Products and methods for inhibition of expression of dynamin-1 variants
Inventors
HARPER, Scott Quenton • Frankel, Wayne N.
Assignees
Columbia University in the City of New York • Nationwide Childrens Hospital Inc
Publication Number
US-12275941-B2
Publication Date
2025-04-15
Expiration Date
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
RNA interference-based methods and products for inhibiting the expression of pathogenic dynamin-1 variants are provided. Delivery vehicles such as recombinant adeno-associated viruses deliver DNAs encoding RNAs that inhibit the expression of the dynamin-1 variants. The methods treat, for example, developmental and epileptic encephalopathies.
Core Innovation
RNA interference-based methods and products for inhibiting the expression of pathogenic dynamin-1 variants are provided. Delivery vehicles such as recombinant adeno-associated viruses deliver DNAs encoding RNAs that inhibit the expression of the dynamin-1 variants. The artificial DNM1 inhibitory RNAs contemplated include, but are not limited to, small interfering RNAs (siRNAs), short hairpin RNAs (shRNAs) and miRNA shuttles (artificial miRNAs) referred to as miDNM1s which target, for example, a coding region or 3′UTR of DNM1 mRNA in a reverse complementary manner resulting in reduced DNM1 mRNA and protein levels and use of the methods and products is indicated, for example, in preventing or treating developmental and epileptic encephalopathies.
DNM1 inhibitory RNAs are provided as well as polynucleotides encoding one or more of the RNAs, and delivery of DNA encoding miDNM1s can be achieved using delivery vehicles including recombinant AAV (rAAV) vectors that lack AAV rep and cap genes, self-complementary AAV (scAAV), other viral vectors, or non-viral vectors such as lipid nanoparticles. The disclosure provides nucleic acids comprising RNA-encoding template DNA sequences and exemplary miDNM1s (full length sequences set out in SEQ ID NOs: 18-34 and processed antisense guide strand sequences set out in SEQ ID NOs: 35-51) and compositions, viral vectors, methods of delivery, and methods of preventing or treating DEE by administering delivery vehicles comprising DNA encoding one or more miDNM1s.
The background identifies that DNM1 encodes dynamin-1 and that individuals with pathogenic DNM1 variants suffer from severe developmental and epileptic encephalopathy syndromes including Lennox-Gastaut Syndrome and Infantile Spasms, with early-onset seizures, global developmental delay, profound intellectual disability and other comorbidities, and that treatment of DEEs is currently limited to treatment of symptoms by antiepileptic drugs that do not address the underlying genetic defect. The disclosure states there exists a need in the art for products and methods for treatment of DEEs and provides RNAi-based approaches and delivery vehicles to specifically induce silencing of deleterious DNM1 isoforms to address that need.
Claims Coverage
The independent inventive features extracted from the claims include three core nucleic acid-related features disclosed in claim 1.
Polynucleotide sequence with specified identity to DNM1 artificial inhibitory RNA-encoding sequences (SEQ ID NOs: 1-17)
A polynucleotide sequence comprising at least 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the dynamin-1 (DNM1) artificial inhibitory RNA-encoding polynucleotide sequence set forth in any one of SEQ ID NOs: 1-17.
Polynucleotide encoding a DNM1 artificial inhibitory RNA (SEQ ID NOs: 18-34)
A polynucleotide sequence encoding a DNM1 artificial inhibitory RNA comprising the artificial inhibitory RNA polynucleotide sequence set forth in any one of SEQ ID NOs: 18-34.
Polynucleotide encoding a mature DNM1 antisense guide strand (SEQ ID NOs: 35-51)
A polynucleotide sequence encoding a mature DNM1 antisense guide strand comprising the polynucleotide sequence set forth in any one of SEQ ID NOs: 35-51.
The claims center on nucleic acids encoding DNM1-targeting artificial inhibitory RNAs defined by sequence identity to specified SEQ ID NOs (1-17, 18-34, 35-51) and further encompass vectors, compositions, delivery methods and administration of those nucleic acids.
Stated Advantages
Inhibition of the expression of pathogenic dynamin-1 variants resulting in reduced DNM1 mRNA and protein levels.
Use of the methods and products is indicated in preventing or treating developmental and epileptic encephalopathies (DEE).
Restoration of DNM1 expression to at least 25% up to 100% or more of normal DNM1 expression in an unaffected subject as described.
Improved physiological and behavioral outcomes demonstrated in examples, including increased survival, increased growth, decreased seizures, improved motor function and grip strength, and diminished gliosis and cellular degeneration.
Sustained gene silencing with vector-expressed artificial inhibitory RNAs enabling long term gene silencing with a single administration while the vector is present and the promoter is active.
Documented Applications
Preventing or treating developmental and epileptic encephalopathies (DEE), including Lennox-Gastaut Syndrome and Infantile Spasms.
Delivering DNA encoding miDNM1s to a subject in need thereof using delivery vehicles such as recombinant AAV (rAAV) or other viral or non-viral vectors for administration to neurons with duplicated and/or mutant DNM1 genes.
Use of at least one nucleic acid, at least one viral vector, or a composition as described in making a medicament for, or in treating a subject suffering from, a pathogenic DNM1 gene variant or in treating DEE in a subject in need thereof.
Prophylactic administration in families known to carry pathological DNM1 gene variants and treatment of subjects at risk for DEE due to a mutation of the DNM1 gene.
Interested in licensing this patent?