MitoEAGLE data: blood and cultured cells

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Working Groups

Short-Term Scientific Missions

Management Committee

Members

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COST Action CA15203 (2016-2021): MitoEAGLE
Evolution-Age-Gender-Lifestyle-Environment: mitochondrial fitness mapping

MitoEAGLE data: blood and cultured cells

WG4

MITOEAGLE data repository for blood cells and clutured cells

WG4 Project application

Description

Biopsy sampling of human muscle and fat tissue for mitochondrial studies is an invasive approach. More recently measurement of mitochondrial function in human blood cells – as well as temporary cryopreservation (Karabatsiakis 2014 Transl Psychiatry) – has become an attractive and by far less invasive alternative for mitochondrial diagnostics. Mitochondrial function of blood cells was linked with numerous diseases and metabolic conditions such as sepsis, neurodegenerative diseases, depression, hypertension, amyotrophic lateral sclerosis, and early-stage heart failure (e.g. Sjoevall 2013 Crit Care; Karabatsiakis 2014 Transl Psychiatry; Hagl 2014 J Alzheimer Dis). The use of human blood cells will thus predictably widen the applicability of respirometry in human physiology and will provide a significant stimulus for the entire field of mitochondrial medicine.

A key towards achieving comparability among data sets and the applicability of human blood cells for the evaluation of mitochondrial fitness in health and disease is the standardization of the procedures to separate or isolate blood cells, the experimental procedure for the evaluation of mitochondrial respiratory characteristics, and the format for reporting. A consensus on protocols should be reached in WG4, calling for a Workshop right at the start of the COST Action. Depending on the WG4 participants input, cell lines will be defined for which harmonization of datasets should be achieved (Fig. 2).

Management

Tasks

1. Kick-off meeting to develop a working plan for the establishment of consensus protocols, reporting schemes, and the assignment of specific study tasks.
2. Development of SOPs for cell separation.
3. Development of laboratory protocols for individual blood cell types (and other cell types) for mitochondrial studies on intact and permeabilized cells.
4. Application of SOPs in experimental studies during Short Term Scientific Missions and feeding of data into the MITOEAGLE data repository.
5. Publication of SOPs and study results.

Milestones

1. Consensus on protocols, reporting schemes and work assignments.
2. Completed SOPs for cell preparation & laboratory protocols.
3. Application study finished and data transmitted to MITOEAGLE data repository.
4. Publication finished.

Deliverables

1. SOPs for blood cell separation and respirometric characterisation open for the research community.
2. MITOEAGLE data repository for comparative data evaluation, planning of future studies, data mining.
3. Publication with a set of reference data.

WG4 Action

WG Leader

Nicoleta Moisoi, Dr. - [email protected] (Cultured cells)

WG Co-Leader

Eskil Elmer, MD, PhD - [email protected] (Blood cells)

