AAV Virions with GH Loop Peptide Insertion
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Solution Overview
Problem
Current adeno-associated virus (AAV) vectors have limited infectivity and transduction efficiency in retinal cells, particularly when administered via intravitreal injection, which hinders their effectiveness in treating ocular diseases.
Innovation Solution
Development of recombinant AAV virions with altered capsid proteins, specifically a variant AAV capsid protein comprising an insertion of a peptide in the GH loop, such as LALGETTRPA, which enhances the infectivity of retinal cells compared to wild-type AAV vectors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type AAV vectors are used for intravitreal injection, then the vector structure remains simple and stable, but the infectivity and transduction efficiency in retinal cells is limited
Solution Approach 1:
The patent applies local quality by making specific localized modifications to the capsid protein structure. A peptide sequence (LALGETTRPA) is inserted at a specific location in the GH loop of the capsid protein, while the rest of the capsid structure remains unchanged. This localized modification enhances retinal cell infectivity without requiring complete restructuring of the entire viral particle.
Solution Approach 2:
The patent employs parameter changes by modifying the amino acid sequence of the capsid protein through peptide insertion. This changes the physical-chemical properties of the capsid surface, enabling improved interaction with retinal cell surfaces and receptors, thereby enhancing transduction efficiency while maintaining overall capsid stability.
2Productivity
If peptide insertion is made in the capsid protein GH loop, then the infectivity of retinal cells is enhanced, but the capsid protein sequence complexity increases
Solution Approach 1:
The patent applies preliminary action by pre-designing and pre-testing specific peptide sequences before full-scale production. The peptide sequence LALGETTRPA was identified and optimized through preliminary experiments, allowing for efficient production of high-performance vectors without requiring extensive trial-and-error during manufacturing.
Solution Approach 2:
The patent uses parameter changes by systematically varying peptide sequences to optimize transduction efficiency. The specific sequence LALGETTRPA represents an optimized parameter set that achieves high productivity while maintaining manageable sequence complexity through rational design rather than random exploration.
3Productivity
If variant AAV capsid protein is used, then the gene delivery capability to retinal cells is improved, but the deviation from wild-type structure increases
Solution Approach 1:
The patent applies local quality by confining structural deviations to a specific localized region (the GH loop) while maintaining the stability and integrity of the rest of the capsid protein. This ensures that the fundamental structural properties and assembly characteristics of the wild-type capsid are preserved, while only the necessary functional region is modified for enhanced gene delivery.
Solution Approach 2:
The patent employs composite materials by combining the proven stable framework of the wild-type AAV capsid with a specially designed peptide sequence. This creates a hybrid structure that leverages the stability of the original capsid while incorporating the enhanced functionality of the inserted peptide, achieving both stability and improved gene delivery.
Data Source
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AI summary
The present disclosure provides adeno-associated virus (AAV) virions with altered capsid protein, where the AAV virions exhibit greater infectivity of retinal cells, when administered via intravitreal injection, compared to wild-type AAV. The present disclosure further provides methods of delivering a gene product to a retinal cell in an individual, and methods of treating ocular disease.