Year: 2020
Type: article
Abstract: The permanent transfer of specific mtDNA sequences into mammalian cells could generate improved models of mtDNA disease and support future cell-based therapies. Previous studies documented multiple biochemical changes in recipient cells shortly after mtDNA transfer, but the long-term retention and function of transferred mtDNA remains unknown. Here, we evaluate mtDNA retention in new host cells using 'MitoPunch', a device that transfers isolated mitochondria into mouse and human cells. We show that newly introduced mtDNA is stably retained in mtDNA-deficient (ρ0) recipient cells following uridine-free selection, although exogenous mtDNA is lost from metabolically impaired, mtDNA-intact (ρ+) cells. We then introduced a second selective pressure by transferring chloramphenicol-resistant mitochondria into chloramphenicol-sensitive, metabolically impaired ρ+ mouse cybrid cells. Following double selection, recipient cells with mismatched nuclear (nDNA) and mitochondrial (mtDNA) genomes retained transferred mtDNA, which replaced the endogenous mutant mtDNA and improved cell respiration. However, recipient cells with matched mtDNA-nDNA failed to retain transferred mtDNA and sustained impaired respiration. Our results suggest that exogenous mtDNA retention in metabolically impaired ρ+ recipients depends on the degree of recipient mtDNA-nDNA co-evolution. Uncovering factors that stabilize exogenous mtDNA integration will improve our understanding of in vivo mitochondrial transfer and the interplay between mitochondrial and nuclear genomes.
Source: Scientific Reports
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,870
Awards: G1523888516, (PQD5) Mass Profiling Melanoma Responses to Improve Therapy Choices and Prognosis, G1820511087, RNA trafficking in mitochondria, Enhancing the Efficiency of Non-REM Sleep Temporal Dynamics to Improve Insight Learning, SPOTs: Optical Technologies for Instantly Quantifying Multicellular Response Profiles, Reverse Mitochondrial Genetics Enabled by Blast, Exogenous mtDNA integration in cardiomyocytes, G6318647045, MARC U*STAR Program at the University of California, Los Angeles, Research Tranining in Cellular and Molecular Biology, UCLA Maximizing Student Development (MSD) Program, (PQ5) Mitochondrial Transfer in Tumor Heterogeneity and Acquired Resistence to Therapy, UCLA Tumor Immunology Institutional Training Grant, Cancer Center Support Grant
OpenAlex ID: W3082823618