AAV with unique capsid protein VP1 and methods of using for treatment
Inventors
Chiorini, John A. • Di Pasquale, Giovanni • Chandler, Randy • Venditti, Charles P.
Assignees
US Department of Health and Human Services
Publication Number
US-11447797-B2
Publication Date
2022-09-20
Expiration Date
2036-05-12
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Abstract
The invention provides an adeno-associated viral (AAV) vector comprising a capsid comprising the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 9, wherein the AAV vector further comprises a heterologous nucleic acid sequence, and wherein the heterologous nucleic acid sequence can encode the NGF-PTH fusion polypeptide or methylmalonyl CoA mutase enzyme. The invention also provides a polypeptide comprising nerve growth factor (NGF) signal peptide and parathyroid hormone (PTH), wherein the polypeptide can comprise, consist essentially of, or consist of the amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 2. The invention provides a nucleic acid encoding the polypeptide, a vector comprising the nucleic acid, and a composition comprising the polypeptide, nucleic acid, or vector, as well as treatment methods comprising the polypeptide, nucleic acid, vector, or composition. The invention further provides a method of treating methylmalonic acidaemia (MMA) in a mammal comprising administering an AAV vector comprising a heterologous nucleic acid sequence encoding methylmalonyl CoA mutase enzyme to the mammal.
Core Innovation
The invention provides an adeno-associated viral (AAV) vector comprising a capsid with the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 9, and further comprising a heterologous nucleic acid sequence. This vector can encode a NGF-PTH fusion polypeptide or a methylmalonyl CoA mutase enzyme. Additionally, it includes a polypeptide combining nerve growth factor (NGF) signal peptide and parathyroid hormone (PTH), along with nucleic acids encoding this polypeptide, vectors containing these nucleic acids, compositions thereof, and related treatment methods.
The background reveals the problem addressed: AAV is useful as a gene therapy vector due to its ability to infect various cells and establish long-term expression without pathogenicity. However, there is a desire for new AAV isolates with different host ranges and improved immunological properties. Moreover, hypoparathyroidism is a hormone deficiency lacking adequate current replacement therapies, demonstrating a need for novel treatment methods.
The invention characterizes a newly isolated AAV termed “44-9,” which shows high gene transfer activity in cell types including salivary gland, liver, and nerve cells despite containing serine residues previously associated with decreased transduction. The invention provides polypeptides corresponding to the VP1 capsid protein of AAV 44-9 and a mutated form with a serine-to-asparagine substitution altering transduction and binding affinity. The invention also details polynucleotide sequences encoding these capsid proteins, AAV vectors comprising these capsids and heterologous nucleic acid sequences, and fusion polypeptides combining NGF signal peptides and PTH for therapeutic use.
Claims Coverage
The patent contains multiple independent claims that define inventive features relating to AAV vectors with specific capsid proteins, vectors encoding therapeutic polypeptides, and methods of treatment using these vectors.
AAV vector comprising specific capsid proteins and heterologous nucleic acid sequences
An adeno-associated viral vector comprising a nucleic acid encoding a capsid protein with the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 9, further comprising a heterologous nucleic acid sequence.
Heterologous nucleic acid sequences encoding therapeutic proteins
The heterologous nucleic acid sequence encodes therapeutic proteins including methylmalonyl CoA mutase enzyme or a polypeptide comprising NGF signal peptide and parathyroid hormone (PTH). The polypeptide comprises the amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 2.
Method of treating methylmalonic acidaemia (MMA)
A method of treating MMA in a mammal comprising administering intravenously an AAV vector encoding methylmalonyl CoA mutase enzyme.
Method of treating hypoparathyroidism
A method of treating hypoparathyroidism in a mammal by administering the AAV vector encoding NGF-PTH fusion polypeptide to the salivary glands of the mammal.
Composition comprising the AAV vector and pharmaceutically acceptable carrier
A pharmaceutical composition comprising the described AAV vector and a pharmaceutically acceptable carrier.
The claims cover AAV vectors with capsids of specific amino acid sequences, vectors encoding heterologous nucleic acid sequences for therapeutic proteins including NGF-PTH fusion and methylmalonyl CoA mutase, methods for treating diseases such as hypoparathyroidism and MMA using these vectors, and compositions containing these AAV vectors.
Stated Advantages
The AAV 44.9 vector demonstrates high gene transfer activity in various cell types including salivary gland, liver, and central nervous system cells, despite structural features typically associated with poor transduction.
The invention enables long-term, endocrine release of biologically active PTH after AAV-mediated delivery to salivary glands, with sustained therapeutic hormone levels for over eight months.
The AAV 44.9 vector allows effective delivery of therapeutic genes to tissues poorly targeted by other AAV serotypes, including CNS and liver, enabling treatment of disorders such as hypoparathyroidism and methylmalonic acidaemia.
The compositions can be formulated to increase stability, reduce degradation, and enhance transduction efficiency, facilitating administration and extending shelf life.
Documented Applications
Treatment of hypoparathyroidism by administering the NGF-PTH fusion polypeptide, nucleic acid, vector, or composition, particularly via delivery to salivary glands.
Treatment of methylmalonic acidaemia (MMA) through delivery of an AAV 44.9 vector encoding methylmalonyl CoA mutase enzyme to the liver of affected mammals.
Gene transfer to salivary gland cells, liver cells, and various brain cell types including cortex, olfactory bulb, brain stem, and Purkinje cells for therapeutic expression of heterologous genes.
Treatment of retinoschisis in the eye using AAV 44.9 vector encoding retinoschisin.
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