Projects / Programmes
Cystatin f as a mediator of immunosuppression in glioblastoma microenvironment
| Code |
Science |
Field |
Subfield |
| 3.01.00 |
Medical sciences |
Microbiology and immunology |
|
| Code |
Science |
Field |
| 3.01 |
Medical and Health Sciences |
Basic medicine |
cystatin F, cytotoxic cells, NK cells, cancer immunotherapy, glioblastoma multiforme, tumor microenvironment, immunosuppression
Researchers (10)
Organisations (3)
Abstract
Glioblastoma multiforme (GBM) is one of the most aggressive and lethal forms of cancer. Regardless of, treatment with maximal surgical resection, radio- and chemotherapy, patients only have a 15- to 19-month median survival rate due to almost-universal tumour recurrence. Therefore, there is an urgent need for developing new therapeutic approaches for GBM treatment. Adoptive immunotherapy, has attracted increasing attention in the past years due to remarkable success in cancer immunotherapies treating advanced solid tumours. For GBM treatment, some promising results have been achieved using adoptive T cell therapies such as tumour-infiltrating lymphocytes (TILs) and chimeric antigen receptor (CAR) T cells. However, effector cells readily lose their functional capacity in suppressive microenvironment of GBM. Therefore, understanding the immune cell composition of the tumour microenvironment and identification of mediators of immune suppression, which can be effectively therapeutically targeted, represents a major challenge in successful treatment of GBM. We have identified a protease inhibitor cystatin F as a potent modulator of cytotoxic efficacy of effector cells such as NK cells and cytotoxic T lymphocytes. Its expression was shown to be increased in GBM patient tissue compared to normal healthy brain tissue and its increased expression correlates with poor patient prognosis. Specific properties of this inhibitor, such as dimeric extracellular protein form, existence of activation peptidase, glycosylation, internalization and transport to endo/lysosomal vesicles, distinguish cystatin F from other related peptidase inhibitors and guarantee selective targeting of processes in cytotoxic cells. Within the proposed project we will characterize the expression, trafficking and main target peptidases of cystatin F within the microenvironment of GBM. We will develop the tools to decrease its availability and activity within tumour microenvironment to show, that by decreasing the content of cystatin F, higher efficiency in tumour cell killing by NK cells can be achieved. The workflow starts with the identification of cellular sources of cystatin F in patient GBM microenvironment (WP1). Based on those results we will start testing the cystatin F expression, trafficking and activation in cell lines representative of cell types expressing cystatin F in GBM microenvironment (WP2). Concurrently, the tools for cystatin F modulation will be developed (WP3, WP4). This will be followed with testing of the effect of modulation in cystatin F intra- and extracellular availability on cytotoxicity of NK cells isolated from healthy donors (WP4). Finally, in vitro model of GBM microenvironment mimicking interactions between NK cells, tumour cells and macrophages will be prepared. On this model we will test the effects of the cystatin F modulation on the cytotoxicity of therapeutic “super charged” cells (WP5). The proposed project will be realized by cooperation of three research groups. The group from Jožef Stefan Institute led by Dr. Milica Perišić Nanut will coordinate the project (WP6) and provide expertise in designing tools for cystatin F manipulation, cell cultivation, glioblastoma stem cells (GSCs) enrichment, generation of primary GBM cell lines from GBM patients and cytotoxicity testing. The research group from National Institute of Biology led by Prof. Tamara Lah Turnšek has extensive expertise in GBM biology and characterization of GSC niche and will be involved in processing patients GBM tissue samples. Assis prof. Urban Švajger from the Blood Transfusion Centre of Slovenia will provide expertise in isolation of NK cells from healthy donors and isolation and expansion of “super charged” NK cells. The research groups have the expertise and access to all of the necessary equipment to successfully finish the proposed project within 3 years.
