The drug-elicitable alternative splicing module for tunable vector expression in the heart.

Area of Science:

• Biotechnology
• Gene Therapy
• Molecular Biology

Background:

• Adeno-associated viruses (AAVs) are crucial for gene therapy but lack methods for fine-tuning expression, limiting clinical applications.
• Current gene therapy approaches face challenges in controlling therapeutic protein levels, impacting safety and efficacy.

Purpose of the Study:

• To develop a novel, drug-inducible system for precise control of AAV-mediated gene expression.
• To enhance the safety, efficacy, and therapeutic scope of AAV gene therapy.

Main Methods:

• Developed the drug-elicitable alternative splicing module (DreAM), responsive to risdiplam.
• Incorporated DreAM into an AAV9 vector targeting cardiomyocytes (AAV9-Tnnt2-miR122TS) for YAP5SA delivery.
• Evaluated DreAM's dose-dependent, temporal, reversible, and repeatable control of gene expression in vivo.

Main Results:

• Risdiplam activated DreAM-regulated AAV expression with a 2,000-fold induction, demonstrating dose-dependency and organ specificity.
• DreAM allowed transient, reversible, and repeatable gene expression control with 2-day temporal resolution.
• Successful cardiac regeneration in a myocardial infarction model via transient YAP5SA activation, with reduced mortality and off-target effects.

Conclusions:

• DreAM offers a clinically relevant method to modulate AAV expression, improving gene therapy control.
• This system enhances therapeutic efficacy and safety by enabling precise temporal and spatial control of gene delivery.
• DreAM holds significant potential for advancing gene therapy applications, particularly in regenerative medicine.