Engineering an immune response in melanoma by targeting cytokine signalling
Lead Research Organisation:
University of Manchester
Department Name: School of Medical Sciences
Abstract
Background
Targeting immunosuppression in the tumour microenvironment allows a latent immune reaction to be unleashed resulting in tumour regression. GM-CSF stimulates the activity of antigen presenting cells and IL12 stimulates cell mediated immunity. Recently, we have observed infiltration of melanoma nodules in zebrafish by macrophages and CD4+ T-cells. Further, we have constructed a vector for inducible gene expression in melanoma nodules and found that induction of IFNy results in the outgrowth of unpigmented tumour cells. As viral vectors (e.g. T-VEC) have now been developed for directing cytokine expression in melanoma tumour nodules that show clinical activity, it would be desirable to have an easily engineered model system to evaluate cytokine efficacy and synergistic action.
Hypothesis
The combinatorial action of cytokines can profoundly modulate the immune response to a melanoma tumour nodule resulting in its rejection
Objectives
To engineer constructs for expression of IL12 and GM-CSF, as well as a combination of these two factors or a combination of IL12 with IFNy or GM-CSF with IFNy and to evaluate the effect of cytokine production on tumour maintenance.
To contrast the expression profiles of mononuclear phagocytes and T cells isolated from control tumours and tumours expressing cytokines and hopefully undergoing regression
To corroborate cytokine cooperation in a mouse transplantation model of melanoma
Methods
Using a tamoxifen-responsive Cre construct, we have been able to induce gene expression in established melanoma tumour nodules in zebrafish. We have also created or acquired fluorescent reporter constructs that allow flow isolation of mononuclear phagocytes and T cells. Combining these reagents we shall induce expression of IL12 or GM-CSF alone, together, or combined with IFNy and evaluate changes in tumour volume. We shall isolate leukocytes from tumour nodules and analyse gene expression by RNA seq. Cytokine combinations will also be evaluated in a mouse melanoma transplantation model by injecting tumour nodule with recombinant protein.
Potential outcome/impact
This study will establish zebrafish as a simple preclinical model for evaluating the efficacy of cytokines and cytokine combinations to induce an immune response to melanoma
Targeting immunosuppression in the tumour microenvironment allows a latent immune reaction to be unleashed resulting in tumour regression. GM-CSF stimulates the activity of antigen presenting cells and IL12 stimulates cell mediated immunity. Recently, we have observed infiltration of melanoma nodules in zebrafish by macrophages and CD4+ T-cells. Further, we have constructed a vector for inducible gene expression in melanoma nodules and found that induction of IFNy results in the outgrowth of unpigmented tumour cells. As viral vectors (e.g. T-VEC) have now been developed for directing cytokine expression in melanoma tumour nodules that show clinical activity, it would be desirable to have an easily engineered model system to evaluate cytokine efficacy and synergistic action.
Hypothesis
The combinatorial action of cytokines can profoundly modulate the immune response to a melanoma tumour nodule resulting in its rejection
Objectives
To engineer constructs for expression of IL12 and GM-CSF, as well as a combination of these two factors or a combination of IL12 with IFNy or GM-CSF with IFNy and to evaluate the effect of cytokine production on tumour maintenance.
To contrast the expression profiles of mononuclear phagocytes and T cells isolated from control tumours and tumours expressing cytokines and hopefully undergoing regression
To corroborate cytokine cooperation in a mouse transplantation model of melanoma
Methods
Using a tamoxifen-responsive Cre construct, we have been able to induce gene expression in established melanoma tumour nodules in zebrafish. We have also created or acquired fluorescent reporter constructs that allow flow isolation of mononuclear phagocytes and T cells. Combining these reagents we shall induce expression of IL12 or GM-CSF alone, together, or combined with IFNy and evaluate changes in tumour volume. We shall isolate leukocytes from tumour nodules and analyse gene expression by RNA seq. Cytokine combinations will also be evaluated in a mouse melanoma transplantation model by injecting tumour nodule with recombinant protein.
Potential outcome/impact
This study will establish zebrafish as a simple preclinical model for evaluating the efficacy of cytokines and cytokine combinations to induce an immune response to melanoma
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| MR/N013751/1 | 01/10/2016 | 30/09/2025 | |||
| 1792521 | Studentship | MR/N013751/1 | 01/10/2016 | 31/03/2020 | Christos Evangelou |
| Description | Collaboration with the startup company Cellular Therapeutics |
| Organisation | Immetacyte Ltd |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Cellular therapeutics is an early stage but fast growing star-up company aiming in developing novel T cell therapies in melanoma patient. Their work involves isolation of melanoma cells from surgically excised tumors, as well as infiltrating T cells (TILs). Their goal is to expand and activate tumor reacting TILs and then administer them back to the patients. My work aims on the identification of novel mechanisms by which the cytokine IFNg drives resistance to immunotherapy in melanoma. Since my research aims in the identification of novel mechanisms by which melanoma cells acquire resistance to immunotherapy, my findings are of great interest and benefit to an Immunotherapy company like Cellular Therapeutics. |
| Collaborator Contribution | Cellular Therapeutics' work involves isolation of melanoma cells from surgically excised tumors, as well as infiltrating T cells (TILs). Their goal is to expand and activate tumor reacting TILs and then administer them back to the patients. Since my work aims on the identification of novel mechanisms by which the cytokine IFNg drives resistance to immunotherapy in melanoma their panel of patient derived short term cultures and their matching T cells is a valuable tool for me, to further translate my preliminary data acquired from cell line models into the more clinically relevant patient derived melanoma cells. They also helped me established protocols for melanoma and T cell co-cultures in order to validate the role of candidate genes in the T cell mediated killing of melanoma cells. |
| Impact | I have shown already that patient derived melanoma cells retain responsiveness to IFNg, similarly to what I have previously shown for melanoma cell lines. We also performed genetic manipulation of our target genes that we believe that impact immunomodulation and therefore are involved in immunotherapy resistance, and we are now at the stage of looking at the effects of these genetic manipulations in the T cell activation and T cell mediated killing of melanoma cells. |
| Start Year | 2018 |
| Description | STEM Ambassador and involvement in some of STEM Ambassador activities in Greater Manchester e.g. Manchester Science Festival |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | STEM Ambassador and involvement in some of STEM Ambassador activities in Greater Manchester e.g. Manchester Science Festival |
| Year(s) Of Engagement Activity | 2018 |