MMACHC improve survival and disease phenotypes in mouse models, offering a potentially improved approach for treating human cblC disease.
Claims Coverage
The patent contains one independent claim that defines the synthetic MMACHC polynucleotide encoding, along with its compositions, expression vectors, and related methods. The inventive features focus on the codon optimization and expression configurations of the MMACHC gene.
Synthetic MMACHC polynucleotide codon-optimized for human expression
A synthetic methylmalonic aciduria cblC type and homocystinuria type C protein (MMACHC) polynucleotide comprising a polynucleotide encoding MMACHC that is codon-optimized for expression in a human, wherein the polynucleotide encoding MMACHC comprises the sequence of SEQ ID NO: 2.
Polynucleotide encoding of detected epitope tags
The synthetic MMACHC polynucleotide can further comprise polynucleotide sequences encoding at least one of a hemagglutinin tag and a 3×FLAG tag for detection and tracking purposes.
Pharmaceutical compositions including synthetic MMACHC polynucleotide
A composition comprising the synthetic MMACHC polynucleotide and a pharmaceutically acceptable carrier, enabling therapeutic administration.
Expression vectors comprising the synthetic MMACHC polynucleotide
Expression vectors comprising the synthetic MMACHC polynucleotide, which can be under the control of a chicken beta actin (CBA) promoter for effective expression.
Expression vectors as viral vectors including AAV vectors with pseudotyped capsids
The expression vector can be a viral vector, specifically an adeno-associated viral (AAV) vector, pseudotyped with capsids such as rh10, type 9, type 8, or 7m8 to facilitate delivery and expression in target tissues.
Specific isolated nucleic acid sequences in expression vectors
Expression vectors comprising isolated nucleic acid sequences selected from SEQ ID NOS: 5-9, covering various MMACHC constructs including wild-type and codon-optimized versions with or without epitope tags.
The claims collectively cover the codon-optimized synthetic MMACHC polynucleotide, tagged variants, their pharmaceutical compositions, and expression vectors including viral vectors, with specific sequences and promoter control. These features enable enhanced expression and delivery of MMACHC for therapeutic use.
Stated Advantages
Improved expression levels of MMACHC via codon optimization as compared to wild-type sequences.
Development of viable mouse models that recapitulate human cblC phenotypes for research and therapeutic assessment.
Gene therapy using AAV vectors expressing codon-optimized MMACHC improves survival, growth, and biochemical abnormalities in mouse models more effectively than conventional hydroxocobalamin treatment.
Potential for less burdensome and more effective treatment of cblC, specifically addressing neurological and ocular manifestations that are not adequately treated by existing therapies.
Documented Applications
Treatment of combined methylmalonic acidemia/aciduria and hyperhomocysteinemia/homocystinuria cobalamin C type (cblC) and deficiency of MMACHC in subjects.
Treatment of vision loss related to cblC by administration of AAV-mediated gene therapy to the eye via subretinal, intravitreal, or retinal artery or vein injection.
Use in treating related disorders associated with MMACHC deficiency, including congenital heart defects, neural tube defects, combined methylmalonic acidemia and homocystinuria X type (cblX), HCFC1 spectrum defects, hyperhomocystinuria, and vitamin B12 deficiency.
Use of codon-optimized MMACHC and tagged alleles in developing cell lines for overexpression and therapeutic applications, including genome editing and homologous recombination.
Methods to detect or track exogenous MMACHC expression in tissues, biospecimens, or body fluids to monitor delivery and expression of gene therapies.
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