Document text
Principal Investigator: Shyam Sunder Chaurasia
Organization: UNIVERSITY OF MISSOURI-COLUMBIA
Fiscal Year: 2019
Award: $193,475
Funding agency: National Eye Institute
PROJECT SUMMARY
Chemical weapons were used in world wars, and more recently in Iraq and Syria. Acrolein, a volatile aldehyde,
used during World War I caused severe problems in eye, skin and lungs in addition to many casualties. At
present, there are no countermeasures available that can prevent, impede or arrest corneal damage and loss
of vision caused by acrolein poisoning. Since acrolein is a vital precursor for variety of commercial goods, over
500,000 tons of acrolein is produced and transported annually in America, which pose a real threat of its use
as a chemical weapon by the terrorists for mass causalities. It is our central hypothesis is that acrolein contact
to eye depletes glutathione and augments various cytokines entry into the corneal stroma resulting in corneal
damage and vision loss. This proposal aims to characterize molecular mechanisms involved in acrolein-
exposed loss of corneal function and test the efficacy and safety of Glutamate Cysteine Ligase Catalytic
subunit (GCLC; a rate-limiting enzyme in the synthesis of GSH) gene therapy given to cornea via our reported
nanoparticlebased gene transfer method to mitigate acrolein-induced toxicity and vision loss in vivo using a
preclinical rabbit model. To the best of our knowledge, no scientific data is currently available on mechanisms
mediating acrolein toxicity and gene-based therapy. Aim-1 will delineate molecular and cellular mechanisms of
acrolein intoxication to the eyes by studying changes in the glutathione content, reactive oxidative stress,
keratocyte death, and transforming growth factor β1 in corneal tissues, and cytokines level in tears after
acrolein contact to eye in a rabbit model. These factors are known to cause ocular irritation, inflammation,
corneal scarring and corneal neovascularization in vivo. Aim-2 will test the hypothesis that nanoparticle-based
GCLC gene therapy enhancing the glutathione redox system will counteract deleterious effects of acrolein in
the cornea by stimulating the synthesis of GSH and increasing the antioxidant index in vivo in a rabbit model.
To accomplish proposed research, we have selected a rabbit model that recapitulates clinical traits of acrolein
toxicity observed in human patients. Rabbit eyes will be exposed to acrolein and ocular health in live rabbits
will be monitored with clinical slit-lamp and confocal microscopes. The changes at the cellular and molecular
levels in tears and corneal tissues will be analyzed using commercial assays such as TUNEL, GSH, ROS etc.,
immunoblotting, immunofluorescence, qPCR etc. techniques. Our team is comprised of corneal researchers,
physician and veterinary ophthalmologists with extensive experience, skills, tools, and joint publications, and is
most suitable for this translational research. Successful completion of this project will lead better understanding
of molecular mechanisms and pathways modulated by acrolein in the eye, and will provide necessary in vivo
data that will lead development of gene-based nanomedicine approaches to treat acrolein-induced ocular
toxicity and blindness. The nanoparticle-based gene transfer method that will be used has showed transgene
expression for two weeks after one topical application in rabbit cornea in vivo.
Terms: <1st World War><2'-deoxy-8-hydroxyguanosine><2-Propenal><4 hydroxynonenal><4-HNE cpd><4-hydroxy-2,3-nonenal><4-hydroxy-2-nonenal><4-hydroxynonen-2-al><8-OH-dG><8-hydroxy-2'-deoxyguanosine><8-hydroxydeoxyguanosine><8-oxodGuo><Acraldehyde><Acrolein><Acrylaldehyde><Acrylic Aldehyde><Aldehydes><Allyl Aldehyde><Americas><Animal Experimental Use><Animal Experimentation><Animal Research><Animals><Antidotes><Antioxidants><Assay><Basic Research><Basic Science><Binding><Bioassay><Biochemical><Biologic Assays><Biological Assay><Blindness><Blood leukocyte><Body Tissues><Bone-Derived Transforming Growth Factor><CCL2><CCL2 gene><CD11b><CD31><CR3A><Causality><Cell Communication and Signaling><Cell Death><Cell Signaling><Cessation of life><Chemical Weapons><Chemokine, CC Motif, Ligand 2><Clinical><Cold-Insoluble Globulins><Collagen><Collagen IV><Collagen Type IV><Cornea><Corneal Angiogenesis><Corneal Neovascularization><Corneal Stroma><Cysteine><DNA><DNA Therapy><Data><Death><Deoxyribonucleic Acid><Development><Diesel Exhaust><Domestic Rabbit><Doppler OCT><Drugs><Dysfunction><EC 2.5.1.18><EC 6.3.2.2><ELISA><Enzyme Gene><Enzyme-Linked Immunosorbent Assay><Enzymes><Erythrocuprein><Ethylene Aldehyde><Etiology><Exposure to><Eye><Eye Exam><Eye Examination><Eye Injuries><Eye pain><Eyeball><Eyelid structure><Eyelids><FN1><Fibronectin 1><Fibronectins><Fibrosis><Fire - disasters><Fires><First World War><Functional disorder><GCLC><GCLC gene><GCLM><GCLM gene><GLCLC><GLCLR><Gamma-Glutamylcysteine Synthetase Catalytic