Master- / Promotionsarbeit

Characterization of Tumor-Derived Extracellular Vesicles in Urological Cancers Using Nanoflow Cytometry

The Laboratory for Cell and Molecular Biology at the Department of Urology and Pediatric Urology, Saarland University, offers a Master’s thesis project in the field of extracellular vesicle (EV) biology and tumor immunology.

Extracellular vesicles (EVs) are membrane-enclosed nanoparticles released by virtually all cell types and have emerged as important mediators of intercellular communication. Tumor-derived EVs contribute to multiple aspects of cancer progression, including immune evasion, metastasis, and therapy resistance. Because EVs carry proteins reflective of their parental cells, they are increasingly recognized as promising biomarker candidates for liquid biopsy applications and personalized medicine. Recent advances in nanoflow cytometry now allow the characterization of individual EVs and provide new opportunities to investigate EV heterogeneity at the single-particle level.

The Department of Urology and Pediatric Urology investigates the molecular mechanisms underlying urological malignancies, including renal cell carcinoma (RCC), bladder cancer, and penile cancer. Several clinically relevant proteins, such as programmed death ligand-1 (PD-L1), carbonic anhydrase IX (CA9), Nectin-4, TROP-2, HER2, and EGFR, have emerged as important biomarkers and therapeutic targets in these diseases. While their expression on tumor cells has been extensively studied, little is known about their distribution among individual tumor-derived EV populations.

This Master project aims to establish and optimize a nanoflow cytometry workflow for the characterization of tumor-associated surface markers on extracellular vesicles derived from urological cancer cell lines. Understanding the heterogeneity of tumor-derived EVs may provide valuable insights into tumor–immune interactions and contribute to the development of future EV-based biomarkers.

In this project, extracellular vesicles will be isolated from established cell culture models of renal cell carcinoma, bladder cancer, and penile cancer. First, the expression of selected tumor-associated markers and classical EV markers will be characterized in parental tumor cells using conventional flow cytometry, immunostaining, and immunoblotting. Subsequently, EVs will be isolated from conditioned cell culture supernatants and characterized according to current extracellular vesicle guidelines using nanoparticle tracking analysis and immunoblotting. Finally, nanoflow cytometry will be established and optimized for the detection of EV-associated surface proteins at the single-particle level. The expression of tumor-associated markers, including PD-L1, CA9, Nectin-4, TROP-2, HER2, and EGFR, will be investigated in EV populations derived from different urological cancer models. Furthermore, the project will characterize the heterogeneous expression of tumor-associated markers on individual EVs and compare EV surface signatures across different urological cancer models.

The project will involve the following techniques:

• Mammalian cell culture
• Extracellular vesicle isolation and characterization
• Nanoparticle tracking analysis (NTA)
• Western blotting
• Conventional flow cytometry
• Nanoflow cytometry
• Immunofluorescence microscopy
• Statistical data analysis

During your Master’s thesis, you will contribute to ongoing research projects within our laboratory. In addition to acquiring expertise in extracellular vesicle biology and advanced flow cytometry technologies, you will gain experience in experimental design, data interpretation, scientific presentation, and critical discussion of current literature. The applicant should hold a Bachelor’s degree in biology, biotechnology, biochemistry, molecular medicine, or a related discipline.

If you are interested in this project, please contact:

Prof. Dr. Kerstin Junker
Head of Clinical and Experimental Research
Department of Urology and Pediatric Urology
Saarland University Medical Center

Kerstin.junker(at)uks.eu

06841-1614734

Dr. Nadia Alawar
Postdoctoral Researcher at the Department of Urology and Pediatric Urology
Saarland University Medical Center

Nadia.alawar(at)uks.eu

06841-1624854