Antisense Oligonucleotides Correct CEP290 Splicing
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Solution Overview
Problem
Current gene augmentation therapies, such as those using adeno-associated viruses, are limited in treating Leber congenital amaurosis caused by CEP290 mutations due to cargo size constraints and difficulty in regulating expression levels, particularly for genes like CEP290 with large cDNAs, and do not effectively address aberrant splicing caused by intronic mutations.
Innovation Solution
Development of specific antisense oligonucleotides (AONs) that bind to and block aberrant splicing of CEP290, inducing exon skipping to exclude the aberrant exon from mRNA, thereby restoring normal splicing and protein synthesis, using sequences complementary to targeted regions within the CEP290 gene.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If gene augmentation therapy using adeno-associated viruses is used to treat CEP290 mutations, then therapeutic effect is improved, but cargo size constraints prevent effective delivery of large CEP290 cDNA
Solution Approach 1:
The invention extracts and corrects only the specific splicing defect (aberrant exon inclusion) rather than attempting to deliver the entire large CEP290 cDNA. By using AONs to target and correct the splicing error at the mRNA level, the therapy achieves therapeutic effect without needing to deliver the full-length gene, thus bypassing cargo size constraints of viral vectors.
Solution Approach 2:
The invention changes the approach from delivering genomic DNA (large cargo) to delivering small oligonucleotide molecules (small cargo). This parameter change in the therapeutic agent's size and molecular form allows effective delivery via AAV vectors while still addressing the underlying splicing defect caused by CEP290 mutations.
2Reliability
If gene replacement therapy is used to treat CEP290 mutations, then functional protein production is improved, but expression levels cannot be tightly controlled
Solution Approach 1:
Instead of replacing the entire gene and relying on complex regulatory elements for controlled expression, the invention uses AONs to copy and correct the splicing pattern of the endogenous gene. The AONs guide the splicing machinery to produce correctly spliced mRNA from the patient's own CEP290 gene, naturally maintaining physiological expression levels without requiring artificial control mechanisms.
Solution Approach 2:
The invention enables the cell's own splicing machinery to perform the correction function. By introducing AONs that bind to aberrant splice sites, the cell's natural splicing apparatus is directed to produce correct mRNA, eliminating the need for external control systems and ensuring expression levels match the cell's natural requirements.
3Ease of operation
If conventional therapy is used for LCA, then treatment simplicity is maintained, but aberrant splicing caused by intronic mutations is not effectively addressed
Solution Approach 1:
The invention introduces AONs as intermediary molecules that mediate between the aberrant splice sites and the splicing machinery. These small oligonucleotide intermediaries bind to specific sequences created by intronic mutations and redirect splicing, providing a simple yet effective mechanism to correct splicing errors without complex therapeutic interventions.
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
The AONs effectively convert aberrantly spliced CEP290 mRNA to correctly spliced mRNA, increasing wild-type CEP290 protein synthesis, demonstrating potential for treating Leber congenital amaurosis by redirecting normal splicing and avoiding issues of expression level control and cargo size limitations.
Implementation Method 1
specific antisense oligonucleotides (AONs) are able to block the aberrant splicing of CEP290 that is caused by the intronic LCA mutation
Data Source
AI summary
The present invention relates to the fields of medicine and immunology. In particular, it relates to novel antisense oligonucleotides that may be used in the treatment, prevention and/or delay of Leber congenital amaurosis.