A molecular mix and match: development and characterisation of 3-Dimensional mini-tissues ex vivo

Dr David Bryant (Cancer Sciences, University of Glasgow/CRUK Beatson Institute) http://www.beatson.gla.ac.uk/david_bryant
Dr Sara Zanivan (CRUK Beatson Institute)

Cancer is a disease where the rules for tissue organization are no longer followed. During cancer progression, a disorganized primary tumour disrupts organ function, while other cancer cells invade into surrounding tissues and spread to distant organs. Despite tissue disorganization being the commonality between most cancers, much of our knowledge comes from studies focused on a single cell type and using culture methods that lack tissue organization. However, a tissue is a complex collection of various cells types, and it is unclear how these each contributes to cancer progression.

This PhD project aims to:
1. Develop 3-Dimensional co-cultures of epithelial, endothelial, and stromal (fibroblast, adipocyte) cells to determine how these influence ex vivo ‘mini-tissue’ architecture.
2. Develop live-imaging methods to quantitatively track and analyse co-cultures.
3. Develop proteomic methods to interrogate the molecular signals between cell types that promote normal tissue organization, and its loss during tumourigenesis.

This PhD project, joint with the Bryant and Zanivan Labs, will allow the student to develop cutting-edge skill sets in 3-Dimensional culture, proteomics, live imaging, genetic manipulations and computational image analysis – all of which are directed towards understanding that molecular mechanisms of epithelial-endothelial-stromal crosstalk in tissue disorganization and tumour progression.

Keywords: Cancer, Mass spectrometry, Proteomics, Live imaging, Image analysis, Signalling, 3D culture, Co-culture, Epithelia, Fibroblast, Endothelia

References:

Román-Fernández A, Bryant DM. (2016) Complex Polarity: Building Multicellular Tissues Through Apical Membrane Traffic. Traffic. doi: 10.1111/tra.12417. [Epub ahead of print] PMID: 27281121.

Yang Z, Zimmerman SE, Tsunezumi J, Braitsch C, Trent C, Bryant DM, et al. (2016) Role of CD34 family members in lumen formation in the developing kidney. Dev Biol. 418(1):66-74. PMID: 27542690.

Bryant DM et al. (2014) A molecular switch for the orientation of epithelial cell polarization. Dev Cell. 31(2):171-87. PMID: 25307480.

Gálvez-Santisteban M, Rodriguez-Fraticelli AE, Bryant DM et al. (2012) Synaptotagmin-like proteins control the formation of a single apical membrane domain in epithelial cells. Nat Cell Biol. 14(8):838-49. PMID: 22820376.

Bryant DM, et al. (2010) A molecular network for de novo generation of the apical surface and lumen. Nat Cell Biol. 12(11):1035-45. PMID: 20890297.

Hernandez-Fernaud JR, Ruengeler E, Casazza A, Neilson LJ, Pulleine E, Santi A, Ismail S, Lilla S, Dhayade S, Macpherson IR, McNeish I, Ennis D, Ali H, Kugeratski F, Al Khamici H, van den Biggelaar M, van den Berghe PVE, Cloix C, McDonald L, Millan D, Hoyle A, Kuchnio A, Carmeliet P, Valenzuela SM, Blyth K, Yin H, Mazzone M, Norman JC and Zanivan S (2016) Secreted CLIC3 drives cancer progression through its glutathione-dependent oxidoreductase activity. Nature Communications (accepted for publication).

Patella F, Neilson LJ, Athineos D, Erami Z, Anderson KI, Blyth K, Ryan KM, Zanivan S (2016) In-Depth Proteomics Identifies a Role for Autophagy in Controlling Reactive Oxygen Species Mediated Endothelial Permeability. J Proteome Res. Jul 1;15(7):2187-97.

Patella F, Schug ZT, Persi E, Neilson LJ, Erami Z, Avanzato D, Maione F, Hernandez-Fernaud JR, Mackay G, Zheng L, Reid S, Frezza C, Giraudo E, Fiorio Pla A, Anderson K, Ruppin E, Gottlieb E, Zanivan S (2015) Proteomics-Based Metabolic Modeling Reveals That Fatty Acid Oxidation (FAO) Controls Endothelial Cell (EC) Permeability. Mol Cell Proteomics. Mar;14(3):621-34.

van den Biggelaar M, Hernández-Fernaud JR, van den Eshof BL, Neilson LJ, Meijer AB, Mertens K, Zanivan S (2014) Quantitative phosphoproteomics unveils temporal dynamics of thrombin signaling in human endothelial cells. Blood. Mar 20; 123(12):e22-36.

Zanivan S*, Maione F, Hein MY, Hernández-Fernaud JR, Ostasiewicz P, Giraudo E, Mann M* (2013) SILAC-based proteomics of human primary endothelial cell morphogenesis unveils tumor angiogenic markers. Mol Cell Proteomics. Dec; 12(12):3599- 611. * Corresponding author.

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