Borooah, Shyamanga

Burr, Karen

Chandran, Siddharthan

Connick, Peter

Hardingham, Giles

Livesey, Matthew

Patani, Rickie

Rzechorzek, Nina Marie

Story, David

Wyllie, David

English

Cooling is the single most effective treatment for acute neuronal injury. Understanding the molecular mechanisms that mediate cooling-induced neuroprotection might yield novel therapeutic targets for neurodegenerative disease. Many of these disorders involve modulation of tau, a protein that is enriched in neurons and becomes hyperphosphorylated in hypothermic, injury-resistant rodent brains as well as in Alzheimer’s disease. We sought to establish a new model for exploring human tau physiology and its role in neuroprotection in the context of therapeutic hypothermia.  Functional cortical neurons (hCNs) were differentiated from three independent human pluripotent stem cell lines and validated for regional identity and tau status. Matched cultures were incubated at 37°C or clinically targeted temperatures for therapeutic hypothermia (32°C) and suspended animation (28°C). Effects of cooling on tau status and neuronal injury elicited by common neurotoxic stressors were established. Injury experiments were repeated while manipulating tau phosphorylation.  hCN differentiation featured transitions in tau status, recapitulating transcriptional and post-translational human in-vivo cortical tau development. Key aspects of this development were reversed by cooling. Notably, cooling induced tau hyperphosphorylation via rapid inhibition of the major tau phosphatase, protein phosphatase 2A (PP2A). Multiplexed injury analysis confirmed that hypothermia robustly protected hCNs from oxidative (100 μM hydrogen peroxide) and excitotoxic (30 μM glutamate) stress (at 28°C, injury was reduced by 78% and 56%, respectively; p&lt;0·0005). Selective block of a major tau kinase reduced hCN tau phosphorylation and abrogated hypothermic protection (p=0·001) against oxidative injury at 28°C (injury reduction diminished to 9% at 100 μM hydrogen peroxide), whereas pharmacological inhibition of PP2A mimicked cooling-induced tau phosphorylation and protected hCNs from oxidative injury at 37°C (up to 22% injury reduction [p=0·008], with no benefit under hypothermic conditions). To our knowledge, this is the first study of human neuronal tau physiology under hypothermic conditions. Although definitive experiments are needed, our findings support previous work linking phospho-tau to neuroprotection. Furthermore, cold-induced tau hyperphosphorylation is a potential trigger for proteostatic priming, a recently discovered mechanism of hypothermic preconditioning in hCNs. Exploiting these cryobiological effects might lead to new treatments for acute and chronic neuronal injury

