Project 6 - Decoding Neuropilin Signaling in Angiogenesis
Applying for Summer 2025
Supervisor: Dr Stephen Robinson
School/Institute: Quadram Institute Bioscience
Introduction: Angiogenesis, the formation of new blood vessels, is fundamental to tissue repair, development, and diseases like cancer and diabetic retinopathy. Neuropilins (NRPs), key co-receptors for vascular endothelial growth factor (VEGF) and semaphorins, integrate signals that regulate endothelial cell behaviour. Understanding how NRPs modulates these processes offers potential for novel cancer therapies. This 8-week project, supervised by Dr. Stephen Robinson at the Quadram Institute, will investigate how NRPs’ regulated angiogenesis, combining molecular techniques and functional assays. Dr. Robinson’s recent work [1,2] highlights the role of NRPs in balancing pro- and anti-angiogenic pathways. While NRPs’ interactions with VEGF and semaphorins are recognised, the specific mechanisms driving differences in pro and anti-angiogenic signalling remain unclear. This project builds on our findings to explore how signalling pathways and functional outcomes during angiogenesis are regulated by NRPs.
The Challenge: NRPs’ ability to promote or inhibit angiogenesis depending on the context presents therapeutic challenges. Key questions include:
How do NRPs regulate endothelial migration and sprouting through VEGF-A and semaphorin signals?
What downstream pathways mediate these effects, and how do they influence angiogenesis?
Can targeted modulation of NRPs create controlled angiogenic responses?
Aims and Objectives: This project aims to elucidate how the two mammalian NRPs (NRP1/NRP2) integrate VEGF and semaphorin signals to regulate endothelial cell function. Objectives include:
Investigating NRP-dependent signalling pathways.
Assessing functional effects on endothelial migration, proliferation, and sprouting.
Exploring NRP’s interactions with ECM components and their role in angiogenesis.
Experimental plan, expected outcomes and potential outcomes: The project is divided into three experimental modules over eight weeks:
1. Molecular Characterization (Weeks 1-3). Employ siRNA knock down of NRP1 and/or NRP2 in endothelial cells and assess changes in downstream signalling pathways, via Western blotting and RT-qPCR to analyse known angiogenic signalling pathways in cells treated with VEGF-A and semaphorins.
2. Functional Assays (Weeks 4-6). Perform scratch wound-healing and transwell assays to measure endothelial migration in cells treated with NRP1/2 siRNA. Evaluate angiogenic sprouting using 3D spheroid assays under VEGF-A and semaphorin gradients.
3. NRP-ECM Interactions (Weeks 7-8). Visualize focal adhesion dynamics with live-cell imaging after siRNA treatment under VEGF-A and semaphorin treatment.
By project completion, the student will have:
• Identified key signalling pathways differentially activated by NRPs.
• Quantified the effects of NRP modulation on endothelial cell function.
• Explored NRP-ECM interactions and their contribution to angiogenesis.
This research could inform therapeutic strategies targeting NRPs in cancer and ischemic diseases. Insights into NRP-ECM interactions may also aid tissue engineering and regenerative medicine.
Skills Gained: The student will gain expertise in molecular biology, angiogenesis assays, and data analysis, enhancing their research and critical thinking skills. This 8-week project offers an exceptional opportunity to contribute to cutting-edge angiogenesis research, providing a solid
References: (1) Benwell, CJ. Johnson, RT, Taylor, JAGE, Lambert, J, and Robinson, SD. (2024). A proteomics approach to isolating neuropilin dependent alpha5 integrin trafficking pathways: neuropilin-1 and 2 co-traffic alpha5 integrin through endosomal p120RasGAP to promote polarised fibronectin fibrillogenesis in endothelial cells. Communications Biology, 7:629. (2) Benwell, CJ, Johnson, RT, Taylor, JAGE, Price, CA, and Robinson, SD. (2022). Endothelial VEGFR coreceptors neuropilin-1 and neuropilin-2 are essential for tumor angiogenesis. Cancer Research Communications, 2:1626-1640.