Participants

Country	City	Contact
Armenia	Yerevan	Trchounian K
Austria	Innsbruck	Karabatsiakis Alexander
Austria	Innsbruck	Fischer Christine
Austria	Innsbruck	Hoppel Florian
Austria	Innsbruck	Meszaros Andras T
Austria	Innsbruck	Torres-Quesada Omar
Austria	Innsbruck	Gnaiger Erich
Austria	Innsbruck	Cardoso Luiza HD
Austria	Innsbruck	Garcia-Souza Luiz F
Austria	Innsbruck	Doerrier Carolina
Belgium	Namur	Renard P
Belgium	Leuven	Bird Matthew
Belgium	Liege	Votion Dominique-Marie
Belgium	Namur	Arnould Thierry
Belgium	Namur	Detraux Damien
Bosnia and Herzegovina	Tuzla	Jusic Amela
Bosnia and Herzegovina	Tuzla	Hajrulahovic Anesa
Brazil	Sao Paulo	Verdaguer Ignasi Bofill
Brazil	Sao Paulo	Dominguez-Amorocho OA
Brazil	Sao Paulo	Alencar Mayke Bezerra
Brazil	São Paulo	Crispim Marcell
Croatia	Zagreb	Rubelj Ivica
Czech Republic	Prague	Pecina Petr
Czech Republic	Prague 4	Bajzikova Martina
Czech Republic	Prague	Mracek Tomas
Czech Republic	Prague	Hansikova Hana
Czech Republic	Pilsen	Kripnerova M
Czech Republic	Vestec	Zobalova Renata
Czech Republic	Pilsen	Markova Michaela
Czech Republic	Prague	Elkalaf Moustafa
Czech Republic	Hradec Kralove	Vernerova Andrea
Czech Republic	Prague	Fisar Zdenek
Czech Republic	Hradec Kralove	Maseko Tumisang Edward
Czech Republic	Pilsen	Kuncova J
Czech Republic and Australia	Prague and Southport	Neuzil Jiri
Czech rep	Hradec Kralove	Falaye T
Denmark	Copenhagen	Desler C
Denmark	Copenhagen N	Mortensen OH
Denmark	Copenhagen	Rasmussen LJ
Denmark	Hvidovre	Nehlin Jan O
Denmark	Aarhus	Rattan Suresh IS
Denmark	Malov	Hey-Mogensen M
Egypt	Cairo	Abdel-Rahman Engy Ali
Egypt	Cairo	Ali Sameh S
Estonia	Tallinn	Truu Laura
Estonia	Tallinn	Klepinina Lyudmila
Estonia	Tartu	Grellier T
Finland	Helsinki	Pirinen E
Finland	Jyvaeskylae	Lehti Maarit
Finland	Turku	Stier Antoine
France	Bordeaux cedex	Mourier A
France	Bron	Rizwan Alam M
France	Paris	Petit Patrice X
Germany	Neuherberg	Borchard Sabine
Germany	Ulm	Schaefer PM
Germany	Ulm	Kolassa Iris
Germany	Gießen	Silaidos Carmina
Germany	Düsseldorf	Prigione Alessandro
Germany	Freising	Klingenspor Martin
Germany	Leipzig	Ost Mario
Germany	Regensburg	Renner-Sattler Kathrin
Germany	Duesseldorf	Piel Sarah
Germany	Freising	Fromme Tobias
Germany	Mainz	Vujacic-Mirski Ksenija
Germany	Freising	Keppner Gloria
Germany	Giessen	Eckert Gunter P
Germany	Mainz	Kalinovic S
Germany	Magdeburg	Vieira Ligo Teixeira Camila
Germany	Munich	Jastroch M
Germany	Hamburg	Singer Dominique
Germany	Freising-Weihenstephan	Musiol Eva
Germany	Freising	Schoettl T
Germany	Biberach an der Riß	Nold V
Germany	Ulm	Hartmann C
Germany	Ulm	Calzia Enrico
Germany	Duesseldorf	Eckers A
Greece	Athens	Trougakos Ioannis P
Hungary	Debrecen	Verebne Tar K
Hungary	Budapest	Tretter Laszlo
Hungary	Budapest	Komlodi Timea
Ireland	Dublin	Karavyraki Marilena
Ireland	Dublin	Porter Richard K
Ireland	Sligo	Murphy J
Israel	Rishon Le Zion	Hachmo Yafit
Israel	Jerusalem	Saada Reisch Ann
Israel	Zrifin	Sova Marina
Italy	Milano	Bottoni P
Italy	Bologna	Gasparre G
Italy	Rome	Scatena Roberto
Italy	Verona	Calabria Elisa
Italy	Bologna	Genova Maria Luisa
Italy	Bolzano	Pichler Irene
Italy	Bolzano/Bozen	Volani Chiara
Italy	Bologna	Paterlini S
Italy	Padova	Fernandez-Vizarra Erika
Italy	Bari	Agrimi G
Italy	Bologna	Porcelli AM
Italy	Bolzano	Castelo Rueda Maria Paulina
Italy	Ancona	Battino Maurizio
Japan	Sapporo	Shirakawa R
Latvia	Riga	Makarova Elina
Latvia	Riga	Kidere Dita
Malaysia	Kuala Lumpur	Abdul Karim Norwahidah
Malta	Msida	Vella Joanna
Netherlands	Nijmegen	Koopman Werner JH
Netherlands	Wageningen	Keijer Jaap
Netherlands	Groningen	Rots MG
Norway	Oslo	Eide L
Norway	Oslo	Lund Jenny
Norway	Oslo	Nikolic N
Norway	Oslo	Rustan AC
Norway	Bergen	Tronstad Karl Johan
Palestine	Ramallah	Stiban Johnny
Poland	Lodz	Labieniec-Watala Magdalena
