Angiogenesis, the growth of new blood vessels, contributes to major pathologies such as blinding ocular disease, inflammation and cancer. Blocking angiogenesis has therefore become a major field of research and an attractive therapeutic strategy. Current anti-angiogenic therapies focus on blockade of pro-angiogenic factors, such as VEGF. However, in cancer, insufficient efficacy, resistance and toxicity restrict the success of anti-VEGF agents. There is thus an urgent unmet need for novel anti-angiogenic strategies. In our ERC Advanced research grant (ECMetabolism), we developed an entirely novel anti-angiogenic concept and strategy, based on targeting key metabolic pathways in endothelial cells (ECs), cells lining blood vessels. More in particular, we identified - for the first time - that PFKFB3, a key glycolytic regulator, as a novel and promising target for anti-angiogenic therapy. Our findings show that glucose metabolism determines vessel sprouting and that lowering glycolysis only partially and transiently (by blocking PFKFB3) sufficed to inhibit pathological angiogenesis without causing systemic effects. In this ERC proof of concept project, PFKFBLOCK, we aim to develop lead small molecule compounds, blocking PFKFB3, and evaluate their potential to block pathological angiogenesis. Currently, no specific and orally available PFKFB3 blocker exists underscoring the value of our proposal and the necessity to develop such blocker for therapeutic applications. Through collaboration with a drug discovery unit we will identify lead molecules with novel intellectual property potential. Those lead compounds will be validated in relevant in vivo models and considered to be patented. The ultimate goal of this project is to get the best PFKFB3 inhibitor in clinical trials.

Plants produce arrays of specialized metabolites that are crucial for plant development, defense and interactions with the ever-changing environment. Discovery of metabolites and metabolic pathways by dissecting the underlying genetic basis is an important challenge, not only to increase our understanding of life, but also to provide indispensable applications for humans such as pharmaceuticals. In this project we will investigate the occurrence and biosynthesis of the 2,5-dihydrochorismates, a novel metabolic class that was recently discovered by the host group in the plant model Arabidopsis (unpublished). Notably, chemically synthesized 2,5-dihydrochorismate and analogues have been shown to be potent in vitro inhibitors of chorismate-utilizing enzymes. This inhibitory activity hints to a potential biological role for 2,5-dihydrochorismates in regulating the flux through these pathways, opening perspectives for applications. The direct objective of my project is to unravel the biosynthetic pathway of 2,5-dihydrochorismates. We therefore aim i) to structurally identify additional pathway intermediates and sinks via LC-MS based metabolite profiling and purification followed by NMR, ii) to test whether chorismate is a precursor of the pathway by feeding plants with Stable Isotope Labeled chorismate, followed by analyzing the MS fragmentation spectra of the labeled molecules, iii) to investigate the role of an operon-like gene cluster involved in the biosynthesis of 2,5-dihydrochorismates via reversed genetics, and iv) to obtain insight in the in planta role of 2,5-dihydrochorismate by feeding experiment followed by metabolic profiling. My experience in reversed genetics and enzyme kinetics in specialized metabolism in rice make me well suited to bring this project to success. My prime objective is to strengthen my skills in metabolite profiling, pathway discovery and structural elucidation, to become an independent plant scientist in plant specialized metabolism.