Investigating the Cellular Effects of Mitochondrial DNA Damage Using Targeted Chemical Probes PDF Download
Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Investigating the Cellular Effects of Mitochondrial DNA Damage Using Targeted Chemical Probes PDF full book. Access full book title Investigating the Cellular Effects of Mitochondrial DNA Damage Using Targeted Chemical Probes by Simon Wisnovsky. Download full books in PDF and EPUB format.
Author: Simon Wisnovsky
Publisher:
ISBN:
Category :
Languages : en
Pages :
Get Book Here
Book Description
Mitochondria are compartments within eukaryotic cells that produce energy to power cellular metabolism. Uniquely amongst mammalian organelles, mitochondria contain a small amount of their own genetic material (mtDNA), the proper maintenance and expression of which is essential for mitochondrial function. Mitochondrial DNA damage has been linked to several pathophysiological processes, but the cellular effects of mtDNA damage remain poorly understood. In order to better understand mtDNA damage, our group has pioneered the development of selective mitochondria-targeted DNA damaging agents using positively charged, hydrophobic delivery vectors known as mitochondria-penetrating peptides (MPPs). In my first study, an analogue of the commonly used anticancer drug cisplatin was conjugated to an MPP, yielding a mitochondria-targeted platinum DNA-damaging agent (mtPt) that induced specific damage to mtDNA. Using mtPt, I showed that selective mtDNA platinum damage induced apoptotic cell death associated with reactive oxygen species production, demonstrating that mtDNA damage is sufficient to mediate the cytotoxic activity of a platinum drug. In another work, I aimed to use mitochondria-targeted DNA damaging agents in a screen to characterize novel factors in mtDNA replication and repair. By knocking down expression of known DNA repair genes, and monitoring synergistic effects of gene knockdown on the activity of an mtDNA-oxidizing agent, I identified a series of genes with novel roles in mtDNA repair and replication. Included amongst these hits was DNA Polymerase Î ̧ (PolÎ ̧), a novel mtDNA polymerase. In a further study, I found that PolÎ ̧ plays an essential role in facilitating mtDNA replication under conditions of oxidative stress. Finally, I conducted a genome-wide CRISPR screen to comprehensively identify genes whose knockout sensitizes cells to mitochondria-localized DNA damage. The hits of this screen were heavily enriched for genes involved in mitochondrial translation, implying the importance of mitochondrial protein synthesis in mediating an adaptive response to mtDNA damage. These studies demonstrate the application of mitochondria-targeted DNA damaging agents to the study of a wide range of significant areas in mitochondrial biology. Going forward, my thesis work shows mtDNA-damaging chemical probes to be essential tools for achieving better understanding of mtDNA damage in health and disease.
Author: Simon Wisnovsky
Publisher:
ISBN:
Category :
Languages : en
Pages :
Get Book Here
Book Description
Mitochondria are compartments within eukaryotic cells that produce energy to power cellular metabolism. Uniquely amongst mammalian organelles, mitochondria contain a small amount of their own genetic material (mtDNA), the proper maintenance and expression of which is essential for mitochondrial function. Mitochondrial DNA damage has been linked to several pathophysiological processes, but the cellular effects of mtDNA damage remain poorly understood. In order to better understand mtDNA damage, our group has pioneered the development of selective mitochondria-targeted DNA damaging agents using positively charged, hydrophobic delivery vectors known as mitochondria-penetrating peptides (MPPs). In my first study, an analogue of the commonly used anticancer drug cisplatin was conjugated to an MPP, yielding a mitochondria-targeted platinum DNA-damaging agent (mtPt) that induced specific damage to mtDNA. Using mtPt, I showed that selective mtDNA platinum damage induced apoptotic cell death associated with reactive oxygen species production, demonstrating that mtDNA damage is sufficient to mediate the cytotoxic activity of a platinum drug. In another work, I aimed to use mitochondria-targeted DNA damaging agents in a screen to characterize novel factors in mtDNA replication and repair. By knocking down expression of known DNA repair genes, and monitoring synergistic effects of gene knockdown on the activity of an mtDNA-oxidizing agent, I identified a series of genes with novel roles in mtDNA repair and replication. Included amongst these hits was DNA Polymerase Î ̧ (PolÎ ̧), a novel mtDNA polymerase. In a further study, I found that PolÎ ̧ plays an essential role in facilitating mtDNA replication under conditions of oxidative stress. Finally, I conducted a genome-wide CRISPR screen to comprehensively identify genes whose knockout sensitizes cells to mitochondria-localized DNA damage. The hits of this screen were heavily enriched for genes involved in mitochondrial translation, implying the importance of mitochondrial protein synthesis in mediating an adaptive response to mtDNA damage. These studies demonstrate the application of mitochondria-targeted DNA damaging agents to the study of a wide range of significant areas in mitochondrial biology. Going forward, my thesis work shows mtDNA-damaging chemical probes to be essential tools for achieving better understanding of mtDNA damage in health and disease.
