Disulfide activated protein kinase G Ialpha as a new therapeutic target in sepsis
Lead Research Organisation:
King's College London
Department Name: Cardiovascular
Abstract
Sepsis, often referred to as blood poisoning, occurs when bacteria get into the blood stream. This is a very common disease and when it occurs the chances of survival are rather poor. Major issues in sepsis are that the patients' blood pressure becomes very low and their blood vessels become leaky. Together this causes a lot of harm and many of the bodies organs are not supplied with enough blood and so they become injured. Recently we identified a new pathway that lowers blood pressure and promotes vessel leakiness in healthy people, and have now we have found this is over-stimulated during sepsis resulting in organ damage. In these studies we hope to develop and test new drugs that prevent this pathway being actuated during sepsis. Such therapies are predicted to prevent organ damage and ultimately enhance patient survival during sepsis.
Technical Summary
Sepsis is a common and often deadly medical condition that exerts an enormous burden on healthcare systems. Previously we showed that PKGIalpha can be activated by oxidative stress independently of the classical NO-cGMP activation pathway. This alternate activation pathway involves oxidants inducing a disulfide bond between the kinases two subunits. This pathway is also relevant to sepsis, consistent with it being a time of oxidative stress that drives formation of the disulfide. This explains in part the hypotension that occurs during sepsis and partly accounts for the associated organ injury as a result of their under-perfusion. Based on the literature and pilot data presented in this application, we think that PKGIalpha oxidation can also directly damage tissue (independently of hypotension and tissue under-perfusion) by inducing apoptosis. These studies proposed here are aimed at defining the relative contributions of these two pathways to injury during sepsis. In addition, as disulfide PKGIalpha formation underlies the damage caused by both these pathways (i.e. both by hypotension as well as apoptosis), we think pharmacological interventions that blocked oxidation would be protective. Consequently, we plan on developing and testing a number of drugs for their ability to prevent PKGIalpha oxidation and limit tissue damage during sepsis.
Planned Impact
Sepsis is a common and often deadly medical condition that exerts an enormous burden on healthcare systems and society. Current therapeutic strategies of administering antibiotics (to combat the underlying infection) or vasopressor (to elevate blood pressure) are often ineffective, with a high risk (approximately 25%) of mortality. In recent years much attention has been paid to the idea that injury during sepsis is driven by over-production of nitric oxide to detrimentally lower blood pressure by activating soluble Guanylate Cyclase and then protein kinase G (PKG). However, clinical trials preventing nitric oxide production during sepsis were ineffective and in fact increased mortality. Clearly our current understanding of the disease is lacking. By better understanding the mechanisms that underlie the dysfunction that occurs in sepsis we improve our chances of implementing or designing rational therapeutic strategies. Our previous work identified an important new way of activating PKG independently of the nitric oxide pathway, involving it becoming oxidised to form an interprotein disulfide. Sepsis is well-known as a time of oxidative stress and so PKG oxidation was anticipated to occur. Indeed, we found PKG oxidative activation would significantly explain the hypotension that occurs during sepsis. Indeed, we found sepsis induces PKG oxidation and this was associated with murine hypotension and organ damage in vivo. In contrast a Cys42Ser PKG knock-in mouse that cannot be activated by disulfide oxidation was resistant in part to sepsis-induced hypotension and ensuing organ damage. However, we now know that additional PKG oxidation signals to tissue apoptosis, and here we will define the contribution of this mechanism to damage during sepsis independently of the role of hypotension. It is evident from our findings so far that PKG oxidation causes damaging hypotension and likely also apoptosis. We hope to identify and test drugs that will prevent PKG oxidation as these should be protective and provide resistance to injury during sepsis - in the same way the knock-in mouse that cannot be oxidised to disulfide is protected. Overall we have identified a potentially important new mechanism that may be injurious during sepsis, and we propose studies that may lead to therapies that help treat this major healthcare problem.
