Mechanisms and therapeutic targeting of the neuronal NMDAR signaling pathway promoting breast cancerpathogenesis
This “ISREC grant” was awarded to Simge Yücel in November 2020 for four years. Simge Yücel is working in the laboratories of Professors Douglas Hanahan and Michele De Palma, EPFL/SV/ISREC.
The research will focus on elucidating pathogenic mechanisms and exploring innovative therapeutic targeting strategies for breast cancer. The focus will be on a neuronal signaling pathway – centered upon the NMDA receptor (NMDAR) – that the Hanahan lab discovered to be hijacked by cancer cells to enhance invasion and metastasis, the hallmarks of malignancy that cause cancer mortality. The NMDAR’s role in cancer pathogenesis was initially discovered and described in 2013 (Li & Hanahan, Cell), and further characterized in 2018 (Li, Zeng et al., Cancer Cell), principally in pancreas cancer. Then in 2019, the role of NMDAR signaling in the metastasis of breast cancer to the brain was established in a major article in Nature (Zeng et al.). Unpublished data from the Hanahan lab suggest that the NMDAR may also be hijacked in certain primary breast tumors and in metastasis to other sites. Simge Yücel will use mouse models of breast cancer in conjunction with analysis of human breast cancer biopsies to pursue the following lines of investigation:
Line of investigation 1
What are the effects of conditional, cancer cell-specific genetic ablation of the key signaling subunit of NMDAR (GluN2B) on the development and lethal progression of invasive primary and lung-metastatic tumors in the MMTV-PymT mouse model of breast cancer? (The 2013 Cell publication showed that the NMDAR is activated in this model.) Other models, e.g. the C3Tag transgenic model of triple negative breast cancer, may also be employed. The plan is to assess not only invasion and metastasis but also effects on the tumor microenvironment, using a portfolio of sophisticated histopathological, cellular and molecular ‘-omic’ technologies, including single cell RNA
Line of investigation 2
Similarly, Simge Yücel will assess the effects of conditional cancer-specific deletion of candidate downstream effectors of NMDAR described in the 2018 Cancer Cell publication, in particular the transcription factor HSF1 and the translational regulator FMRP. She will use engineered breast cancer cell lines in culture and as transplant tumors to further investigate these effectors and to identify key genes amongst the myriad of genes regulated by each one.
Line of investigation 3
She will work with collaborators in Lund, Sweden, and Bern to assess tissue microarrays of human breast cancer for expression of components of the NMDAR signaling pathway, and in particular diagnostic pathway activity, as revealed by immunostaining for phosphorylated GluN2B. She will look for correlations with histological and molecular subtypes of human breast cancer, and associations with poor survival. She will study both primary breast tumors and metastases to other organs, such as lung, liver, and bone. She will develop hypotheses which she will then test in appropriate mouse models of breast
Line of investigation 4
Simge Yücel will explore mechanism-guided therapeutic targeting strategies aimed to pharmacologically inhibit either NMDAR signaling or genetically validated downstream effectors of its malignancy-enhancing program, in particular to impair invasion and metastasis. She will test rational combinations with drugs that disrupt other key tumor progression pathways, or that modulate the immune system so as to promote the efficacy of
These lines of investigation are likely to reveal new opportunities, which will be incorporated in the research.
References
- Li, L., & Hanahan, D. (2013). Hijacking the neuronal NMDAR signaling circuit to promote tumor growth and invasion. Cell. 153: 86-100.
- Li, L., Zeng, Q., Bhutkar, A., Galvan, J., Karamitopoulou, E., Noordermeer, D., Peng, M.W., Piersgilli, A., Perren, A., Zlobec, I., Robinson, H., Iruela-Arispe, M.L., & Hanahan D. (2018) GKAP acts as a genetic modulator of NMDAR signaling to govern invasive tumor growth. Cancer Cell, 33: 736-751.
- Zeng, Q., Michael, I.P., Zhang, P. Saghafini, S., Knott, G., Jiao, W., Brian D. McCabe, B.D., José A. Galván, J.A., Robinson, H.P.C., Zlobec, I., Ciriello, G., and Hanahan, D. (2019). Synaptic proximity enables NMDAR signaling to promote brain metastasis. Nature, 573: 526-531.
Transcriptomic and phenotypic profiling of the white blood cells in breast cancer
This “allocated fund” was awarded to Prof. Curzio Rüegg (University of Fribourg) in October 2020 for 4 years.
Despite considerable improvements in breast cancer management, approximately one in four patients still die due to metastases. In order to decrease this mortality, the disease needs to be diagnosed as early as possible, and metastases must be prevented or treated effectively. Studies we have conducted show that the presence of breast cancer alters quantifiable phenotypic and transcriptomic characteristics of white blood cells (leucocytes) circulating in the blood. These results suggest that it may be possible to exploit these changes in the leucocytes circulating in the blood to reveal the presence of primary breast cancer (screening) or a relapse (monitoring).
