Role of the mitochondrial Translocator Protein (mTSPO) in cell homeostasis: a molecular pathway in signalling and self-conservation mechanisms

Lead Research Organisation: Royal Veterinary College
Department Name: Comparative Biomedical Sciences CBS

Abstract

In search of the molecules that may set the pace of cell metabolism and define its healthy homeostasis, we are now interested in elucidating the cell biological role of the mTSPO. Although associated with many pathophysiological conditions, its contribution to processes guarding cell integrity such as autophagy and apoptosis is obscure, and its role in mitochondrial performance ill-defined. In order to shed light on these two fundamental processes, it will be instructive to ascertain how mTSPO ratio of expression with that of the VDAC, and its pharmacological modulation, defines the signalling of Ca2+, dynamics of the ROS, ATP rationing and Redox State. The physiology of cellular Ca2+, the ATP balance, the programmed apoptotic cell death as well as the Autophagy all depend on the efficient handling of Ca2+ by the mitochondrion. Despite some minor attempts, however, the contribution of mTSPO to this pathway of signalling has been largely ignored, and we now aim to explore and elucidate it, considering this as the hub in the mTSPO contribution to mitochondrial coupling by which ATP synthesis, ROS generation and network remodelling, via targeted autophagy, originate. However, modifications in ROS signalling would impinge also on downstream effectors: the kinases which promote the unphysiological phosphorylation of VDAC. This indirect pathway, as well as the direct physical interaction between mTSPO and VDAC, will therefore be addressed as potential underlying mechanisms. The following experiments will clarify the above points using Mouse Embryonic Fibroblasts (MEF) cell lines: 1. Live imaging, luminescent and fluorescence based methods will be used to assess mTSPO role in cell autopagy and signalling. a) The mTSPO protein will be transiently manipulated to increase (+) or decrease (-) its expression via conventional techniques of molecular biology; the outcome of this manipulation will then be ascertained on b) Autophagy, c) Ca2+ signalling d) ROS generation and e) Autophagy in conditions of buffered Ca2+ and ROS. 2. Confocal Microscopy, fluorescence and biochemical based methods will be used to determine the role of mTSPO in mitochondrial coupling efficiency and remodelling. After transient modulation of mTSPO or pharmacological binding the following will be addressed: a) The mitochondrial redox state, b) ATP generation, and c) Mitochondrial membrane potential, will be evaluated; d) The volume of the mitochondrial network will be calculated; e) The expression of genes associated with the organelle's genesis will be profiled, and f) The activity of mitochondria-triggered autophagy will be investigated. 3. Live cell imaging and standard assays will test the kinases' activity, VDAC phosphorylation which this generates, and the outcome in mitochondrial regulated apoptotic demise. Following molecular and pharmacological modulation of mTSPO: a) The efficiency of translocation and activity of PKA and PKCepsilon will be evaluated; b) The phosphorylation state of VDAC revealed; and c) The efficiency of mitochondrial-dependent apoptosis assessed. This object will also see the utilization of MEF cell lines from PKCepsilon KO mice. 4. Immunohistochemical, biochemical and yeast based methods will be employed to identify the molecular interplay between mTSPO and VDAC: a) Cellular co-localization between mTSPO and VDAC isoforms will be monitored; b) The interaction and its type studied via two-hybrid strategy and co-immunoprecipitation; and c) The homo-oligomers of VDAC evaluated. All this will reveal the molecular and cellular physiology of an important but so far neglected regulatory pathway. The results of our studies will form a conceptual and experimental framework for future investigations aiming to identify in mTSPO a novel bio-marker and target for pharmacological intervention of conditions associated with remodelling of metabolism and proliferation.