Organisations
People |
ORCID iD |
Philip Eaton (Principal Investigator) |
Publications
Eaton P
(2014)
Preface for redox signalling in the cardiovascular system.
in Journal of molecular and cellular cardiology
Stubbert D
(2014)
Protein kinase G Ia oxidation paradoxically underlies blood pressure lowering by the reductant hydrogen sulfide.
in Hypertension (Dallas, Tex. : 1979)
Charles R
(2014)
Gel-based methods in redox proteomics.
in Biochimica et biophysica acta
Rudyk O
(2014)
Biochemical methods for monitoring protein thiol redox states in biological systems.
in Redox biology
Charles RL
(2014)
Protection from hypertension in mice by the Mediterranean diet is mediated by nitro fatty acid inhibition of soluble epoxide hydrolase.
in Proceedings of the National Academy of Sciences of the United States of America
Hohl M
(2014)
Meeting highlights from the 2013 European Society of Cardiology Heart Failure Association Winter Meeting on Translational Heart Failure Research.
in European journal of heart failure
Lorenz JE
(2014)
Oxidant-induced activation of cGMP-dependent protein kinase Ia mediates neuropathic pain after peripheral nerve injury.
in Antioxidants & redox signaling
Kohl T
(2015)
Intensity matters: Ryanodine receptor regulation during exercise.
in Proceedings of the National Academy of Sciences of the United States of America
Cameron JM
(2015)
Polarized cell motility induces hydrogen peroxide to inhibit cofilin via cysteine oxidation.
in Current biology : CB
Nakamura T
(2015)
Prevention of PKG1a oxidation augments cardioprotection in the stressed heart.
in The Journal of clinical investigation
Prysyazhna O
(2015)
Redox regulation of cGMP-dependent protein kinase Ia in the cardiovascular system.
in Frontiers in pharmacology
Burgoyne JR
(2015)
Deficient angiogenesis in redox-dead Cys17Ser PKARIa knock-in mice.
in Nature communications
Mistry RK
(2016)
Transcriptional Regulation of Cystathionine-?-Lyase in Endothelial Cells by NADPH Oxidase 4-Dependent Signaling.
in The Journal of biological chemistry
Fernandez-Caggiano M
(2016)
Oxidant-induced Interprotein Disulfide Formation in Cardiac Protein DJ-1 Occurs via an Interaction with Peroxiredoxin 2.
in The Journal of biological chemistry
Prysyazhna O
(2016)
Phosphodiesterase 5 Inhibition Limits Doxorubicin-induced Heart Failure by Attenuating Protein Kinase G Ia Oxidation.
in The Journal of biological chemistry
Fernández-Caggiano M
(2016)
Analysis of Mitochondrial Proteins in the Surviving Myocardium after Ischemia Identifies Mitochondrial Pyruvate Carrier Expression as Possible Mediator of Tissue Viability.
in Molecular & cellular proteomics : MCP
Stathopoulou K
(2016)
S-glutathiolation impairs phosphoregulation and function of cardiac myosin-binding protein C in human heart failure.
in FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Khavandi K
(2016)
Pressure-induced oxidative activation of PKG enables vasoregulation by Ca2+ sparks and BK channels.
in Science signaling
Akashi S
(2016)
Persistent Activation of cGMP-Dependent Protein Kinase by a Nitrated Cyclic Nucleotide via Site Specific Protein S-Guanylation.
in Biochemistry
Bassi R
(2017)
Redox-dependent dimerization of p38a mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3
in Journal of Biological Chemistry
Valek L
(2017)
Redox-guided axonal regrowth requires cyclic GMP dependent protein kinase 1: Implication for neuropathic pain.
in Redox biology
Donzelli S
(2017)
Oxidant sensor in the cGMP-binding pocket of PKGIa regulates nitroxyl-mediated kinase activity.
in Scientific reports
Burgoyne JR
(2017)
Proof of Principle for a Novel Class of Antihypertensives That Target the Oxidative Activation of PKG Ia (Protein Kinase G Ia).
in Hypertension (Dallas, Tex. : 1979)
Rudyk O
(2017)
Examining a role for PKG Ia oxidation in the pathogenesis of cardiovascular dysfunction during diet-induced obesity.
