DOCSLIB.ORG

Inhibiting Mitochondrial Phosphate Transport As an Unexploited Antifungal Strategy

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Inhibiting Mitochondrial Phosphate Transport As an Unexploited Antifungal Strategy ARTICLE PUBLISHED ONLINE: 11 DECEMBER 2017 | DOI: 10.1038/NCHEMBIO.2534 Inhibiting mitochondrial phosphate transport as an unexploited antifungal strategy Catherine A McLellan1,2,10, Benjamin M Vincent1,3,8,10, Norma V Solis4, Alex K Lancaster1,8 , Lucas B Sullivan5 , Cathy L Hartland6,8, Willmen Youngsaye6,8, Scott G Filler4,7 , Luke Whitesell1,8* & Susan Lindquist1,2,9 The development of effective antifungal therapeutics remains a formidable challenge because of the close evolutionary rela- tionship between humans and fungi. Mitochondrial function may present an exploitable vulnerability because of its differential utilization in fungi and its pivotal roles in fungal morphogenesis, virulence, and drug resistance already demonstrated by others. We now report mechanistic characterization of ML316, a thiohydantoin that kills drug-resistant Candida species at nanomolar concentrations through fungal-selective inhibition of the mitochondrial phosphate carrier Mir1. Using genetic, biochemical, and metabolomic approaches, we established ML316 as the first Mir1 inhibitor. Inhibition of Mir1 by ML316 in respiring yeast diminished mitochondrial oxygen consumption, resulting in an unusual metabolic catastrophe marked by citrate accumulation and death. In a mouse model of azole-resistant oropharyngeal candidiasis, ML316 reduced fungal burden and enhanced azole activity. Targeting Mir1 could provide a new, much-needed therapeutic strategy to address the rapidly rising burden of drug- resistant fungal infection. health crisis from fungal diseases is looming, as there is a mitochondria, where it is used by ATP synthase to generate ATP. marked increase in at-risk populations and a rising preva- Guided by this insight, we used the compound as a probe to inves- A lence of drug resistance. The limited number of antifungal tigate the surprising biological consequences of disrupting fungal classes available and the unacceptably high mortality rate from respiration at its terminal step and to explore the translational ther- invasive infections make the need for new antifungals dire. Fungi apeutic potential of inhibiting Mir1 in culture and in mice. are eukaryotes, and as such share many conserved proteins and pro- cesses with humans, which presents a daunting challenge for the RESULTS development of new, mechanistically distinct antifungals1,2. With ML316 is fungicidal under respiratory conditions this challenge in mind, we executed a high-throughput phenotypic As previously reported5, ML316 (1) exhibited more potent antifungal screen for compounds that could reverse resistance to the widely activity against azole-resistant C. albicans than our original screen used antifungal fluconazole in a clinical isolate of Candida albi- hit (2), whereas substitution of oxygen for sulfur in the thiohy- cans (National Center for Biotechnology Information. PubChem dantoin core (3) greatly diminished activity (Fig. 1a). Although Bioassay Database; AID 2007, https://pubchem.ncbi.nlm.nih. ML316 was equally effective at inhibiting the growth of wild-type gov/bioassay/2007). A particularly intriguing hit to emerge from C. albicans in media with either glucose or nonfermentable glyc- this screen was a thiohydantoin (PubChem CID 3889161), which erol as the sole carbon source, it only inhibited growth of the model exerted potent antifungal activity as a single agent with no apparent fungal organism, Saccharomyces cerevisiae, when it was grown in toxicity to mammalian cells. Substructure searches did not reveal glycerol to enforce respiration (Fig. 1b). C. albicans is a facultative any assay interference alerts3. Its thiohydantoin core is present in aerobe and respires when oxygen is present, regardless of the carbon enzalutamide, an androgen-receptor antagonist used in the treat- source. S. cerevisiae, on the other hand, solely ferments glucose even ment of prostate cancer4. Follow-up