Technical Summary

By manipulating mTSPO expression or pharmacologically inhibiting its function in Murine Fibroblasts we aim to assess: 1) Cell autophagy and signalling; 2) Mitochondrial homeostasis and remodelling; 3) Kinase-mediated phosphorylation of VDAC and the result on apoptosis, and 4) Nature of mTSPO and VDAC interaction. Seventeen experiments were designed which break down as follows: I. 1. By techniques of molecular biology modify mTSPO up and down regulating its expression with recombinant protein and short hairpin RNA; 2. By combining means of imaging and biochemistry define the outcome of this on cell autophagy; 3. Evaluate the homeostasis of Ca2+ with fluorescence and luminescence tools. Monitor 4. The intracellular level of ROS; 5. Autophagic activity after Ca2+ and ROS neutralization. II. 6. Mitochondrial pools of NAD will be evaluated via confocal analysis. 7. Both cell imaging and luminescent techniques will be used to assess ATP generation. 8. Inner mitochondrial membrane potential will be imaged and 9. Via combined fluorescent probes and dyes the volume fraction of the organelles pondered. 10. The genes responsible for the biogenesis of the mitochondria expansion will be mapped with RT-QPCR and Immunoblotting as well as 11. Targeted autophagosomes investigated. III. 12. Test PKA and PKCepsilon activity and mitochondrial localization following activation in cells manipulated for mTSPO expression in response to PK11195 treatment; in the same cells 13. Analyze by immunoblotting VDAC phosphorylation and 14. Exploit the results of this conformational change on the intrinsic pathway of apoptosis. IV. 15. The co-localization of mTSPO and VDAC will be scrutinized by immunofluorescence and immunoblotting. 16. The interaction between the two proteins profiled by conventional immunoprecipitation and yeast two-hybrid system. Lastly, 17. Clarify via biochemical analysis if VDAC homo-oligomers are affected by mTSPO modified expression or pharmacological modulation.

Planned Impact

The cell biology of the mTSPO remains enigmatic and unexplored although the literature provides evidence for its involvement in cholesterol transport, synthesis of steroid hormones, apoptosis, cell proliferation and immunomodulation. Binding of the mTSPO with [11C](R)-PK11195 via positron emission tomography (PET) is routinely employed to diagnose inflammatory states of the Central Nervous System (CNS) as mTSPO level of expression is increased in activated microglia. mTSPO ligands are also used in experimental protocols to mark cancerous cells where mTSPO is remarkably over-expressed. In spite of that, the actual contribution of mTSPO in regulation of metabolism and mechanisms to avoid cellular pathologies remains unexplained. This proposal aims to address these fundamental aspects of cell physiology associated with mTSPO and to inform novel approaches in which mTSPO can be considered in the context of the diagnosis and treatment of pathologies. We have therefore identified the following principal points of impact for this study: I. The growing interest by pharmaceutical and biotechnological companies in the outcomes of our study will be further exploited, and strategies to foster collaborative work established. The first aim is to create a bond with partners for the creation of protocols for clinical purposes in order to consolidate mTSPO as read-out of pathological conditions, as well as functional assays to assess the efficacy of therapeutic agents acting on it. Nevertheless, we are currently working to identify the ideal partner to present a case for a PhD studentship to increase the number of people involved in the subject. II. The data generated will be pubblished by various activities set ad hoc for the project. The relevance of this fundamental research to major diseases means it has general and popular interest, contributing to greater public awareness and understanding of science. III. A thorough understanding of the protein's cell biology will assist specialists developing therapies directed to this protein. The eventual patenting of the intellectual property that may arise from this project will be provided by the appointed organisation of the college. This will be made possible by the close involvement of the Enterprise Team of the Royal Veterinary College, from the earliest stages in research project development, which will provide assistance in impact planning, identification of and liaison with company partners as well as in formalizing commercial contracts. Pro-active market research and marketing are used to identify and develop relationships with potential licensee businesses. Proof-of-concept funding will be accessed, whenever appropriate and available, for market research and technical development to enhance the appeal of new developments for commercial partners. IV. The major findings from this project will be disseminated accordingly and people involved in the research will contribute to that by providing interviews or press quotes as appropriate. To this end, we will also benefit from the Communications Management Team who will advise on the creation of web-based material for publicising the work to a general audience. V. Staff appointed to run this project will benefit from highly qualified training, becoming skilled in cutting-edge techniques for cell-based investigation. In conclusion, the results of this work will lead to a greater understanding of molecular regulators of cell energy and quality. A deeper comprhension of the mTSPO role will enable novel approaches to diagnose and manage diseases related to alterations of pathways associated to malfunctions of autophagy, metabolism and apoptosis. Hence the economy (e.g. Pharmaceutical and Biotechnological companies) may benefit from this study as well as the quality of life of cohorts of patients by the development of novel therapeutics.

