Covalent Linkage of Multiple AAV Vectors for Gene Therapy
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Solution Overview
Problem
Current methods for delivering multiple adeno-associated virus (AAV) vectors into a single cell are inefficient, requiring high doses and leading to immunogenic responses due to the limited packaging size and low success rate of AAV particles reaching the cell nucleus, making multi-gene delivery and large transgene delivery challenging.
Innovation Solution
Creating a physical linkage between multiple AAV vectors using covalent bonds formed by functionalizing surface moieties and reacting them with biorthogonal reactions such as SPAAC, CuAAC, or SPANC, allowing linked vectors to enter cells together and increase delivery efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If high doses of AAV vectors are administered to improve delivery of multiple genes to the same cell, then the likelihood of multi-gene delivery increases, but severe immunogenic responses occur
Solution Approach 1:
The patent merges multiple separate AAV vectors into a single linked complex structure where multiple vectors are covalently connected through surface moieties. This allows multiple genes to be delivered together in one administered unit, achieving multi-gene delivery reliability without requiring high doses that would cause immunogenic responses. The linked structure ensures that if one vector enters the cell, the others follow, improving delivery consistency.
2Adaptability or versatility
If separate AAV vectors are used to deliver large transgenes exceeding packaging limits, then the transgene can be delivered, but both vectors must enter the same cell which requires high doses
Solution Approach 1:
Multiple AAV vectors carrying different transgene fragments are covalently linked into a single administered complex. This merging allows the fragments to be delivered together as one unit, eliminating the need for high doses to ensure both vectors reach the same cell. The linked structure physically ensures co-entry into the cell, enabling delivery of large transgenes that would exceed single-vector packaging limits.
3Reliability
If multiple separate AAV vectors are administered to achieve multi-gene delivery, then gene therapy effectiveness may improve, but the complexity of ensuring co-delivery to the same cell increases
Solution Approach 1:
The patent combines multiple AAV vectors into a single linked complex structure, simplifying the delivery system from multiple separate vectors to one administered unit. This merging reduces the complexity of ensuring co-delivery to the same cell, as the linked structure physically ensures that all vectors enter together. The approach maintains gene therapy effectiveness while reducing the logistical and biological complexity of multi-vector coordination.
4Quantity of substance
If AAV vectors are used to deliver genes, then gene expression can be altered, but the small packaging size limits the genetic cargo capacity
Solution Approach 1:
The patent merges multiple AAV vectors into a linked complex, effectively pooling their cargo capacities. This allows the delivery of large transgenes that would exceed the packaging limits of a single AAV vector. The linked structure enables the system to accommodate larger genetic cargo while maintaining the benefits of AAV delivery, such as nuclear targeting and gene expression capability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enhances the delivery of multiple AAV vectors into a single cell, reducing the overall dose required and minimizing immunogenic responses, thereby improving the effectiveness of gene therapy by ensuring that linked vectors enter cells together.
Implementation Method 1
the first surface moiety and the second surface moiety react to form a covalent linkage, thereby resulting in the physical linkage of both AAV vectors to each other
Implementation Method 2
the reaction that forms the covalent linkage comprises a strain-promoted azide-alkyne click cycloaddition (SPAAC) reaction
Data Source
AI summary
Methods and structures are disclosed for creating a physical linkage between two or more viral particles, which can covalently link the viral particles together. The methods and structures described herein are designed for purposes of improving efficiency and effectiveness of vector delivery into cells and tissues for purposes of gene therapy. Methods for linking two or more viral vectors comprise functionalizing a first vector with a first surface moiety and functionalizing a second vector with a second surface moiety. Thereafter, the first functionalized vector and second functionalized vector are combined so that the two surface moieties can react. During this reaction, the first surface moiety and the second surface moiety form a covalent linkage, thereby resulting in the physical linkage of both vectors to each other.