synthesis and testing of 51 ana- in the presence of abundant oxygen6. A distantly related Candida logs in a limited SAR study yielded the designated probe compound species, Candida glabrata, also shuts off respiration in the presence ML316, a para-fluoro phenyl derivative with improved potency of glucose. As demonstrated in the probe report, toxicity in this fun- and selectivity5. gal species was similarly dependent on growth in glycerol, confirm- The promising attributes of ML316 encouraged further inves- ing a requirement for respiration in ML316 toxicity and suggesting a tigation, and we now report discovery of its molecular mecha- mode of action targeting some aspect of mitochondrial function. © 2018 Nature America, Inc., part of Springer Nature. All rights reserved. All rights part Nature. of Springer Inc., America, Nature © 2018 nism of action through a combination of genetic and biochemical In addition to effectively inhibiting wild-type Candida growth, approaches. This drug-like compound, distinct from all previously ML316 exhibited potent antifungal activity against the moderately- characterized mitochondrial poisons, selectively inhibits Mir1, azole-resistant C. albicans strain CaCi-2 (ref. 7) as a single agent the major mitochondrial phosphate carrier protein (PiC). Mir1 (MIC 0.05 μg/ml; Fig. 1c). Despite its clinically acquired azole transports inorganic phosphate into the inner matrix of fungal resistance, when CaCi-2 was simultaneously exposed to ML316 1Whitehead Institute for Biomedical Research, Cambridge, Massachusetts, USA. 2Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. 3Microbiology Graduate Program, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. 4Division of Infectious Diseases, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, California, USA. 5Koch Institute for Integrative Cancer Research at Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. 6Chemical Biology Platform and Probe Development Center, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA. 7David Geffen School of Medicine at UCLA, Los Angeles, California, USA. 8Present addresses: Benjamin M. Vincent, Yumanity Therapeutics Cambridge, Massachusetts, USA (B.M.V.); Ronin Institute, Montclair, N.J., USA (A.K.L.); Center for Developmental Therapeutics, Broad Institute, Cambridge, Massachusetts, USA (C.L.H.); Ra Pharmaceuticals, Cambridge, Massachusetts, USA (W.Y.) Department of Molecular Genetics,University of Toronto, Toronto, Ontario, Canada (L.W.). 9Deceased. 10These authors contributed equally to this work. *e-mail: [email protected] NATURE CHEMICAL BIOLOGY | VOL 14 | FEBRUARY 2018 | www.nature.com/naturechemicalbiology 135 ARTICLE NAture CHemiCAl BiOlOGY Doi: 10.1038/NchEmbio.2534 a b S. cerevisiae, Glu C. albicans, Glu strains (Fig. 1d). For these experiments, a series of clinical isolates (1) (2) S. cerevisiae, Gly C. albicans, Gly EtO O EtO cultured from a single HIV-infected patient over a 2-year period O O 7,9 O 1.00 μ N was assessed . ML316 remained very active (MIC 0.5 g/ml) as a N N O N O single agent against all the isolates tested, including the most flu- S S 0.75 F ML316 Screen hit Me conazole-resistant strain in the series, CaCi-17 (fluconazole MIC MIC = 0.06 µg/ml MIC = 0.3 µg/ml 0.50 >128 μg/ml). ML316 retained activity against this strain despite the (3) EtO Relative growth fact that a major contributor to the high-level azole-resistance of O 0.25 O N CaCi-17 is upregulation of both the ATP-binding cassette (ABC) N O 0.00 and multidrug resistance (MDR) efflux pump systems7, which con- O 0.001 0.01 0.1 1 Me Inactive ML316 (µg/ml) fer pleiotropic resistance to many small molecules by limiting their MIC > 7 µg/ml d Fluconazole (mg/ml) intracellular accumulation. To confirm that activity was not limited 10 c 1.00 0 0.1250.25 0.5 1 2 4 8 16 32 64 128 to one patient series, we tested other azole-resistant strains and ML 316 CaCi2 ML 316 + Flu CaCi8 found that they also remained sensitive to ML316 (Supplementary 0.75 CaCi12 CaCi15 Table 1). Four strains of the emerging pathogen Candida auris, a CaCi17 1.0 0.50 0.8 cause of increasing alarm because of its frequent resistance to known ML316 (µg/ml) 0.6 0.4 antifungals and potential for epidemic spread in hospital settings, 11 Relative growth 0.25 0.2 0 0.0160.0310.0620.1250.25 0.5 1 2 4 8 16 0 were also tested , and were all found to be sensitive to ML316 as CaCi2 Relative growth well (Supplementary Table 1). 