Poland	Poznań	Ludziejewska A
Poland	Lodz	Siewiera Karolina
Poland	Poznan	Swiniuch Daria
Poland	Poznan	Michalak Slawomir
Poland	Warsaw	Wasilewski M
Poland	Krakow	Kaczara P
Poland	Warsaw	Dymkowska Dorota
Poland	Warsaw	Kampa Rafal Pawel
Poland	Warsaw	Sek Aleksandra
Poland	Poznan	Rybacka-Mossakowska Joanna
Poland	Lodz	Watala Cezary
Portugal	Cantanhede	Pinho Sonia A
Portugal	Porto	Beleza Jorge
Portugal	Catanhede	Pereira da Silva Grilo da Silva Filomena
Portugal	Porto	Carrageta D
Portugal	Porto	Magalhaes Jose
Portugal	Porto	Martins Ana Dias
Portugal	Porto	Crisostomo Luis
Portugal	Coimbra	Carvalho Eugenia
Portugal	Porto	Moreira Bruno P
Portugal	Cantanhede	Bento Guida Jose Freitas
Portugal	Porto	Oliveira Pedro Fontes
Portugal	Coimbra	Palmeira Carlos
Portugal	Covilhã	Dias Tania R
Portugal	Porto	Bernardino RL
Portugal	Porto	Alves Marco G
Portugal	Coimbra	Pereira Susana P
Republic of Korea	Yuseong-gu Daejeon	Quang Don T
Republic of Korea	Daejeon	Park SH
Republic of Serbia	Belgrade	Pavlovic Kasja
Republic of Serbia	Belgrade	Krako Jakovljevic Nina
Romania	Timisoara	Lighezan R
Romania	Timisoara	Betiu AM
Romania	Timisoara	Avram Vlad F
Romania	Timisoara	Muntean Danina M
Romania	Timisoara	Danila Maria-Daniela
Romania	Timisoara	Lelcu Theia
Russia	Moscow	Itkis YS
Russia	Moscow	Tsygankova P
Russia	Moscow	Krylova T
Saudi Arabia	Giza	Kotb IA
Serbia	Belgrad	Markovic Ivanka
Serbia	Belgrade	Misirlic-Dencic S
Serbia	Belgrade	Jeremic M
Serbia	Belgrade	Isakovic A
Serbia	Belgrade	Ivanovic Tijana
Serbia	Belgrade	Vukojevic M
Slovakia	Košice	Cizmarova Beata
Slovakia	Bratislava	Ukropcova B
Slovakia	Bratislava	Maderova D
Slovakia	Bratislava	Ukropec J
Slovakia	Bratislava	Nemec Michal
Slovakia	Bratislava	Sumbalova Zuzana
Slovenia	Ljubljana	Pirkmajer Sergej
Slovenia	Ljubljana	Mars T
Slovenia	Ljubljana	Kopitar-Jerala Natasa
Spain	Valencia	Casado Pinna Marta
Spain	Barcelona	Tura Ceide O
Spain	Barcelona	Zorzano Antonio
Spain	Sevilla	Bustos Matilde
Spain	Salamanca	Almeida Angeles
Spain	Granada	Quiles JL
Spain	Barcelona	Arago Marc
Spain	Barcelona	Hyrossova Petra
Spain	Barcelona	Juarez D
Spain	Barcelona	Smolders V
Spain	Armilla, Granada	Escames Germaine
Spain	A Coruña	Mayan MD
Spain	Armilla (Granada)	Fernandez-Ortiz Marisol
Spain	Barcelona	Gil J
Spain	Madrid	De los Rios C
Spain	Barcelona	Gama Perez Pau
Spain	Valencia	Victor Victor Manuel
Spain	A Coruña	Blanco FJ
Spain	Ourense	Reboredo-Rodriguez Patricia
Spain	Lleida	Gonzalo Hugo
Spain	Madrid	Cano-Abad MF
Spain	Madrid	Cadenas S
Spain	Madrid	Monsalve Maria
Spain	Lleida	Nogueras L
Spain	Valencia	Borras Consuelo
Spain	Madrid	Lopez MG
Spain	Barcelona	Dahdah Norma Ramon
Spain	Valencia	Fuertes Agudo Marina
Spain	Sevilla	Muntane Jordi
Spain	Barcelona	Gonzalez Casacuberta I
Sweden	Linköping	Fontes-Oliveira CC
Sweden	Lund	Chamkha Imen
Sweden	Södertälje	Alfaro-Moreno E
Sweden	Lund	Simon Serrano Sonia
Sweden	Lund	Leffler M
Sweden	Lund	Abliva
Sweden	Lund	Karlsson M
Sweden	Lund	Westerlund E
Sweden	Lund	Elmer Eskil
Sweden, SE	Lund	Aasander Frostner Eleonor
Switzerland	Lausanne	Zanou Nadege
Switzerland	Geneva	Martinou JC
The Netherlands	BT Amsterdam	Wuest Rob CI
Turkey	Tekirdag	Aral Cenk
Turkey	Izmir	Aydin HH
Turkey	Ankara	Tuncay Erkan
Turkey	Ankara	Engin Ayse Basak
Turkey	Istanbul	Baykal AT
USA	Philadelphia	Janowska Joanna
USA	Iowa City	Buettner Garry R
USA	Fayetteville	Rao RR
USA	St Louis	Welch R
USA	Philadelphia	Kilbaugh Todd
USA	Baton Rouge	Irving Brian A
USA	Orlando	Coen Paul M
USA	Philadelphia	Starr Jonathan
USA	Fayetteville	Iyer Shilpa
United Kingdom	Brighton	Moore Anthony L
United Kingdom	Stoke-on-Trent	McStay Gavin P
United Kingdom	Oxford	Makrecka-Kuka Marina
United Kingdom	Leicester	Moisoi Nicoleta
United Kingdom	Birmingham	Thakker A
United Kingdom	Birmingham	Joanisse S
Uruguay	Montevideo	Radi R

