Live-Imaging Readouts and Cell Models for Phenotypic Profiling of Mitochondrial Function

Iannetti, Eligio F.; Prigione, Alessandro; Smeitink, Jan A.M.; Koopman, Werner J.H.; Renkema, G. Herma

doi:10.3389/fgene.2019.00131

Cited by 16 publications

(15 citation statements)

References 103 publications

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“…To test this idea, we measured the mitochondrial inner membrane potential, generated by the ETC complexes. Mitochondria membrane potential is used for the production of ATP being therefore an indicator of mitochondrial respiration (reviewed in Iannetti et al, 2019 ). Mitochondrial inner membrane potential was measured as the ratio of TMRM (an established indicator of mitochondrial membrane potential) to MitoTracker Deep Red (a marker of mitochondrial mass) (Zhang et al, 2019 ).…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial Dynamics in the Drosophila Ovary Regulates Germ Stem Cell Number, Cell Fate, and Female Fertility

Garcez

Branco-Santos

Grácio

et al. 2021

Front. Cell Dev. Biol.

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The fate and proliferative capacity of stem cells have been shown to strongly depend on their metabolic state. Mitochondria are the powerhouses of the cell being responsible for energy production via oxidative phosphorylation (OxPhos) as well as for several other metabolic pathways. Mitochondrial activity strongly depends on their structural organization, with their size and shape being regulated by mitochondrial fusion and fission, a process known as mitochondrial dynamics. However, the significance of mitochondrial dynamics in the regulation of stem cell metabolism and fate remains elusive. Here, we characterize the role of mitochondria morphology in female germ stem cells (GSCs) and in their more differentiated lineage. Mitochondria are particularly important in the female GSC lineage. Not only do they provide these cells with their energy requirements to generate the oocyte but they are also the only mitochondria pool to be inherited by the offspring. We show that the undifferentiated GSCs predominantly have fissed mitochondria, whereas more differentiated germ cells have more fused mitochondria. By reducing the levels of mitochondrial dynamics regulators, we show that both fused and fissed mitochondria are required for the maintenance of a stable GSC pool. Surprisingly, we found that disrupting mitochondrial dynamics in the germline also strongly affects nurse cells morphology, impairing egg chamber development and female fertility. Interestingly, reducing the levels of key enzymes in the Tricarboxylic Acid Cycle (TCA), known to cause OxPhos reduction, also affects GSC number. This defect in GSC self-renewal capacity indicates that at least basal levels of TCA/OxPhos are required in GSCs. Our findings show that mitochondrial dynamics is essential for female GSC maintenance and female fertility, and that mitochondria fusion and fission events are dynamically regulated during GSC differentiation, possibly to modulate their metabolic profile.

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Section: Resultsmentioning

confidence: 99%

Mitochondrial Dynamics in the Drosophila Ovary Regulates Germ Stem Cell Number, Cell Fate, and Female Fertility

Garcez

Branco-Santos

Grácio

et al. 2021

Front. Cell Dev. Biol.

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“…Decreased oxidative capacity due to mitochondrial respiratory chain impairment is associated with an increased release of reactive oxygen species, leading to a reduction of calcium retention, and apoptosis [ 32 ]. However, detailing the intricacy of mitochondrial dysfunction is time-consuming and expensive, which may limit its utility in precision diagnosis and treatment [ 33 , 34 ].…”

Section: Discussionmentioning

confidence: 99%

Fibrosis Distinguishes Critical Limb Ischemia Patients from Claudicants in a Transcriptomic and Histologic Analysis

Cong

Cui

Ferrari

et al. 2020

JCM

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Most patients with critical limb ischemia (CLI) from peripheral arterial disease (PAD) do not have antecedent intermittent claudication (IC). We hypothesized that transcriptomic analysis would identify CLI-specific pathways, particularly in regards to fibrosis. Derivation cohort data from muscle biopsies in PAD and non-PAD (controls) was obtained from the Gene Expression Omnibus (GSE120642). Transcriptomic analysis indicated CLI patients (N = 16) had a unique gene expression profile, when compared with non-PAD controls (N = 15) and IC (N = 20). Ninety-eight genes differed between controls and IC, 2489 genes differed between CLI and controls, and 2783 genes differed between CLI and IC patients. Pathway enrichment analysis showed that pathways associated with TGFβ, collagen deposition, and VEGF signaling were enriched in CLI but not IC. Receiver operating curve (ROC) analysis of nine fibrosis core gene expression revealed the areas under the ROC (AUC) were all >0.75 for CLI. Furthermore, the fibrosis area (AUC = 0.81) and % fibrosis (AUC = 0.87) in validation cohort validated the fibrosis discrimination CLI from IC and control (all n = 12). In conclusion, transcriptomic analysis identified fibrosis pathways, including those involving TGFβ, as a novel gene expression feature for CLI but not IC. Fibrosis is an important characteristic of CLI, which we confirmed histologically, and may be a target for novel therapies in PAD.

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“…Mitochondrial respiration is the primary source of cellular ROS and both cell lines under investigation are oxidative. Therefore, we analyzed mitochondrial and cytoplasmatic ROS levels by flow cytometry, and quantitative imaging (Operetta CLS™) on living cells stained with MitoSOX and 2′,7′-dichlorofluorescein diacetate (H2DCFDA), respectively [ 64 ]. MitoSOX is a mitochondrion selective dye which is oxidized by superoxide but not by other reactive oxygen species (ROS) and reactive nitrogen species (RNS), while H2DCFDA is a fluorogenic dye that allow measuring hydroxyl, peroxyl and other ROS species within the cell.…”

Section: Resultsmentioning

confidence: 99%

Profiling and Targeting of Energy and Redox Metabolism in Grade 2 Bladder Cancer Cells with Different Invasiveness Properties

Pasquale

Ducci

Campioni

et al. 2020

Cells

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Bladder cancer is one of the most prevalent deadly diseases worldwide. Grade 2 tumors represent a good window of therapeutic intervention, whose optimization requires high resolution biomarker identification. Here we characterize energy metabolism and cellular properties associated with spreading and tumor progression of RT112 and 5637, two Grade 2 cancer cell lines derived from human bladder, representative of luminal-like and basal-like tumors, respectively. The two cell lines have similar proliferation rates, but only 5637 cells show efficient lateral migration. In contrast, RT112 cells are more prone to form spheroids. RT112 cells produce more ATP by glycolysis and OXPHOS, present overall higher metabolic plasticity and are less sensitive than 5637 to nutritional perturbation of cell proliferation and migration induced by treatment with 2-deoxyglucose and metformin. On the contrary, spheroid formation is less sensitive to metabolic perturbations in 5637 than RT112 cells. The ability of metformin to reduce, although with different efficiency, cell proliferation, sphere formation and migration in both cell lines, suggests that OXPHOS targeting could be an effective strategy to reduce the invasiveness of Grade 2 bladder cancer cells.

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Live-Imaging Readouts and Cell Models for Phenotypic Profiling of Mitochondrial Function

Cited by 16 publications

References 103 publications

Mitochondrial Dynamics in the Drosophila Ovary Regulates Germ Stem Cell Number, Cell Fate, and Female Fertility

Mitochondrial Dynamics in the Drosophila Ovary Regulates Germ Stem Cell Number, Cell Fate, and Female Fertility

Fibrosis Distinguishes Critical Limb Ischemia Patients from Claudicants in a Transcriptomic and Histologic Analysis

Profiling and Targeting of Energy and Redox Metabolism in Grade 2 Bladder Cancer Cells with Different Invasiveness Properties

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