Gene Splice-Switching Technology for Treating CACNA1A-Associated Neurological Diseases

SUMMARY

This technology regulates the expression of the CACNA1A gene, which encodes a neuronal calcium channel subunit, by redirecting alternative splicing to treat related neurological diseases. This approach can potentially correct gene expression variations linked to disorders including episodic ataxia, migraines, epilepsy, autism, and spinocerebellar ataxia.

The Unmet Need: Effective approaches in treating CACNA1A-associated diseases

• The field of alternative splicing and precision medicine is increasingly important in tackling genetic disorders. CACNA1A, encoding the pore-forming subunit of the neuronal P/Q type calcium channel, plays a critical role in neurotransmission. Mutations in this gene lead to various neurological conditions such as episodic ataxia type 2 (EA2), developmental and epileptic encephalopathy 42 (DEE42), familial hemiplegic migraine 1 (FHM1), and spinocerebellar ataxia 6 (SCA6). Current treatment options are limited, and there is an urgent demand for targeted medical therapies that can precisely regulate the gene's expression.

• Current approaches to treating diseases associated with CACNA1A mutations face significant challenges. Existing methods lack specificity in targeting the gene or its splicing mechanisms to up- or down-regulate its expression based on the mutation type. This lack of precision results in suboptimal therapeutic outcomes and potential side effects. Splice-switching oligonucleotides (SSOs) have shown promise, but their application to CACNA1A remains underexplored. Moreover, the complexity of alternative splicing mechanisms and the need for translation-dependent degradation of mRNAs with premature termination codons add layers of difficulty in developing effective treatments. There is a clear need for innovations that can harness these mechanisms to correct or mitigate the effects of CACNA1A mutations.

The Proposed Solution: Technique to regulate CACNA1A mRNA and protein expressions by redirecting alternative splicing of an identified nonsense-mediated mRNA decay (NMD) exon

• The faculty inventor leveraged Alternative Splicing coupled with Nonsense-Mediated mRNA Decay (AS-NMD), which degrades mRNAs with premature termination codons, to identify an alternatively spliced exon in CACNA1A that triggers NMD when included in transcripts. Targeting this exon using splice-switching oligonucleotides (SSOs) could regulate CACNA1A expression, offering new treatments for these diseases.

• This approach to regulate CACNA1A mRNA and protein expression by redirecting alternative splicing represents a significant innovation. For the first time, an NMD-exon in CACNA1A has been identified and validated in both mouse and human models. By including alternative exon transcripts, the process induces premature stop codons leading to mRNA decay. This splicing redirection’s promising results suggest the potential to control gene expression precisely, catering to both upregulation and downregulation needs. Notably, the modification hinges on already FDA-approved SSO chemistry, ensuring feasibility and a streamlined path for therapeutic development against CACNA1A-associated diseases.

ADVANTAGES

ADVANTAGES

• First time regulating CACNA1A mRNA and protein expressions by redirecting alternative splicing

• Splice-switching oligonucleotides (SSOs) as a viable method for regulating gene expression and treating neurological disorders

• Potential for developing novel precision medicines to treat CACNA1A-associated diseases like EA2, DEE42, FHM1, and SCA6

• ASOs that suppress the NMD exon can treat loss-of-function CACNA1A mutations (e.g., EA2, DEE42, ASD)

• ASOs that promote NMD exon inclusion can treat gain-of-function CACNA1A mutations (e.g., FHM1, seizures, SCA6)

APPLICATIONS

• Genetic disease treatments

• Gene expression regulation

• RNA-targeted drugs

• Neurological disorders treatments