in Free radical biology & medicine
Wolhuter K
(2017)
How widespread is stable protein S-nitrosylation as an end-effector of protein regulation?
in Free radical biology & medicine
Description | Disulfide activated protein kinase G Ialpha as a new therapeutic target in sepsis |
Amount | £502,927 (GBP) |
Funding ID | MR/L009684/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2014 |
End | 02/2018 |
Description | Does redox state of PKG1alpha control hypoxic vasoconstriction and remodelling in the pulmonary vasculature? |
Amount | £381,870 (GBP) |
Funding ID | FS/14/57/31138 |
Organisation | British Heart Foundation (BHF) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2015 |
End | 03/2019 |
Description | Does redox state of PKG1alpha control hypoxic vasoconstriction and remodelling in the pulmonary vasculature? |
Amount | £381,870 (GBP) |
Funding ID | FS/14/57/31138 |
Organisation | British Heart Foundation (BHF) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2015 |
End | 03/2019 |
Description | Does redox state of PKG1alpha control hypoxic vasoconstriction and remodelling in the pulmonary vasculature? |
Amount | £381,870 (GBP) |
Funding ID | FS/14/57/31138 |
Organisation | British Heart Foundation (BHF) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2015 |
End | 03/2019 |
Description | ERC Advanced - CARDIOREDOX |
Amount | € 2,255,659 (EUR) |
Funding ID | 339095 |
Organisation | European Research Council (ERC) |
Sector | Public |
Country | Belgium |
Start | 12/2013 |
End | 11/2018 |
Description | Expanding our understanding of the redox regulation of soluble Epoxide Hydrolase in cardiovascular health and disease |
Amount | £253,143 (GBP) |
Funding ID | PG/15/26/31373 |
Organisation | British Heart Foundation (BHF) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2015 |
End | 03/2018 |
Description | MRC project grant |
Amount | £502,927 (GBP) |
Funding ID | MR/L009684/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 02/2014 |
End | 01/2018 |
Description | project grant |
Amount | £289,772 (GBP) |
Funding ID | MGU0581 |
Organisation | Queen Mary University of London |
Sector | Academic/University |
Country | United Kingdom |
Start | 06/2021 |
End | 05/2024 |
Title | PEG-switch |
Description | Allso reversible proetin oxidation in candidate proteins to be monitored |
Type Of Material | Technology assay or reagent |
Year Produced | 2013 |
Provided To Others? | Yes |
Impact | People can more easily and routeinely monitor protein thiol redox state |
Title | anti-sulforaphane protein adduct antibody |
Description | Allows sulforaphane-modified proteins to be monitored and identified. |
Type Of Material | Antibody |
Year Produced | 2013 |
Provided To Others? | Yes |
Impact | Has allowed novel targets of sulforaphane to be identified. |
Description | Media interest |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Media (as a channel to the public) |
Results and Impact | After we published a paper titled: Protection from hypertension in mice by the Mediterranean diet is mediated by nitro fatty acid inhibition of soluble epoxide hydrolase, we had a lot of press interest and phonecalls with jouralsits froma round the work. Here are a very few examples of the news agencies that cited the work. http://www.dailymail.co.uk/health/article-2632932/Tossing-lettuce-olive-oil-sprinkling-nuts-avocado-boosts-heart-health.html http://www.bbc.co.uk/news/health-27470115 http://www.express.co.uk/news/health/476945/Why-a-Mediterranean-diet-is-good-for-your-health http://www.telegraph.co.uk/health/healthnews/10840283/Olive-oil-on-salad-may-save-your-life.html http://www.independent.co.uk/life-style/health-and-families/health-news/why-is-a-mediterranean-diet-so-good-for-you-scientists-think-they-know-9400889.html Apart from substantive interest from all types of news agencies, we even had members of the public get in contact to discuss the findings. |
Year(s) Of Engagement Activity | 2014 |
URL | http://www.bbc.co.uk/news/health-27470115 |