CoEN 4007- Is prion like propagation of alpha synuclein aggregation associated with a ferroptotic cell death

Lead Research Organisation: Newcastle University
Department Name: Translational and Clinical Res Institute

Abstract

This project proposes an innovative study that could rapidly lead to neuroprotective treatments
against Parkinson's disease, a disorder that affects millions of people within our ageing population.
Current available drugs attempt to only treat the disease symptoms not its etiology. We base our
project upon recent findings concerning two main features involved in the targeted death of
specialized neurons in the brain that secrete a neurotransmitter called dopamine. The first is the
abnormal aggregation of a protein called alpha-synuclein that progressively spreads throughout the
brain in correlation with the dopaminergic cell death. The second is the involvement of a newly
discovered pathway of cell death that we have observed in dopaminergic neurons called ferroptosis.
Both phenomena are related with iron; alpha-synuclein is involved in cellular iron balance with its
aggregation state modified by the presence of iron, and ferroptosis is an iron dependent process.
Thus this project proposes to elucidate how the modifications to alpha-synuclein observed in Parkinson's
disease can promote ferroptosis to exacerbate the spreading of neuron death throughout the brain.
Outcomes expected from this work are innovative therapeutic strategies based upon recently
generated anti-ferroptotic drugs.

Technical Summary

Cytoplasmic inclusions of aggregated alpha-synuclein (alpha-syn) are the major constituent of Lewy bodies;
a pathological hallmark of synucleinopathies including Parkinson's disease (PD). Within certain
biological environments alpha-syn self promotes aggregation and neuronal death in a prion-like mode of
action. In alpha-syn mutations (i.e. familial PD) and various posttranslational modifications increase the
propensity for aggregation, preventing the monomer from functioning as a key regulator of synaptic
and endosomal vesicle trafficking. This disrupts dopamine compartmentalization and iron
importation, leading to harmful dopamine auto-oxidation and iron accumulation; both prevalent
features in PD. Recently, we have shown that a novel regulated necrotic cell death pathway called
ferroptosis, defined by iron-dependent lipid peroxidation, is predominant in pro-oxidant models. Our
current hypothesis is that alpha-syn dysfunction promotes ferroptosis via iron and dopamine
dyshomeostasis, leading to a cellular environment that generates a conformational form of alpha-syn
required for prion-like propagation. We aim to demonstrate a pivotal interplay between alpha-syn, iron
and ferroptosis using translational studies to genetically and pharmacologically modulate ferroptosis
together with clinical longitudinal studies in patients. Outcomes are expected to identify relevant wet
(cerebro-spinal fluid) and dry biomarkers (MRI) as well as pave the way to future innovative
therapeutic strategies based upon upcoming anti-ferroptotic drugs.

Planned Impact

Recently, we have shown that a novel regulated necrotic cell death pathway called
ferroptosis, defined by iron-dependent lipid peroxidation, is predominant in pro-oxidant models. Our
current hypothesis is that alpha-syn dysfunction promotes ferroptosis via iron and dopamine
dyshomeostasis, leading to a cellular environment that generates a conformational form of alpha-syn
required for prion-like propagation.
This project will validate our novel hypothesis on the implications of ferroptosis in the
synucleinopathy process (i.e. oligomers formation, aggregation and propagation). We then expect to
demonstrate ferroptosis as a viable therapeutic target as well as identify suitable therapeutic
candidates in protection against synucleinopathy. Clinical, biological and imaging readouts will
identify changes during natural progression associated with ferroptosis and synucleionopathy. As alpha-
syn, iron, glutathione and dopamine are biochemically linked with ferroptosis, combined detection
holds significant potential as a non-invasive biomarker panel of severity. These findings could be
clinically useful in predicting disease trajectory and provide a foundation for anti-ferroptotic drug
administration to subjects with risk factors in the disease. Promising outcomes from our pre-clinical
studies will lead to future clinical trials similar to those led by the consortium. Encouragement for
the manufacturer (ApoPharma) to pursue registration of this compound in such trials (i.e.
Alzheimer: 3D trial, ALS: FAIR-ALS-II), possibly in collaboration with big pharma, will also
encourage the development of other novel iron chelators (i.e. hydroxypyridinones) and ferroptosis
inhibitors (i.e. liproxstatin-1, Ebselen) as next generation therapeutics with a better safety profile.
HCS pharma (Lille, France), the University of Munich and their industrial transfers are currently
developing a range of new ferroptosis inhibitors with potential neuroprotective capabilities that will
be suitable for patients with Parkinson disease.

