Mutated AAV Capsid Enhancing Transduction Efficiency
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Solution Overview
Problem
Current adeno-associated virus (AAV) technologies face limitations in efficiently transducing muscle, airway, liver, central nervous system, retina, or lung cells, and in providing effective delivery of therapeutic agents for infectious and genetic diseases, particularly for viruses like Ebola and Marburg.
Innovation Solution
A recombinant AAV particle with a mutated capsid protein, specifically with amino acid substitutions at positions 129, 445, and 731, enhances transduction efficiency and mediates expression of therapeutic agents, including monoclonal antibodies, to provide protection against infectious diseases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If wild-type AAV capsid is used, then the virus can naturally transduce cells, but transduction efficiency is limited and tissue tropism is constrained
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions (F129L, Y445F, Y731F) in the AAV6 capsid protein sequence. These mutations alter the physical and chemical properties of the capsid surface, enabling enhanced binding affinity to cellular receptors and improved transduction efficiency across multiple tissue types including muscle, airway, liver, and central nervous system cells.
Solution Approach 2:
The patent creates a composite capsid structure by combining multiple mutation elements (F129L, Y445F, Y731F) within the AAV6 capsid framework. This composite approach integrates the beneficial properties of each mutation: F129L enhances receptor binding, while Y445F and Y731F mutations prevent phosphorylation and degradation, resulting in a capsid with superior overall performance compared to wild-type AAV.
2Reliability
If tyrosine residues on AAV capsid are left unmutated, then the capsid structure is simpler, but the vector undergoes phosphorylation leading to ubiquitination and degradation
Solution Approach 1:
The patent converts the harmful phosphorylation event into a beneficial outcome by mutating tyrosine residues to phenylalanine. This prevents phosphorylation-mediated degradation while maintaining capsid integrity. The mutations at positions Y445F and Y731F specifically block EGFR-PTK phosphorylation sites, thereby preventing ubiquitination and proteasomal degradation, leading to enhanced vector stability and prolonged transgene expression.
3Reliability
If AAV mediates expression of therapeutic agents, then protection against disease is provided, but transduction efficiency to target cells remains insufficient
Solution Approach 1:
The patent applies parameter changes to the capsid protein sequence to enhance transduction efficiency, which directly improves the delivery of therapeutic agents. The F129L mutation enhances cellular uptake, while Y445F and Y731F mutations increase vector stability, collectively enabling more efficient transduction of target cells and thereby improving therapeutic agent delivery to muscle, airway, liver, and central nervous system tissues.
Data Source
AI summary
A recombinant adeno-associated virus (rAAV) particle with a mutated capsid protein is provided. In particular, the present disclosure provides methods of delivering a therapeutic agent to a muscle, airway, liver, central nervous system, retina or lung cell in a subject, and methods of treating or preventing infectious, acquired or genetic disease, with said rAAV particle.