Project 6 - Decoding Neuropilin Signaling in Angiogenesis

Applying for Summer 2025

Apply Now

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:

  1. Investigating NRP-dependent signalling pathways.

  2. Assessing functional effects on endothelial migration, proliferation, and sprouting.

  3. 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.

Project 6 - Decoding Neuropilin Signaling in Angiogenesis