HIV-1 protease inhibitors and cytomegalovirus vMIA induce mitochondrial fragmentation without triggering apoptosis

Roumier, Thomas; Szabadkai, György; Am, Simoni; Perfettini, Jean-Luc; Al, Paulau; Castedo, Maria; Métivier, Didier; Badley, Andrew D.; Rizzuto, Rosario; Kroemer, Guido

doi:10.1038/sj.cdd.4401750

Cited by 23 publications

(21 citation statements)

References 36 publications

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“…These findings support earlier evidence in the literature where increased ROS levels or mitochondrial dysfunction were also observed individually upon treatment with type 1 HIV protease inhibitors (Estaquier et al 2002;Matarrese et al 2003;Roumier et al 2006;Chandra et al 2009;Touzet and Philips 2010;Bociąga-Jasik et al 2013;Xiang et al 2015), but it is the first time these parameters were measured simultaneously. The major advantage is that an unambiguous determination of both factors together in space and time allows pinpointing causal relationships.…”

Section: Discussionsupporting

confidence: 91%

“…HIV protease inhibitors (HIV PI) have been shown to induce increased basal ROS levels (Chandra et al 2009;Touzet and Philips 2010), and lowered antioxidant defenses (represented by lowered expression of SOD2 (Xiang et al 2015)). Independently, other reports have linked HIV PI to changed mitochondrial morphofunction (Estaquier et al 2002;Matarrese et al 2003;Roumier et al 2006;Bociąga-Jasik et al 2013;Xiang et al 2015). As a case study, we have used the established workflow to assess the effect of HIV PI on both parameters.…”

Section: Biological Applicationsmentioning

confidence: 99%

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Integrated High-Content Quantification of Intracellular ROS Levels and Mitochondrial Morphofunction

Sieprath

Corne

Willems

et al. 2016

Advances in Anatomy, Embryology and Cell Biology

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Oxidative stress arises from an imbalance between the production of reactive oxygen species (ROS) and their removal by cellular antioxidant systems. Especially under pathological conditions, mitochondria constitute a relevant source of cellular ROS. These organelles harbor the electron transport chain, bringing electrons in close vicinity to molecular oxygen. Although a full understanding is still lacking, intracellular ROS generation and mitochondrial function are also linked to changes in mitochondrial morphology. To study the intricate relationships between the different factors that govern cellular redox balance in living cells, we have developed a high-content microscopy-based strategy for simultaneous quantification of intracellular ROS levels and mitochondrial morphofunction. Here, we summarize the principles of intracellular ROS generation and removal, and we explain the major considerations for performing quantitative microscopy analyses of ROS and mitochondrial morphofunction in living cells. Next, we describe our workflow and finally, we illustrate that a multiparametric readout enables the unambiguous classification of chemically perturbed cells as well as laminopathy patient cells. Principles of intracellular ROS generation and removalReactive oxygen species (ROS) are small, short-lived derivatives of molecular oxygen (O 2 ) of radical and non-radical nature (Halliwell and Gutteridge 2007 • ), nitrate radical (NO 3 • ) and many other nitrogen derivatives. ROS were originally described as molecular constituents of the defense system of phagocytic cells, but it has become clear that besides their damaging properties, they also function as signaling molecules and mediate a variety of other cellular responses including cell proliferation, differentiation, gene expression and migration (Lambeth 2004;Bartz and Piantadosi 2010). Intracellular ROS metabolismROS can be generated at various sites in the cell (Fig. 1A). This can be either deliberately, e.g. by NADPH oxidases (NOX), or as a byproduct, e.g. during normal cellular respiration in mitochondria (Babior 1999;Turrens 2003;Murphy 2009). The NOX family of NADPH oxidases (NOX1, NOX2, NOX3, NOX4, NOX5, DUOX1 and DUOX2) are proteins that transport electrons (e -) from NADPH across biological membranes (plasma or endomembranes) (Bedard and Krause 2007;Dupre-Crochet et al. 2013). The activation mechanisms and tissue distribution of the isoforms differ but they all use O 2 as e --acceptor, producing O 2•-. Through ROS generation, they play a role in many cellular processes including host defense, regulation of gene expression and cell differentiation (Bedard and Krause 2007). Despite their sometimes significant contribution to the global ROS pools, NOX are not the predominant source of intracellular ROS. Mitochondria are considered the major culprit, in particular under pathological conditions. Mitochondrial ROS are generated as a byproduct of the oxidative phosphorylation (OXPHOS, cf. below). Irrespective of its source, ROS production generally sta...

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Section: Discussionsupporting

confidence: 91%

Section: Biological Applicationsmentioning

confidence: 99%

Integrated High-Content Quantification of Intracellular ROS Levels and Mitochondrial Morphofunction

Sieprath

Corne

Willems

et al. 2016

Advances in Anatomy, Embryology and Cell Biology

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show abstract

“…It is also possible to reversibly fragment mitochondria with uncouplers or other drugs, showing that fragmentation need not cause apoptosis. 24,25 Rather than the morphology of mitochondria per se, it appears that what is important is the mitochondrial recruitment, or the nonrecruitment, of proteins involved in the fission and fusion of the organelle during cell death. So rather than looking at the morphology of Figure 1 Mitochondrial fission together with cristae remodelling contribute to the dynamics of cytochrome c release.…”

Section: When Does Mitochondria Fission Occur?mentioning

confidence: 99%

Which came first, the cytochrome c release or the mitochondrial fission?

2006

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“…These data underline the notion that mitochondrial fragmentation and initiation of the mitochondrial pathway of apoptosis do not correlate in an obligatory fashion. 13,15 This holds true also at a molecular level, as recently demonstrated for the protein hFis1. 16 Notably, hFis1 (that normally promotes both apoptosis and mitochondrial fragmentation) loses selectively its lethal effects (not the ability to fragment mitochondria) in cells lacking the proapoptotic modulators Bax and Bak, or upon different mutations in the intermembrane region.…”

Section: Macrofragm Normalmentioning

confidence: 56%

“…8 We have recently reported that, similar to Bax, vMIA can stimulate the fragmentation of the mitochondrial network. 13 This is intriguing and counterintuitive because vMIA is a strong inhibitor of apoptosis, 9 while mitochondrial fragmentation is often associated with apoptosis induction. 14 Since vMIA-mediated mitochondrial fragmentation occurs even in Bax-depleted cells, 8 this effect must be independent of the vMIA-mediated recruitment of Bax to the outer mitochondrial membrane.…”

mentioning

confidence: 99%