0.00 CaCi8 0.001 0.01 0.11CaCi12 Because respiration was required for toxicity in fungi, we evalu- ML316 (µg/ml) CaCi15 CaCi17 e 1.00 ated the selectivity of ML316 using human cells grown under con- f ML316 (µg/ml) ditions enforcing respiration. As expected, a classical inhibitor of 0.75 respiration, antimycin A, was highly toxic to both 293T (kidney- 3.5 1.75 0.86 0.44 0.22 0.11 0.05 0 12 0.50 293T ML316 derived) and HepG2 (liver-derived) cells . Under these same con- HepG2 ML316 30 °C Relative growth ditions, however, ML316 had no effect on the growth and survival 0.25 293T antimycin 10 mm HepG2 antimycin of these cells (Fig. 1e). To rule out potential effects of serum on 0.00 0.01 0.1137 °C the bioavailability or stability of ML316, experiments were repeated Concentration (µg/mL) under serum-free growth conditions in which ML316 remained nontoxic to respiring human cells (Supplementary Fig. 2). Figure 1 | ML316 is a potent, highly selective fungicide. (a) Structures of Many widely used antifungals are cytostatic, stopping the ML316, the original screen hit and the inactive analog. Minimal inhibitory growth of yeast but not killing them. To characterize the nature of concentrations (MICs) are indicated for the C. albicans strain CaCi-2. ML316’s antifungal effects, we performed clonogenic survival assays Conversion factor for ML316, 1 μM = 0.35 μg/ml. (b) Glucose-grown using the CaCi-2 strain and a broad range of ML316 concentra- S. cerevisiae are not sensitive to ML316. Standard antifungal susceptibility tions (Fig. 1f). Fungicidal amphotericin and fungistatic fluconazole testing was performed in YNB-CSM media with either glucose (Glu, 2% served as controls (Supplementary Fig.
Load more

Recommended publications
  • Electron Transport Discovery Four Complexes Complex I: Nadhà Coqh2
    BI/CH 422/622 OUTLINE: Pyruvate pyruvate dehydrogenase Krebs’ Cycle How did he figure it out? Overview 8 Steps Citrate Synthase Aconitase Isocitrate dehydrogenase Ketoglutarate dehydrogenase Succinyl-CoA synthetase Succinate dehydrogenase Fumarase Malate dehydrogenase Energetics Regulation Summary Oxidative Phosphorylation Energetics (–0.16 V needed for making ATP) Mitochondria Transport (2.4 kcal/mol needed to transport H+ out) Electron transport Discovery Four Complexes Complex I: NADHà CoQH2 Complex II: Succinateà CoQH2 2+ Complex III: CoQH2à Cytochrome C (Fe ) 2+ Complex IV: Cytochrome C (Fe ) à H2O Electron Transport à O2 Inhibitors of Electron Transport Big Drop! • Inhibitors all stop ET and ATP synthesis: very toxic! Spectral work Big Drop! NADH Cyto-a3 Cyto-c1 Big Drop! Cyto-b Cyto-c Cyto-a Fully reduced Flavin Cyto-c + rotenone + antimycin A 300 350 400 450 500 550 600 650 700 1 Electron Transport Electron-Transport Chain Complexes Contain a Series of Electron Carriers • Better techniques for isolating and handling mitochondria, and isolated various fractions of the inner mitochondrial membrane • Measure E°’ • They corresponded to these large drops, and they contained the redox compounds isolated previously. • When assayed for what reactions they could perform, they could perform certain redox reactions and not others. • When isolated, including isolating the individual redox compounds, and measuring the E°’ for each, it was clear that an electron chain was occurring; like a wire! • Lastly, when certain inhibitors were added, some of the redox reactions could be inhibited and others not. Site of the inhibition could be mapped. Electron Transport Electron-Transport Chain Complexes Contain a Series of Electron Carriers • Better techniques for isolating and handling mitochondria, and isolated various fractions of the inner mitochondrial membrane • Measure E°’ • They corresponded to these large drops, and they contained the redox compounds isolated previously.