The aim of our project is to generate additional data supporting these observations. To this end, we will use novel approaches and technologies (single cell RNA sequencing and multi-parameter analysis of the cell surface). We plan to compare first-diagnosis female patients with healthy women, and patients at the time of the first relapse after therapy with patients not having suffered a relapse. We will investigate the three biological subtypes of breast cancer (ER+, HER2+, triple negative). This multicentric study, conducted in the Lake of Geneva area, will be coordinated at the CHUV in Lausanne.
A 20 ml blood sample will be taken for the following lab tests: i) leucocyte phenotyping by flow cytometry; ii) transcriptomic analysis and phenotyping by single cell RNA sequencing, followed by bioinformatic analyses. The lab tests will be conducted at the University of Fribourg, the sequencing at the Genomic Technologies Facility of the University of Lausanne (LGTF) and the bioinformatic analyses at the Swiss Institute of Bioinformatics (SIB).
We hope that these analyses will help us validate our preliminary observations more precisely and that we will be able to identify candidate biomarkers (cells, phenotypes, gene expression) and combinations associated with primary breast cancer or the first relapse.
This study is part of a long-term goal to develop a cancer screening blood test and a monitoring blood test for early detection of relapses. The practical implications of these tests are potentially significant: the tests could contribute to changing the way women are screened and patients monitored, thereby greatly improving the quality of life of women in general (screening) and of breast cancer patients (monitoring). From a clinical point of view, the monitoring test would be a valuable new tool that could help support therapeutic choices in the treatment of breast cancer.
This “allocated fund” derived from a donation of the Biltema Foundation was granted in June 2020 to Prof. Jean Bourhis (CHUV) for two and a half years.
Development of a novel B cell-based vaccine for metastatic solid cancers
This allocated fund for immunotherapies was awarded in September 2020 to the research group of Prof. Lana Kandalaft (Oncology Department UNIL/CHUV).
It is now well established that the immune system plays a very important role in controlling tumor growth; in fact, the remarkable results achieved in the past few years with the arrival of cancer immunotherapies and checkpoint inhibitors have clearly revolutionized the field of oncology and have dramatically changed the therapeutic scenario in many tumor types. Among the different cancer immunotherapies now available, cancer vaccines focus on the therapeutic use of a special type of immune cells called antigen-presenting cells (APCs). The main physiological role of these cells is to intercept and recognize pathogen-associated and foreign material (called antigens), and to subsequently initiate an immune response aimed at clearing the original antigen-containing threat from the host. Given the pivotal role played by APCs in orchestrating an immune response, it comes as no surprise that this type of cells has been employed since the infancy of modern cancer immunotherapy in the fight against cancer.
Canonical approaches have so far relied on the use of a special type of APCs called dendritic cells (DCs), which have long been considered the most potent type of APCs in the human body. However, despite proving generally safe and being associated with very mild collateral effects, DC-based therapeutic vaccines have so far demonstrated limited therapeutic efficacy. Mounting evidence now suggests that a second type of APCs called B cells present a valid alternative with several advantages. First of all, B cells can be obtained and expanded in large quantities from the human body, while cell availability often constitutes a limiting factor with DC-based vaccines. Contrary to DCs, B cells are also resistant to functional inhibition induced by tumor-produced factors, another aspect often limiting DC therapeutic efficacy. Several previous studies have also shown that B cells are indeed able to induce a potent and cancer-specific immune response.
Based on this collective evidence, our project focuses on developing a novel therapeutic vaccine against cancer, based on B cells. One of the first project goals is to manipulate and engineer B cells to express special receptors that increase their accumulation at the tumor site after infusion into the patient. This aspect is obviously crucial in improving their efficacy against their tumor targets and in limiting off-side and systemic effects. In a second phase, we will test our B cell vaccine formulation in combination with checkpoint blockade inhibitors, another promising immunotherapy with a different and complementary mode of action. Checkpoint blockade refers to specific pathways normally developed by tumors that inhibit T cell activities, exerting a sort of “break” on the function of the immune system and anti-cancer properties. Thus, combining these two therapies constitutes an interesting potential therapeutic option, in which B cells would activate the immune system (more specifically T cells) against tumor targets, whilst checkpoint blockade inhibition would remove inhibitory “breaks” on T cell function and unleash these cells’ true anti-cancer potential.
In the last part of this project and thanks to the infrastructure for translational studies available at the University Hospital of Lausanne and at the Ludwig Institute (Lausanne Branch), we will also develop protocols and tests for an efficient production of our B cell final formulation in a good manufacturing practices (GMP) context, so as to enable its future application in clinical studies and patient tests. The requirements for testing cellular therapies in patients (usually referred to as GMP conditions) are obviously very different and stricter than laboratory bench and animal research settings, and a great deal of work is usually needed to adapt therapy production in order to meet such requirements. Thus, the validation of the findings of this project in a GMP context will pave the way for future translational studies in cancer patients, and will potentially help advance both the therapeutic scenario and clinical outcomes.
This “ISREC grant” was awarded to Silvia Podavini in August 2019 for four years. Silvia Podavini is working in the group of Professor Margot Thome Miazza (UNIL, Biochemistry Department).
This « ISREC grant » was awarded to Andrea Agnoletto in September 2019 for four years. Andrea Agnoletto is working in the lab of Prof. Cathrin Brisken, EPFL/SV/ISREC.