Publications

10 25 50
 
Description Mitochondria play a vital role in the wellbeing of mammals. Quality of their function and hence their ability to master signaling events occurring in their hosting environment (the cell) are at the basis of any biological function which impacts at systemic level.
With the completion of this program of research we have demonstrated that TSPO expression ration in mitochondria contributes the quality of vital intracellular signals such as Ca2+, the Reactive Oxygen Species (ROS) and that of the whole pace of mitochondrial bio-energy. In addition TSPO accumulation alters the morphological aspects of the mitochondria.
The scientific evidences collected during the grant have demonstrated that TSPO can really be seen as a molecular regulator of the mitochondrial channel (VDAC) via which metabolites pass from the cytosol into the organelle: a vital step of the organelle's regulation. We have indeed been able to demonstrate in the physical interaction between TSPO and VDAC the drive of the outcome on mitochondrial and cellular physiology.
TSPO, when accumulated in the mitochondria, prevents the execution of organelle-mediated apoptosis thus underlying a pathological cellular proliferation sustained by the oxidative stress caused by the parallel inhibition of mitophagy.


In detail we have been able to demonstrated that TSPO:
1- Plays part in cell autophagy regulating the mitochondrial type of it.
2- Affects mitochondrial metabolism and structure reducing the efficient coupling of the organelle as well as the network dynamics.
3- Prevents mitochondrial apoptosis causing the phosphorylation of VDAC via the activation of the cellular Kinases.
4- Directly and indirectly affects the degree of VDAC phosphorylation regulating its molecular physiology.

We are therefore confident that our work has brought to light a regulatory molecular pathway which, by defining cell metabolism as well as the fundamental mechanisms of cell integrity preservation, impacts the wellbeing of tissues and mammals outlying strategies of intervention.
Exploitation Route Based on the work produced and results obtained we are highlighting below the specific aspects in which we can predict further development in the years to come

(1) Mitochondrial Quality control and Ageing. The biological evidences obtained with the support of this grant have lead towards the exploitation of TSPO as a potential time-reader of mitochondrial ageing thus making the molecule a valuable target for cutting-edge diagnostic of cell and tissues senescence. This is part of the work we are keen to progress with colleagues based at Imanova, King's College London and Imperial College active in the clinical diagnostic.

(2) Cell Metabolism and Deterioration. The knowledge we have obtained with this grant will be of assistance to perfect the tracers currently used in Positron Emission Tomography studies an din this way improve the quality of their reading. Specifically, the accumulation of cellular free species of the oxygen (ROS), which we found depending on TSPO accumulation, may be the cause of the unclear signal recorded in occasions and therefore stimulate the investigation of protocols to combine anti-oxidant treatment to standards means of PET tracers infusion.

(3) Apoptosis Regulation. The efficacy of chemotherapy could be improved by combining the treatment with standard anti-cancer drugs with that of compounds active on TSPO. The pharmacological "masking" of TSPO could indeed represent a means to improve the efficacy of cancer treatment. Pilot studies are now running to maximise the fundamental discoveries matured from this grant.