    [Show full text]
  • Seahorse XF Cell Mito Stress Test Kit User Guide 3 4 Agilent Seahorse XF Cell Mito Stress Test Kit User Guide Agilent Seahorse XF Cell Mito Stress Test Kit User Guide
    Agilent Seahorse XF Cell Mito Stress Test Kit User Guide Kit 103015-100 Agilent Technologies Notices © Agilent Technologies, Inc. 2019 Warranty (June 1987) or DFAR 252.227-7015 (b)(2) (November 1995), as applicable in any No part of this manual may be reproduced The material contained in this docu- technical data. in any form or by any means (including ment is provided “as is,” and is sub- electronic storage and retrieval or transla- ject to being changed, without notice, tion into a foreign language) without prior Safety Notices agreement and written consent from in future editions. Further, to the max- Agilent Technologies, Inc. as governed by imum extent permitted by applicable United States and international copyright law, Agilent disclaims all warranties, CAUTION laws. either express or implied, with regard to this manual and any information A CAUTION notice denotes a contained herein, including but not hazard. It calls attention to an oper- Manual Part Number limited to the implied warranties of ating procedure, practice, or the merchantability and fitness for a par- like that, if not correctly performed 103016-400 ticular purpose. Agilent shall not be or adhered to, could result in liable for errors or for incidental or damage to the product or loss of Kit Part Number consequential damages in connection important data. Do not proceed 103015-100 with the furnishing, use, or perfor- beyond a CAUTION notice until the mance of this document or of any indicated conditions are fully Edition information contained herein. Should understood and met. Agilent and the user have a separate Second edition, May 2019 written agreement with warranty Revision G0 terms covering the material in this WARNING Printed in USA document that conflict with these terms, the warranty terms in the sep- A WARNING notice denotes a Agilent Technologies, Inc.
    [Show full text]
  • Pregnancy-Associated Plasma Protein-Aa Supports Hair Cell
    RESEARCH ARTICLE Pregnancy-associated plasma protein-aa supports hair cell survival by regulating mitochondrial function Mroj Alassaf1,2, Emily C Daykin1, Jaffna Mathiaparanam1, Marc A Wolman1* 1Department of Integrative Biology, University of Wisconsin, Madison, United States; 2Neuroscience Training Program, University of Wisconsin, Madison, United States Abstract To support cell survival, mitochondria must balance energy production with oxidative stress. Inner ear hair cells are particularly vulnerable to oxidative stress; thus require tight mitochondrial regulation. We identified a novel molecular regulator of the hair cells’ mitochondria and survival: Pregnancy-associated plasma protein-aa (Pappaa). Hair cells in zebrafish pappaa mutants exhibit mitochondrial defects, including elevated mitochondrial calcium, transmembrane potential, and reactive oxygen species (ROS) production and reduced antioxidant expression. In pappaa mutants, hair cell death is enhanced by stimulation of mitochondrial calcium or ROS production and suppressed by a mitochondrial ROS scavenger. As a secreted metalloprotease, Pappaa stimulates extracellular insulin-like growth factor 1 (IGF1) bioavailability. We found that the pappaa mutants’ enhanced hair cell loss can be suppressed by stimulation of IGF1 availability and that Pappaa-IGF1 signaling acts post-developmentally to support hair cell survival. These results reveal Pappaa as an extracellular regulator of hair cell survival and essential mitochondrial function. DOI: https://doi.org/10.7554/eLife.47061.001 Introduction *For correspondence: Without a sufficient regenerative capacity, a nervous system’s form and function critically depends [email protected] on the molecular and cellular mechanisms that support its cells’ longevity. Neural cell survival is Competing interests: The inherently challenged by the nervous system’s high energy demand, which is required to support authors declare that no basic functions, including maintaining membrane potential, propagating electrical signals, and coor- competing interests exist.
