Anti-sense Oligonucleotides Modulating NOTCH3 Expression for CADASIL

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Solution Overview

Problem

CADASIL, caused by NOTCH3 gene mutations leading to misfolding and accumulation of the NOTCH3 protein, results in vascular smooth muscle cell degeneration and impaired cerebral blood flow, with current understanding of pathogenesis being poorly understood and lacking effective treatments.

Innovation Solution

The method involves using anti-sense oligonucleotides to reduce or modify NOTCH3 mRNA expression by excluding exon 4 and 5, thereby restoring an even number of cysteine residues in the EGFL-repeats, reducing the toxic effect of mutant NOTCH3 protein and delaying its accumulation on vascular smooth muscle cells.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If mutant NOTCH3 protein is expressed, then vascular smooth muscle cell degeneration occurs, but the toxic effect of the protein cannot be eliminated

Engineering Contradiction:

Improvecell survivalVSAvoidtoxic effect of mutant NOTCH3

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent extracts and removes the harmful exon 4 from the NOTCH3 mRNA transcript using anti-sense oligonucleotides. This selective removal of the mutated exon eliminates the toxic effect of the mutant protein while preserving the rest of the functional protein sequence, thereby resolving the contradiction between maintaining cell survival and eliminating harmful protein effects.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the molecular parameter of the mRNA transcript by altering its structure through exon skipping. By modifying which exons are included in the final mRNA product, the patent transforms the harmful mutant protein into a functional protein, thereby changing the protein's properties from toxic to beneficial while maintaining cell viability.

Inventive Principle:

Principle #35Parameter changes

2Quantity of substance

If NOTCH3 mRNA expression is reduced, then accumulation of NOTCH3 ectodomain is decreased, but the protein function may be compromised

Engineering Contradiction:

ImproveNOTCH3 ectodomain accumulationVSAvoidprotein function

Core Design Contradiction:

Quantity of substanceVSReliability

Solution Approach 1:

The patent segments the NOTCH3 mRNA transcript into functional and non-functional portions. By selectively skipping exon 4, the patent separates the harmful mutated region from the functional regions, allowing the protein to maintain its essential functions while reducing the accumulation of toxic ectodomain material.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent converts the harmful effect of reduced NOTCH3 expression into a benefit by using exon skipping to remove only the toxic portion while preserving function. The reduction in total protein expression is reframed as a selective removal of harmful elements, transforming a potential detriment into a therapeutic advantage.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

3Stability of the object's composition

If anti-sense oligonucleotides are used to exclude exon 4 and 5, then cysteine residue balance is restored, but the complexity of treatment increases

Engineering Contradiction:

Improvecysteine residue balance in EGFL-repeatsVSAvoidtreatment complexity

Core Design Contradiction:

Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The patent designs anti-sense oligonucleotides that can target multiple mutations within exon 4 and 5 simultaneously. These oligonucleotides serve multiple functions: they skip the harmful exons, restore cysteine balance, and can be applied to different mutation types, thereby reducing treatment complexity despite the sophisticated mechanism.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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

This approach decreases the accumulation of NOTCH3 ectodomain and reduces vascular smooth muscle cell degeneration, potentially alleviating symptoms of CADASIL by producing a modified NOTCH3 protein with even cysteine residues, thereby reducing the risk of ischemic stroke and dementia.

Implementation Method 1

providing a cell with an anti-sense oligonucleotide that is complementary to and capable of specifically hybridizing to a specific sequence of a polymorphic site on a pre-mRNA encoding a NOTCH3 protein

Methodology Applied

Scientific EffectHybridization:

Data Source

PatentUS9062311B2Means and methods for modulating NOTCH3 protein expression and/or the coding region of NOTCH3; compositions and use thereof in the treatment of CADASIL

Publication Date: 2015.06.23 ACADEMISCH ZIEKENHUIS LEIDEN (H O D N LUMC)

AI summary

The invention among other provides means and methods for modulating NOTCH3 expression and/or protein coding domain. In one aspect the invention provides a method for at least reducing an elevated level of NOTCH3 protein in a NOTCH3 expressing cell or the immediate vicinity thereof said method comprising providing said cell with an anti-sense oligonucleotide specific for NOTCH3 m RNA or pre-m RNA thereby decreasing production of said NOTCH3 protein or thereby altering the protein coding region in said NOTCH3 m RNA or pre-m RNA.