Isolation, cloning and characterization of new adeno-associated virus (AAV) serotypes
Inventors
Schmidt, Michael • Chiorini, John A.
Assignees
National Institutes of Health NIH • US Department of Health and Human Services
Publication Number
US-8283151-B2
Publication Date
2012-10-09
Expiration Date
2026-05-01
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Abstract
The present invention provides new adeno-associated virus (AAV) viruses and vectors, and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV vectors and particles.
Core Innovation
The invention provides new adeno-associated viruses (AAVs), vectors derived therefrom, and viral particles comprising those vectors. These new AAVs, designated as AAV-X1, AAV-X1b, AAV-X5, AAV-X19, AAV-X21, AAV-X22, AAV-X23, AAV-X24, AAV-X25, and AAV-X26, exhibit unique nucleic acid sequences, capsid proteins, and Rep proteins. The vectors can contain AAV inverted terminal repeats (ITRs), promoters, as well as nucleic acids encoding the capsid and Rep proteins specific to these new serotypes. The invention also provides methods of delivering nucleic acids to cells using these AAV-derived vectors and particles, including novel combinations of ITRs, Rep proteins, and capsids, or chimeric capsids, for efficient gene transfer.
The problem being solved involves limitations in existing AAV serotypes used for gene therapy, including preexisting immunity in human populations, limited tissue tropism, and inefficient transduction of certain cell types. Existing AAV serotypes such as AAV1-11 and others may not efficiently transduce all desired cell or tissue types and can be neutralized by human antibodies, limiting their therapeutic use. The novel AAV serotypes disclosed here provide distinct immunological profiles, receptor binding characteristics, and tissue tropisms, allowing transduction of cell types or subjects not amenable to current AAV vectors, thereby overcoming these challenges.
Claims Coverage
The patent claims include independent claims directed to isolated nucleic acid molecules encoding AAV-X26 polypeptides, isolated nucleic acid sequences, isolated proteins, isolated antibodies, and AAV particles containing these nucleic acids or proteins. There are 5 main inventive features identified from the independent claims.
Isolated nucleic acid encoding an AAV-X26 polypeptide
An isolated nucleic acid molecule encodes a polypeptide consisting of at least 95% amino acid identity to SEQ ID NO:30, which binds an antibody raised against the protein of SEQ ID NO:30.
Isolated nucleic acid molecule comprising sequences related to SEQ ID NO:20
An isolated nucleic acid sequence is at least 95% identical to SEQ ID NO:20 or consists of at least 50 contiguous nucleotides thereof or is fully complementary to such sequences, wherein the encoded polypeptide binds an antibody raised against a protein of SEQ ID NO:30.
Isolated AAV-X26 capsid protein
An isolated protein comprising at least 50 contiguous amino acids or at least 95% amino acid identity to SEQ ID NO:30, which selectively binds an antibody raised against SEQ ID NO:30.
Isolated antibody selectively binding AAV-X26 capsid protein
An isolated antibody selectively binds the AAV-X26 protein as defined in the preceding feature.
AAV particle comprising the isolated nucleic acid or protein
An isolated AAV particle comprising the isolated nucleic acid or protein as described above.
The claims focus on nucleic acid molecules encoding AAV-X26 capsid proteins and related sequences, isolated proteins and antibodies specific to AAV-X26, and AAV particles comprising these nucleic acids or proteins, emphasizing antigenic distinctness and sequence identity as inventive features.
Stated Advantages
Vectors based on new AAV serotypes have different host ranges and distinct immunological properties, allowing more efficient transduction in certain cell types.
Characterization of new serotypes aids in identifying viral elements that confer altered tissue tropism.
The new AAV serotypes are serologically distinct from known AAVs and may allow transduction in subjects with neutralizing antibodies against other AAV serotypes.
The disclosed AAVs exhibit varied tropisms across human cancer cell lines and are not inhibited by common receptor analogs used by other AAVs, indicating unique cell entry pathways.
Recombinant AAV12 is highly resistant to neutralization by pooled human immunoglobulins, enabling potential use in gene therapy for patients with preexisting immunity to other AAVs.
Documented Applications
Gene delivery to a variety of human cell types, including cancer cells (non-small cell lung cancer, colon, CNS derived, ovarian, prostate, breast, cervical cord), kidney cells, salivary gland cells, and muscle cells.
Gene delivery to inner ear epithelial and hair cells, including auditory and vestibular hair cells.
Delivery of nucleic acids ex vivo to cells for transplantation into subjects.
In vivo gene transfer to salivary glands and skeletal muscles.
Use as gene therapy vectors in subjects with neutralizing antibodies to existing AAV serotypes.
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