    [Show full text]
  • Mitochondrial Electron Transport Chain Complex III Is Required for Antimycin a to Inhibit Autophagy
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Chemistry & Biology Article Mitochondrial Electron Transport Chain Complex III Is Required for Antimycin A to Inhibit Autophagy Xiuquan Ma,1,4 Mingzhi Jin,1,4 Yu Cai,1 Hongguang Xia,1 Kai Long,1 Junli Liu,1 Qiang Yu,2,* and Junying Yuan1,3,* 1State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Ling-ling Road, Shanghai 200032, China 2Department of Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China 3Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA 4These authors contributed equally to this work *Correspondence: [email protected] (Q.Y.), [email protected] (J.Y.) DOI 10.1016/j.chembiol.2011.08.009 SUMMARY starvation conditions (Hailey et al., 2010; McEwan and Dikic, 2010), but the detailed mechanism is unclear. Autophagy is a cellular lysosome-dependent catabolic Antimycin A (AMA) is a chemical compound produced by mechanism mediating the turnover of intracellular Streptomyces kitazawensis (Nakayama et al., 1956). AMA is organelles and long-lived proteins. We show that anti- known to bind to the Qi site of cytochrome c reductase in the mi- mycin A, a known inhibitor of mETC complex III, can tochondrial complex III to inhibit the oxidation of ubiquinol in the inhibit autophagy. A structural and functional study electron transport chain, which blocks the mitochondrial electron b c shows that four close analogs of antimycin A that transfer between cytochrome and (Alexandre and Lehninger, 1984; Campo et al., 1992; Maguire et al., 1992; Pham et al., have no effect on mitochondria inhibition also do not 2000; Xia et al., 1997).
    [Show full text]
  • Elevated Levels of Mitochondrial Coq10 Induce ROS-Mediated
    www.nature.com/scientificreports OPEN Elevated levels of mitochondrial CoQ10 induce ROS‑mediated apoptosis in pancreatic cancer Tulin Dadali1,3, Anne R. Diers1,3, Shiva Kazerounian1, Senthil K. Muthuswamy2, Pallavi Awate1, Ryan Ng1, Saie Mogre1, Carrie Spencer1, Katerina Krumova1, Hannah E. Rockwell1, Justice McDaniel1, Emily Y. Chen1, Fei Gao1, Karl T. Diedrich1, Vijetha Vemulapalli1, Leonardo O. Rodrigues1, Viatcheslav R. Akmaev1, Khampaseuth Thapa1, Manuel Hidalgo2, Arindam Bose2, Vivek K. Vishnudas1, A. James Moser2, Elder Granger1, Michael A. Kiebish1, Stephane Gesta1, Niven R. Narain1 & Rangaprasad Sarangarajan1* Reactive oxygen species (ROS) are implicated in triggering cell signalling events and pathways to promote and maintain tumorigenicity. Chemotherapy and radiation can induce ROS to elicit cell death allows for targeting ROS pathways for efective anti‑cancer therapeutics. Coenzyme Q10 is a critical cofactor in the electron transport chain with complex biological functions that extend beyond mitochondrial respiration. This study demonstrates that delivery of oxidized Coenzyme Q10 (ubidecarenone) to increase mitochondrial Q‑pool is associated with an increase in ROS generation, efectuating anti‑cancer efects in a pancreatic cancer model. Consequent activation of cell death was observed in vitro in pancreatic cancer cells, and both human patient‑derived organoids and tumour xenografts. The study is a frst to demonstrate the efectiveness of oxidized ubidecarenone in targeting mitochondrial function resulting in an anti‑cancer efect. Furthermore, these fndings support the clinical development of proprietary formulation, BPM31510, for treatment of cancers with high ROS burden with potential sensitivity to ubidecarenone. Dysregulated metabolism is considered a hallmark of cancer and is a consequence of adaptive responses to sup- port unregulated proliferation.
