Mitochondria-dependent molecular mechanisms in cardioprotection
Universita degli Studi di Ferrara
Dep. of Morphology, Surgery and Experimental Medicine. Section of Pathology, Oncology and Experimental Biology. Via Fossato di Mortara, 64, 44121 Ferrara
Dr. Giampaolo Morciano
Dr. Giampaolo Morciano
English; Italian; French
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No Yes No Yes No No No Yes Yes Yes No
Type of Research Project
- Basic science
What is the background of the project?
Mitochondria are not isolated organelles but form complex networks which are under strict control by two distinct processes. The first one is mitochondrial fusion, which forms long filamentous mitochondria, and the second one is mitochondrial fission, which generates small spherical mitochondria. Both processes depend on the metabolic needs of the cell. Proper mitochondrial function is associated with a balance between the two previous processes. Additionally, another source of mitochondrial quality control is the selective degradation of the dysfunctional organelles through autophagy which is named as mitophagy. Mitochondria are the site of reactive oxygen species (ROS) generation. Uncontrolled production of ROS and impairment of mitochondrial dynamics result in mitochondrial dysfunction and ultrastructural changes of cellular lipids, proteins, enzymes and DNA. There’s increasing the attention towards the permeability transition pore complex (PTPC) that determines mitochondrial swelling and cell death in several cardiovascular diseases. All the above-mentioned players are the pathophysiologic background for the development of multiple cardiac diseases. Therefore, targeting of mitochondrial dysfunction is a crucial step in the treatment of many cardiac diseases and several approaches have been tested in experimental and clinical studies with, controversial findings.
What is the aim of the project?
To investigate molecular mechanisms involving mitochondrial players (ROS, morphology, PTPC, proteins involved in mitophagy) for cardioprotection purposes in myocardial infarction, aortic stenosis and heart failure
What techniques and methods are used?
Step 1: to study background and mitochondrial proteins of interest by in silico approaches (both literature and bioinformatics tools); Step 2: to study cell phenotype upon either overexpression or silencing of the proteins of interest in a cardiomyocyte cell line by biomolecular and biochemical approaches (cell subfractionation, western blot, cell culture, transfections and infections); Step 3: to study the effects in terms of altered molecular pathways due to either overexpression or silencing of the proteins of interest in a cardiomyocyte cell line by biochemical and bioluminescence approaches (cell calcium and ATP (Adenosine Triphosphate) measurements in living cells population); Step 4: to study cell death and and their involvement in an ischemic pathological context by applying confocal microscopy in living cells population. Step 5: analysis of data
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
- To understand the theory of the project (state of the art of the background); - To handle cells and plasmids in sterility conditions; - To perform experiments aimed at evaluating protein expression in biological samples (biological fluids, cells, tissues) and to measure intracellular calcium fluxes, ATP content and its production; - To understand the concept of confocal microscpopy for research purposes and to use it for protein (co)localization assessment, to study kinetics of calcium fluxes and PTPC activity, mitochondrial morphology.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Teaching about theory and experimental plans before each experiment
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Desire to learn and to do research. Minimum skills about laboratory equipment. Subjects passed: Pathology, Cardiology. Previous experience with: Laboratory
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- Graduated students (less than 6 months)
- Pre-Medical students from the American-British system
- Morciano G; Giorgi C; Bonora M; Punzetti S; Pavasini R; Wieckowski MR; Campo G; Pinton P (2015) Molecular identity of the mitochondrial permeability transition pore and its role in ischemia-reperfusion injury. J Mol Cell Cardiol 78:142-153
- Marchi S; Patergnani S; Missiroli S; Morciano G; Rimessi A; Wieckowski MR; Giorgi C; Pinton P (2018) Mitochondrial and Endoplasmic Reticulum Calcium Homeostasis and Cell Death. Cell Calcium
- Campo G; Pavasini R; Morciano G; Lincoff AM; Gibson CM; Kitakaze M; Lonborg J; Ahluwalia A; Ishii H; Frenneaux M; Ovize M; Galvani M; Atar D; Ibanez B; Cerisano G; Biscaglia S; Neil BJ; Asakura M; Engstrom T; Jones DA; Dawson D; Ferrari R; Pinton P; Ottani F (2017) Clinical Benefit of Drugs Targeting Mitochondrial Function as an Adjunct to Reperfusion in ST-segment Elevation Myocardial Infarction: a Meta-Analysis of Randomized Clinical Trials. Int J Cardiol 244:59-66
- Morciano G; Pedriali G; Sbano L; Iannitti T; Giorgi C; Pinton P (2016) Intersection of mitochondrial fission and fusion machinery with apoptotic pathways: role of Mcl-1. Biol Cell 108:279-293