(4) Methodological Improvements. The degree of the mitochondrial accumulation of TSPO could per se represent a methodological advancement to score mitochondria that have evaded the process of cell mitophagy. However, the characterization of the pro-mitophagic tool PMI will become vastly utilized by colleagues engaged in the understanding of mitophagy and its exploitation for therapeutic purposes. This is another great merit of this research-grant.
Sectors Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology,Other

 
Description The research-work made possible via the support of the BBSRC BB/I013695/1 grant has taken to a new level of general understanding of the functional role played by TSPO in mitochondrial cell homeostasis. Till the assignment of this award, TSPO has prelvantly gained attention by clinically orientated segments of science, and its core cell biology has remained ill-defined thus limiting the full comprehension of its actual contribution to cell physiology and its exploitation for diagnosis and treatment of cell and tissues conditions. Having discovered hidden aspects of TSPO physiology such as its contribution to quality control of mitochondria (mitophagy) and cell apoptosis - both core regulatory processes of cellular wellbeing- we have obtained attention by colleagues (both at national and international level) active in these fields of science which stemmed in collaborations and greater interest for TSPO. Equal degree of interest was also granted by colleagues exploiting TSPO in clinical scenarios with whom we are now constantly collaborating towards a more immediate and tangible impact on healthcare diagnostic and treatment. This award proved therefore instrumental to permit the stabilization of my group of research via an expertise renown around the globe focused on TSPO and pathways for the determination of mitochondrial function. Our findings on fundamentally relevant aspects of TSPO biology are now permitting the outline of an experimental model in which decay of mitochondrial autophagy can be scored by the accumulation of this protein. In parallel, being the expression level of TSPO increased during ageing (a very recent discovery) we are now in the position to focus to corroborate a framework in which TSPO represents a molecular read-out for mitochondrial function deregulation which underlies ageing. The notions obtained and shared till this point have been used both by academic colleagues and industry to input their protocols and particularly fine-tune TSPO-focused products which will eventually impact health-care via improved diagnostic and targeting approaches. Work published (see publications returned), together with talks and presentations in international meetings and symposia have had the merit to pave the way to future prospective for TSPO science as well as places UK at the far front of TSPO experimental biology. The BBSRC sparking impact meeting -chaired by us- and to be held in December 2014 at the Royal Veterinary College will represent the pinnacle of this common endeavour, as it will summon expertise from different fields of basic and clinical science aligning a common strategy to impact at various levels via the improved TSPO knowledge. The last two years have seen invitations to learn more of the subject incrementing exponentially suggesting how topical and relevant our endeavours are becoming for the whole community.
First Year Of Impact 2013
Sector Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology,Other
Impact Types Cultural

 
Description BBSRC Project Grant
Amount £450,000 (GBP)
Funding ID BB/N007042/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2016 
End 08/2019
 
Description BBSRC iCase Studentship
Amount £150,000 (GBP)
Funding ID Bioactive targets of mitochondrial quality control and function. 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2016 
End 09/2020
 
Description BBSRC-LiDo Studentship
Amount £105,000 (GBP)
Funding ID Imaging the Ageing Brain: Cellular and PET Tracing of the Neuroinflammatory Protein TSPO 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2014 
End 09/2018
 
Description BBSRC-iCase Studentship
Amount £150,000 (GBP)
Funding ID Focusing on the mitochondrial expression of TSPO as a marker and promoter of neuroinflammation 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2015 
End 09/2019
 
Description Bloomsbury Colleges-PhD Studentship
Amount £65,000 (GBP)
Organisation Bloomsbury Colleges 
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 10/2016
 
Description Form and Function of the Mitochondrial Retrograde Response
Amount £1,400,000 (GBP)
Funding ID European Research Council Consolidator Grant 
Organisation European Research Council (ERC) 
Sector Public
Country European Union (EU)
Start 11/2019 
End 11/2024
 
Description Internal-PhD Studentship
Amount £65,000 (GBP)
Organisation Royal Veterinary College (RVC) 
Sector Academic/University
Country United Kingdom
Start 10/2011 
End 10/2014
 
Description Keystone Symposia Future of Science Fund
Amount $1,200 (USD)
Organisation Keystone Symposia on Molecular and Cellular Biology 
Sector Charity/Non Profit
Country United States
Start 02/2014 
End 02/2014
 
