GENE THERAPY ENGINEERING STRATEGIES FOR LONG-TERM CORRECTION OF HEMOPHILIA-ASSOCIATED MUTATIONS

Abstract

Background: Hemophilia is an inherited bleeding disorder, predominantly caused by mutations in the F8 and F9 genes that lead to deficiencies of coagulation factors VIII and IX. Standard therapies replace the faulty genes on a temporary basis, relieving symptoms but not permanently correcting the genetic defect.

Objective: The objective of this study was to evaluate gene therapy engineering strategies for long-term correction of hemophilia-associated mutations by advanced genome engineering and viral delivery systems.
Methodology: CRISPR-Cas9 genome editing, adeno-associated viral (AAV) vectors, lentiviral delivery systems, and base-editing technologies were used for experimental and computational analyses of pathogenic F8 and F9 mutations. The therapeutic efficiency was assessed by the levels of factor expression, the rates of mutation correction, the restoration of clotting activity and the analysis of immune response.

Results: Gene therapy approaches showed up to 78% mutation correction efficiency, and 65% increase of the clotting factor expression in engineered cellular models . CRISPR editing enhanced coagulation activity and markedly reduced bleeding-associated phenotypes. AAV-based delivery systems also exhibited increased stability of the transgene and extended therapeutic persistence.

Conclusions: Engineering strategies for gene therapy provide promising long-term therapeutic approaches to correct hemophilia-associated mutations. Precision genome editing, improved delivery technologies and personalized medicine could greatly enhance the future treatment of hemophilia.