Task groups

TG leaders: suggestions discussed at the MitoFit Science Camp 2016 Kuehtai AT.

TG participants: listed as a summary from the feedback received.

TG4.1 Blood

TG leaders: Calabria Elisa IT / Calzia Enrico DE

TG participants: Aasander Frostner Eleonor SE, Elmer Eskil SE, Krako Nina RS, Labieniec-Watala Magdalena PL, Lehti Maarit FI, Monsalve Maria ES, Muntean Danina M RO, Renner-Sattler Kathrin DE, Michalak Slawomir PL, Westerlund Emil SE

TG4.2 Cultured cells

TG leader: Renner-Sattler Kathrin DE

TG participants: Borras Consuelo ES, Komlodi Timea HU, Krako Nina RS, Labieniec-Watala Magdalena PL, Manuela Lopez ES, Mars Tomaz SI, Palmeira Carlos PT, Pirkmajer Sergej SI, Rustan Arild NO, Scatena Roberto IT, Wuest Rob C NL

Comments and discussion

1. Dr. Ali will work within the realms of WG4 (blood and cultured cells) and potentially WG1 (protocol development) as well as WG5 (dissemination and training). He will participate in data generation, quality control, and repository efforts of protocols recommended by these working groups. Dr. Ali will contribute to training workshops and conferences that aim at defining standard procedures and terminology in the field of mitochondria physiology. - Sameh S Ali (2016).
2. I am very interested in being part of the management committee either as a MC member for managment tasks or in working group 4 MITOEAGLE data repository for blood cells and cultured cells. I have experience in analyzing mitochondrial function in different human tissues as cultured cells and blood cells (also subsets of specific blood populations, see list of publications), muscle and isolated mitochondria. This together with my experience in organizing summer schools and conferences could be helpful as MC member taking care of management tasks. However, if you think my expertise is more helpful in working group 4, I am looking forward to support this working group. - Kathrin Renner-Sattler (2016).
3. Specifically, we are studying mitochondrial metabolism in cancer cells. At this aim we are also participating to mt-HPP project, an Italian consortium whose target is a definition of mitochondrial proteome in health and disease. Briefly, my specific activity is to compare mitochondrial functional status with the mitochondrial proteome picture in different human cell lines. I do think that a "contact" between these two important initiatives should be useful for both. - Roberto Scatena (2016).
4. My role in Mito-EAGLE would be mainly in WG 2 and 4, with a focus on skeletal and cardiac muscle and cultured cardiomyocytes and endothelial cells. I would particularly like to bridge the gap between the group leaders and young investigators, trying to represent the young people who are the active users of the OROBOROS equipment. This will fit in nicely with the goals in WG1. ... Major congratulations on obtaining this beautiful collaborative grant! Very glad to see this network getting the recognition it deserves! Looking forward to being part of this! - Rob C Wuest (2016).
5. We mostly work with cultured human skeletal muscle cells, from different donor groups such as lean, obese, type 2 diabetics, as well as muscle cells from athletes. We are interested in collaboration with our cells and measurements on energy metabolism and mitochondral function. - Arild Rustan (2016).
6. Based on your previous suggestions, with a great pleasure I will take up some managements duties, i.e. Short-Term Sicentific Missions organizations/coordinatings, and/or so on. I choose WG 4 (blood cells and cultured cells) eventually also WG 3 (other tissues than fat) but this will depend on grant funds receving(s) in the near future. ... I have just read the COST evaluation form and I congratulate you this beautiful/great success!!!! This news makes me very happy and I am proud of becoming a member of this "mitochondrial family. - Magdalena Labieniec-Watala (2016).
7. Please, find enclosed my declaration to participate with my team in the following workgroups: WG1, WG2, WG3, WG4. We are now involved and have been in not a distant past – in studying of the following cell /tissue types:
   1. Blood cells, with particular emphasis on platelets and lymphocytes – isolated from animals and humans.
   2. Endothelial cells, fibroblasts, neurocytes (neuroblastoma), astrocytes, cancer cell lines – predominantly cultured.
   3. Hepatocytes, cardiomyocytes, cell isolated from brain – isolated from lab animals. - Cezary Watala (2016).
8. I would like to participate in WG4 - MITOEAGLE data repository for blood cells and clutured cells. We are already working with cryopreserved PBMC from lung cancer, breast cancer and colon cancer patients. Thus, I would like to participate in WG4 tasks. - Slawomir Michalak (2016).
