Utjecaj utišavanja integrina ? V?3 i ? V?5 na vezanje, internalizaciju i transdukciju adenovirusnim vektorima

master's thesis

• Title (english): Influence of ?v?3 and ?v?5 integrin silencing on attachment, internalization and transduction with adenoviral vectors
• Author: Irena Bardak
• Mentor: Andreja Ambriović Ristov (mentor)
• Committee member: Mladen Krajačić (predsjednik povjerenstva)
• Committee member: Andreja Ambriović Ristov (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Dubravka Hranilović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2011-02-18, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Integrini su heterodimeri sastavljeni od α i β podjedinica koji se nalaze na površini stanica. Integrini αvβ3 i αvβ5 su receptori za proteine izvanstaničnog matriksa i sekundarni receptori pri infekciji adenovirusom tip 5 (Ad5). U ovom smo radu željeli istražiti ulogu integrina αvβ3 i αvβ5 u Ad5 infekciji i to metodom utišavanja ekspresije transfekcijom specifičnih siRNA. Pokazali smo da utišavanje β3 podjedinice integrina u stanicama MDA-MB-435S snižava ekspresiju αvβ3 i povećava ekspresiju integrina αvβ5, što rezultira povećanom transdukcijom divljeg tipa Ad5RSVβgal i Ad5 preusmjerenog na integrine αv Ad5RGD4CRSVβgal. Utišavanje β5 podjedinice snižava ekspresiju integrina αvβ5 i dovodi do smanjivanja transdukcije spomentih Ad5. Nakon utišavanja β3 i β5 podjedinice integrina zapazili smo malo smanjeno vezanje i internalizaciju Ad5RGD4CRSVβgal. Prema tome povećana transdukcija Ad5RSVβgal i Ad5RGD4CRSVβgal u stanicama transficiranima sa β3-specifičnom siRNA, nije rezultat promjena u vezanju i internalizaciji, nego boljeg oslobađanja iz endosoma zahvaljujući povećanoj ekspresiji integrina αvβ5. Zaključujemo da je ekspresija integrina αvβ5 ključni faktor u predviđanju djelotvornosti trandukcije Ad5 jer regulira izlazak iz endosoma.

Abstract (english)

Integrins are cell surface heterodimers composed of α and β subunits. Integrins αvβ3 and αvβ5 bind to extracellular matrix proteins and serve as a secondary receptors in adenovirus type 5 (Ad5) infection. The aim of this work was to investigate the role of αvβ3 and αvβ5 integrins in Ad5 infection using siRNA-mediated gene silencing. We have shown that silencing of β3 integrin subunit in MDA-MB-435S cells decreases the expression of αvβ3, but increases the expression of αvβ5 integrin, which results in increased transduction by wild type Ad5RSVβgal and adenovirus retargeted to αv integrins Ad5RGD4CRSVβgal. Silencing of β5 integrin subunit decreases the expression of αvβ5 integrin and leads to decreased transduction by both, abovementioned Ad5. Silencing of β3 and β5 integrin subunits slightly decreased Ad5RGD4CRSVβgal attachment and internalization. According to that, the observed increased Ad5RSVβgal and Ad5RGD4CRSVβgal transduction in cells transfected with specific β3 siRNA, is not a result of changes in attachment and internalization, but is the consequence of more efficient endosome release. Therefore, expression of αvβ5 integrin is the key factor to predict Ad5 transduction efficacy because it regulates escape from the endosome.

• Keywords: αvβ3; αvβ5; adenovirus; genska terapija tumora
• Keywords (english): αvβ3; αvβ5; adenovirus; tumor gene therapy
• Language: croatian
• URN:NBN: urn:nbn:hr:217:183179
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-02 12:38:02