Galgano Luca
Doctoral Student
Struttura di afferenza: CLASSE SUV


October 2014 – July 2017: Graduation in Biological Sciences at University of Pavia and discussion of the project with title Role of Focal Adhesion Kinase Pyk2 in Neutrophil Extracellular Trap Formation with Prof.ssa Canobbio as supervisor
October2016 – July 2017: Internship for experimental thesis in Laboratory of Biochemistry of Prof. Torti, Department of Biology and Biotechnology Lazzaro Spallanzani, University of Pavia
October 2017 – July 2019: Graduation in Molecular Biology and Genetics at University of Pavia discussion of the project with title Role of the Proline-rich Tyrosine Kinase-2 Pyk2 in Neutrophil Activation with Prof.ssa Canobbio as supervisor
October 2017 – July 2019: Internship for experimental thesis in Laboratory of Biochemistry of Prof. Torti, Department of Biology and Biotechnology Lazzaro Spallanzani, University of Pavia

PhD research will be about the analysis of the possible role(s) of Pyk2 (proline-rich tyrosine kinase-2) and FAK (focal adhesion kinase) in platelet and neutrophil activation and in platelet/neutrophil crosstalk leading to thromboinflammation. Thromboinflammation results from the coordinate activation of inflammatory and haemostatic responses and leads to impaired inflammation and haemostasis. Neutrophils and platelets are the two key circulating cells implicated in thromboinflammation. The crosstalk between those two cell types leads to reciprocal activation under several pathological conditions. Platelets and neutrophils present several membrane receptors implicated in their own activities. Some of these receptors are involved in the interaction between those two cell types. Activated platelets expose P-selectin that binds to resting neutrophil PSGL-1 (P-selectin glycoprotein ligand-1). This first unstable adhesion is stabilised by the activation of neutrophil 2 integrins (M2 and L2 integrins) after PSGL-1 activation. M2 integrin binds to platelet IIb3 integrin-bound fibrinogen, GPIb (glycoprotein-Ib), and JAM3 (junctional adhesion molecule-3). Instead, L2 integrin binds to platelet ICAM-2 (intercellular adhesion molecule-2). The firm adhesion is favoured by the activation of SFKs (Src family kinases), and other effectors. These platelet-neutrophil ligand-receptor interactions activate neutrophil signalling cascades, leading to tissue factor expression (coagulation factor III), ROS production, degranulation, and NETs formation, contributing to thrombus formation. However, even activated neutrophils are able to interact with resting platelets. In this condition, neutrophil M2 integrin binds to platelet GPIb, constituting the firm adhesion and bypassing P-selectin involvement. Activated neutrophils themselves undergo degranulation and cathepsin-G secretion. Neighbour platelets bind cathepsin-G through PAR4 (protease-activated receptor-4), being activated and leading to thrombus formation. Among different signalling proteins that regulate platelet/neutrophil interaction, the cytosolic non-receptor tyrosine kinases Pyk2 and FAK may be relevant, since they participate to cell-cell adhesion. In particular, Pyk2 plays multiples roles in neutrophils and platelets activities as it is required for neutrophil degranulation as well as for platelet activation and thrombus formation. The exact contribution of Pyk2 and FAK in platelet/neutrophil crosstalk is not known, and the present project will decipher the possible contribution of Pyk2 and FAK in the signalling pathways activated in platelets and in neutrophils that coordinate their interaction and finally result in thromboinflammation.
The whole work will be based on the use of both human and murine neutrophils and platelets. I will take advantage of two different tools: kinase inhibitors (Pyk2 inhibitors PF-4594755 and PF-4520440; FAK inhibitor GSK2256098) and transgenic Pyk2 knockout (KO) mice. The experiments will be about the analysis of the localisation of Pyk2 and FAK in both platelets and neutrophils and the role(s) of Pyk2 and FAK in neutrophil and platelet cytoskeleton remodelling, neutrophil adhesion signalling and transmigration, neutrophil and platelet ROS production and signalling pathways for ROS formation, NETs formation, and platelet-neutrophil aggregate (PNA) formation.
The research has the final goal to discover new potential pharmaceutical targets for thromboinflammation, improving the quality of life of a large number of patients affected by several diseases triggered by thromboinflammatory reactions.