Description Morrison Animal Foundation
Amount $100,000 (USD)
Organisation Morris Animal Foundation 
Sector Charity/Non Profit
Country United States
Start 12/2013 
End 12/2015
 
Description PPCT-Full Research Grant
Amount £75,000 (GBP)
Organisation Petplan Charity 
Sector Private
Country United Kingdom
Start 02/2013 
End 03/2015
 
Description PhD Studentship
Amount £100,000 (GBP)
Funding ID Imaging the Ageing Brain: Cellular and PET Tracing of the Neuroinflammatory Protein TSPO 
Organisation Bloomsbury Colleges 
Sector Academic/University
Country United Kingdom
Start 10/2016 
End 09/2019
 
Title Cancer Biomarker 
Description We are devising a way via which we can score tumor stages in animals by checking TSPO expression 
Type Of Material Biological samples 
Provided To Others? No  
Impact A potential way to image diagnostically tumor stage and progression via TSPO 
 
Title Mitophagy Inducer 
Description A new tool to physiologically activate mitophagy which we called PMI as working via the p62 pathway. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Mitochondrial quality control is a fundamental process in cellular homeostasis, and its deficiency is linked to several neurodegenerative diseases and cancers. Despite this, the discrete mechanisms, regulatory pathways, and impact oncellular physiology are still far from being elucidated. The current canonical methods to trigger mitophagy in vitro typically involve the abrupt depolarization of mitochondrial membrane potential (DYm) using ionophores such as carbonyl cyanide m-chlorophenyl hydraz 
 
Title TSPO marking to detect mito-nuclear contact sites 
Description TSPO is found to be required for the establishment of contact sites between mitochondria and nucleus, This has allowed us to enrol the protein to assess the frequency of these contacts. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2018 
Provided To Others? No  
Impact Impact is potentially multiple given the ground-breaking nature of the discovery and ability gained to map this interaction which bears implications for inter-organelles communication and core functions regulation. 
 
Description Cell Biology and Pharmacology Elucidation of Mitophagy Activators 
Organisation University College London
Department School of Pharmacy
Country United Kingdom 
Sector Academic/University 
PI Contribution My team assessed the in cell function of chemicals devised by colleague of the UCL SoP (Dr. Geoff Wells).
Collaborator Contribution Our work proved instrumental to move forward the development of chemicals synthesised by Geoff Wells and collaborators as well as to consent an actual technological improvement in the field of mitophagy activation/regulation.
Impact One Manuscript published
Start Year 2013
 
Description Elucidate the Neuronal Cell Biology of the Neuroimaging Tool TSPO 
Organisation King's College London
Department Brain Bank
Country United Kingdom 
Sector Academic/University 
PI Contribution Coordination of the research work on functional role of the PET tracer TSPO in neuronal cell physiology and toxicity determination.
Collaborator Contribution Research Grant Share to support a PDRA and consumable.
Impact A manuscript is now submitted for final revision which will outline TSPO role in neurotoxicity besides characterising its function in cell signalling homeostasis.
Start Year 2014
 
Description TSPO role in Cancer Cell Biology and Oncogenesis 
Organisation Institute Oncology Veneto
Country Italy 
Sector Hospitals 
PI Contribution We received extra charity funds to run parallel work on the comparative relevance of TSPO role in cancer biology and oncogenesis.
Collaborator Contribution We designed, developed and completed the experimental work and immunohistochemial analysis on TSPO on breast cancer tissues. An accurate evaluation of the underlying regulatory pathways is currently progressing which should lead to an high impact revaluation of the mitochondria-nucleus signalling.
Impact There are two manuscripts finalised to be submitted in the incoming months and another grant application under evaluation.
Start Year 2012
 
