SAINTargs and SAINT-O-Somes

novel lipid-based targeting devices for siRNA delivery

Niek G.J. Leus, Piotr S. Kowalski, Grietje Molema, and Jan A.A.M. Kamps

University Medical Center Groningen, Department of Pathology & Medical Biology, Laboratory of Endothelial Medicine & Vascular Drug Targeting, Hanzeplein 1, 9713 GZ Groningen

The endothelium represents an important therapeutic target because its pivotal role in many diseases such as chronic inflammation (e.g. Rheumatoid Arthritis) and cancer. Furthermore the endothelium is fully accessible for systemically administered therapeutics. RNA interference by small interfering RNA (siRNA) has become in the last decade a very powerful tool in basic research, and has huge potential to become an important new class of therapeutics for humans. However, due to their size and charge, siRNAs have no bioavailability to enter unperturbed cells. To overcome this problem, our laboratory developed two lipid-based targeting devices for siRNA delivery, so called SAINTargs and SAINT-O-Somes, which efficiently and specifically deliver siRNA into endothelial cells. Molecular determinants expressed on the surface of inflammation-activated endothelial cells, like certain adhesion molecules and receptors involved in endocytosis, are excellent candidates to increase carrier-mediated siRNA uptake. Therefore we conjugated monoclonal anti-E-selectin or anti-VCAM-1 antibodies to these SAINTargs and/or SAINT-O-Somes, resulting in two targeted siRNA delivery devices (Ásgeirsdóttir et al., 2010; Adrian et al., 2010).

Taken together, SAINTargs and SAINT-O-Somes demonstrate specific and efficient targeting to inflammation-activated difficult-to-transfect primary endothelial cells and results in strong siRNA specific gene silencing. We are now further developing these two carriers for in vivo application. Within this research we have challenging opportunities for motivated HLO students to do an internship in our laboratory.

Techniques:

-  eukaryotic cell culturing

-  eukaryotic transfections

-  liposome technology

-  RNA isolation, integrity check (electrophoresis), cDNA synthesis

-  (quantitative) PCR

-  flow cytometry

-  immunohistochemistry

-  western blotting

-  fluorescence and/or confocal and/or laser-dissection microscopy

Contact: Niek Leus

Phone: 050-3610943 or 06-30329596

Email:

http://www.rug.nl/umcg/faculteit/disciplinegroepen/plg/medbiol/research/ec/index