AAV Capsid Amino Acid Substitutions for Higher AAVR Affinity
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Solution Overview
Problem
Existing adeno-associated viruses (AAVs) face challenges in transduction efficiency and immunogenicity, which are linked to their affinity for the AAV receptor (AAVR), necessitating improved capsid modifications to enhance targeting and reduce dose requirements.
Innovation Solution
Engineering AAV capsids with specific amino acid substitutions in major and minor capsid proteins, such as VP1, VP2, and VP3, to increase affinity for AAVR, thereby improving transduction efficiency and reducing immunogenicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the affinity of AAV capsid for AAVR is increased through amino acid substitutions, then transduction efficiency is improved, but the complexity of capsid structure increases
Solution Approach 1:
The patent applies local quality by introducing specific amino acid substitutions (A472H, A273N, Q387K, E500R, E500P) at targeted positions within the capsid structure, particularly at the receptor binding interface. These localized modifications enhance affinity for AAVR without requiring global structural changes, thereby improving transduction efficiency while minimizing overall structural complexity
Solution Approach 2:
The patent employs parameter changes by systematically varying amino acid residues at specific positions in the capsid proteins (VP1, VP2, VP3). Through rational design and screening of substitutions at positions such as 472, 273, 387, and 500, the invention optimizes binding parameters to AAVR, achieving enhanced transduction efficiency through controlled parameter modification rather than structural redesign
2Object-affected harmful factors
If the dose of AAV is reduced to mitigate immunogenicity, then safety is improved, but transduction efficiency decreases
Solution Approach 1:
The patent applies preliminary anti-action by pre-modifying the capsid structure with amino acid substitutions that enhance affinity for AAVR before administration. This preliminary optimization of binding capability allows the virus to achieve effective transduction at lower doses, thereby preventing immunogenicity issues that would arise from high-dose administration while maintaining therapeutic efficacy
3Reliability
If multiple amino acid substitutions are introduced in capsid proteins, then affinity for AAVR is increased, but manufacturing complexity increases
Solution Approach 1:
The patent applies segmentation by dividing the capsid modification strategy into discrete, independently optimizable amino acid substitution sites (positions 472, 273, 387, 500 in VP1, VP2, and/or VP3). Each substitution can be independently introduced and screened, allowing systematic optimization of AAVR affinity while maintaining manageable manufacturing complexity through modular genetic engineering approaches
Data Source
AI summary
Provided herein are engineered adeno-associated virus (AAV) capsids having altered (e.g., increased) affinity for AAV receptor (AAVR); methods of using the same; and methods of determining the affinity of an AAV capsid for a query protein using virus-like particles (VLPs) having a capsid consisting of the VP3 subunit of the AAV capsid.