Background
Radiotherapy is a cornerstone in cancer treatment, not only inducing DNA damage and direct tumor cell death but also potentially eliciting anti-tumor immune responses. Recent studies have shown that combining radiotherapy with DNA damage response inhibitors can enhance immune signaling in tumor cells. For example, our recent work demonstrated that the type I interferon (IFN-I) response is increased in glioblastoma and lung cancer cells following treatment with X-ray or proton radiation combined with inhibitors of the DNA repair kinases ATR and ATM. However, the downstream effects of this immune signaling on human immune cells remain largely unknown.
The project
The aim of this project is to study whether immune signaling from lung cancer cells treated with ionizing radiation (X-rays or protons) combined with inhibitors of the DNA damage response (e.g. ATR, ATM) has the potential to activate human dendritic cells (DCs). DCs are immune cells that play a crucial role in bridging innate and adaptive immune responses. Functional assays measuring DC activation are therefore important tools for assessing the potential of a treatment to stimulate overall anti-tumor immunity. The specific objectives of the project are to i) Set up assays to measure DC activation. ii) Assess whether DCs can be activated by culture medium harvested from treated cancer cells.
The results will increase our knowledge of the immunogenic potential of combined treatment with ionizing radiation and inhibitors of the DNA damage response.
Methods
The student will isolate peripheral blood mononuclear cells (PMBCs) from blood donors and differentiate them into immature DCs (iDCs) with cytokine treatment. These iDCs will then be exposed to culture medium harvested from treated cancer cells, and the transformation of iDCs into active DCs will be measured. DC activation will be analyzed in two ways:
- Live cell flow cytometry of activation markers on the cell surface of DCs.
- Boyden chamber assay to assess DC migratory capacity.
The student will also perform cell culturing, irradiation and drug treatment of the cancer cells. Thus, the major methods to be used in this project will be: Cell culturing techniques (both suspension and adherent human cells), DNA damage induction by X-ray and proton irradiation (to be employed at the new proton therapy facility at OUH located in close proximity to our lab), PMBC isolation and differentiation, multiparameter live cell flow cytometry, and Boyden chamber cell migration assay. Additional techniques include enzyme-linked immunosorbent assay (ELISA) to quantify immune signaling in cancer cells and microscopy (phase contrast and/or fluorescence microscopy) to study morphological changes in DCs. Altogether, we offer a great opportunity for a master student to learn how to plan, optimize and carry out experiments in a state-of-the-art laboratory.
We provide a friendly and stimulating working environment and seek a highly motivated and dedicated student who is interested in an exciting project in molecular cancer research.
Supervisors
Main supervisor: Scientist Sissel Hauge 22781460; sissel.hauge@ous-hf.no
Co-supervisor: Group Leader Randi G. Sylju?sen 22781468; randi.syljuasen@ous-hf.no
Research Group:
Radiation Biology and DNA damage signaling
Department of Radiation Biology
Institute for Cancer Research
Norwegian Radium Hospital
Oslo University Hospital
Ullernchausseen 70, 0379 Oslo
http://www.ous-research.no/syljuasen/