rAAV Vector Packaging Efficiency for Cardiac Gene Therapy
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Solution Overview
Problem
Current treatments for dilated cardiomyopathy (DCM) primarily focus on calcium handling abnormalities, but fail to address mitochondrial dysfunction and apoptosis, which are critical in disease progression, especially at advanced stages.
Innovation Solution
The use of recombinant adeno-associated virus (rAAV) vectors to deliver transgenes encoding S100A1 and ARC proteins, which improve calcium handling and inhibit apoptosis, respectively, thereby addressing all drivers of DCM onset and progression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If traditional gene delivery vectors are used to deliver transgenes, then the vector capacity is limited, but delivering multiple transgenes (S100A1 and ARC) is necessary to address all drivers of DCM
Solution Approach 1:
The patent combines two separate transgenes (S100A1 and ARC) into a single self-complementary AAV vector construct. The vector contains both transgenes flanked by AAV ITRs, with appropriate promoters and polyadenylation signals, allowing simultaneous delivery of multiple therapeutic genes to address calcium handling dysfunction and apoptosis/mitochondrial dysfunction together in one vector particle.
2Productivity
If the transgene sequence length is reduced to improve packaging efficiency, then more transgenes fit in the vector, but the coding sequence may become incomplete
Solution Approach 1:
The patent modifies the transgene sequences by removing introns and optimizing the coding sequences to encode only the essential protein regions. The S100A1 transgene encodes the mature protein without intronic sequences, and the ARC transgene similarly provides a streamlined coding sequence. These parameter changes in sequence composition and structure reduce the overall vector length while preserving the functional integrity of the encoded proteins.
Data Source
AI summary
The present disclosure is related to compositions and methods useful in treating heart conditions. The disclosed compositions and methods are based on gene therapies comprising a recombinant AAV vector for delivering two or more transgenes into the heart of a subject, wherein the transgenes encode an S100A1 protein and a cardiac Apoptosis Repressor with caspase recruitment Domain (cARC) apoptotic inhibitor, respectively. In various embodiments, the compositions and methods disclosed herein comprise vectors comprising S100A1 and/or cARC cDNA sequences that are codon-optimized for expression in humans. In various embodiments, the compositions and methods disclosed herein comprise vectors with improved packaging efficiencies. In some aspects, targeting multiple sources of one or more heart conditions can provide synergistic benefits during treatment.