Adeno-associated virus with engineered capsid
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Publication Number
US-12104165-B2
Publication Date
2024-10-01
Expiration Date
Abstract
The present disclosure provides recombinant adeno-associated virus (rAAV) virions with an engineered capsid protein identified using a high-throughput sequencing screen or rational design. In particular, the disclosure provides AAV5 virions and capsid proteins with increased transduction efficiency in cardiac cells, increased cell-type selectivity, and/or other desirable properties.
Core Innovation
The invention provides a method for treating arrhythmogenic right ventricular cardiomyopathy (ARVC) in a human subject by intravenously administering a therapeutically effective amount of a composition comprising a recombinant adeno-associated virus (rAAV) AAV9 virion. The AAV9 virion comprises a heterologous nucleic acid encoding a plakophilin 2 (PKP2) polypeptide operatively linked to a cardiac-specific promoter, a Woodchuck Post-transcriptional Regulatory Element (WPRE), and a growth hormone polyA signal.
The document describes the therapeutic strategy as delivering PKP2 via an AAV9 virion under control of a cardiac-specific promoter. It further specifies use of WPRE and a growth hormone polyA signal as part of the heterologous nucleic acid expression cassette, with a pharmaceutically acceptable carrier or excipient including buffer and/or salt.
The invention further describes engineered recombinant adeno-associated virus (rAAV) AAV5 virions with capsid modifications, including surface-exposed GH loop insertions of 5–11 amino acids and/or substitutions at residues such as S651, T578, and T582. These engineered capsids are configured to improve transduction efficiency and cell-type selectivity in cardiac cells, including human cardiac fibroblasts and induced pluripotent stem cell-derived cardiomyocytes.
The document further describes an approach for identifying capsids and capsid variants using a library-based capsid identification workflow. The workflow involves cap polynucleotide library variants and enrichment for target-cell transduction followed by sequencing to identify capsid variants, and the engineered capsids are also described as part of pharmaceutical compositions for delivering gene products to cardiac cells.
Claims Coverage
The claims cover 6 inventive features.
Intravenous AAV9 delivery of cardiac-PKP2 expression cassette for ARVC
A method for treating arrhythmogenic right ventricular cardiomyopathy (ARVC) in a human subject by intravenously administering a therapeutically effective amount of a composition comprising an rAAV AAV9 virion comprising a heterologous nucleic acid encoding a plakophilin 2 (PKP2) polypeptide operatively linked to a cardiac-specific promoter, a Woodchuck Post-transcriptional Regulatory Element (WPRE), and a growth hormone polyA signal.
Cardiac-specific promoter option in the PKP2 cassette
The method wherein the cardiac-specific promoter is an MHC promoter, an MLC-2 promoter, or a cTnT promoter.
Carrier or excipient defined as buffer and/or salt
The method wherein the pharmaceutically acceptable carrier or excipient includes a buffer, a salt, or a combination of a buffer and a salt.
Engineered AAV5 capsids with GH loop insertions and residue substitutions
Recombinant adeno-associated virus (rAAV) AAV5 virions with capsid modifications including surface-exposed GH loop insertions of 5–11 amino acids and/or substitutions at residues such as S651, T578, and T582.
Improved transduction efficiency and cell-type selectivity in cardiac cells
The engineered capsids are configured to improve transduction efficiency and cell-type selectivity in cardiac cells, including human cardiac fibroblasts (hCF) and induced pluripotent stem cell-derived cardiomyocytes (iPS-CM).
Library-based capsid identification workflow
An approach for identifying capsids and capsid variants using cap polynucleotide library variants and enrichment for target-cell transduction followed by sequencing to identify capsid variants.
Overall claim coverage centers on intravenously administering an AAV9 rAAV virion carrying a heterologous nucleic acid encoding PKP2 with a cardiac-specific promoter, WPRE, and a growth hormone polyA signal, and also includes engineered AAV5 capsids, cardiac-cell transduction/selectivity features, and a library-based capsid identification workflow.
Stated Advantages
Treats arrhythmogenic right ventricular cardiomyopathy (ARVC) in a human subject in need thereof.
Improved transduction efficiency in cardiac cells.
Altered cell-type selectivity between human cardiac fibroblasts (hCF) and iPSC-derived cardiomyocytes (iPS-CM).
Documented Applications
Treatment of arrhythmogenic right ventricular cardiomyopathy (ARVC) in a human subject in need thereof.
Delivering gene products to cardiac cells.
Transduction of human cardiac fibroblasts (hCF) and induced pluripotent stem cell-derived cardiomyocytes (iPS-CM).
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