Myosin Heavy Chain Base Editing for MYH7 R403Q Correction
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current treatments for cardiomyopathies, such as hypertrophic cardiomyopathy (HCM), are limited to heart transplants and there is a need for effective genetic mutation correction to address the underlying causes of these heart muscle diseases.
Innovation Solution
The use of guide RNAs (gRNAs) with CRISPR-Cas9 systems and fusion proteins comprising deaminases and Cas9 nickases or deactivated endonucleases for base-pair editing to correct genetic mutations in the MYH7 gene, specifically targeting the R403Q mutation, is employed to reverse cardiomyopathy phenotypes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If heart transplant is used to treat cardiomyopathy, then patient survival is improved, but the treatment is limited and does not address the underlying genetic cause
Solution Approach 1:
The patent replaces the mechanical intervention of heart transplant with a molecular editing approach using base editors. The base editor system (comprising deaminase enzyme and guide RNA) directly corrects the genetic mutation in the MYH7 gene, substituting the need for organ replacement with genetic correction, thereby addressing both survival and underlying cause.
Solution Approach 2:
The patent introduces base editing as an intermediary mechanism between the genetic mutation and the disease phenotype. The base editor acts as a mediator that converts the mutant DNA sequence back to the wild-type sequence, enabling correction of the underlying cause without requiring transplant.
2Reliability
If base editing is used to correct genetic mutations, then the underlying cause of cardiomyopathy is addressed, but the complexity of the treatment system increases
Solution Approach 1:
The patent merges the deaminase enzyme and guide RNA into a single base editing system that can be delivered together via viral vectors. This integration simplifies the treatment approach by combining multiple functional elements into a unified system, reducing the complexity of coordinating separate interventions.
Solution Approach 2:
The patent utilizes changes in the chemical state of DNA bases through deamination to achieve editing. By converting cytosine to uracil (or adenine to inosine) through enzymatic action, the system creates a chemical parameter change that leads to permanent genetic correction, simplifying the overall mechanism compared to more complex editing approaches.
3Reliability
If base editing corrects the MYH7 gene mutation, then cardiomyopathy phenotype is reversed, but off-target effects may occur
Solution Approach 1:
The patent employs guide RNA sequences that are specifically designed to target only the mutant MYH7 gene sequence. The guide RNA binds with high specificity to the unique sequence context surrounding the mutation, ensuring that the base editing activity is localized exclusively to the intended target site, thereby minimizing off-target effects while achieving phenotype reversal.
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 effectively corrects genetic mutations in the MYH7 gene, improving heart function and potentially extending lifespan and reducing heart failure symptoms in cardiomyopathy models.
Implementation Method 1
fusion proteins comprising a deaminase and an Cas9 nickase or deactivated Cas9 endonuclease for base-pair editing
Implementation Method 2
guide RNAs (gRNAs) for use with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associate protein 9 (Cas9) systems that successfully reverse phenotypes associated with familial cardiomyopathies
Data Source
AI summary
Disclosures herein are directed to compositions comprising single guide RNA (sgRNA) and fusion proteins comprising a Cas9 nickase and deaminase designed for a CRISPR-Cas9 system and method of using thereof for preventing, ameliorating or treating one or more cardiomyopathies.