Development and optimization of human glucocerebrosidase-encoding mRNA for Gaucher disease therapy

Area of Science:

• Biochemistry
• Genetics
• Molecular Biology

Background:

• Gaucher disease (GD) is a rare genetic disorder caused by GBA1 gene mutations, leading to glucosylceramide accumulation.
• Current treatments for GD have limitations, necessitating novel therapeutic strategies.

Purpose of the Study:

• To design and optimize human GBA1 mRNA (hGBA1-mRNA) for enhanced expression and stability.
• To evaluate the in vitro and in vivo efficacy of optimized hGBA1-mRNA as a potential therapy for Gaucher disease.

Main Methods:

• hGBA1-mRNA constructs were engineered by modifying untranslated regions (UTRs), codon usage, and poly(A) tails.
• Performance was assessed in HEK293T and RAW264.7 cells, and in vivo studies were conducted in FVB mice using mRNA-lipid nanoparticles (LNPs).

Main Results:

• Optimized mRNA constructs yielded over 6-fold higher glucocerebrosidase (GCase) activity with an average half-life exceeding 54 hours.
• Functional GCase was delivered in vitro to GBA1-knockout cells, restoring normal morphology and reducing substrate accumulation.
• In vivo studies showed detectable GCase activity in mouse liver and spleen within 72 hours after a single LNP administration.

Conclusions:

• Optimized hGBA1-mRNA-LNPs effectively deliver functional human GCase in vivo.
• This mRNA-based approach represents a promising and efficient therapeutic strategy for Gaucher disease.