This thesis centers on the mechanisms of estrogen action and the effects on the development of atherosclerosis. We have focused on the liver as central organ in lipid and glucose metabolism and the... Show moreThis thesis centers on the mechanisms of estrogen action and the effects on the development of atherosclerosis. We have focused on the liver as central organ in lipid and glucose metabolism and the vessel wall as the actual site where the injury occurs. To gain insight in tissue-specific actions of estrogens, we have spent considerable effort to develop tools for liver and blood vessel specific modulation of the estrogen receptor (ER) signaling cascade. The generation, characterization and application of these tools in vitro and in vivo will be described in the different chapters of this thesis. Show less
The adenoviral vector is one of the most used viral vectors for gene therapy applications. The specificity towards its primary receptor is a limitation of the use of HAdV as therapeutical vector,... Show moreThe adenoviral vector is one of the most used viral vectors for gene therapy applications. The specificity towards its primary receptor is a limitation of the use of HAdV as therapeutical vector, as not all cells have these primary receptors on their surface. To overcome this problem scientists have started to change the receptor specificity of the HAdV vectors by introducing heterologous polypeptides. One of the most promising candidates to function as anchor for these heterologous polypeptides is the minor capsid protein IX (pIX). Alpha-helical spacers of different lengths have been inserted between pIX and a polypeptide ligand. The results showed that an increase in spacer length positively correlated with the accessibility of the peptide on the capsid surface for antibodies and cellular membrane proteins. Next, a hyper stable scFv directed against __-galactosidase was incorporated in the virion via fusion to pIX. These data were the first to show incorporation of a hyper-stable scFv into the HAdV, and that was functional on the capsid surface. Beside the technical approach to use pIX as anchor for heterologous polypeptides here also the function of pIX in the capsid has been studied. This study has led to the model in which the capsid stability mediated by pIX might not be the result of homo-multimerization of this molecule. Show less
