Mitobetes · The role of Von Willebrand Domain-containing Protein 8 in mitochondrial physiology
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2020-04-01 → 2022-03-31
- EU contribution
- €183,473
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
Lines connect the coordinator with its partners.
Results in brief
The role of Von Willebrand Domain-containing Protein 8 in mitochondrial physiology
Mitobetes project aimed at describing the role of Vwa8 protein in mitochondrial physiology on cellular and whole body level. We proposed to study this protein based on the evidence that increase in Vwa8 protein levels have been associated with increased obesity and diabetes. Our preliminary data has been also showing the correlation with Opa1 (comigration on native gels and coexpression during Norepinephrine-induced browning of mesenchymal cells). We expected, that studying this protein would allow us to understand how Vwa8 protein can contribute to obesity and diabetes progression since both diseases belong amongst most abundant in European population. Therefore, also the dissemination activity was focused on obesity and diabetes prevention. I took the advantage of Crispr Cas9 technology and created Vwa8 knock-out in Hek, HepG2 and MEF cells to understand the role of Vwa8 in mitochondrial physiology (WP1). In addition, we have established the collaboration Prof. Massimo Zeviani, who has the fibroblasts from Vwa8 patient. We did not observe significant changes in mitochondrial respiration using permeabilized Hek cells, but we have noticed changes in intact respiration on different substrates indicating the role of Vwa8 in mitochondrial substrate selection for respiration. To understand the molecular mode of Vwa8 action we performed several immunoprecipitations followed by mass spectrometry quantification and found potential interacting partners. Based on immunoprecipitation data and mitochondrial morphology analysis we were able to assess the Vwa8 and Opa1 relationship (WP2). I also characterised the Vwa8 KO mouse model, which showed increased fatty acid oxidation in plasma and increased browning of subcutaneous adipose tissue (WP3).
Data: CORDIS, © European Union
Project objective
Mitobetes project brings a new research on poorly characterized mitochondrial protein involved in important cellular andphysiological processes such diabetes, kidney failure and cancer. It also aims on the role of mitochondria in diabetes, thedisease which affects almost 10% of European population. The mitobetes project has a potential to help to find a newmitochondrial protein involved in diabetes, find a mechanism of diabetes development, increase the public knowledge aboutdiabetes and thus helps to decrease EU healthcare costs.Mitochondria are crucial organelles not only in energy conversion, but also in a plethora of other biological processes. Theirfunction is closely related to their dynamics, controlled by the pro-fusion proteins Mitofusin (Mfn) 1 and 2 and Optic atrophy 1(Opa1); and by the fission proteins mitochondrial fission factor (Mff) and dynamin related protein 1 (Drp1). Opa1 not onlycontrols mitochondrial fusion, but also shape of the mitochondrial cristae, a crucial parameter in determining mitochondrialfunction and participation in apoptosis. Opa1 exists in high molecular weight complexes of unknown composition that aredynamically modulated during cristae remodeling.Recently, the host laboratory completed a proteomic catalogue of proteins associating with Opa1 in intact cristae and leavingthe complex only when cristae shape was disrupted. Among the hits associating with Opa1, the host lab discovered VonWillebrand Domain-containing Protein 8 (Vwa8), a mitochondrial protein of unknown function whose higher levels correlatewith worse prognosis of Acute Myeloid Leukemia (AML). Here, I propose to understand the function of Vwa8 in mitochondrialnetwork and cristae shape, Opa1 function, apoptosis and bioenergetics. This project will not only characterize the role of anovel protein in involved mitochondrial morphology and function, but also verify if a link between mitochondrial morphologyand AML exists.
Original text from CORDIS.
Participants
- UNIVERSITA DEGLI STUDI DI PADOVA · PadovaCoordinatorItaly
Links
Data: CORDIS, © European Union