Metabolic effects of the HIV protease inhibitor – saquinavir in differentiating human preadipocytes

Bociąga‐Jasik, Monika; Polus, Anna; Góralska, Joanna; Czech, Urszula; Gruca, Anna; Śliwa, Agnieszka; Garlicki, Aleksander; Mach, Tomasz; Dembińska-Kieć, A.

doi:10.1016/s1734-1140(13)71075-2

Cited by 16 publications

(13 citation statements)

References 60 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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“…PIs are toxic via different pathways, in particular by inducing mitochondrial dysfunction and oxidative stress and by promoting inflammation (Gougeon et al 2004;Caron et al 2007;Caron-Debarle et al 2010;Mencarelli et al 2012;Bociaga-Jasik et al 2013;Cassol et al 2013;Reyskens et al 2013). Several studies also reported that some PIs are able to induce ER stress in different types of cells (e.g., macrophages, adipocytes, hepatocytes) and tissues (e.g., liver, intestine) (Zhou et al 2005;Gupta et al 2007;Cao et al 2010;Touzet and Philips 2010;Wu et al 2010;Zha et al 2010Zha et al , 2013Br€ uning 2011;Zhou 2011;Br€ uning et al 2012;Apostolova et al 2013;Taura et al 2013;Wang et al 2013b).…”

Section: Protease Inhibitorsmentioning

confidence: 99%

Role of endoplasmic reticulum stress in drug‐induced toxicity

Foufelle

Fromenty

2016

Pharmacology Res & Perspec

194

161

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Drug‐induced toxicity is a key issue for public health because some side effects can be severe and life‐threatening. These adverse effects can also be a major concern for the pharmaceutical companies since significant toxicity can lead to the interruption of clinical trials, or the withdrawal of the incriminated drugs from the market. Recent studies suggested that endoplasmic reticulum (ER) stress could be an important event involved in drug liability, in addition to other key mechanisms such as mitochondrial dysfunction and oxidative stress. Indeed, drug‐induced ER stress could lead to several deleterious effects within cells and tissues including accumulation of lipids, cell death, cytolysis, and inflammation. After recalling important information regarding drug‐induced adverse reactions and ER stress in diverse pathophysiological situations, this review summarizes the main data pertaining to drug‐induced ER stress and its potential involvement in different adverse effects. Drugs presented in this review are for instance acetaminophen (APAP), arsenic trioxide and other anticancer drugs, diclofenac, and different antiretroviral compounds. We also included data on tunicamycin (an antibiotic not used in human medicine because of its toxicity) and thapsigargin (a toxic compound of the Mediterranean plant Thapsia garganica) since both molecules are commonly used as prototypical toxins to induce ER stress in cellular and animal models.

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“…Saquinavir and its nitric oxide-derivative have been shown to trigger or foster apoptosis [22][23][24][25][26][27][28][29][30], an effect paralleled by inhibition of AKT-phosphorylation [22], downregulation of NF-κB [24,28,29], proteotoxic stress [22], endoplasmic reticulum stress [25], autophagy [25] and mitochondrial depolarization [31].…”

Section: Introductionmentioning

confidence: 99%

Saquinavir Induced Suicidal Death of Human Erythrocytes

Waibel¹,

Bissinger²,

Bouguerra³

et al. 2015

Cell Physiol Biochem

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Background/Aims: The antiretroviral protease inhibitor saquinavir is used for the treatment of HIV infections. Effects of saquinavir include induction of apoptosis, the suicidal death of nucleated cells. Saquinavir treatment may further lead to anemia. In theory, anemia could result from accelerated erythrocyte loss by enhanced suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Stimulators of eryptosis include Ca²⁺ entry with increase of cytosolic Ca²⁺ activity ([Ca²⁺]_i), oxidative stress with increase of reactive oxygen species (ROS) and ceramide. The present study explored, whether and how saquinavir induces eryptosis. Methods: To this end, flow cytometry was employed to estimate erythrocyte volume from forward scatter, phosphatidylserine exposure at the cell surface from annexin-V-binding, [Ca²⁺]_i from Fluo3-fluorescence, ROS abundance from DCFDA fluorescence and ceramide abundance utilizing specific antibodies. Results: A 48 hours exposure of human erythrocytes to saquinavir significantly decreased forward scatter (≥ 5 µg/ml), significantly increased the percentage of annexin-V-binding cells (≥ 10 µg/ml), significantly increased Fluo3-fluorescence (15 µg/ml), significantly increased DCFDA fluorescence (15 µg/ml), but did not significantly modify ceramide abundance. The effect of saquinavir on annexin-V-binding was significantly blunted, but not abolished by removal of extracellular Ca²⁺. Conclusions: Saquinavir triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect in part due to stimulation of ROS formation and Ca²⁺ entry.

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Metabolic effects of the HIV protease inhibitor – saquinavir in differentiating human preadipocytes

Abstract: The results indicate that SQV affects not only metabolic (mitochondrial) activity of adipocytes, but affects the expression of genes related to differentiation and to a lesser extent to cell apoptosis.

Cited by 16 publications

References 60 publications

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

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

Role of endoplasmic reticulum stress in drug‐induced toxicity

Saquinavir Induced Suicidal Death of Human Erythrocytes

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