Molecular characterisation of RNA-seq identified changes in TCR signalling in prostate cancer disease
Lead Research Organisation:
University of Glasgow
Department Name: College of Medical, Veterinary, Life Sci
Abstract
Keywords: Prostate cancer, immune cells, TCART
Background: Increasing evidence suggests that both CD4+ and CD8+ T cells play a critical role in cancer immunosurveillance & anti-tumor immunity, and manipulation of these immune cells to recognize and eradicate tumor cells is a promising strategy for treating patients with invasive & metastatic cancers. It has become clear that the tumor suppressive microenvironments created by malignant tumors are a major obstacle for effective anti-tumor immunity and successful tumor immunotherapy (1,2).
Tumor cells can utilize different strategies to expand & recruit various types of suppressive tumor-infiltrating lymphocytes (TILs), including naturally occurring & adaptively induced regulatory T (Treg) cells, tolerogenic dendritic cells (DCs), tumor-derived macrophages and myeloid suppressor cells (MSCs). In addition, tumor cells can secrete suppressive factors (IL-10, TGF-b and IDO), express immune inhibitory molecules (FasL & PD-L1), directly inhibit tumor-specific T-cell expansion and proliferation, or induce T-cell apoptosis (1). A better understanding of the molecular mechanisms involved in creating and sustaining the tumor-induced immune suppressive microenvironment is critical for the development of novel tumor vaccines & therapeutic strategies active against human cancers.
We have recently described the identification of PDE4D7 as novel putative tumor suppressor gene which is significantly associated with aggressive disease and poor patient outcome (3). Based on correlation studies performed on RNAseq data of >500 human prostate cancer tumor tissues derived from resected patient material we identified a range of genes that are involved in the regulation of T-cell activation & are disturbed in prostate tumors with aggressive disease characteristics. We hypothesize that the gene expression changes that we observe in the RNAseq data is originating from tumor infiltrating lymphocytes, more specifically tumor infiltrating T-cells. The activation potential of these T-cells might be impaired by the change in the transcription of these genes.
Aims: The goal of this project is to understand whether and to what extent the RNAseq identified genes of interest are dis-regulated in tumor infiltrating lymphocytes, whether we can determine the type of affected lymphocyte (e.g., CD4+, CD8+) and whether we can measure any effects on the activation of downstream transcription factors. For this we will develop assays to measure the identified genes of interest on pathology tissue sections with RNAscope, and/or immuno-histochemistry. qPCR assays will be used to determine how the expression of these genes is affected in tumor-infiltrating T-cells isolated from patient tissue and blood samples from patients with prostate cancer. Patients are selected from different stages of the disease (primary, advanced, metastatic, castration-resistant) & after various sorts of therapies (radiation, hormones, chemotherapy). Functional proliferation assays will be used to co-culture purified T-cells from human donors with different prostate cancer cell lines in vitro to determine the level of tumor suppression by purified T-cells depending on their activation status of the genes of interest. This will allow us to correlate the expression status of the genes of interest with the level of immune cell induced suppression of growth prostate cancer cells in vitro.
References:1) Croci DO et al (2007).Dynamic cross-talk between tumor and immune cells in orchestrating the immunosuppressive network at the tumor microenvironment. Cancer Immunol Immunother 56: 1687-1700
2) Whiteside TL(2008).The tumor microenvironment and its role in promoting tumor growth. Oncogene 27: 5904-5912
3) Henderson et al (2019).Creating a potential diagnostic for prostate cancer risk stratification (InformMDxTM by translating novel scientific discoveries concerning cAMP degrading phosphodiesterase-4D7 (PDE4D7). Clinical Science 133(2): 269-28
Background: Increasing evidence suggests that both CD4+ and CD8+ T cells play a critical role in cancer immunosurveillance & anti-tumor immunity, and manipulation of these immune cells to recognize and eradicate tumor cells is a promising strategy for treating patients with invasive & metastatic cancers. It has become clear that the tumor suppressive microenvironments created by malignant tumors are a major obstacle for effective anti-tumor immunity and successful tumor immunotherapy (1,2).
Tumor cells can utilize different strategies to expand & recruit various types of suppressive tumor-infiltrating lymphocytes (TILs), including naturally occurring & adaptively induced regulatory T (Treg) cells, tolerogenic dendritic cells (DCs), tumor-derived macrophages and myeloid suppressor cells (MSCs). In addition, tumor cells can secrete suppressive factors (IL-10, TGF-b and IDO), express immune inhibitory molecules (FasL & PD-L1), directly inhibit tumor-specific T-cell expansion and proliferation, or induce T-cell apoptosis (1). A better understanding of the molecular mechanisms involved in creating and sustaining the tumor-induced immune suppressive microenvironment is critical for the development of novel tumor vaccines & therapeutic strategies active against human cancers.
We have recently described the identification of PDE4D7 as novel putative tumor suppressor gene which is significantly associated with aggressive disease and poor patient outcome (3). Based on correlation studies performed on RNAseq data of >500 human prostate cancer tumor tissues derived from resected patient material we identified a range of genes that are involved in the regulation of T-cell activation & are disturbed in prostate tumors with aggressive disease characteristics. We hypothesize that the gene expression changes that we observe in the RNAseq data is originating from tumor infiltrating lymphocytes, more specifically tumor infiltrating T-cells. The activation potential of these T-cells might be impaired by the change in the transcription of these genes.
Aims: The goal of this project is to understand whether and to what extent the RNAseq identified genes of interest are dis-regulated in tumor infiltrating lymphocytes, whether we can determine the type of affected lymphocyte (e.g., CD4+, CD8+) and whether we can measure any effects on the activation of downstream transcription factors. For this we will develop assays to measure the identified genes of interest on pathology tissue sections with RNAscope, and/or immuno-histochemistry. qPCR assays will be used to determine how the expression of these genes is affected in tumor-infiltrating T-cells isolated from patient tissue and blood samples from patients with prostate cancer. Patients are selected from different stages of the disease (primary, advanced, metastatic, castration-resistant) & after various sorts of therapies (radiation, hormones, chemotherapy). Functional proliferation assays will be used to co-culture purified T-cells from human donors with different prostate cancer cell lines in vitro to determine the level of tumor suppression by purified T-cells depending on their activation status of the genes of interest. This will allow us to correlate the expression status of the genes of interest with the level of immune cell induced suppression of growth prostate cancer cells in vitro.
References:1) Croci DO et al (2007).Dynamic cross-talk between tumor and immune cells in orchestrating the immunosuppressive network at the tumor microenvironment. Cancer Immunol Immunother 56: 1687-1700
2) Whiteside TL(2008).The tumor microenvironment and its role in promoting tumor growth. Oncogene 27: 5904-5912
3) Henderson et al (2019).Creating a potential diagnostic for prostate cancer risk stratification (InformMDxTM by translating novel scientific discoveries concerning cAMP degrading phosphodiesterase-4D7 (PDE4D7). Clinical Science 133(2): 269-28
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| MR/R01566X/1 | 01/10/2018 | 30/09/2025 | |||
| 2287896 | Studentship | MR/R01566X/1 | 01/10/2019 | 31/03/2023 | Emma Parsons |