AAV Vector Backbone with TLR-Modulating Sequences
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Solution Overview
Problem
Current viral vectors, such as AAV, face challenges including low efficiency, packaging capacity limitations, lack of specificity, and elicited inflammatory responses, which can reduce transgene expression and introduce contaminants, necessitating improved methods for gene therapy and biological product production.
Innovation Solution
A vector construct comprising a polynucleotide with a promoter, terminal repeats, and a backbone polynucleotide that modulates Toll-like receptors (TLRs), specifically packaged in an AAV capsid to reduce inflammatory responses and enhance immunogenicity, utilizing nucleic acid sequences that inhibit or activate TLR responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral vectors are used to deliver exogenous DNA into eukaryotic cells, then delivery efficiency is improved, but inflammatory responses are elicited that reduce transgene expression
Solution Approach 1:
The patent incorporates TLR-modulating sequences into the viral vector backbone to convert the harmful inflammatory response into a beneficial control mechanism. These sequences specifically modulate Toll-like receptor responses to reduce unwanted inflammation while maintaining delivery efficiency, transforming the vector's immunogenicity from a harmful side effect into a controlled feature that enhances therapeutic outcome
Solution Approach 2:
The patent modifies the viral vector by incorporating specific nucleic acid sequences that alter the immunogenic parameters of the vector. By changing the backbone sequence composition to include TLR-modulating elements, the vector's interaction with the immune system is parameterically adjusted to reduce inflammatory responses while preserving transduction efficiency
2Productivity
If viral vectors are used for gene therapy, then transgene delivery is achieved, but contaminants are packaged alongside the expression cassette
Solution Approach 1:
The patent extracts and removes contaminating sequences from the viral vector preparation by optimizing the backbone design to exclude extraneous DNA. The streamlined backbone sequence minimizes the packaging of contaminating plasmid DNA or chromosomal DNA from producer cell lines, thereby extracting unwanted elements while retaining essential transgene delivery functionality
3Object-generated harmful factors
If immunosuppressive drugs are used with viral vector based gene therapy, then inflammatory responses are reduced, but the patient's immune system is compromised
Solution Approach 1:
The patent introduces TLR-modulating sequences as an intermediary mechanism that mediates between the viral vector and the immune system. These sequences specifically target Toll-like receptor pathways to reduce inflammation locally at the site of vector interaction, providing immune modulation without requiring systemic immunosuppressive drugs that would compromise overall immune function
Data Source
AI summary
The present disclosure provides the vector constructs comprising (a) a polynucleotide comprising a promoter operably linked to a nucleic acid of interest; (b) a first terminal repeat and a second terminal repeat; and (c) a backbone polynucleotide comprising a nucleic acid sequence that modulates a Toll-like receptor (TLR). Some aspects of the disclosure are related to methods for packaging the nucleic acid of interest and the nucleic acid sequence that modulates the TLR in an AAV particle, and some aspects are related to methods of modulating an immune response in a subject, comprising administering to said subject an effective amount of such AAV particles.