9. I have already contacted the portuguese CNC to nominate me for the MC of theaction. Concerning my active participation, taking into account the type of work we do in my lab I could be included in all WGs. As you know we work with mitochondria from liver, muscle and fat, as well as cultured cells from different origins (WG2-4). The elaboration of SOPs (WG1) we can certainly contribute (see the book I edited on Mitochondrial Bioenergetics and in which you contribute a chapter). I believe that everybody can be part of the WG5, but as the coordinators of the action, you and Sandra should decide what is best for the success of the action. - Carlos Palmeira (2016).
10. While going through MITOEAGLE pages on bioblast, I am realizing how substantial this your project is going to be and I will be happy to assist at my best and to grow as a scientist through this exciting collaboration. As stated in MITOEAGLE, I find it urgent to standardise the procedures to separate or isolate blood cells, experimental procedure for the evaluation of mitochondrial respiratory characteristics, and the format for reporting obtained data in order to exploit them toward future clinical and diagnostic application. I will be happy to share with MITOEAGLE network of experts, my experience and my data beyond the published records on evaluation of mitochondrial physiology in different cell lines in order to establish and publish SOPs, apply them during Short Term Scientific Missions and during my every-day scientific practice, as well as to feed the obtained data into the MITOEAGLE data repository. Even if I have no experience with data management and public relations on a large scale, I am ready to learn some new skills if necessary in order to answer your requests for balancing of MC members and management tasks, so feel free to contact me with any other suggestions regarding other tasks or different WGs. Please consider, that I have done 3-months training on evaluation of mitochondrial function in muscle biopsies in the OROBOROS reference lab of Prof. Roden in Germany - Nina Krako (2016).
11. As you know, I have received the MC nomination from our CNC in Bucharest (Ms Elena Dinu) for my colleague, Rodica Lighezan and myself. My expertise is most on muscle tissue (in particular, heart) and also, liver - so I will go for WG2 if this is fine with you. However, Both my colleague and I are mostly interested in performing respirometry in blood cells (and this is what we will start to try in the near future); thus, you decide whether we can fit both of us in WG4, or only her. Also, in order to extend the Network in other Romanian cities with powerful state medical universities, Cluj-Napoca (as you know) and Tg Mures - 2 young men, Bela Kiss and Ovidiu Cotoi have been proposed as substitute members (for gender balance too). In particular, we want to jointly ”explore” the exciting fields of mitochondrial ”hidden toxicity” and dynamics, respectively since the former colleague is a toxicologist and the latter a pathologist. I wish you good luck and I am indeed happy and proud to be part of this growing network. - Danina M Muntean (2016).
12. We work mainly on the cardiovascular system (endothelium) and in the liver, we also work with human blood cells and tissues, so we would fit bets in WG4. - Maria Monsalve (2016).
13. Congratulations on the considerable progress with MITOEAGLE! I am keen to participate in this exciting project and I’m sure I can make a valuable contribution to it's success. I have extensive experience performing respirometry assays in human muscle biopsies, myoblasts/tubes and mouse tissues in the contexts of aging, obesity, exercise and inactivity. I feel that I could contribute to any of the five working groups, but particularly to WG2, WG4, and WG5. I am also happy and willing to serve on the Management Committee if needed. Again, congratulations on organizing such an exciting project. - Paul M Coen (2016).
14. As research group working in the field of skeletal muscle research (development,function and regeneration, metabolic pathways, signalling, and energy metabolism and skeletal muscle in special condition-ischemia, hypoxia, sepsis) we found several topics in the COST Action particularly interesting. Indeed, we are highly interested in effects of media and protocols on experimental readouts in cell cultures (please see Pirkmajer 2011 Am J Physiol Cell Physiol and Rajh 2016 PLOS ONE). We therefore find the focus on harmonization and validation of protocols and terminology especially important. - Tomaz Mars (2016).

Projects in progress

» MitoFit-Bloodcells

» MiPMap

Next steps

» COST 2016 Verona IT