Background
Immunotherapy has revolutionized cancer treatment. However, most patients with advanced cancer do not respond to current immunotherapies or develop secondary resistance. Current regimens for cancer immunotherapy are mostly focused on using one type of immune cells called CD8 T cells. The Corthay lab (https://ous-research.no/corthay) has previously reported that another type of immune cells named macrophages may also be very efficient at fighting cancer. The main objective of this master project is to demonstrate in vitro the potential of macrophages for eliminating cancer cells.
In a tumor, the cancerous cells are surrounded by a tumor microenvironment typically consisting of extracellular matrix molecules, blood vessels, fibroblasts, and numerous types of immune cells. To mimic the tumor environment in 3D, we will take advantage of the tumor on chip technology.
Tumor on chip technology. (A) Example of a microfluidic device (tumor on chip). A complex tumor microenvironment is recreated in the tumor (red) chambers. (B) T-cell migration on chip. Tumor-specific CAR T cells (green) loaded in the immune chamber (left) are seen migrating towards the tumor chamber (right) via micro-channels (center). In the tumor chamber (right), a basic human tumor micro-environment was created in a 3D collagen gel. Upon migration CAR T cells make contacts with clusters of cancer cells (blue) and macrophages (white).
The master project
Time-lapse microscopy will be used to investigate the ability of mouse and human macrophages to eliminate cancer cells by cytotoxicity and phagocytosis. A critical R&D challenge is that the functions of macrophages in tumors are negatively influenced by the immunosuppressive tumor microenvironment. To solve this, we will optimize the activation of macrophages in vitro in a complex tumor microenvironment using microfluidic devices and 3D co-cultures (tumor on chip technology).
Specific aims of the master project
- To establish a microfluidic chip-based assay to visualize by time-lapse microscopy the killing of cancer cells by activated macrophages.
- To demonstrate the ability of mouse and human macrophages to kill cancer cells in a complex and immunosuppressive tumor microenvironment.
- To decipher the cellular and molecular mechanisms whereby macrophages kill cancer cells.
Methods and tools that will be used in the master project:
- Isolation of cells from blood and tissues
- Culturing of cell lines and primary cells in vitro
- Labeling of cells and cellular functions (such as apoptosis) with various fluorescent probes
- Tumor on-chip technology in close collaboration with the Hybrid Technology Hub, a centre of excellence supported by the Research Council of Norway (https://www.med.uio.no/hth/english/)
- Fluorescence microscopy techniques and time-lapse microscopy
- Image analysis by use of computer algorithms
- Optimization of macrophage activation using various molecules (e.g., toll-like receptor agonists, cytokines, antibodies, polysaccharides, and recombinant viral particles)
- Flow cytometry
- Luminex technology for quantification of cytokines
Additional learning outcomes
- Learning how to plan and perform experiments
- Data analysis and figure design
- Participation is scientific meetings and immunology journal club
- Presentation techniques (orally and in writing)
Working environment
The master project will be performed in the Tumor Immunology Lab (https://www.ous-research.no/corthay) which is located at Rikshospitalet (Department of Pathology). The research focus of the group is cancer immunology and immunotherapy. The Tumor Immunology Lab is also full member of the Centre of Excellence Hybrid Technology Hub (The Corthay Group). The master student will be closely supervised by Alexandre Corthay (main supervisor) and Inger ?ynebr?ten (co-supervisor). Finn-Eirik Johansen will serve as internal supervisor from IBV. In our lab, we use standard molecular biology techniques, perform advanced cellular functional assays, do immunostaining of mouse and human cells and tissue, flow cytometry, ELISA, multiplex analysis of cytokines, and microscopy. We use mouse models of cancer to test if strategies and compounds have potential in vivo for cancer immunotherapy. We have a broad network of collaborators nationally and internationally.