    [Show full text]
  • Minimal Mitochondrial Respiration Is Required to Prevent Cell Death by Inhibition of Mtor Signaling in Coq-Deficient Cells ✉ Ying Wang 1 and Siegfried Hekimi1
    www.nature.com/cddiscovery ARTICLE OPEN Minimal mitochondrial respiration is required to prevent cell death by inhibition of mTOR signaling in CoQ-deficient cells ✉ Ying Wang 1 and Siegfried Hekimi1 © The Author(s) 2021 Coenzyme Q (CoQ) is a lipid-like mobile electron transporter of the mitochondrial respiratory chain. Patients with partial loss-of- function mutations in the CoQ biosynthesis pathway suffer from partial primary CoQ deficiency (MIM 607426). This leads to mitochondrial dysfunction, which presents like mitochondrial disease syndrome (MDS). In addition, many other conditions, including MDS itself, lead to secondary CoQ deficiency. We sought to identify drugs that can alleviate the consequences of the mitochondrial dysfunction that is associated with CoQ deficiency. Loss of the CoQ-biosynthetic enzyme COQ7 prevents CoQ synthesis but leads to the accumulation of the biosynthetic intermediate demethoxyubiquinone (DMQ). Coq7-knockout mouse embryonic fibroblasts (MEFs) die when rapid ATP generation from glycolysis is prevented. We screened for drugs that could rescue cell death under these conditions. All compounds that were identified inhibit mTOR signaling. In the CoQ-deficient cells, the beneficial action mTOR inhibition appears to be mediated by inhibition of protein translation rather than by stimulation of autophagy. We further studied the Coq7-knockout cells to better determine under which conditions mTOR inhibition could be beneficial. We established that Coq7-knockout cells remain capable of a low level of mitochondrial respiration mediated by DMQ. To obtain more profound mitochondrial dysfunction, we created double-knockout mutant MEFs lacking both Coq7, as well as Pdss2, which is required for sidechain synthesis. These cells make neither CoQ nor DMQ, and their extremely small residual respiration depends on uptake of CoQ from the culture medium.
    [Show full text]
  • Role of Mitochondrial Complex I and Protective Effect of Coq10 Supplementation in Propofol Induced Cytotoxicity
    J Bioenerg Biomembr (2016) 48:413–423 DOI 10.1007/s10863-016-9673-9 Role of mitochondrial complex I and protective effect of CoQ10 supplementation in propofol induced cytotoxicity Christian Bergamini1 & Noah Moruzzi2 & Francesco Volta2 & Laura Faccioli1 & Jantje Gerdes2 & Maria Cristina Mondardini3 & Romana Fato1 Received: 16 June 2016 /Accepted: 2 August 2016 /Published online: 15 August 2016 # Springer Science+Business Media New York 2016 Abstract Propofol (2,6-diisopropylphenol) is an anaesthetic of the enzyme. Cells supplemented with CoQ10 are more resis- widely used for human sedation. Due to its intrinsic antioxidant tant to propofol toxicity. Propofol exposure induces cellular properties, rapid induction of anaesthesia and fast recovery, it is damages due to mitochondrial impairment. The site of propofol employed in paediatric anaesthesia and in the intensive care of inhibition on Complex I is the Q module. CoQ10 supplementa- premature infants. Recent studies have pointed out that expo- tion protects cells against the loss of energy suggesting its pos- sure to anaesthesia in the early stage of life might be responsible sible therapeutic role to minimizing the detrimental effects of of long-lasting cognitive impairment. The apoptotic neurode- general anaesthesia. generation induced by general anaesthetics (GA) involves mi- tochondrial impairment due to the inhibition of the OXPHOS machinery. In the present work, we aim to identify the main Keywords Propofol . Complex I . Mitochondria . mitochondrial respiratory chain target of propofol toxicity and Anaesthesia . Oxygen consumption to evaluate the possible protective effect of CoQ10 supplemen- tation. The propofol effect on the mitochondrial functionality Abbreviations was assayed in isolated mitochondria and in two cell lines mPTP Mitochondrial permeability transition pore (HeLa and T67) by measuring oxygen consumption rate.
    [Show full text]
  • Ca2+-Induced Mitochondrial ROS Regulate the Early Embryonic Cell Cycle
    bioRxiv preprint doi: https://doi.org/10.1101/223123; this version posted November 21, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Ca2+-induced mitochondrial ROS regulate the early embryonic cell cycle Yue Han1, 2, 5, Shoko Ishibashi1, 5, Javier Iglesias-Gonzalez1, Yaoyao Chen1, 3, Nick R. Love1, 4 and Enrique Amaya1,6, * 1 Division of Cell Matrix Biology & Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UK 2 Institute of Stem Cell and Regenerative Medicine, Medical College, Xiamen University, Xiamen, Fujian, 361102 P. R. China 3 Current address: STEMCELL Technologies UK ltd, Building 7100, Cambridge Research Park, Beach Drive, Waterbeach, Cambridge, UK, CB25 9TL 4 Current address: Stanford University School of Medicine, Stanford, CA 94305, USA 5Co-first author 6Lead Contact *Correspondence: [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/223123; this version posted November 21, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. SUMMARY While it has long been appreciated that reactive oxygen species (ROS) can act as second messengers in both homeostastic and stress response signaling pathways, potential roles for ROS during early vertebrate development have remained largely unexplored.