Publications

10 25 50
 
Description CoEN Funding Partners 
Organisation French National Research Agency
Country France 
Sector Public 
PI Contribution Each of these funding partners have collaborated to secure the funds for the PRION-IRON project. Each of the CoEN funding partners is represented by a lead applicant, who has expertise in the field of Parkinson's Disease and responsibility for the individually awarded grant. In each case, the partnership works to ensure the highest level of expertise in the field and sharing of data to move the project forward, including education and training of staff at the relevant sites. Each partner will have responsibility for measuring outcomes relevant to the primary objective, i.e, to determine the role of the abnormal form of the protein alpha-synuclein in Parkinson's disease in promoting ferroptosis in the brain and the resulting neuronal degeneration. This will be achieved by collecting clinical data and through the identification of relevant biomarkers, including cerebrospinal fluid and MRI.
Collaborator Contribution The collaboration with the named partners has ultimately lead to the implementation of additional work packages for the identification of relevant biomarkers, including cerebrospinal fluid and MRI, within the FAIRPARK II trial, investigating the effect of deferiprone, a drug which binds excess iron/anti-ferroptotic drug, in a double-blind placebo controlled phase II European trial. Each site has recruited Parkinson's patients, not yet on dopaminergic therapy, to assess the potentially disease-modifying effect of deferiprone on the disease process underlying Parkinson's disease. The implementation and running of this trail and the additional work packages is made possible through the collaborative partnerships highlighted and the main objective is the development of a neuroprotective treatment for Parkinson's disease.
Impact The FAIRPARK II trial is an ongoing phase II trial. This trial is still actively recruiting and currently there has not been any quantitative data as the trial is still in the early stage. We would hope to have results in the near future, which may confirm the potential for slowing progression in Parkinson's disease by targeting excess iron in relevant parts of the brain. In this way, the trial through this collaborative partnership would lead to a neuroprotective treatment against Parkinson's disease. For Work package 1. Does treatment with preformed oligomeric a-syn and/or Fe2+ activate the ferroptosis pathway, resulting in lipid peroxidation? Considerable work has been undertaken and demonstrates that 3D differentiated SH-SY5Y are less susceptible to iron-induced cell death than undifferentiated cells. Cell viability assay of LUHMES with Erastin demonstrates dramatic cell death in contrast to SH-SY5Y and Erastin. VDAC expression is negligible in SH-SY5Y in comparison to LUHMES cells indicating the ferroptosis pathway is down-regulated. Metal uptake in 3D differentiated SH-SY5Y increases the intracellular concentration of a-syn and decreases GPX4, with a significant reduction in the ratio of GSH:GSSG being observed. The decrease in the GSH:GSSG ratio would indicate that oxidative stress is occurring in the cell lines following metal treatment. Although markers of oxidative stress are observed iron treatment shows defined cells that are positive for lipid peroxidation although the number of cells is few. Activation of the ferroptosis pathway and the effects of oxidative stress following oligomer treatment are now ongoing in both 3D differentiated SH-SY5Y and LUHES cells.
Start Year 2018
 
Description CoEN Funding Partners 
Organisation German Centre for Neurodegenerative Diseases
Country Germany 
Sector Public 
PI Contribution Each of these funding partners have collaborated to secure the funds for the PRION-IRON project. Each of the CoEN funding partners is represented by a lead applicant, who has expertise in the field of Parkinson's Disease and responsibility for the individually awarded grant. In each case, the partnership works to ensure the highest level of expertise in the field and sharing of data to move the project forward, including education and training of staff at the relevant sites. Each partner will have responsibility for measuring outcomes relevant to the primary objective, i.e, to determine the role of the abnormal form of the protein alpha-synuclein in Parkinson's disease in promoting ferroptosis in the brain and the resulting neuronal degeneration. This will be achieved by collecting clinical data and through the identification of relevant biomarkers, including cerebrospinal fluid and MRI.
Collaborator Contribution The collaboration with the named partners has ultimately lead to the implementation of additional work packages for the identification of relevant biomarkers, including cerebrospinal fluid and MRI, within the FAIRPARK II trial, investigating the effect of deferiprone, a drug which binds excess iron/anti-ferroptotic drug, in a double-blind placebo controlled phase II European trial. Each site has recruited Parkinson's patients, not yet on dopaminergic therapy, to assess the potentially disease-modifying effect of deferiprone on the disease process underlying Parkinson's disease. The implementation and running of this trail and the additional work packages is made possible through the collaborative partnerships highlighted and the main objective is the development of a neuroprotective treatment for Parkinson's disease.
Impact The FAIRPARK II trial is an ongoing phase II trial. This trial is still actively recruiting and currently there has not been any quantitative data as the trial is still in the early stage. We would hope to have results in the near future, which may confirm the potential for slowing progression in Parkinson's disease by targeting excess iron in relevant parts of the brain. In this way, the trial through this collaborative partnership would lead to a neuroprotective treatment against Parkinson's disease. For Work package 1. Does treatment with preformed oligomeric a-syn and/or Fe2+ activate the ferroptosis pathway, resulting in lipid peroxidation? Considerable work has been undertaken and demonstrates that 3D differentiated SH-SY5Y are less susceptible to iron-induced cell death than undifferentiated cells. Cell viability assay of LUHMES with Erastin demonstrates dramatic cell death in contrast to SH-SY5Y and Erastin. VDAC expression is negligible in SH-SY5Y in comparison to LUHMES cells indicating the ferroptosis pathway is down-regulated. Metal uptake in 3D differentiated SH-SY5Y increases the intracellular concentration of a-syn and decreases GPX4, with a significant reduction in the ratio of GSH:GSSG being observed. The decrease in the GSH:GSSG ratio would indicate that oxidative stress is occurring in the cell lines following metal treatment. Although markers of oxidative stress are observed iron treatment shows defined cells that are positive for lipid peroxidation although the number of cells is few. Activation of the ferroptosis pathway and the effects of oxidative stress following oligomer treatment are now ongoing in both 3D differentiated SH-SY5Y and LUHES cells.
Start Year 2018
 
