3D Quantitative and Ultrastructural Analysis of Mitochondria in a Model of Doxorubicin Sensitive and Resistant Human Colon Carcinoma Cells

Moscheni, Claudia; Malucelli, Emil; Castiglioni, Sara; Procopio, Alessandra; Palma, Clara De; Sorrentino, Andrea; Sartori, Patrizia; Locatelli, Laura; Pereiro, Eva; Maier, Jeanette A. M.; Iotti, Stefano

doi:10.3390/cancers11091254

Cited by 17 publications

(18 citation statements)

References 58 publications

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“…On the other hand, the use of X-ray based imaging can provide information about thick and large area biological samples and under cryogenic conditions to avoid the radiation damage, for instance the modalities of X-ptychography of vegetal tissues 18 and soft X-rays of whole cells 19 . Mitochondria has been already explored by X-ray imaging to be identified in the cellular context of wild-type single cells 19,20 , in cancer cells 21,22 and isolated from the cells aiming to unveil its internal structure 23 . However, there are no reports of X-ray imaging-based analysis focusing on extracting 3D morphological and quantitative information about the OMM and IMM organization.…”

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confidence: 99%

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Polo

Fonseca‐Alaniz

Chen

et al. 2020

Sci Rep

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Mitochondria are dynamic organelles that change morphology to adapt to cellular energetic demands under both physiological and stress conditions. Cardiomyopathies and neuronal disorders are associated with structure-related dysfunction in mitochondria, but three-dimensional characterizations of the organelles are still lacking. In this study, we combined high-resolution imaging and 3D electron density information provided by cryo-soft X-ray tomography to characterize mitochondria cristae morphology isolated from murine. Using the linear attenuation coefficient, the mitochondria were identified (0.247 ± 0.04 µm−1) presenting average dimensions of 0.90 ± 0.20 µm in length and 0.63 ± 0.12 µm in width. The internal mitochondria structure was successfully identified by reaching up the limit of spatial resolution of 35 nm. The internal mitochondrial membranes invagination (cristae) complexity was calculated by the mitochondrial complexity index (MCI) providing quantitative and morphological information of mitochondria larger than 0.90 mm in length. The segmentation to visualize the cristae invaginations into the mitochondrial matrix was possible in mitochondria with MCI ≥ 7. Altogether, we demonstrated that the MCI is a valuable quantitative morphological parameter to evaluate cristae modelling and can be applied to compare healthy and disease state associated to mitochondria morphology.

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confidence: 99%

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Polo

Fonseca‐Alaniz

Chen

et al. 2020

Sci Rep

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“…A bMSC-4D 3D central slice reconstruction, constituted by four stitched tomograms, is shown in Figure 1 a. It displays the nucleus at the center and the surrounding cytoplasm showing vacuoles, a variety of vesicles, cytoskeleton, and other cellular structures commonly visible in TEM resin-embedded thin sections [ 15 ]. The automatic segmentation of all the dense objects, which are abundant in this kind of cell, is reported in Figure 1 b,c for bMSC-4D.…”

Section: Resultsmentioning

confidence: 99%

Calcite as a Precursor of Hydroxyapatite in the Early Biomineralization of Differentiating Human Bone-Marrow Mesenchymal Stem Cells

Sorrentino

Malucelli

Rossi

et al. 2021

IJMS

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Biomineralization is the process by which living organisms generate organized mineral crystals. In human cells, this phenomenon culminates with the formation of hydroxyapatite, which is a naturally occurring mineral form of calcium apatite. The mechanism that explains the genesis within the cell and the propagation of the mineral in the extracellular matrix still remains largely unexplained, and its characterization is highly controversial, especially in humans. In fact, up to now, biomineralization core knowledge has been provided by investigations on the advanced phases of this process. In this study, we characterize the contents of calcium depositions in human bone mesenchymal stem cells exposed to an osteogenic cocktail for 4 and 10 days using synchrotron-based cryo-soft-X-ray tomography and cryo-XANES microscopy. The reported results suggest crystalline calcite as a precursor of hydroxyapatite depositions within the cells in the biomineralization process. In particular, both calcite and hydroxyapatite were detected within the cell during the early phase of osteogenic differentiation. This striking finding may redefine most of the biomineralization models published so far, taking into account that they have been formulated using murine samples while studies in human cell lines are still scarce.

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“…In EM studies, vehicle-treated cells exhibited tubular mitochondria, however individual cristae were hardly discernable, suggestive of deranged mitochondrial respiration 46 (Figure 7C, red arrows). Furthermore, some mitochondria were in various stages of membrane fusion/fission as marked by "tethered" structures indicative of dynamic remodeling (Figure 7C, blue arrow).…”

Section: Vitamin D Modulates Mitochondrial Structure and Dynamics In Mg-63 Cancer Cellsmentioning

confidence: 99%

Vitamin D modulation of mitochondrial oxidative metabolism and mTOR enforces stress tolerance and anti-cancer responses

Quigley

Rieger

Capobianco³

et al. 2021

Preprint

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The relationship between vitamin D and reactive oxygen species (ROS), two integral signaling and damaging molecules of the cell, is poorly understood. This is striking, given that both factors are involved in cancer cell regulation and metabolism. Mitochondria (mt) dysfunction is one of the main drivers of cancer, producing higher cellular energy and ROS that can enhance oxidative stress and stress tolerance responses. To study the effects of vitamin D on metabolic and mt dysfunction, we used the vitamin D receptor (VDR)-sensitive MG-63 osteosarcoma cell model. Using biochemical approaches, active vitamin D (1,25-dihydroxyvitamin D, 1,25(OH)2D3) decreased mt ROS levels, membrane potential (ΔΨmt), biogenesis, and translation, while enforcing endoplasmic reticulum/mitohormetic stress tolerance responses. Using a mitochondria-focused transcriptomic approach, gene set enrichment and pathway analyses show that 1,25(OH)2D3 lowered mt fusion/fission and oxidative phosphorylation (OXPHOS). By contrast, mitophagy, ROS defense, and epigenetic gene regulation were enhanced after 1,25(OH)2D3 treatment, as well as key metabolic enzymes that regulate fluxes of substrates for cellular architecture and a shift toward non-oxidative energy metabolism. ATACseq revealed putative oxi-sensitive and tumor-suppressing transcription factors that may regulate important mt functional genes such as the mTORC1 inhibitor, DDIT4/REDD1. DDIT4/REDD1 was predominantly localized to the outer mt membrane in untreated MG-63 cells yet sequestered in the cytoplasm after 1,25(OH)2D3 and rotenone treatments, suggesting a level of control by membrane depolarization to facilitate its cytoplasmic mTORC1 inhibitory function. The results show that vitamin D activates distinct adaptive metabolic responses involving mitochondria to regain redox balance and control the growth of cancer cells.

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3D Quantitative and Ultrastructural Analysis of Mitochondria in a Model of Doxorubicin Sensitive and Resistant Human Colon Carcinoma Cells

Cited by 17 publications

References 58 publications

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Three-dimensional imaging of mitochondrial cristae complexity using cryo-soft X-ray tomography

Calcite as a Precursor of Hydroxyapatite in the Early Biomineralization of Differentiating Human Bone-Marrow Mesenchymal Stem Cells

Vitamin D modulation of mitochondrial oxidative metabolism and mTOR enforces stress tolerance and anti-cancer responses

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