    [Show full text]
  • Cellular Consequences of Coenzyme Q10 Deficiency In
    International Journal of Molecular Sciences Review Cellular Consequences of Coenzyme Q10 Deficiency in Neurodegeneration of the Retina and Brain Haider Manzar 1, Dalia Abdulhussein 1, Timothy E. Yap 1 and M. Francesca Cordeiro 1,2,* 1 Imperial College Ophthalmology Research Group, Western Eye Hospital, 153-173 Marylebone Road, Marylebone, London NW1 5QH, UK; [email protected] (H.M.); [email protected] (D.A.); [email protected] (T.E.Y.) 2 Glaucoma & Retinal Neurodegeneration Research Group, Institute of Ophthalmology, University College London, London EC1V 9EL, UK * Correspondence: [email protected] Received: 7 November 2020; Accepted: 4 December 2020; Published: 6 December 2020 Abstract: Coenzyme Q10 (CoQ10) is a ubiquitous cofactor in the body, operating in the inner mitochondrial membrane, where it plays a vital role in the generation of adenosine triphosphate (ATP) through the electron transport chain (ETC). In addition to this, CoQ10 serves as an antioxidant, protecting the cell from oxidative stress by reactive oxygen species (ROS) as well as maintaining a proton (H+) gradient across lysosome membranes to facilitate the breakdown of cellular waste products. Through the process of ageing, the body becomes deficient in CoQ10, resulting in several systemic manifestations. On a cellular level, one of the consequences of CoQ10 deficiency is apoptosis, which can be visualised in tissues of the central nervous system (CNS). Diseases affecting the retina and brain such as age-related macular degeneration (AMD), glaucoma, Alzheimer’s disease (AD) and Parkinson’s disease (PD) have shown defects in cellular biochemical reactions attributed to reduced levels of CoQ10.
    [Show full text]
  • Across: Two Neurotoxicity AOP-Based IATA Case Studies*
    ALTEX, accepted manuscript published June 10, 2021 doi: 10.14573/altex.2103051 Research Article New Approach Methods (NAMs) Supporting Read- Across: Two Neurotoxicity AOP-based IATA Case Studies* Wanda van der Stel1, Giada Carta2, Julie Eakins3, Johannes Delp4, Ilinca Suciu4,5, Anna Forsby6, Andrea Cediel-Ulloa7, Kristina Attoff6, Florentina Troger8, Hennicke Kamp9, Iain Gardner10, Barbara Zdrazil8, Martijn J. Moné1, Gerhard F. Ecker8, Manuel Pastor11, Jose Carlos Gómez-Tamayo11, Andrew White12, Erik H.J. Danen1, Marcel Leist 4, Paul Walker 3, Paul Jennings 2, Susanne Hougaard Bennekou13 and Bob van de Water1 1Division of Drug Discovery and Safety, Leiden Academic Centre of Drug Research, Leiden University, Leiden, The Netherlands; 2Division of Molecular and Computational Toxicology, Department of Chemistry and Pharmaceutical Sciences, AIMMS, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; 3Cyprotex Discovery Ltd., Alderley Park, Macclesfield, Cheshire, United Kingdom; 4Chair for In vitro Toxicology and Biomedicine, Department of Inaugurated by the Doerenkamp-Zbinden Foundation, University of Konstanz, Germany; 5Konstanz Research School Chemical Biology, University of Konstanz, Konstanz, Germany; 6Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden; 7Department of Organismal Biology, Uppsala University, Uppsala, Sweden; 8Department of Pharmaceutical Chemistry, University of Vienna, Austria; 9BASF, Ludwigshafen, Germany; 10Certara UK Limited, Sheffield, United Kingdom; 11Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Spain; 12Unilever, Bedfordshire, United Kingdom; 13National Food Institute Technical University of Denmark (DTU), Lyngby, Denmark Abstract Read-across approaches are considered key in moving away from in vivo animal testing towards addressing data-gaps using new approach methods (NAMs). Ample successful examples are still required to substantiate this strategy. Here we present and discuss the learnings from two OECD IATA endorsed read-across case studies.
