Mitochondrial Transfer by Photothermal Nanoblade Restores Metabolite Profile in Mammalian Cells

• Published in: Cell metabolism Vol. 23; no. 5; pp. 921 - 929
• Main Authors: Wu, Ting-Hsiang, Sagullo, Enrico, Case, Dana, Zheng, Xin, Li, Yanjing, Hong, Jason S., TeSlaa, Tara, Patananan, Alexander N., McCaffery, J. Michael, Niazi, Kayvan, Braas, Daniel, Koehler, Carla M., Graeber, Thomas G., Chiou, Pei-Yu, Teitell, Michael A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 10.05.2016
• Subjects: Animals; Base Sequence; Cell Line, Tumor; Clone Cells; DNA, Mitochondrial - genetics; Energy Metabolism; Gene Expression Regulation; Humans; Light; Mammals - metabolism; Metabolome - genetics; Metabolomics; Mitochondria - metabolism; Nanoparticles - chemistry; Reproducibility of Results; Temperature
• ISSN: 1550-4131; 1932-7420
• DOI: 10.1016/j.cmet.2016.04.007

mtDNA sequence alterations are challenging to generate but desirable for basic studies and potential correction of mtDNA diseases. Here, we report a new method for transferring isolated mitochondria into somatic mammalian cells using a photothermal nanoblade, which bypasses endocytosis and cell fusion. The nanoblade rescued the pyrimidine auxotroph phenotype and respiration of ρ0 cells that lack mtDNA. Three stable isogenic nanoblade-rescued clones grown in uridine-free medium showed distinct bioenergetics profiles. Rescue lines 1 and 3 reestablished nucleus-encoded anapleurotic and catapleurotic enzyme gene expression patterns and had metabolite profiles similar to the parent cells from which the ρ0 recipient cells were derived. By contrast, rescue line 2 retained a ρ0 cell metabolic phenotype despite growth in uridine-free selection. The known influence of metabolite levels on cellular processes, including epigenome modifications and gene expression, suggests metabolite profiling can help assess the quality and function of mtDNA-modified cells. [Display omitted] •Proof-of-principle photothermal nanoblade transfer of mitochondria is reported•Transfer into 143BTK− ρ0 cells generated rescue clones with recovered respiration•Mitochondrial transfer reset metabolic enzyme gene expression patterns•Two of three rescue clones showed metabolite profiles similar to 143BTK− parent cells Optimizing mtDNA transfer into mammalian cells is an important step for basic studies and mitochondrial disease therapies. Using a photothermal nanoblade, Wu et al. are able to deliver isolated mitochondria into respiration-deficient cells. Rescued cell lines recover mitochondrial respiration and reset cellular metabolism to the parental cell level.