The Princess Máxima Center for pediatric oncology is a new nationwide research hospital concentrating healthcare, research and education with regard to cancer in children and is located at the Utrecht Science Park de Uithof. The institute has the ambition to give the best treatment to children with cancer with the highest cure rate and the least side effects of (often complex) anti-cancer treatment. The center brings together the best possible care and scientific research, creating a unique comprehensive childhood cancer center in Europe. Communication is key: * Design of a high dimensional flow cytometry panel to understand macrophage – T cell therapy crosstalk in the context of DMG targeting* DREAM3DLAB / Rios group, Princess Máxima Center for pediatric oncology Supervisor: Amber Wezenaar Background: As part of the innate immune system, macrophages are present in all human tissues where they maintain tissue homeostasis and initiate an immune response to pathogens. Additionally, macrophages have also been described to have a key role in tumour survival and progression. Once infiltrated into tumours, macrophages can be polarized to a tumour-promoting, immunosuppressive state by multiple tumour types. In this state, tumour-associated macrophages (TAMs) have been shown to negatively impact T cell responses, either through direct interactions or through modulation of the tumour microenvironment (TME) (DeNardo & Ruffell, 2019). Despite the significant role of TAMs in T cell modulation, this phenomenon has been less well studied in the context of T cell therapy (Lindo, Wilkinson & Hay, 2021). A potential negative impact of the immune-suppressive myeloid compartment on GD2 CAR-T cell efficacy has recently been suggested in diffuse midline glioma (DMG), were an immunosuppressive signature of the myeloid compartment appeared to coincide with poor treatment outcome in the first in-human clinical trial with GD2 CAR-T cells (Majzner et al, 2022). Therefore, we aim to unravel the cellular crosstalk between macrophages and therapeutic T cells to identify potential targets to overcome negative modulation of T cell activity and improve T cell therapy efficacy for DMG. Project description: To understand the interactions that occur between macrophages and therapeutic T cells in DMG, during the internship project a high dimensional flow cytometry panel will be established for phenotyping co-cultures of macrophages, therapeutic T cells and DMG organoids. The candidate will learn to design, optimize and establish a high dimensional flow cytometry panel on a spectral flow cytometer. The panel will comprise at least 35 markers to enable assessment of cell type, subset and state, as well as functional readouts that include tumor reactivity, immune suppression and cellular interaction. The high dimensional flow cytometry dataset generated will be analyzed with R using analyses such as UMAP dimensionality reduction, FlowSOM clustering and Diffcyt differential expression analysis. In addition, the student will receive training in the basics of organoid culturing and immune co-cultures. With this technology we will address key questions, such as: 1) How does co-culture with DMG organoids change the polarization state of macrophages? 2) Which macrophage population interacts with therapeutic T cells? 3) How does interaction change the state and function of therapeutic T cells? We are asking for We are looking for a highly motivated and enthusiastic master student with a strong interest in immune-oncology research for a 9-month internship. Experience with flow cytometry, cell culture and R programming is an advantage, but not required. We offer The student will be working in the DREAM3DLAB led by Dr. Anne Rios. We are a multidisciplinary team focussed on advancing high-dimensional 3D imaging and human in vitro modeling technologies to decipher human biology and design innovative treatments against cancer. Interested? You can apply for this position by pressing the “apply” button on this screen. For further information about the position, please send an email to Amber Wezenaar, A.K.L.Wezenaar@prinsesmaximacentrum.nl Literature of interest:
DeNardo, D. G., & Ruffell, B. (2019). Macrophages as regulators of tumour immunity and immunotherapy. Nature Reviews Immunology, 19(6), 369–382. doi: 10.1038/s41577-019-0127-6
Lindo, L., Wilkinson, L. H., & Hay, K. A. (2021). Befriending the Hostile Tumor Microenvironment in CAR T-Cell Therapy. Frontiers in Immunology, 11, 618387. doi: 10.3389/fimmu.2020.618387
Majzner, R. G. et al. (2022) GD2-CAR T cell therapy for H3K27M-mutated diffuse midline gliomas. Nature 1–10. doi: 10.1038/s41586-022-04489-4
Park, L. M., Lannigan, J. & Jaimes, M. C. (2020). OMIP‐069: Forty‐Color Full Spectrum Flow Cytometry Panel for Deep Immunophenotyping of Major Cell Subsets in Human Peripheral Blood. Cytometry, 97, 1044–1051. doi: 10.1002/cyto.a.24213
Palit, S. et al. (2019). Meeting the Challenges of High-Dimensional Single-Cell Data Analysis in Immunology. Frontiers in_ Immuno_logy,10, 1515. doi: 3389/fimmu.2019.01515
Acquisitie naar aanleiding van deze vacature wordt niet op prijs gesteld.