Mitochondrial Transfer by Photothermal Nanoblade Restores Metabolite Profile in Mammalian Cells

Abstract

SUMMARY 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, suggest metabolite profiling can help assess the quality and function of mtDNA modified cells.

Horizontal transfer of whole mitochondria restores tumorigenic potential in mitochondrial DNA-deficient cancer cells

“…Our findings are consistent with the emerging notion of the essential role of respiration in cancer cell proliferation and tumor progression (LeBleu et al, 2014; Viale et al, 2014; Birsoy et al, 2015; Sullivan et al, 2015; Cardaci et al, 2015; Lussey-Lepoutre et al, 2015; Berridge et al, 2015; Viale et al, 2015), and of a role for mitochondrial transfer in maintaining the bioenergetics balance (Sinha et al, 2016; Wu et al, 2016). From translational angle, recent studies on horizontal mitochondrial transfer indicate two tantalising, novel approaches to cancer therapy: targeting mitochondrial respiration and blocking transfer of mitochondria from stromal cells to cancer cells.…”

Transit and integration of extracellular mitochondria in human heart cells

“…Our earlier experiments, and those of others, established that endocytosis of extracellular mitochondria was dependent on actin polymerization and that internalisation of these organelles resulted in long-term replenishment of mtDNA in HeLa cell-derived ρ 0 cells, which are otherwise devoid of mtDNA 9,11,14,29–31 . Restoration of the ρ 0 mitochondrial genome through uptake of mitochondria isolated from HeLa cells led to enhancement of intracellular ATP levels and oxygen consumption rates 9 .…”

Defining the momiome: Promiscuous information transfer by mobile mitochondria and the mitochondrial genome

“…These approaches include peptide-mediated mitochondrial delivery [332], cell-cell fusion [335], co-culture [236], microinjection [336], photothermal nanoblade [337], and import of mitochondria via liposome-modified RNA import complex [338]. It is now reasonable to assume that these methods can be used as tools to further examine the benefits of internalization of healthy mitochondria by diseased or damaged cells, to understand the impact that intercellular transfer of mitochondria or mtDNA might have on healthy cells, and to assess the practical applicability of intercellular transfer of mitochondria or mtDNA in the treatment and prevention of diseases involving heritable or acquired defects in mitochondria.…”