Preeclampsia occurs in 3-8% of all pregnancies causing rapid hypertension and kidney dysfunction in pregnant woman which is difficult to predict and untreatable. Moreover, it predisposes both mother and baby to lifelong risk of cardiovascular disease. Recent advances identified an imbalance of angiogenic factors as pivotal to the development of preeclampsia. Oxidative stress and endothelial dysfunction are major factors that and strongly associated with preeclampsia, however, the molecular pathways associated with oxidative stress are unknown. We recently linked oxidative signalling with regulation of angiogenic factors in ischemia1,2. Oxidative stress is sensed within the cell by modulating redox-sensitive cysteines on proteins forming diverse oxidative post-translational modifications which regulate intracellular signalling. Dr Murdoch is coordinating a Horizon 2020 grant, Innovative Modelling of Placenta for Foetal Maternal Health (iPLACENTA.eu)3, collaborating with industry to develop new innovative ways to model and understand preeclampsia (generating 3D placenta-on-a-chip and an integrative systems biology approach).
This PhD will aim to establish how oxidative stress regulates the production of antiangiogenic factors in preeclampsia using multicellular and physiological models. The project will combine cutting edge multidisciplinary techniques including CRISPr/CAS9 genetic engineering, 3D-cell-modelling, microfluidics, stem cells, cardiovascular physiology and proteomics.
A combination of iPSc and CRISPr/CAS9 gene editing technology will be integrated into a 3D placenta-on-a-chip model to mimic oxidative stress observed in preeclampsia. Validation and identification of target pathways will take place by coupling with advanced assay systems for analytical measurement of vascular factors and function (angiogenesis). Proteomic screening (Iodo-TMT) developed by Dr Henderson will determine targets that undergo oxidative post-translational modifications4. To establish proof of concept, comprehensive physiological assessment will take place to phenotype the disease state using cell specific genetically modified mice and surgical models. Cutting-edge methodology assessing; cardiac function via pressure volume loops and echocardiography; vascular function via laser Doppler and blood pressure via telemetry 5. Imaging of novel reporter mice6 (Henderson) will assess the location of oxidative signalling.
The project provides exposure for the student to translational research spanning cell based complex models to physiological studies and has the potential to convert to iCASE award through collaborations with industry partners.
1. Watanabe Y, Murdoch CE, Sano S, et al. Glutathione adducts induced by ischemia and deletion of glutaredoxin-1 stabilize HIF-1α and improve limb revascularization. Proc Natl Acad Sci. 2016;(13):201524198. doi:10.1073/pnas.1524198113
2. Murdoch CE, Shuler M, Haeussler DJ, et al. Glutaredoxin-1 up-regulation induces soluble vascular endothelial growth factor receptor 1, attenuating post-ischemia limb revascularization. J Biol Chem. 2014. doi:10.1074/jbc.M113.517219
3. Murdoch CE. The Science Of Safer Pregnancy: How Mini “Placentas On A Chip” May Help Us Understand Pre-eclampsia. Huffington Post UK. http://www.huffingtonpost.co.uk/dr-colin-murdoch/the-science-of-safer-preg_b_18285082.html?guccounter=1&guce_referrer_us=aHR0cHM6Ly93d3cuZ29vZ2xlLmNvLnVrLw&guce_referrer_cs=rs5DsrcAw67MaXxuR1G7hQ
. Published November 17, 2017.
4. McGarry DJ, Chakravarty P, Wolf CR, Henderson CJ. Altered protein S-glutathionylation identifies a potential mechanism of resistance to acetaminophen-induced hepatotoxicity. J Pharmacol Exp Ther. 2015;355(2):137-144. doi:10.1124/jpet.115.227389
5. Murdoch CE, Chaubey S, Zeng L, et al. Endothelial NADPH oxidase-2 promotes interstitial cardiac fibrosis and diastolic dysfunction through pro-inflammatory effects and endothelial-mesenchymal transition. J Am Coll Cardiol. 2014;63(24):2734-2741. doi:10.1016/j.jacc.2014.02.572
6. McMahon M, Ding S, Acosta-Jimenez LP, Frangova TG, Henderson CJ, Wolf CR. Measuring in vivo responses to endogenous and exogenous oxidative stress using a novel haem oxygenase 1 reporter mouse. J Physiol. 2018;596(1):105-127. doi:10.1113/JP274915