Description TSPO role in Cancer Cell Biology and Oncogenesis 
Organisation Royal Veterinary College (RVC)
Country United Kingdom 
Sector Academic/University 
PI Contribution We received extra charity funds to run parallel work on the comparative relevance of TSPO role in cancer biology and oncogenesis.
Collaborator Contribution We designed, developed and completed the experimental work and immunohistochemial analysis on TSPO on breast cancer tissues. An accurate evaluation of the underlying regulatory pathways is currently progressing which should lead to an high impact revaluation of the mitochondria-nucleus signalling.
Impact There are two manuscripts finalised to be submitted in the incoming months and another grant application under evaluation.
Start Year 2012
 
Description TSPO role in Cancer Cell Biology and Oncogenesis 
Organisation University College London
Department Biosciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We received extra charity funds to run parallel work on the comparative relevance of TSPO role in cancer biology and oncogenesis.
Collaborator Contribution We designed, developed and completed the experimental work and immunohistochemial analysis on TSPO on breast cancer tissues. An accurate evaluation of the underlying regulatory pathways is currently progressing which should lead to an high impact revaluation of the mitochondria-nucleus signalling.
Impact There are two manuscripts finalised to be submitted in the incoming months and another grant application under evaluation.
Start Year 2012
 
Description TSPO role in Cancer Cell Biology and Oncogenesis 
Organisation University of Padova
Country Italy 
Sector Academic/University 
PI Contribution We received extra charity funds to run parallel work on the comparative relevance of TSPO role in cancer biology and oncogenesis.
Collaborator Contribution We designed, developed and completed the experimental work and immunohistochemial analysis on TSPO on breast cancer tissues. An accurate evaluation of the underlying regulatory pathways is currently progressing which should lead to an high impact revaluation of the mitochondria-nucleus signalling.
Impact There are two manuscripts finalised to be submitted in the incoming months and another grant application under evaluation.
Start Year 2012
 
Description TSPO role in Hormonal Chemotherapy Resistance 
Organisation Imperial College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We are defining the link between cholesterol mediated resistance to hormonal chemotherapy and TSPO triggered expression in mammary cancer models.
Collaborator Contribution The main goal is trap the definition of pro-survival genes transcription mediated by TSPO and its potential intracellular relocation from mitochondrial outer membrane when pathologically overexpressed. Concomitantly TSPO ligands are being tested as pharmacological means to facilitate/promote chemotherapy induced cell death and prevent long-term resistance.
Impact Grants is being drafted
Start Year 2016
 
Description TSPO role in Osteoarthritis (OA) and Bone Remodelling 
Organisation Royal Veterinary College (RVC)
Department Comparative Biomedical Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We are dissecting the contribution by TSPO in Osteoarthritis (OA) and mediated bone pathology. We are doing so in respect of the role played by TSPO in general and targeted autophagy (mitophagy).
Collaborator Contribution Our contribution consisted in defining the pattern of expression and role in mitochondrial morphology and quality control mediation.
Impact We have a drafted manuscript to be soon submitted
Start Year 2014
 
Description Testing mitochondrial mechanisms as early marker of Drugs Attrition 
Organisation Royal Veterinary College (RVC)
Country United Kingdom 
Sector Academic/University 
PI Contribution The Royal Veterinary, University of London
Collaborator Contribution By the work on TSPO we succeeded in devising a suitable protocol via which we could detect mitochondrial toxicity and therefore provide early detection of pharmaceuticals which might reveal in vivo toxicity.
Impact We hope to gain sufficient evidences to secure valuable Intellectual Property.
Start Year 2018
 
Title Invention in the field of mitochondrial function and structure 
Description Characterisation of a sentinel molecule to mitochondrial toxins. 
IP Reference RVCBY/P58617GB 
Protection Patent application published
Year Protection Granted 2015
Licensed No
Impact The long-term impact will be on drugs attritions since many of these fail for associated mitochondrial toxicity.The impact envisaged shall therefore be at industrial level in the drugs discovery sector.
 
