Modified AAV9 Capsid Peptide Insertion for Endothelial Transduction
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Solution Overview
Problem
Current AAV vectors, such as AAV2, have low transduction efficiency and are hindered by neutralizing antibodies, making efficient gene transfer into endothelial cells challenging, particularly in animal models, which limits their therapeutic use in vascular diseases.
Innovation Solution
A polynucleotide encoding an AAV capsid polypeptide with an inserted peptide at a site exposed on the capsid, such as amino acid 588 or 591, is used to enhance the transduction efficiency and specificity of gene transfer, specifically for AAV9 vectors, by modifying the capsid to improve viral attachment and infection processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If AAV2 vectors are used for gene transfer, then long-term expression of therapeutic genes is achieved, but transduction efficiency into endothelial cells is too low for therapeutic use
Solution Approach 1:
The patent applies local quality by inserting a specific peptide sequence (RGD motif) at a defined location within the AAV2 capsid protein (amino acids 585-592). This localized modification at the capsid surface specifically enhances endothelial cell binding without altering the overall capsid structure or long-term expression capability, thereby resolving the contradiction between reliable long-term expression and sufficient transduction efficiency.
2Reliability
If AAV2 vectors are used, then gene transfer capability is maintained, but neutralizing antibodies in most individuals prevent efficient transduction
Solution Approach 1:
The patent changes the biochemical parameters of the AAV2 capsid by incorporating an RGD peptide motif, which alters the surface properties and binding specificity of the virus. This parameter change enables the vector to bind to endothelial cells via integrin receptors, providing an alternative entry mechanism that is not blocked by pre-existing neutralizing antibodies against wild-type AAV2, thus maintaining gene transfer capability while overcoming antibody-mediated inhibition.
3Productivity
If peptide is inserted into AAV capsid, then transduction efficiency is enhanced, but capsid assembly activity may be interfered with
Solution Approach 1:
The patent applies partial action by inserting a relatively short peptide sequence (7 amino acids including the RGD motif) into a specific region of the capsid protein rather than making extensive modifications. This limited, targeted insertion is sufficient to enhance transduction efficiency through improved endothelial cell binding while minimizing disruption to the overall capsid assembly process, thereby maintaining reliable capsid formation.
Data Source
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AI summary
The present invention is concerned with the provision of a polynucleotide encoding an AAV capsid polypeptide comprising an inserted peptide and a vector comprising said polynucleotide. Moreover, contemplated is a host cell comprising said polynucleotide or vector, a method for the manufacture of said capsid polypeptide as well as said polypeptide. Further included is an antibody specifically binding to said polypeptide and a medicament comprising said polynucleotide, vector, polypeptide, or antibody. Also contemplated are the use of said polynucleotide, vector, polypeptide, or antibody for the manufacture of a medicament for the treatment of vascular disease and a method for the identification of a compound binding to said polypeptide.