Ovarian cancer (OC) is one of the most lethal cancers in the world due to late-stage disease at diagnosis. Standard therapy consists of debulking surgery and chemotherapy. However, despite this aggressive treatment, recurrent disease almost invariably occurs resulting in a five-year survival rate of approximately 30%.
Immunotherapy could be a way to increase survival in OC patients. However, a major barrier to a successful deployment of cancer immunotherapy for ovarian cancer patients is the immunosuppressive tumor microenvironment.
Envisioned solution/research direction
Tumor-related inflammation is one of the hallmarks of cancers in general. Innate immunity specifically is a common denominator that is involved in the pathogenesis of OC. To improve the patient's outcome and identify novel therapeutic targets, one needs a deeper understanding of the tumor-induced changes in the bone marrow myeloid progenitor cells. Furthermore, treatment of these cells by nanoparticles or other agents that induce a program of 'trained immunity' may be a novel way to re- educate myeloid cells and their bone marrow progenitors in OC patients.
Hypothesis
We hypothesize that by exposing myeloid cells or their progenitors to various agents that induce trained immunity (e.g. trained immunity-inducing agents: BCG, heat-killed Candida,), these immune cells will undergo functional reprogramming to induce a tumor-suppressive phenotype. In the future, this could be explored as a novel immunotherapy for tumors that are refractory to conventional treatment.
Objective
To characterize and phenotype the immune state of OC patients compared to controls without cancer with a focus on the hematopoietic organs and the immune cells originating from these organs. In addition, the effect of established trained immunity-inducing agents on these cells will be evaluated in vitro, potentially providing new therapies.
This will be executed by assessing the transcriptional, epigenetic, and functional reprogramming of circulating monocytes and myeloid progenitor cells in OC and by assessing the in vitro effect of trained immunity inducers on the reprogramming of circulating monocytes and myeloid progenitor cells.
Study design: investigator-initiated, multi-center explorative cross-sectional study at the Catharina hospital Eindhoven, Radboud University Medical Center and Eindhoven University of Technology.
Subjects should be at least 18 years old and mentally competent;
Newly diagnosed patients with OC who go for primary debulking surgery or patients with OC who are scheduled for interval debulking;
Controls: women who undergo surgery for benign gynaecological conditions under general anaesthesia.
Mentally incompetent;
Pregnant or breastfeeding;
Known inflammatory of infectious diseases or an immunosuppressive status;
Using medication interfering with the immune system;
Parallel
Bone marrow aspiration
Procedure/SurgeryBone marrow samples will be obtained from the sternum or the iliac crest according to standard practice by experienced operators during the surgery patients will undergo as part of their medical treatment. BM mononuclear cells will be isolated using Ficoll-Paque and progenitor cells will be enriched using positive selection with CD34 beads by MACS. Progenitor cell composition will be identified using flow cytometry. HSPC proliferation assays (standard CFU assays) to assess myeloid colony-forming potential will be performed. In addition, single-cell RNA- sequencing and epigenetic assessment (ATAC-seq, ChIP-seq, DNA methylation) will be performed.
Vena puncture
Procedure/SurgeryWhole blood will be collected and peripheral blood mononuclear cells (PBMC) will be isolated using Ficoll-Paque. Cellular subpopulations will be purified using negative selection of monocytes with PanMonocyte beads by MACS.
Peritoneal fluid
Procedure/SurgeryPeritoneal fluid will be collected during the surgery.
Spleen biopsy
Procedure/SurgerySpleen samples will be obtained by experienced surgeons during the debulking surgery. Spleen mononuclear cells will be isolated using Ficoll-Paque and progenitor cells will be enriched using positive selection with CD34 beads by MACS. Cellular subpopulations will be further purified using negative selection of monocytes with PanMonocyte beads by MACS. Progenitor cell composition will be identified using flow cytometry. HSPC proliferation assays (standard CFU assays) to assess myeloid colony-forming potential will be performed. In addition, single- cell RNA-sequencing and epigenetic assessment (ATAC-seq, ChIP-seq, DNA methylation) will be performed.
Tumor biopsy
Procedure/SurgeryDuring debulking surgery, all visible tumor tissue will be removed by experienced surgeons.
Omental biopsy
Procedure/SurgeryDuring debulking surgery, an omental biopsy will be done.
Cell composition of immune organs using flow cytometry
The number and ratios of different types of stem and immune cells will be determined using general cell protein markers in combination with flow cytometry. Analysis will be performed on blood, spleen, bone marrow and the intraperitoneal fluid.
Cell composition and epigenetic status of cells of immune organs using ATAC and RNA sequencing
Cell ratios and epigenetic profile of immune cells in the blood, tumor, bone marrow, and spleen will be analyzed using single-cell RNA and single-cell ATAC sequencing.
Trained immunity response
We focus on the degree of trained immunity response upon ex vivo stimulation with trained immunity inducers and measured as the concetration of inflammatory cytokines and ROS production.
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Gynaecologisch Oncologisch Centrum Zuid
Lead sponsor
Eindhoven University of Technology
Collaborator
Radboud University Medical Center
Collaborator