Author: Linsey Robertson
Publisher:
ISBN:
Category :
Languages : en
Pages :
Get Book Here
Book Description
The implication of oxidative stress in ageing, and various disease states, has resulted in great interest in understanding the processes associated with it. Mitochondria are the major source of reactive oxygen species (ROS) via the electron transport chain. ROS may be free radicals, or molecules that rapidly form free radicals. These free radicals can undergo various rapid reactions with cellular components resulting in cell damage, and eventually cell death. Therefore, there is much interest in studying ROS production particularly from mitochondria. In this thesis three novel classes of chemical probes for the study of oxidative stress are presented. Firstly, progress was made towards a novel mitochondria-targeted hydroxylamine iii based on the 1,1,3,3-tetraethylisoindoline (TEIO) scaffold. This hydroxylamine iii should undergo reaction with radicals to form a stable nitroxide which gives a simple, persistent, quantifiable EPR (electron paramagnetic resonance) signal. Phosphonium salt i was prepared and EPR spectroscopy studies in a trial oxidation reaction indicated the formation of the late stage nitroxide precursor ii. Three new pyridinium salt nitrone spin traps iv-vi, designed to accumulate in mitochondria, were synthesised using the Zincke reaction as the key pyridinium salt forming reaction. The compatibility of nitrones with the Zincke reaction was demonstrated for the first time. The capability of spin traps iv-vi to trap methyl radicals was demonstrated using EPR spectroscopy. Two novel selective uncoupling molecules (SUMs), DNP-SUM vii and FCCP-SUM viii, were designed to react with H2O2 and release a mitochondrial uncoupler. These were synthesised and their reaction with H2O2 was examined using UV spectrophotometry. DNP-SUM vii reacted slowly with a second order rate constant of 0.20 M-1s-1 and was deemed unsuitable for further investigation. However, the reaction of H2O2 and FCCP-SUM viii displayed good pseudo-first order kinetics when FCCP-SUM viii was used in excess and a second order rate constant of 64 M-1s-1 for the release of the mitochondrial uncoupler FCCP (carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone) in its ionised form. New analogues of FCCP-SUM viii are now in development to enable targeting to mitochondria.
Author: Henry F. Epstein
Publisher: Academic Press
ISBN: 0125641494
Category : Caenorhabditis elegans
Languages : en
Pages : 687
Get Book Here
Book Description
The first of its kind, this laboratory handbook emphasizes diverse methods and technologies needed to investigate C. elegans, both as an integrated organism and as a model system for research inquiries in cell, developmental, and molecular biology, as well as in genetics and pharmacology. Four primary sections--Genetic and Culture Methods, Neurobiology, Cell and Molecular Biology, and Genomics and Informatics--reflect the cross-disciplinary nature of C. elegans research. Because C. elegans is a simple and malleable organism with a small genome and few cell types, it provides an elegant demonstr.