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     Edinburgh Research Explorer                                      Hypothermic modulation of human cortical neurons to explore arole for tau protein in neuroprotectionCitation for published version:Rzechorzek, NM, Connick, P, Livesey, M, Borooah, S, Patani, R, Burr, K, Story, D, Wyllie, D, Hardingham,G & Chandran, S 2016, 'Hypothermic modulation of human cortical neurons to explore a role for tau proteinin neuroprotection', The Lancet, vol. 387, pp. S88.Link:Link to publication record in Edinburgh Research ExplorerDocument Version:Publisher's PDF, also known as Version of recordPublished In:The LancetGeneral rightsCopyright for the publications made accessible via the Edinburgh Research Explorer is retained by the author(s)and / or other copyright owners and it is a condition of accessing these publications that users recognise andabide by the legal requirements associated with these rights.Take down policyThe University of Edinburgh has made every reasonable effort to ensure that Edinburgh Research Explorercontent complies with UK legislation. If you believe that the public display of this file breaches copyright pleasecontact openaccess@ed.ac.uk providing details, and we will remove access to the work immediately andinvestigate your claim.Download date: 23. Jul. 2021Poster Abstracts88 www.thelancet.com   Hypothermic modulation of human cortical neurons to explore a role for tau protein in neuroprotectionNina M Rzechorzek, Peter Connick, Matthew R Livesey, Shyamanga Borooah, Rickie Patani, Karen Burr, David Story, David J A Wyllie, Giles E Hardingham, Siddharthan ChandranAbstract Background Cooling is the single most eff ective treatment for acute neuronal injury. Understanding the molecular mechanisms that mediate cooling-induced neuroprotection might yield novel therapeutic targets for neurodegenerative disease. Many of these disorders involve modulation of tau, a protein that is enriched in neurons and becomes hyperphosphorylated in hypothermic, injury-resistant rodent brains as well as in Alzheimer’s disease. We sought to establish a new model for exploring human tau physiology and its role in neuroprotection in the context of therapeutic hypothermia.Methods Functional cortical neurons (hCNs) were diff erentiated from three independent human pluripotent stem-cell lines and validated for regional identity and tau status. Matched cultures were incubated at 37°C or clinically targeted temperatures for therapeutic hypothermia (32°C) and suspended animation (28°C). Eff ects of cooling on tau status and neuronal injury elicited by common neurotoxic stressors were established. Injury experiments were repeated while manipulating tau phosphorylation.Findings hCN diff erentiation featured transitions in tau status, recapitulating transcriptional and post-translational human in-vivo cortical tau development. Key aspects of this development were reversed by cooling. Notably, cooling induced tau hyperphosphorylation via rapid inhibition of the major tau phosphatase, protein phosphatase 2A (PP2A). Multiplexed injury analysis confi rmed that hypothermia robustly protected hCNs from oxidative (100 μM hydrogen peroxide) and excitotoxic (30 μM glutamate) stress (at 28°C, injury was reduced by 78% and 56%, respectively; p<0·0005). Selective block of a major tau kinase reduced hCN tau phosphorylation and abrogated hypothermic protection (p=0·001) against oxidative injury at 28°C (injury reduction diminished to 9% at 100 μM hydrogen peroxide), whereas pharmacological inhibition of PP2A mimicked cooling-induced tau phosphorylation and protected hCNs from oxidative injury at 37°C (up to 22% injury reduction [p=0·008], with no benefi t under hypothermic conditions).Interpretation To our knowledge, this is the fi rst study of human neuronal tau physiology under hypothermic conditions. Although defi nitive experiments are needed, our fi ndings support previous work linking phospho-tau to neuroprotection. Furthermore, cold-induced tau hyperphosphorylation is a potential trigger for proteostatic priming, a recently discovered mechanism of hypothermic preconditioning in hCNs. Exploiting these cryobiological eff ects might lead to new treatments for acute and chronic neuronal injury.Funding Wellcome Trust, Anne Rowling Regenerative Neurology Clinic, Euan MacDonald Centre for Motor Neurone Disease Research.ContributorsNMR conceived the study, devised the methodology, conducted investigations, analysed the data, acquired funding and resources, and wrote the fi rst draft of the abstract. PC devised the methodology, analysed the data, acquired funding and resources, and reviewed and edited the abstract. MRL conducted investigations, analysed the data, and reviewed and edited the abstract. RP, DS, and KB reviewed and edited the abstract. SB and DJAW acquired resources and reviewed and edited the abstract. GEH and SC acquired funding and resources, reviewed and edited the abstract, and supervised the study.Declaration of interestsWe declare no competing interests.Published OnlineFebruary 25, 2016Poster 44Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh, UK (N M Rzechorzek PhD, Prof S Chandran PhD); MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK (N M Rzechorzek, S Borooah MRCP(UK), K Burr PhD, D Story BSc, Prof S Chandran); Anne Rowling Regenerative Neurology Clinic, University of Edinburgh, Edinburgh, UK (P Connick PhD, S Borooah, R Patani PhD, Prof S Chandran); Euan MacDonald Centre for MND Research, University of Edinburgh, Edinburgh, UK (N M Rzechorzek, M R Livesey PhD, S Borooah, K Burr, D Story, Prof D J A Wyllie PhD, Prof S Chandran); Centre for Integrative Physiology, University of Edinburgh, Edinburgh, UK (M R Livesey, Prof D J A Wyllie, Prof G E Hardingham PhD); and Institute of Neurology, University College London, London, UK (R Patani)Correspondence to: Dr Nina M Rzechorzek, Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor’s Building, Edinburgh EH16 4BS, UKnina.rzechorzek@ed.ac.ukThe full paper is published in eBiomedicine http://dx.doi.org/10.1016/j.ebiom.2015.12.010

Hypothermic modulation of human cortical neurons to explore a role for tau protein in neuroprotection

https://www.research.ed.ac.uk/files/24712462/Rzechorzek_et_al_Hypothermic_modulation_of_human_cortical_neurons_to_explore_a_role_for_tau_protein_in_neuroprotection.pdf

Hypothermic modulation of human cortical neurons to explore a role for tau protein in neuroprotection

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