Dipartimento di Scienza e Tecnologia del Farmaco

Descrizione del progetto

Sponsors:

University of Turin, Italy

Description

Objectives:
The study of the effect of post-squalene deficiencies and cholesterol intermediates accumulation (mainly those dependent on malfunctioning of C-4 demethylation complex) on  (i) other enzymes of post-squalene cholesterol biosynthesis, (ii) the mammalian embryo development and  (iii)  the Hedgehog protein-dependent tumors.

This study will be achieved by:

• enzymatic assays on 3-ketosteroid-reductase deficient mouse embryos;
• toxicity assays of 3-ketosteroids (prepared in our lab by fermentation procedure with engineered yeast strains) on Hedgehog-dependent tumor cell lines;
• sterol profile determination and enzymatic assays on mouse embryo cell line lacking C-3 sterol dehydrogenase (C-4 decarboxylase), an enzyme belonging to the sterol C-4 demethylase complex; deficiency mimics human CHILD syndrome;
• sterol profile determination and enzymatic assays on mouse embryo cell line lacking Erg 28p, the non-enzymatic scaffold protein of the sterol C-4 demethylase complex.

The study of the antiangiogenic and antitumoral activity of the sterol biosynthesis inhibitors and biochemical mechanism involved.

 Techniques and facilities:

• human cell culture and in vitro enzymatic tests
•  fully equipped laboratory for radioactive molecules and tracing of intermediate metabolites with radiolabelled precursors
• fully equipped biochemistry laboratory for cell/tissue homogenization and protein purification
•  protein electrophoresis and western blot
•  fermentation laboratory for production of intermediates of cholesterol biosynthesis by recombinant yeast strains

 Collaborations:
- Matti Poutanen, Turku Center for Disease Modelling, Turku, Finland.
- Gail Herman, Herman Lab. Center for Human and Molecular Genetics, the Research Institute at Nationwide Children's Hospital, Columbus).
- Donald Poirier, Department of Molecular Medicine, Research Center and Laval University, Quebec City, Canada.
- Martin Bard, IUPU University, Indianapolis, USA

Keywords:
Cholesterol, Cholesterol biosynthesis, Inhibitors, C-4 sterol demethylase, Postsqualene sterol biosynthesis, Antitumoral activity, Antiangiogenic activity, Enzymatic activity, Cholesterol disorders, Hedgehog proteins, Cell proliferation, Signalling pathway

Links:
Turku Center for Disease Modelling