|Tipo di tesi||Tesi di dottorato di ricerca|
|Titolo||Studi proteomici per l'identificazione e validazione delle alterazioni dei profili cellulari e molecolari indotte da nuovi candidati farmaci e in risposta alla terapia|
|Titolo in inglese||Proteomics studies for the identification and validation of alterations of molecular cellular profiles induced by new drug candidates and in response to therapy|
|Settore scientifico disciplinare||CHIM/08 - CHIMICA FARMACEUTICA|
|Corso di studi||CLINICAL AND EXPERIMENTAL MEDICINE (CEM) - MEDICINA CLINICA E SPERIMENTALE|
|Data inizio appello||2018-03-22|
|Disponibilità||Accessibile via web (tutti i file della tesi sono accessibili)|
Spettrometria di massa e si è evoluta in una potente piattaforma bioanalitica per la misurazione simultanea di un gran numero di proteine espresse in cellule e tessuti ed è diventato uno strumento importante per la scoperta di nuovi biomarcatori utili per l'applicazione clinica.
Mass spectrometry (MS) Proteomics has evolved into a very powerful bioanalytical platform for the simultaneous measurement of a large number of proteins expressed in cells and tissues, and has become an important tool for the discovery of new biomarkers useful for clinical application. My work is focused on the evaluation of folate dependent proteins as drug targets and biomarkers. In particular, I followed two types of projects: a) studies of protein modulations following drug therapy in clinical samples from patients affected by ovarian cancer. The aim was to track the protein set associated to the drug mechanism of action and to be used as an early pharmacodynamics biomarker. b). tracking of the protein modulations associated with anti-Leishmania drug treatments for the characterization of a relevant protein set. The aim of both projects was to understand more of the mechanism of action for drug combinations and identify the connected relevant proteins, as well as discover novel targets for drug development. In the first project, Pemetrexed was used after the first-line therapy on ovarian tissue samples from patients affected by platinum-resistant epithelial ovarian cancer (PR-EOC). A differential proteomic approach was developed in order to identify the Differentially Expressed Proteins (DEPs). A combined proteomic and bioinformatic approach was employed. After network data analysis, we identified a panel of 24 proteins that characterize the response to Pemetrexed. The method applied and workflow developed included a protein set of reference as a new tool to help protein validation. A second project of the first type was based on the analysis of 5-FU-modulated proteins to track the drug efficacy in therapy within a retrospective study on serum samples from recurrent epithelial ovarian cancer (EOC) patients treated with the drug after multiple other treatments and to evaluate the early response for each patient. More than 200 proteins were differentially modulated after 5-FU treatment. Comparing the basal serum samples, three DEPs between responder and not-responder patients were discovered. A second differential analysis between post and pre-treatment on each patient was performed on samples collected at different times of chemotherapy schedule and allowed the identification of 14 proteins that could represent a potential pharmacodynamic biomarker of drug efficacy. Among them, the FHR1 protein showed a specific behavior. It is involved in complement regulation and may play a role on the protection of ovarian tumor cells against humoral immune attack. Project b) focused on the identification of the protein profile associated with the action of two anti-Leishmania agents. Parasitic proteins availability show a limited characterization and annotation in databases and often their function is not classified. In our case, 406 proteins were identified and quantified. 17 modulated proteins were obtained and their involvement in the mechanism of action compound H0080 is under investigation. In a second study upon Miltefosine, milte1 and milte2 treatments, we identified 35 differentially expressed proteins that are modulated by three treatments in the same direction and are involved in several biological processes. This suggested which components were relevant for drug resistance development and the identification of new targets. A third sub-project was related to the biological screening of new thymidylate synthase inhibitors in which new leads were identified.