Creation of cybrid cultures containing the mitochondrial genome mutation m.1555A>G (MT-RNR1 gene), which has a protective effect in atherosclerosis

Abstract

Introduction. Cybrid cell models are one of the best objects for studying pathological processes in the human body, and they are of increasing interest to scientists worldwide. Our laboratory was the first to create such models for studying the protective effect of mutations in the mitochondrial genome that protect the human body from mitochondrial dysfunction and atherosclerotic lesions. Aim: To create cybrid cultures with a high heteroplasmy level for the mitochondrial genome mutation m.1555A>G localized within the coding region of the human mitochondrial genome in the MT-RNR1 gene. Preliminary studies showed that the threshold heteroplasmy level for the m.1555A>G mutation has a protective effect in atherosclerosis. Methods. Cybrid cultures were created by fusion of rho0 (mtDNA-depleted) cells and mitochondria from platelets with a high heteroplasmy level for the studied mutations. To obtain mtDNA-free cells, a culture of monocytic origin, THP-1, was used. Results. We obtained four cybrid cell lines containing the m.1555A>G mutation with a heteroplasmy level above the threshold value. Conclusion. Four cybrid cultures with a high heteroplasmy level for the mtDNA mutation m.1555A>G were created. These cybrid cell lines can serve as models for developing methods of gene therapy for patients with atherosclerosis. In addition, using these cybrid cell models, it will be possible to study molecular and cellular mechanisms that protect cells from mitochondrial dysfunction.