The mission of the Angiogenesis Laboratory, which is part of the Department of Pathology, is two-fold. Firstly, it is aimed to unravel fundamental processes and mechanisms of angiogenesis. Secondly, it is aimed to apply research and new technology for development of novel (cancer) treatment modalities in the clinic. Four different interconnective research lines are currently ongoing.
I. Genetic profiling of tumor endothelium.
This research line focuses on the identification of specific targets on tumor endothelium. We compared the transcriptional profile, by suppression subtractive hybridization analysis, of tumor endothelial cells with that of normal resting endothelial cells, normal but angiogenically activated placenta endothelial cells, and tumor-conditioned cultured endothelial cells. Though the majority of transcripts were classified as general angiogenesis markers, we identified 17 genes that show specific overexpression in tumor endothelium. We recently showed that targeting of four cell-surface expressed or secreted products with antibodies inhibited angiogenesis in vitro and in vivo. In collaboration with Dr. Van Engeland we also identified a series of genes that are silenced in tumor endothelium. The role of these genes in angiogenesis is currently being investigated.
This research is supported by GROW and by substantial support from industry.
II. Identification and development of new angiostatic agents.
The angiostatic designer peptide anginex and its mimetic KM0118 have been developed in the Angiogenesis Laboratory in collaboration with the University of Minnesota (Prof. K.H. Mayo). The research to identify the mechanisms of action has revealed galectin-1 as the receptor that transduces the signaling by anginex. It was found that this molecule is of extreme importance in development of vasculature as well. This was demonstrated by morpholino knock-down experiments in zebrafish.
The potential application of angiostatic agents in therapy is studied by gene expression profiling. This includes known molecules involved in angiogenesis as well as newly discovered ones. The prognostic value of such angiogenesis parameters is studied in lung cancer and in pediatric tumors.
Research line II is supported by KWF, NWO/STW and NIH.
III. Study of the cross-talk between the vasculature and the immune system. Current research has revealed that angiogenic potential, as measured by proliferating endothelial cells, in colorectal cancer is a prognostic factor (see figure). Similarly, leukocyte infiltration is of prognostic value. This has therapeutic implications and is in support to future combination therapies of anti-angiogenesis and immunotherapy. The novel angiogenesis inhibitors as identified in research line II were found to improve vascular adhesion molecule expression and leukocyte infiltration into the tumor. Immunotherapy protocols will be tested in mice to develop these compounds for clinical testing.
This research line is supported by KWF/NKB and by the Coenegracht Stichting.
IV. Tumor cell plasticity and vasculogenic mimicry.
Recent work has indicated that blood lakes in Ewing sarcoma contribute to circulation and can be regarded as a sign of vasculogenic mimicry. We demonstrated that hypoxia is a driving force behind this phenomenon.
This work is financially supported by GROW.
Selected publications
Thijssen, V.L., Postel, R., Brandwijk, R.J., Dings, R.P., Nesmelova, I., Satijn, S.A., Verhofstad, N., Nakabeppu, Y., Baum, L. G., Bakkers, J., Mayo, K.H., Poirier, F., and Griffioen, A.W.
Galectin-1 is essential in angiogenesis and is a target for anti-angiogenesis therapy. Proc.Natl.Acad.Sci.USA, 103:15975-15980, 2006
Derksen, P.W., Liu, X., Saradin, F., Evers, B., Van Beijnum, J.R., Griffioen, A.W., Van der Gulden, H., Zevenhoven, J., Peterse, H., Cardiff, R., Vink, J., Krimpenfort, P., Berns, A., and Jonkers, J.
Somatic inactivation of E-cadherin and p53 in mice leads to metastatic lobular mammary carcinoma through induction of anoikis resistance and angiogenesis.
Cancer Cell, 10:437-449, 2006
Dings, R.P., Chen, X., Nesmelova, I., Haseman, J., Hellebrekers, D.M.E.I., Maxwell, J., Van Eijk, L.I., Hoye, T.R., Griffioen, A.W., Mayo, K.H.,
Design of non-peptidic helix/sheet topomimetics: applications to inhibition of angiogenesis and tumor growth in mice.
J Natl Cancer Inst, 98:932-936, 2006.
Van Beijnum, J.R., Dings, R.P.M., Zwaans, B., Van der Linden, E., Ramaekers, F.C.S., Mayo, K.H. and Griffioen, A. W.
Gene expression of tumor angiogenesis dissected; specific targeting of colon cancer angiogenic vasculature.
Blood, 108:2339-2348, 2006.
Mulder, W.J.M., Koole, R., Brandwijk, R.J., Storm, G., Chin, P., Strijkers, G.J., Celso de Mello Donega, Nicolay, K., Griffioen, A.W.
Paramagnetic quantum dots as a bimodal molecular imaging probe for angiogenesis.
Nano Letters. 6:1-6, 2006.
Figure: Loss of galectin-1 L2 and L3 results in hemorrhaging and defective vessel formation in the zebrafish brain.
(A-D) Staining for blood (arrows). (A) wild type control (B) morpholino knock- down lgals1 L2 AT-MO, (C) lgals-1 L3 ATG-MO, (D) both lgals-1 L2 and L3 ATG- MOs. Co-injection of L2 and L3 ATG-MO results in severe hemorrhaging in the brain region (arrowheads). (E) Schematic drawing of blood vessels in the dorsal brain at 2.5 day of development. Confocal microscopy from Tg(fli1:egfp)y1 transgenic embryos at the level of the dorsal brain vessels. (F) wild type control embryo.
(G) Embryos co-injected with lgals-1 L2 and -L3 ATG-MO display aberrant sprouting and misguidance of the middle cerebral vein (MCeV) into the dorsal longitudinal vein (DLV) (arrowheads). Defective angiogenic sprouting is also observed in the mesencephalic vein (arrow). (H) Co-injection of the lgals-1 L2 and L3 splice-MO shows similar defects in angiogenic sprouting of the brain vessels. DLV, dorsal longitudinal vein; MCeV, l vein; MsV mesencephalic vein; MtA, metencephalic artery; PCeV, posterior cerebral vein.
Research group
Dr. Arjan W. Griffioen, cell biologist, immunologist, project leader
Dr. Judy R. van Beijnum, molecular biologist
Dr. Freek Bot, MD, pathologist
Dr. Anne-Marie Dingemans, MD, pulmonologist
Dr. Ruud P.M. Dings, cell biologist
Dr. Bernd Granzen, MD, pediatrician
Dr. Daisy W.J. van der Schaft, cell biologist
Dr. Sebastien Tabruyn, molecular biologist.
Dr. Victor Thijssen, molecular biologist
PhD Students
Bisan Ahmed
Marcella Baldewijns
Vivian van den Boogaart
Ricardo Brandwijk
Karolien Castermans
Debby Hellebrekers
Femke Hillen
Veerle Melotte
Willem Mulder
Technicians
Loes van Eijk
Petra Hautvast
Sarah Hulsmans
Edith van der Linden
Sietske Satijn
Students
Rene Marx