Author: Lawrence H. Lash
Publisher: Elsevier
ISBN: 1483218619
Category : Science
Languages : en
Pages : 527
Get Book Here
Book Description
Methods in Toxicology, Volume 2: Mitochondrial Dysfunction provides a source of methods, techniques, and experimental approaches for studying the role of abnormal mitochondrial function in cell injury. The book discusses the methods for the preparation and basic functional assessment of mitochondria from liver, kidney, muscle, and brain; the methods for assessing mitochondrial dysfunction in vivo and in intact organs; and the structural aspects of mitochondrial dysfunction are addressed. The text also describes chemical detoxification and metabolism as well as specific metabolic reactions that are especially important targets or indicators of damage. The methods for measurement of alterations in fatty acid and phospholipid metabolism and for the analysis and manipulation of oxidative injury and antioxidant systems are also considered. The book further tackles additional methods on mitochondrial energetics and transport processes; approaches for assessing impaired function of mitochondria; and genetic and developmental aspects of mitochondrial disease and toxicology. The text also looks into mitochondrial DNA synthesis, covalent binding to mitochondrial DNA, DNA repair, and mitochondrial dysfunction in the context of developing individuals and cellular differentiation. Microbiologists, toxicologists, biochemists, and molecular pharmacologists will find the book invaluable.
Author: Steven Howard
Publisher: Royal Society of Chemistry
ISBN: 1782625658
Category : Medical
Languages : en
Pages : 314
Get Book Here
Book Description
Fragment-based drug discovery is a rapidly evolving area of research, which has recently seen new applications in areas such as epigenetics, GPCRs and the identification of novel allosteric binding pockets. The first fragment-derived drug was recently approved for the treatment of melanoma. It is hoped that this approval is just the beginning of the many drugs yet to be discovered using this fascinating technique. This book is written from a Chemist's perspective and comprehensively assesses the impact of fragment-based drug discovery on a wide variety of areas of medicinal chemistry. It will prove to be an invaluable resource for medicinal chemists working in academia and industry, as well as anyone interested in novel drug discovery techniques.
Author: Lihe Zhang
Publisher: Springer Nature
ISBN: 9811995850
Category :
Languages : en
Pages : 72
Get Book Here
Book Description
Author: Sabzali Javadov
Publisher:
ISBN: 9783039363841
Category :
Languages : en
Pages : 434
Get Book Here
Book Description
Mitochondria are subcellular organelles evolved by the endosymbiosis of bacteria with eukaryotic cells. They are the main source of ATP in the cell and engaged in other aspects of cell metabolism and cell function, including the regulation of ion homeostasis, cell growth, redox status, and cell signaling. Due to their central role in cell life and death, mitochondria are also involved in the pathogenesis and progression of human diseases/conditions, including neurodegenerative and cardiovascular disorders, cancer, diabetes, inflammation, and aging. However, despite the increasing number of studies, precise mechanisms whereby mitochondria are involved in the regulation of basic physiological functions, as well as their role in the cell under pathophysiological conditions, remain unknown. A lack of in-depth knowledge of the regulatory mechanisms of mitochondrial metabolism and function, as well as interplay between the factors that transform the organelle from its role in pro-survival to pro-death, have hindered the development of new mitochondria-targeted pharmacological and conditional approaches for the treatment of human diseases. This book highlights the latest achievements in elucidating the role of mitochondria under physiological conditions, in various cell/animal models of human diseases, and in patients.
Author:
Publisher:
ISBN: 9780815332183
Category : Cells
Languages : en
Pages : 0
Get Book Here
Book Description
Author: Harpreet Singh
Publisher: Springer
ISBN: 3319573136
Category : Medical
Languages : en
Pages : 491
Get Book Here
Book Description
This special book is conceived to highlight mitochondrial structural and functional integrity and how they are associated with several human diseases such as cardiovascular, cancer, renal, neurological disorder, and genetic disorders. The chapters contributed by leading mitochondrial researchers in the handbook will take us through the novel pharmacological strategies via mitochondria to understand their physiological and pathological role as well as present them as therapeutic targets.
Author: United States. Public Health Service. Office of the Surgeon General
Publisher:
ISBN:
Category : Government publications
Languages : en
Pages : 728
Get Book Here
Book Description
This report considers the biological and behavioral mechanisms that may underlie the pathogenicity of tobacco smoke. Many Surgeon General's reports have considered research findings on mechanisms in assessing the biological plausibility of associations observed in epidemiologic studies. Mechanisms of disease are important because they may provide plausibility, which is one of the guideline criteria for assessing evidence on causation. This report specifically reviews the evidence on the potential mechanisms by which smoking causes diseases and considers whether a mechanism is likely to be operative in the production of human disease by tobacco smoke. This evidence is relevant to understanding how smoking causes disease, to identifying those who may be particularly susceptible, and to assessing the potential risks of tobacco products.