    [Show full text]
  • Mechanisms of Mitochondrial ROS Production in Assisted Reproduction: the Known, the Unknown, and the Intriguing
    antioxidants Review Mechanisms of Mitochondrial ROS Production in Assisted Reproduction: The Known, the Unknown, and the Intriguing James N. Cobley Redox Biology Group, Institute for Health Sciences, University of the Highlands and Islands, Old Perth Road, Inverness IV2 3JH, UK; [email protected] Received: 21 July 2020; Accepted: 21 September 2020; Published: 29 September 2020 Abstract: The consensus that assisted reproduction technologies (ART), like in vitro fertilization, to induce oxidative stress (i.e., the known) belies how oocyte/zygote mitochondria—a major presumptive oxidative stressor—produce reactive oxygen species (ROS) with ART being unknown. Unravelling how oocyte/zygote mitochondria produce ROS is important for disambiguating the molecular basis of ART-induced oxidative stress and, therefore, to rationally target it (e.g., using site-specific mitochondria-targeted antioxidants). I review the known mechanisms of ROS production in somatic mitochondria to critique how oocyte/zygote mitochondria may produce ROS (i.e., the unknown). Several plausible site- and mode-defined mitochondrial ROS production mechanisms in ART are proposed. For example, complex I catalyzed reverse electron transfer-mediated ROS production is conceivable when oocytes are initially extracted due to at least a 10% increase in molecular dioxygen exposure (i.e., the intriguing). To address the term oxidative stress being used without recourse to the underlying chemistry, I use the species-specific spectrum of biologically feasible reactions to define plausible oxidative stress mechanisms in ART. Intriguingly, mitochondrial ROS-derived redox signals could regulate embryonic development (i.e., their production could be beneficial). Their potential beneficial role raises the clinical challenge of attenuating oxidative damage while simultaneously preserving redox signaling.
    [Show full text]
  • Synthesis and Cellular Effects of a Mitochondria-Targeted Inhibitor of the Two-Pore Potassium Channel TASK-3
    Pharmacological Research 164 (2021) 105326 Contents lists available at ScienceDirect Pharmacological Research journal homepage: www.elsevier.com/locate/yphrs Synthesis and cellular effects of a mitochondria-targeted inhibitor of the two-pore potassium channel TASK-3 Magdalena Bachmann a, Andrea Rossa b, Giuseppe Antoniazzi b, Lucia Biasutto c,d, Andrea Carrer a,d, Monica Campagnaro a, Luigi Leanza a, Monika Gonczi e, Laszlo Csernoch e, Cristina Paradisi b, Andrea Mattarei f, Mario Zoratti c,d, Ildiko Szabo a,c,* a Department of Biology, University of Padua, Italy b Department of Chemical Sciences, University of Padua, Italy c CNR Institute of Neuroscience, Padua, Italy d Department of Biomedical Sciences, University of Padua, Italy e Department of Physiology, Faculty of Medicine, University of Debrecen, Hungary f Department of Pharmaceutical and Pharmacological Sciences, University of Padua, Italy ARTICLE INFO ABSTRACT Keywords: The two-pore potassium channel TASK-3 has been shown to localize to both the plasma membrane and the Mitochondria mitochondrial inner membrane. TASK-3 is highly expressed in melanoma and breast cancer cells and has been TASK-3 potassium channel proposed to promote tumor formation. Here we investigated whether pharmacological modulation of TASK-3, Pharmacological targeting and specifically of mitochondrial TASK-3 (mitoTASK-3), had any effect on cancer cell survival and mitochon­ Melanoma drial physiology. A novel, mitochondriotropic version of the specific TASK-3 inhibitor IN-THPP has been syn­ Chemical compounds studied in this article: thesized by addition of a positively charged triphenylphosphonium moiety. While IN-THPP was unable to induce Staurosporin (PubChem CID: 44259) FCCP (PubChem CID: 3330) apoptosis, mitoIN-THPP decreased survival of breast cancer cells and efficientlykilled melanoma lines, which we Antimycin A (PubChem CID: 16218979) show to express mitoTASK-3.
    [Show full text]