Award Title: Identification of novel druggable targets in canine insulinoma through single-cell transcriptomic analysis
Funder: The Academy of Medical Sciences

Abstract: Aims and objectives: 
The key aim of this project is to identify novel therapeutic targets in canine insulinoma (INS). 

I hypothesise that single-cell transcriptomic analysis (scRNA-Seq) will reveal novel druggable targets with the potential to inhibit growth or function of INS, ultimately enabling improvement of treatment modalities for both canine and human INS patients.

To achieve this aim, I will pursue the following objectives:
1)Undertake scRNA-Seq on fresh and frozen surgically-excised canine INS, to understand their transcriptomic landscape and to determine whether cryo-archiving of surgical samples is feasible in future studies without data loss.
2)Compare scRNA-Seq of the canine INS cell line canINS to that of surgically-excised canine INS and human INS cell line CM, to understand whether this is an appropriate model.
3)To utilise these data to identify potential novel druggable targets in canine INS.

INS is the most common pancreatic neuroendocrine tumour in dogs. Despite current multimodal treatment protocols, the prognosis for INS is poor in 95% of the cases, because primary INS is frequently inoperable and/or micrometastases are present during surgery. 

This funding will allow me to optimise the scRNA-Seq protocol and workflow to process canine INS patient samples. Optimising cryopreservation holds strong appeal as it will allow me to archive samples more effectively in future, for batch processing.  

To identify candidate druggable targets, I will select differentially expressed genes with a plausible effect on INS proliferation and progression, for in vitro functional validation experiments and immunohistochemistry on a canine INS tissue microarray.

Lay Abstract: Dogs and humans can develop a tumour of the pancreas that produces too much insulin, called an ‘insulinoma’. These tumours cause signs such as collapse at exercise and very low blood sugar. Insulinomas are difficult to cure by surgery because they spread to other tissues very commonly, and current drug treatments do not work very effectively.  New treatments are needed to improve patient survival.

This project will study canine insulinomas using a new technology that allows each cell to be examined individually, to see which genes are turned on and which are turned off. This will allow us to identify which genes are crucially important for survival of insulinoma cells, and then to design new treatments targeted at these genes specifically.

We will use tumour tissue which is surplus to diagnostic requirements, taken from pet dogs undergoing surgical removal of insulinomas at the Royal Veterinary College. An important part of this project will also be to understand whether it will be possible in the future to study tumours that have been frozen after surgery. If this is the case, it will greatly improve our ability to collect samples, because they can be frozen on the day of surgery and then studied in batches.

We will also be using a technique called tissue culture to grow insulinoma cells in the laboratory, and studying these to see how closely they resemble insulinoma cells in actual tumours. We hope that this work will improve treatment options for both dogs and humans with insulinoma.

Research Activity Codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.1 PHARMACEUTICALS
Health Categories: CANCER AND NEOPLASMS