Subunit><Gamma-Glutamylcysteine Synthetase, Regulatory Subunit><Gamma-Glutamylcysteinyl Synthetase><Gasoline><Gene Delivery><Gene Transfer><Gene Transfer Clinical><Genes><Genetic Intervention><Glutamate-Cysteine Ligase><Glutamate-Cysteine Ligase, Catalytic Subunit><Glutamate-Cysteine Ligase, Modifier Subunit><Glutamate-Cysteine Ligase, Regulatory><Glutamylcysteine Synthetase><Glutathione><Glutathione Organic Nitrate Ester Reductase><Glutathione S-Alkyltransferase><Glutathione S-Aralkyltransferase><Glutathione S-Aryltransferase><Glutathione S-Epoxidetransferase><Glutathione S-Transferase><Glutathione Transferase><Great War><Half-Cystine><Half-Life><Health><Heme Transfer Protein><Hemocuprein><Herbicides><Histidine><Histologic Technics><Histologic Techniques><Histological Technics><Histological Techniques><Human><ITGAM><ITGAM gene><Immunoblotting><Immunofluorescence><Immunofluorescence Immunologic><In Situ Nick-End Labeling><Infection><Inflammation><Intoxication><Intracellular Communication and Signaling><Investigators><Iraq><Irritants><Joints><Knowledge><L-Cysteine><L-Lysine><LETS Proteins><Large External Transformation-Sensitive Protein><Lead><Lectin><Leukocytes><Leukocytes Reticuloendothelial System><Ligandins><Lipid Peroxidation><Literature><Lung><Lung Respiratory System><Lysine><MAC-1><MAC1A><MCAF><MCP-1><MCP1><MO1A><Marrow leukocyte><Measures><Mediating><Medication><Mercaptans><Mercapto Compounds><Messenger RNA><Methods><Microscope><Milk Growth Factor><Modeling><Modern Man><Molecular><Molecular Interaction><Monitor><Monocyte Chemoattractant Protein-1><Monocyte Chemotactic Protein-1><Monocyte Chemotactic and Activating Factor><Monocyte Chemotactic and Activating Protein><Monocyte Chemotactive and Activating Factor><Monocyte Secretory Protein JE><OCT Tomography><Ocular Injury><Ophthalmic examination and evaluation><Ophthalmologist><Opsonic Glycoprotein><Opsonic alpha(2)SB Glycoprotein><Optical Coherence Tomography><Oryctolagus cuniculus><Outcome><Oxidation-Reduction><Oxidative Stress><PECAM1><PECAM1 gene><Palpebra><Pathologic><Pathway interactions><Patients><Pb element><Pesticides><Pharmaceutic Preparations><Pharmaceutical Preparations><Physicians><Physiopathology><Platelet Transforming Growth Factor><Play><Poisoning><Proteins><Publications><Publishing><R-Series Research Projects><R01 Mechanism><R01 Program><Rabbits><Rabbits Mammals><Redox><Reduced Glutathione><Reporting><Research><Research Grants><Research Personnel><Research Project Grants><Research Projects><Researchers><Role><S-Hydroxyalkyl Glutathione Lyase><SCYA2><Safety><Scientific Publication><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skin><Small Inducible Cytokine A2><Smooth Muscle Actin><Smooth Muscle Actin Staining Method><Stromal Cells><Sulfhydryl Compounds><Superoxide Dismutase><Swelling><Syria><Syrian Arab Republic><System><TGF B><TGF-beta><TGF-β><TGFbeta><TGFβ><TUNEL><TUNEL Assay><TdT-Mediated dUTP Nick End Labeling Assay><Techniques><Testing><Theriacs><Thiols><Time><Tissues><Topical Drug Administration><Topical application><Toxic effect><Toxicities><Transforming Growth Factor beta><Transforming Growth Factor-Beta Family Gene><Transforming Growth Factors><Translational Research><Translational Science><Tumor Growth Factors><Type IV (Basement Membrane) Collagen><VEGF><VEGFs><Vascular Endothelial Growth Factors><War><White Blood Cells><White Cell><Wood><Wood material><World War I><acryaldehyde><alpha 2-Surface Binding Glycoprotein><anti-oxidant><base><biological signal transduction><catalase><causation><cicatrix corneal><cigarette smoke><corneal><corneal scar><cytocuprein><cytokine><developmental><disease causation><drug/agent><efficacy testing><experience><experiment><experimental research><experimental study><fight against><fire><forest><gamma-Glutamyl-Cysteine Synthetase><gamma-L-Glu-L-Cys-Gly><gamma-L-Glutamyl-L-Cysteinylglycine><gene therapy><gene-based therapy><genetic therapy><genomic therapy><glutathione aralkyltransferase><glutathione aryltransferase><glutathione peroxidase><heavy metal Pb><heavy metal lead><in vivo><indexing><innovate><innovation><innovative><irritation><mRNA><nano medicinal><nano medicine><nano particle><nano-sized particle><nanomedicinal><nanomedicine><nanoparticle><nanosized particle><necrocytosis><neovascularization><new approaches><novel><novel approaches><novel strategies><novel strategy><ocular pain><optical Doppler tomography><optical coherence Doppler tomography><oxidation reduction reaction><pathophysiology><pathway><poisoned><pre-clinical><preclinical><prevent><preventing><pulmonary><response><skills><social role><sulfhydryl group><terminal nick end labeling><tonometry><tool><topical administration><topical delivery><topical drug application><topical treatment><topically applied><trait><transforming growth factors Animal growth regulators><transgene expression><translation research><vapor><vision loss><visual loss><white blood cell><white blood corpuscle>