Description EMBO workshop on Autophagy 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Plenary selected talk to share our latest on the topic begun and currently developed with BBSRC support
Year(s) Of Engagement Activity 2015
 
Description 59th Biophysical Society Meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Invited Talk within ad hoc symposium on TSPO
Year(s) Of Engagement Activity 2015
 
Description 60th Biophysical Society Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Data presentation on the past, current and future TSPO related activities of the laboratory of research lead by MC
Year(s) Of Engagement Activity 2016
 
Description Anglo-Israeli Workshop 18-21 February 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Key activity to build up collaborations and consequent exchange of reagents with experts based abroad.

Merging forces and hence maximize the impact on mitochondrial science via the novel level of understanding of TSPO functional biology.
Year(s) Of Engagement Activity 2013
 
Description Autophagy Workshop 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Workshop on latest development in the field of Autophagy, its applications and modulators.
Year(s) Of Engagement Activity 2017
URL https://www.unifesp.br/campus/sao/hidden/eventos-e-cursos/658-i-workshop-molecular-mechanisms-of-aut...
 
Description Biochemical Society-BBSRC Sparking Impact Focused Meeting on TSPO 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Two days focused meeting
Year(s) Of Engagement Activity 2014
 
Description European Cell Death Organization Annual Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The work on TSPO role in apoptotic cell death regulation grasped the interest of many senior and affirmed colleagues which awarded the work of a fellowship covering the expenses of meeting participation by the poster's presenter (my PhD Student).

The presentation of the paper made me known at the ECDO executives and leading figures in the organization with whom I started to collaborate and consequently published in the subsequent years.
Year(s) Of Engagement Activity 2012
 
Description Fusion Conference Talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited Presentation in a meeting on Cancer Metabolism
Year(s) Of Engagement Activity 2016
URL https://www.fusion-conferences.com/conference43.php
 
Description Invited Seminar Talk at specialised audience 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Invited Seminar at the Beatson Institute for Cancer Research on September the 6th 2018
Year(s) Of Engagement Activity 2018
 
Description Invited Talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited Talk to update on our current work on TSPO
Year(s) Of Engagement Activity 2017
URL http://meetings.embo.org/event/17-mito-cancer
 
Description Keystone Symposia-Autophagy 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Decisive moment to broadcast our science on the regulation of mitochondrial autophagy by TSPO and convince on the solidity of the model.

Impact was quite immediate as right after the presentation of the work orally to a competent audience our manuscript at the moment pending was finally accepted by the journal of Autophagy (11.35IF).
Year(s) Of Engagement Activity 2014
 
Description Keystone Symposia: Mitochondrial Dynamics and Physiology 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact Poster presentation stimulated many questions and fruitful discussion during and after the poster session.

Constructive discussion of the work assisted in the finalisation and publication of a research manuscript. Greater awareness of the research also promoted increased collaborative interest.
Year(s) Of Engagement Activity 2014
 
Description MitOX 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited Plenary Talk on TSPO
Year(s) Of Engagement Activity 2015
 
Description Mitochondria, metabolic regulation and the Biology of Aging 13th - 16th February 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The talk sparked major interest on the line of research in the community of experts active in the field of mitochondrial role in cell physiology and pathology.

The topic of research core of my talk TSPO begun to receive essential attention by colleagues.
Year(s) Of Engagement Activity 2013
 
Description Neuroinflammation workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited Plenary Talk in focused event on TSPO use in diagnostic and possibly treatment of neuroinflamation.
Year(s) Of Engagement Activity 2014
 
Description Rotary International Seminars 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Attention by general public on the activities within research laboratories and practice of research was raised.

Continuos request for updates on science progress and activities run in the laboratory of research I am leading.
Year(s) Of Engagement Activity 2012,2013,2014
 
Description World Congress of Pharmacology 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact The latest results presented at the congress sparked genuine interest in the community of colleagues working in the field of cell sigalling and pharmacology.

Given the appreciation and interest for our work colleagues active at the far front of signal transduction and tools devising to monitor key cellular parameters offered collaboration and their latest probes to continue in our endeavors with their support at the far front of the field.
Year(s) Of Engagement Activity 2014