Description 2. Y-Move, Yorkshire Movement Disorder Group. "Treatment of Parkinson's Disease. A Practical Update" Lees, UK, 5th December 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact A Practical Update on Treatment of Parkinson's Disease to consultant Neurologists. We discussed recent clinical trial, including preliminary results of the effect of deferiprone in patients with Parkinson. Discussed MRI biomarkers of Parkinson's.
Year(s) Of Engagement Activity 2019
 
Description Education Afternoon for patents recently diagnosed with Parkinson's disease 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact Description of research projects running at the Clinical Ageing Research Unit. Dissemination of preliminary results with Deferiprone in Parkinson's disease.
Year(s) Of Engagement Activity 2019
 
Description MSA Trust Study Day, Newcastle 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact This was an event to discuss the condition and clinical treatment of MSA, including a multi-disciplinary approach. We were invited to discuss the current state of research in MSA and associated conditions, and specifically Clinical Research Fellow, Dr Laura Best, did a presentation specifically on this project. This invited much discussion and increased interest in the application, and emphasised importance of developing neuroimaging in these conditions. We also were able to highlight the study to the other healthcare professionals who were present and worked in the field of Parkinsonism, which we hope will be beneficial with regard to recruitment.
Year(s) Of Engagement Activity 2018
 
Description PD UK research roadshow: Penrith, Cumbria 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact The purpose of this event was primarily for the education of patient, care-givers and professionals involved with Parkinson's disease. It consisted of brief presentations by invited speakers, which included specialist clinicians and researchers, followed by discussions in small groups. This allowed the audience, primarily patients and their families, to directly ask the experts questions about ongoing research in Parkinson's disease. It also afforded the opportunity for patients to become involved and express an interest in being involved in studies such as our own. We were able to chat about our study in detail and its rationale, and get a sense that patients felt it was important in moving the field of Parkinson's research forward.
Year(s) Of Engagement Activity 2018
 
Description Parkinson's UK research roadshow: Durham 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact This was again an event designed to promote research in Parkinson's Disease. This included formal presentations by experts in this field, as well as, informal discussion in small groups with patients, affiliated professionals and carers regarding current projects funded by PD UK and how to get involved. This sparked a lot of questions and discussions with the audience with regard to our project, with many patients interested in getting involved and putting their names forward for further information and potential recruitment.
Year(s) Of Engagement Activity 2018
 
Description Participation in New PD patient education afternoons, based at the CRESTA clinic, Newcastle Upon Tyne 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Patients, carers and/or patient groups
Results and Impact The Regional Movement Disorder clinic, based at the CRESTA clinic in Newcastle Upon Tyne, organises a 'New Patient Education Afternoon' every 2-3 months. This is ran by the movement disorder Specialist Consultants, Neurology Registrars and PD Specialist Nurses. It is also attended by the Speech and Language Team, who work closely with service, and Specialist Physiotherapists with expertise in PD management. A member of our research team, Dr Laura Best, attends these meetings to educate patients and their carers regarding active research in the until, as well as some of the science underpinning the PD diagnosis and its treatment. These educational afternoons are also an opportunity for patients to express an interest in becoming involved in active research and has been one avenue for recruitment to this study.
Year(s) Of Engagement Activity 2018,2019
 
Description Presentation at invited talk Copenhagen University 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Invited talks on neuroimaging findings in prodromal Parkinson's
Year(s) Of Engagement Activity 2020
 
Description Presentation at invited talk First Parkinson's disease and movement disorders Congress organised in Romania. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation at invited talk First Parkinson's disease and movement disorders Congress organised in Romania.
Year(s) Of Engagement Activity 2020
 
Description Royal College of Paediatrics and Child Health - President's Day 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Research in the Northeast: Ageing. Presentation of preliminary findings of Deferiprone in Parkinson's disease. MRI biomarker of Parkinson's.
The audience consisted of Consultants paediatrician. Interesting discussion on disease modifying agents in Parkinson's disease.
Year(s) Of Engagement Activity 2019