Platelet Biochemistry Laboratory

Seat: Department of Biology and Biotechnology - Laboratory of Biochemistry
Cascina Cravino - via Bassi 21, 27100 Pavia, Italy
Referents: Prof. Mauro Torti (Full Professor), Prof. Ilaria Canobbio (Associate Professor), Prof. Gianni Guidetti (Associate Professor)
Co-workers: Marta Zarà (researcher RTT), Michela Gemme (Research fellow), Serena Rustichelli (PhD student), Valentina Bonaldi (PhD student), Giosia Rossignod (PhD student).
Research activity encompasses fundamental aspects of signaling in platelets. Circulating blood platelets have a crucial role in hemostasis, thrombosis, and in many other processes, such as inflammation, neurodegeneration, and cancer metastasis. Understanding the exact contribution of platelets to these events is a big challenge for the future and may provide promising opportunities in biomedicine.
1) Analysis of platelet signal transduction pathways in thrombosis, hemostasis, and thromboinflammation
The understanding of the mechanisms supporting platelet activation is critical for the identification of novel pharmacological targets for thrombotic diseases, which represent a major cause of death in western countries. In this context, our research is focused on the following aspects:
- the C-type lectin receptor CD93 in platelet function and thrombosis
- role of tyrosine kinase Pyk2 in thrombosis and thromboinflammation
- the contribution of different phosphatidylinositol 3-kinase (PI3K) isoforms in the release of platelet-derived extracellular vesicles (PEVs).
2) Analysis of Metabolism and Function of Amyloid Precursor Protein (APP) and Amyloid Aβ Peptide in Platelets: A Connection Between Hemostasis, Thrombosis, and Alzheimer’s Disease
Alzheimer’s disease is associated with the accumulation of amyloid peptides in the brain and cerebral vessels. These amyloid peptides are derived from the metabolism of the amyloid precursor protein (APP). Platelets express APP and release amyloid Aβ peptides into the bloodstream when activated. In our laboratory, we are interested in:
- Studying the effect of amyloid peptide accumulation in peripheral circulation. We have demonstrated that amyloid peptides promote platelet adhesion and activation and induce a state of chronic inflammation and oxidative stress. The molecular mechanisms underlying these events are not yet fully understood and are the subject of our ongoing research.
- Amyloid peptides have been shown to play an antimicrobial role in brain infections. We are investigating whether amyloid peptides present in circulation may have antimicrobial effects on systemic infections and sepsis.
3) Analysis of the contribution of platelet-derived extracellular vesicles to cancer progression
Cancer progression is strongly influenced by host-cancer interactions, and it has been demonstrated that platelets are key players in cancer spread, as their depletion, in thrombocytopenic patients or animal models, is associated to reduced metastasis. This suggests that also the pharmacological inhibition of platelet activation could represent a novel approach to control metastatic diffusion.
The goal of our study is the understanding of the molecular mechanisms behind the contribution of platelets to cancer, with a specific interest on glioblastoma multiforme (GBM). Our attention is focused on the role of platelet-derived extracellular vesicles (PEVs), which are small vesicles released upon platelet activation and represent important carriers of biological signals. With this research we aim to:
- analyze the ability of PEVs to alter the progression of GBM and its response to drugs
- investigate the molecular mechanisms through which PEVs modulate the cancer cells behavior
- develop engineered PEVs with antitumoral activity.
Research group collaborations
- Giampaolo Minetti (Department of Biology and Biotechnology, University of Pavia, Italy)
- Federico Galvagni, Maurizio Orlandini (Department of Biotechnology, Chemistry and Pharmacy, University of Siena, Italy).
- Silvia Barbieri, Marina Camera, Andrea Baggiano, Sebastiano Gili (Istituto Cardiologico Monzino, Milano, Italy)
- Marcello Manfredi (Department of Translational Medicine, University of Eastern Piedmont, Novara, Italy)
- Sonia Severin (CRCN Inserm, Toulouse, France)
- Paolo Gresele (Department of Internal Medicine, University of Perugia)
- Federico Bertoglio (Abteilung Medizinische Biotechnologie, Technical University of Braunschweig)
Methods and Techniques
Blood cell purification (platelets and neutrophils). Isolation and characterization of extracellular vesicles. Protein analysis (SDS-PAGE, western blotting, immunoblotting, chemiluminescence, spectrophotometry, spectrofluorometric analysis, flow cytometry). Cell adhesion assays under static and flow conditions. Microscopy (phase contrast and fluorescence microscopy). Molecular biology techniques, DNA and RNA extraction and analysis, PCR. Protein purification (expression, ion-exchange, affinity and size-exclusion chromatography, HPLC, FPLC). Cell culture and transfection, analysis of cell migration, invasion, proliferation, metalloproteinases release. Genetically modified animal models (Mus musculus): Pyk2 KO, APP KO, APP23, CD93 KO.
Updated: November, 15th, 2024