Proteomic analysis of platelets from deep venous thrombosis patients

• Grant number: 11/05996-1
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2011
• End date: September 30, 2013
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Joyce Maria Annichino-Bizzacchi
• Grantee: Joyce Maria Annichino-Bizzacchi
• Host Institution: Centro de Hematologia e Hemoterapia (HEMOCENTRO). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Deep venous thrombosis (DVT) is a multifactorial disease, and is resulted from the inappropriated activation of the hemostatic response and can occur in any part of the venous system. It has a high incidence and is associated to a high morbi-mortality by complications, as the pulmonary embolism and the post-flebitic syndrome. From 20 to 30% of the patients do not show any initiate factor, and is defined as spontaneous venous thrombosis. Besides, the venous thrombosis is considered a chronic disease, because in 5 years approximately 25% of the patients will show recurrence. So, the identification of new risk factors that could prevent the recurrence has high relevance on clinical practice. So, proteins from platelets from patients with DVT are interesting targets of research and object of this study. Blood samples will be collected and the platelets will be separated. The platelets will be washed by a methodology that minimizes their activation and then they will be lyzed. After that, the proteins will be digested by trypsin and the salt will be removed applying C18 columns (Sep-Pak®). The peptides will be fractionated by FPLC and the mass spectrometry will be performed applying the hybrid spectrometer LTQ-Orbitrap. The spectra will be analyzed by Sorcerer-SEQUEST, and the semi-quantitative analysis of the proteins will be performed applying the Apex Quantitative Proteomics Tool software. The results will be validated by Western Blotting applying specific monoclonal and polyclonal antibodies against the differentially expressed proteins. (AU)