S-adenosylhomocysteine hydrolase (AHCY) deficiency is an ultra-rare metabolic disease discovered in Croatia in 2002, with muscle weakness (myopathy) among its main clinical features. R3forAHCY builds the first patient-cell model of this myopathy and tests whether the causative mutation can be corrected.

We will reprogram patient cells into induced pluripotent stem cells (iPSCs) and differentiate them into muscle cells — the first such model of the disease worldwide. On this model we carry out multi-omics profiling: the first telomere-to-telomere (T2T) long-read genome sequencing in AHCY deficiency, native DNA methylation, RNA sequencing and episignatures. We then correct the pathogenic variant with CRISPR base and prime editing and test whether normal muscle-cell function is restored.

Objectives

• validated patient iPSC-derived myoblasts and isogenic CRISPR-corrected controls
• the first multi-omics dataset for AHCY deficiency-mediated myopathy (T2T long-read sequencing, methylation, RNA-seq)
• correction of the pathogenic variant by genome editing at over 60% efficiency
• functional evidence of phenotypic rescue and a mechanistic model of the disease

Collaboration

Copenhagen Neuromuscular Center, Rigshospitalet, Copenhagen, Denmark

Funded by the Croatian Science Foundation.