rAAV RNA Interference for Pathogenic DNM1 Variant Silencing

Solution Overview

Problem

Current treatments for developmental and epileptic encephalopathies (DEEs) caused by pathogenic dynamin-1 variants are limited to symptom management with antiepileptic drugs, which do not address the underlying genetic defect and offer no hope of stopping or slowing disease progression.

Innovation Solution

RNA interference-based methods using recombinant adeno-associated viruses (rAAV) to deliver DNAs encoding artificial inhibitory RNAs, such as small interfering RNAs (siRNAs) and microRNAs (miRNAs), specifically targeting and silencing pathogenic DNM1 isoforms to inhibit their expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Object-affected harmful factors

If antiepileptic drugs are used to treat DEEs, then seizure symptoms are managed, but the underlying genetic defect is not addressed and disease progression continues

Engineering Contradiction:

Improveseizure symptomsVSAvoiddisease progression control

Core Design Contradiction:

Object-affected harmful factorsVSReliability

Solution Approach 1:

The invention extracts and removes the harmful pathogenic DNM1 protein from the system using RNA interference technology. The rAAV-delivered shRNA specifically targets and silences the mutant DNM1 gene, extracting the harmful genetic element while preserving normal cellular function.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention introduces an intermediary mechanism (RNA interference pathway) between the genetic defect and its harmful effects. The shRNA acts as a mediator that intercepts the pathogenic DNM1 mRNA before it can be translated into harmful protein, blocking the disease pathway without directly modifying the original gene.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If RNA interference-based methods are used to inhibit pathogenic DNM1 variants, then disease progression is addressed, but treatment complexity increases compared to conventional drug therapy

Engineering Contradiction:

Improvedisease progression controlVSAvoidtreatment method complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The invention employs self-service by utilizing the cell's own RNA interference machinery to carry out therapeutic function. Once the rAAV delivers the shRNA, the cell's natural cellular processes handle the rest—processing the shRNA, loading it into RISC complexes, and executing gene silencing—without requiring external intervention for each therapeutic action.

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The treatment applies preliminary action by delivering the shRNA sequence in advance via rAAV, establishing a persistent silencing mechanism before the pathogenic protein can cause extensive damage. The therapeutic effect is preemptively established at the genetic level, preventing future protein production rather than reacting to symptoms as they occur.

Inventive Principle:

Principle #10Preliminary action

3Ease of operation

If conventional antiepileptic drugs are used, then treatment is simple to administer, but they do not address the underlying genetic defect

Engineering Contradiction:

Improvetreatment administrationVSAvoidgenetic defect correction

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

The invention introduces an intermediary mechanism (RNA interference pathway) between the genetic defect and its harmful effects. The shRNA acts as a mediator that intercepts the pathogenic DNM1 mRNA before it can be translated into harmful protein, blocking the disease pathway without directly modifying the original gene.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention extracts and removes the harmful pathogenic DNM1 protein from the system using RNA interference technology. The rAAV-delivered shRNA specifically targets and silences the mutant DNM1 gene, extracting the harmful genetic element while preserving normal cellular function.

Inventive Principle:

Principle #2Taking out (Extraction)

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 methods significantly reduce DNM1 mRNA and protein levels, leading to improved survival, growth, seizure control, and developmental outcomes in animal models of DEEs, restoring normal DNM1 expression and alleviating disease symptoms.

Implementation Method 1

Delivery vehicles such as recombinant adeno-associated viruses deliver DNAs encoding RNAs that inhibit expression of the dynamin-1 variants

Methodology Applied

Scientific EffectViral vector delivery:

Implementation Method 2

The artificial inhibitory RNAs are referred to as miDNM1s herein. The miDNM1s are small regulatory sequences that act post-transcriptionally by targeting, for example, a coding region or 3′UTR of DNM1 mRNA in a reverse complementary manner resulting in reduced DNM1 mRNA and protein levels

Methodology Applied

Scientific EffectRNA interference:

Data Source

PatentUS20250230443A1Products and methods for inhibition of expression of dynamin-1 variants

Publication Date: 2025.07.17 THE TRUSTEES OF COLUMBIA UNIV IN THE CITY OF NEW YORK

AI summary

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.