Histopathologic Effect of Calcium Electroporation on Cancer in the Skin

The primary endpoint of this study is to evaluate differences in TIL population in tissue samples from treated cancer tumours two days after calcium electroporation treatment compared to before treatment (biopsy taken on the day of treatment before the calcium electroporation procedure). TIL content in biopsies will be evaluated by pathological examination and expressed as percent of cells.

Protein expression levels of immune markers e.g. markers for the innate and adaptive immune system and markers of the STING pathway compared from before treatment and at different timepoints up to 3 months.

Gene expression signatures including the 18-gene TIS will be calculated as a weighted linear average of the constituent genes.

Gene expression profiling according to tumour histology.

To clinically measure changes in lesion size 1, 2 and 3 months after treatment using caliper measurement. Changes in size due to biopsies will be accounted for.

To describe any relation between change in TIL population (percentage of cells) and tumour type before and after calcium electroporation.

To describe presence of regressive changes including necrosis at different timepoints (percentage of tissue).

To describe presence of residual tumour (yes/no) and description of topographical location.

To investigate vascular effects of calcium electroporation including changes in capillary structures by histochemical staining for endothelial biomarkers CD31 and/or CD34.

To document evolution of tumours before and after treatment using digital photography including a ruler.

To sum number of patients with complete response after one or two treatments, respectively. Complete response will be defined as disappearance of all target lesions.

To sum number of lesions across all patients with complete remission after one or two treatments, respectively (expressed at percentage of all treated lesions).

To establish number of treated tumours with complete response depending on tumour type.

To detect signs of systemic immunologic response from any routine scans before and after treatment in the inclusion period.

To investigate differences in effect depending whether the treated tumour was in a previously irradiated area.

To evaluate effect on adjacent non-tumour tissue.

To assess PD-L1 expression over time by biopsy.

To investigate any relation between change in PD-L1 expression and tumour response.

To describe PD-L1 expression in relation to cell types found in the tumour environment

To describe PD-L1 expression on different cell types of the different tumour histologies investigated.

To examine frozen tissues samples by western blotting in order to support any of the above mentioned endpoints.

Blood samples may be analyzed for NK cell- and T-cell gene expression levels. Levels before and after treatment will be compared.

To measure current during treatment as indicated by the pulse generator.

To examine frozen tissues samples by PCR. Relevant gene expression will be compared before and after treatment in breast cancer and non breast cancer samples.