Investigation of nanoscale RAS polarizations in instructing cancer cell stemness

• Applicant: Professor Dr. Christian Eggeling
• Subject Area: Cell Biology ; Biophysics ; Hematology, Oncology
• Term: from 2020 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 432186367

• Final Report Year: 2024

Final Report Abstract

The original motivation of the this project was in the area of cancer research and investigations of mechanisms that could lead to improved treatments. Currently major drug development efforts focus on one of the major cancer proteins called Ras, which has evaded drug targeting for 30 years. The main aim was to understand, what makes Ras so special that it is mutationally exploited in 30% of all cancers, including major cancer types, such as colon and lung. We were interested in the role of Ras to drive aberrant dedifferentiation that closely correlates with cancer severity. Our approach contrasted with the predominant focus on cell proliferation in cancer research. Using sophisticated microscopic methods that can reveal details down to the molecular scale, we investigated how Ras controls the normal organisation of cells. Our insights impacted on understanding not only the biology of Ras, but also of similar cancer associated proteins, as well as brought up novel technological microscopy advancements on the investigation of the membrane organization in polarized cells. Finally, our results inspired us to start investigations on the role of Ras proteins in the cilium, which laid ground for newly submitted applications of the same team.

Publications

• International Conference on the Bioscience of Lipids (ICBL) – Poster: Fluorescence microscopy studies of polarized cells - 12-15 October 2021 – Utrecht, Netherlands
  Rouzbahani, Y.
  
• Promotion of cancer cell stemness by Ras. Biochemical Society Transactions, 49(1), 467-476.
  Chippalkatti, Rohan & Abankwa, Daniel
  
• FEBS Practical Course on functional imaging of cellular dynamics – Poster and Talk: Advanced microscope studies of the cell polarization instructing activity of Ras proteins - 12- 18 June 2022 – Amsterdam, Netherlands
  Rouzbahani, Y.
  
• A flow-cytometry-based pipeline for the rapid quantification of C2C12 cell differentiation. STAR Protocols, 4(4), 102637.
  Parisi, Bianca; Sünnen, Maxime; Chippalkatti, Rohan & Abankwa, Daniel Kwaku
  
• EMBO workshop on cell polarity and membrane dynamics – Poster: Adaptation of microscopy for studying cell polarization - 21-25 May 2023 – Girona, Spain
  Rouzbahani, Y.
  
• Influence of different surface cleaning methods on STED‐FCS and scanning STED‐FCS calibration measurements. Journal of Microscopy, 296(2), 150-153.
  Koerfer, Agnes; Reina, Francesco & Eggeling, Christian
  
• International Conference on the Bioscience of Lipids (ICBL) – Poster: Adaptation of microscopy for studying cell polarization - 2-5 October 2023 – Palma de Mallorca, Spain
  Rouzbahani, Y.
  
• Leibniz Association PhD Retreat – Poster: Advanced microscopy studies of the cell polarization instructing activity of Ras proteins - 6-8 March 2023, Dornburg, Germany
  Rouzbahani, Y.
  
• Photonics West - Poster: Adaptation of microscopy for studying cell polarization - 28 Jan - 2 Feb 2023 - San Francisco, USA
  Rouzbahani, Y.
  
• Quantitative analysis of peroxisome tracks using a Hidden Markov Model. Scientific Reports, 13(1).
  Svensson, Carl-Magnus; Reglinski, Katharina; Schliebs, Wolfgang; Erdmann, Ralf; Eggeling, Christian & Figge, Marc Thilo
  
• Effects and avoidance of photoconversion-induced artifacts in confocal and STED microscopy. Nature Methods, 21(7), 1171-1174.
  Dasgupta, Anindita; Koerfer, Agnes; Kokot, Boštjan; Urbančič, Iztok; Eggeling, Christian & Carravilla, Pablo
  
• Nanobody-mediated neutralization of candidalysin prevents epithelial damage and inflammatory responses that drive vulvovaginal candidiasis pathogenesis. mBio, 15(3).
  Valentine, Marisa; Rudolph, Paul; Dietschmann, Axel; Tsavou, Antzela; Mogavero, Selene; Lee, Sejeong; Priest, Emily L.; Zhurgenbayeva, Gaukhar; Jablonowski, Nadja; Timme, Sandra; Eggeling, Christian; Allert, Stefanie; Dolk, Edward; Naglik, Julian R.; Figge, Marc T.; Gresnigt, Mark S. & Hube, Bernhard
  
• Neural network informed photon filtering reduces fluorescence correlation spectroscopy artifacts. Biophysical Journal, 123(6), 745-755.
  Seltmann, Alva; Carravilla, Pablo; Reglinski, Katharina; Eggeling, Christian & Waithe, Dominic
  
• RAS isoform specific activities are disrupted by disease associated mutations during cell differentiation. European Journal of Cell Biology, 103(2), 151425.
  Chippalkatti, Rohan; Parisi, Bianca; Kouzi, Farah; Laurini, Christina; Ben, Fredj Nesrine & Abankwa, Daniel Kwaku