- Investigating the effect of calcium electroporation on the inflammatory microenvironment and crosstalk to immune cell activation across the Barrett's-dysplasia-adenocarcinoma disease sequence.
Prof. Jacintha O’Sullivan
AllCaN Oesophageal Lead & Principal Investigator
Ms. Lorraine Smith
AllCaN Oesophageal PhD Candidate
- September 2023 (4 years)
- Industry Partners
Dr Declan Soden
Mirai Medical
Research question
Can electroporation alter the inflammatory features of non-dysplastic and dysplastic BO tissue environment and how does this treatment affect immune cell activation in BO patients?
Overview – Part 1
In this project, our industry collaborators Mirai Medical are a company with over 30 years’ experience in the field of electroporation.
Electroporation is the term used when electrical pulses are applied to a cell rendering its membrane porous as shown in the figure across. Their unique ePORE technology is a precision therapy platform that targets disease tissue, whilst preserving surrounding healthy tissue structure. Proven to be safe, simple and cost effective, it greatly reduces side effects, it is less invasive and allows for faster treatment delivered as a day case procedure (https://mirai-medical.com). As the electroporation pulses vastly increase the porosity of the cell membrane, different drugs, including certain immunotherapies which require internalisation, can be used in combination to facilitate their targeted absorption. Electroporation has been used for many years to allow different substances to enter the cell. However, to date electroporation has resulted in powerful muscular contractions causing significant discomfort for the patient, limiting its clinical adoption with the procedure having to be performed under general anaesthetic. The background IP, which has been developed by Mirai overcomes this issue via a filed patented design which increases the pulse frequency up to 500KHz.
In this project, using Mirai Medical’s ePORE technology, we will examine the effect of electroporation on the inflammatory microenvironment in Barrett’s non dysplastic and dysplastic ex vivo human tissues and the effect of this secretome on immune cell activation (Dendritic cells and T cells). We will also acquire additional biopsies (pre and post electroporation) and specifically assess DC’s and T cell biology also in this ex vivo digested material. If successful, we will be able to determine if electroporation treatment alters the inflammatory secretome and compare the responses in Barrett’s non dysplastic and dysplastic tissues from Barrett’s patients. This data will also be extensively correlated with the detailed clinical and pathology data already inputted on our National Barrett’s Registry (from work package 1).
Overview – Part 2
The second part of this project will examine if the tissue conditioned media and its secreteome collected in part 1 can influence immune cell biology. There is an urgent need to determine how the tissue microenvironment (pre and post electroporation) cross talks to immune cells. We hypothesise that electroporation will boost the immunity profile. Dendritic cells (DCs) are present in tissues in an immature state which recognise and capture tumour antigens. As DCs mature they present the tumour antigen to T cells to initiate T cell killing to eradicate the disease cell/tumour. Importantly, we have previously shown in our group using tumour conditioned media from cultured ex vivo human colorectal cancer (CRC) explant tissue, patients that do not respond to Avastin (the licenced anti-angiogenic therapy) and have poor survival and show significant inhibition of DC maturation induced by the treated CRC tumour microenvironment. We aim to examine similar processes across the Barrett’s-dysplasia-adenocarcinoma disease sequence.
Moreover, in collaboration with Dr John Mackrill (UCC), the Ph.D. student (Ms. Lorraine Smith) will be seconded to Dr Mackrill’s lab to study the effects of calcium-electroporation on calcium homeostasis in BO and OAC. We will examine the effects of extracellular fluid from electroporated cells and tissues on calcium handling in BO and OAC cells. A further secondment will be carried out in collaboration with Dr Sharon McKenna (UCC), an internationally recognised lab in autophagy. We will assess pre and post electroporation if autophagy is activated. Autophagy is a survival mechanism that is linked with treatment resistance and selective inhibition of autophagic regulators has the potential to improve treatment regimes. These two secondments for the student will enhance our functional work linked to this electroporation programme of research. Once completed, we will be able to determine the significance of the cross talk between the electroporated Barrett’s tissue microenvironment and immune cell activation.
Anticipated impact of this AllCaN Oesophageal research project
This translational project will lead to the first human ex vivo study to examine how human Barrett’s tissues respond to electroporation therapy and if this treatment boosts immunity, alters autophagic and calcium responses. This could have significant implications in the clinic where the addition of electroporation treatment may be applied in outpatients. This pre-clinical study will generate this preliminary data for evaluation. We will also measure the impact of these industry and academic partnerships to accelerate our translational findings.