Single-nucleus RNA sequencing of OCTN2-defective engineered heart tissues and isogenic control

Primary carnitine deficiency (PCD) is an autosomal recessive disorder caused by mutations in the gene SLC22A5, encoding for the plasmalemmal carnitine transporter OCTN2. PCD patients suffer from muscular weakness and dilated cardiomyopathy (DCM). However, currently available PCD models were unable to distinguish causative from secondary pathomechanisms. To further understand the contribution of cellular subclusters to the PCD disease phenotype we analyzed engineered heart tissues from OCTN2-defective genotype in comparison to isogenic control using single-nucleus RNA sequencing. Two OCTN2-defective human induced pluripotent stem cell lines were generated from a healthy control line, carrying a full OCTN2-knockout and a homozygous OCTN2 (N32S) loss of function mutation. Cardiomyocytes were differentiated and the engineered heart tissue (EHT) technology served as a three-dimensional disease model. EHTs were cultured for 14 days and subsequently analyzed by single-nucleus RNA sequencing. **RAW data not provided due to GDPR regulations of this cell line**