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Research summary

Our overall aim is to determine how EVs facilitate communication between cells within the human tumor microenvironment. The primary goal is to identify new EV-mediated mechanisms that contribute to cancer growth and progression, ultimately creating a knowledge base to support future advancements in cancer diagnostics and therapeutics. Our research group primarily focuses on investigating EVs in the context of melanoma, with additional ongoing projects examining the role of EVs in ovarian, pancreatic, breast, and lung cancers.

EVs are a heterogeneous group of bilayer membrane-enclosed nanoparticles released by all cells, and seemingly even more so by tumor cells. They are recognized as a crucial means of intercellular communication due to their ability to transfer cargo, including lipids, proteins, and nucleic acids.

The tumor microenvironment is established by cell-to-cell and cell-to-matrix interactions, and one of the ways cells interact is through the secretion and uptake of EVs that convey a vast array of signals between cells. The interstitial tissue of tumors contains large amounts of EVs that are released from both cancer and non-cancer cells into the tumor microenvironment.

Our research builds on a breakthrough method recently developed by the Crescitelli group that allows us to isolate highly purified EVs directly from human tumor tissues (Figure 1). Using this technique, EVs have been isolated from a range of tumors, including skin, breast, ovarian, colon, and pancreatic cancers, revealing disease-specific biomarkers within EVs. (Crescitelli, R., et al. Nat Protoc 2021, Crescitelli, R., et al. J Extracell Vesicles 2020, Jang, S.C. & Crescitelli R. et al. J Extracell Vesicles 2019).

Most studies focusing on EVs in the tumor microenvironment are in vitro studies. For the first time, we are able to visualize 3D reconstructions of EVs in an in vivo environment, and we were the first to present a 3D model highlighting the presence of EVs in the extracellular space. An electron microscopy-based tomographic reconstruction of liver melanoma metastatic tissue was created, and the 3D model clearly showed the presence of spherical structures in the extracellular space surrounded by five individual cells (Figure 2). Watch the video of the . (Olofsson Bagge, R. et al. J Extracell Vesicles 2023)

We have recently set up an innovative slicing machine to study the fate and biological functions of tissue-derived EV subpopulations in complex tumor tissues. The “Krumdieck slicer” is a tool that can produce high-precision tissue slices. Briefly, a punched cylindrical core of tissue is cut into 250 μm slices that are then cultured in regular tissue culture plates (Figure 3). The slices represent a miniature model of the studied organ because they leave the intercellular and cell-matrix interactions intact.

We strongly believe that this system will prove useful in studying EV subpopulation dynamics within tumor tissues.

Research tools and resources

• EV isolation from tissues, cell lines, and serum/plasma using the most advanced techniques in the field (differential centrifugation, iodixanol density gradients, and size exclusion chromatography)
• EV characterization using transmission electron microscopy, Nanoparticle Tracking Analysis (ZetaView), single-vesicle surface analysis (ExoView), and flow cytometry for single nanoparticle characterization (nanoFCM)
• Functional assays using tumor and non-tumor tissues from humans, mice, and rats
• The use of a Krumdieck slicer to produce precision tissue slices
• 3D tomographic reconstructions using the IMOD program
• Proteomics analyses using mass spectrometry

Selected publications

1. 
   *Crescitelli R#, Lässer C, Lötvall J. Nature Protocols, 2021 Mar;16(3):1548-1580
    
2. 
   Olofsson Bagge R, Berndtsson J, UrzìR, Lötvall J, Micaroni M, Crescitelli R^#. Journal of extracellular vesicles 2023Dec;12(12):e12380.
    
3. 
   Urzì O, Bergqvist M, Lässer C, Moschetti M, Johansson J, D´Arrigo D, Olofson Bagge R, Crescitelli R^#. J Extracell Vesicles. 2024 Jan;13(1):e12408. doi: 10.1002/jev2.12408.
    
4. 
   Crescitelli R*, Lässer C, Jang SC, Cvjetkovic A, Malmhäll C, Karimi N, Höög JL, Johansson I, Fuchs J, Thorsell A, Gho YS, Olofsson Bagge R, Jan Lötvall J (2020) Journal of Extracellular Vesicles, 2020 Feb 11;9(1), 1722433
    

5. 
   *Crescitelli R#, Filges S, Karimi N, Urzì O, Alonso-Agudo T, Ståhlberg A, Lötvall J, Lässer C, Bagge R.O. Front. Cell Dev. Biol. 2022 Dec; 10:1028854
    

6. 
   Jang SC*, Crescitelli R*, Cvjetkovic A, Belgrano V, Olofsson Bagge R, Sundfeldt K, Ochiya T, Kalluri R, Lötvall J. Journal of Extracellular Vesicles 2019 Aug 27;8(1):1635420. 
    

7. Ultrastructure of multi-vesicular bodies in fission yeast.
   Zabeo D*, Crescitelli R*, O’Toole E*, Roque H*, Höög J. 3D 91̽�� Matters Mar 2017 10.19185/matters.201702000007
    

8. 
   Cvjetkovic A, Karimi N, Crescitelli R, Thorsell A, Taflin H, Lässer C, Lötvall J Journal of Extracellular Biology 2024 Feb 6;3(2):e127.
    

9. 
   Park KS, Svennerholm K, Crescitelli R, Lässer C, Gribonika I, Andersson M, Boström J, Alalam H, Ali M Harandi AM, Farewell A, Lötvall J. Journal of Nanobiotechnology (2023) 19;21(1):156.
    

10. 
    Urzì O, Olofsson Bagge R, Crescitelli R#. Journal of Extracellular Vesicles, 2022, 11, e12271. 
     

11. 
    Crescitelli R*, Lässer, C, Szabó TG, Kittel A, Eldh M, Dianzani I, Buzás EI, Lötvall J. J Extracellular Vesicles 2013 Sep 12;2.

Research Opportunities in the Laboratory

We have openings for MSc thesis projects.