The project will focus upon the use of small animal optical imaging to monitor in vivo molecular expression of pH and the transportome in the PDAC tumour cell compartment. One of the first tasks will be to optimize our near infrared pH activatable probe to image the PDAC tumour. Then we will utilise in vivo dual coloured bioluminescence constructs to express two luciferase reporter genes to monitor transporter protein expression and tumour growth simultaneously. The first reporter will be a knock-in of the luciferase gene (onto the end of the endogenous gene using CRISPR-CAS) or by using a promoter construct for the selected transporter. The second reporter will be a constitutive promoter with near infrared luciferase to determine tumour size. Cell lines expressing the gene reporters will be transplanted into nude mice.

Live imaging will be complemented by immunohistochemical analysis. The use of optoacoustic imaging will be used to analyse the tumour microenvironment in order to derive quantitative information (pH, pO2). Modification by CD40 agonists and hyaluronidase will be measured and we will assess the complex array of components that indicate physiological status, phenotype and functionality as a function of PDAC progression/pH landscape by flow cytometry /CyTOF (Cytometry by Time-Of-Flight).