| Grant number: | 16/18066-6 |
| Support type: | Regular Research Grants |
| Duration: | November 01, 2016 - April 30, 2019 |
| Field of knowledge: | Biological Sciences - Biochemistry - Molecular Biology |
| Principal Investigator: | Leny Toma |
| Grantee: | Leny Toma |
| Home Institution: | Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil |
| Assoc. researchers: | Helena Bonciani Nader ; Nora Manoukian Forones |
Abstract
Molecules from tumor microenvironment has become the purpose of intense research in cancer treatment. Proteoglycans (PGs) has been the target of these studies due to their location, as well as their structural features which contributes to their regulatory and multifactorial functions. In the previous project, we have shown the importance of 6-O-sulfation in heparan sulfate proteoglycans through the extracellular enzymes 6-O-endosulfatases, Sulf 1 and Sulf 2, which remove sulfate from the 6-O position, editing these extracellular domains. In the present project, the aim is to further investigate these enzymes expression, now in vivo, analyzing human colorectal cancer tissues and to compare to the normal adjacent ones. A second aim of this project will be to study proteoglycans from colorectal adenocarcinoma cell lines, which have also 6-O-sulfated domains, however in chondroitin sulfate chains. These molecules are denominated chondroitin sulfate E (CS-E), related in the literature with higher motility of neural cells. For this purpose, knockdown of the N-acetylgalactosamine-4-sulfate 6-O-sulfotransferase (GalNAc4S-6ST), enzyme from the biosynthesis of the CS-E will be achieved by interference RNA using shRNA. Resistant clones will be isolated and selected by Real Time PCR and western blot. The effects of GalNAc4S-6ST knockdown will be investigated by structural studies with specific enzymes, and biological assays analyzing cell proliferation, adhesion, migration and invasion. Furthermore, the involvement of CS-E in Wnt signaling will be studied because some evidence has shown recognition of CS-E and Wnt3a ligant, although this signaling pathway needs to be further investigated. Proving these results we hope to advance towards the elucidation of the role these charged molecules play in cancer biology and disease progression. (AU)