Split-intein protein ligation: enabling AAV delivery of large genes for clinical translation in genetic cardiac diseases

The National Heart, Lung, and Blood Institute awarded $599,669 to Kinea Bio Inc. on September 1, 2026, under the Cardiovascular Diseases Research program (CFDA 93.837) to develop and validate KNA-123, an innovative gene therapy for Duchenne muscular dystrophy targeting cardiac muscle degeneration. The award is classified as a Project Grant.

Kinea Bio will leverage the split intein-mediated protein ligation for gene therapy (SIMPLI-GT) platform to enable delivery of a larger, more functional midi-dystrophin transgene using dual AAV vectors, addressing size limitations of current gene therapies. The research comprises three specific aims: evaluating cardiac biodistribution, dystrophin expression, and functional rescue following systemic delivery of AAVmyo-midi-dystrophin in dystrophin-deficient rats; demonstrating rescue of dystrophic phenotypes in human iPSC-derived DMD cardiomyocytes; and developing and qualifying a functional potency assay using 3D engineered heart tissue from DMD patients. The work integrates molecular tools and preclinical models to advance therapeutic strategies for genetic disorders involving large genes.

The recipient is Kinea Bio Inc., located in Seattle, Washington. Performance extends through August 31, 2027.

Abstract

/summary duchenne muscular dystrophy (DMD) is a severe genetic disorder caused by mutations in the DMD gene, leading to progressive skeletal and cardiac muscle degeneration. While advancements in gene therapy have shown promise, existing approaches like micro-dystrophins provide incomplete restoration of function, particularly in cardiac muscle. This project aims to develop and validate KNA-123, an innovative gene therapy leveraging the split intein-mediated protein ligation for gene therapy (SIMPLI-GT) platform. This approach enables the delivery of a larger, more functional midi-dystrophin transgene using dual AAV vectors, overcoming the size limitations of current gene therapies. The research will focus on three specific aims: (1) evaluate cardiac biodistribution, dystrophin expression, and functional rescue following systemic delivery of aavmyo-midi-dystrophin in dystrophin-deficient rats (2) demonstrate rescue of dystrophic phenotypes in human ipsc-derived DMD cardiomyocytes.; and (3) develop and qualify a functional potency assay using 3D engineered heart tissue from DMD patients. This study integrates cutting-edge molecular tools and physiologically relevant preclinical models to advance therapeutic strategies for DMD. By addressing critical gaps in current treatments, this project seeks to improve cardiac outcomes, enhance patient quality of life, and establish a roadmap for tackling genetic disorders involving large genes.