Liver cancer is one of the most frequently diagnosed and fatal
cancers worldwide, with
hepatocellular carcinoma (HCC) being the most common primary
liver cancer. Hundreds of studies involving thousands of patients have now been analysed across different
cancer types, including HCC, regarding the effects of immune infiltrates on the prognosis of
cancer patients. However, for these analyses, an unambiguous delineation of the
cancer area is paramount, which is difficult due to the strong heterogeneity and considerable inter-operator variability induced by qualitative visual assessment and manual assignment. Nowadays, however, multiplex analyses allow the simultaneous evaluation of multiple
protein markers, which, in conjunction with recent machine learning approaches, may offer great potential for the objective, enhanced identification of
cancer areas with further in situ analysis of prognostic immune parameters. In this study, we, therefore, used an exemplary five-marker multiplex immunofluorescence panel of commonly studied markers for prognosis (CD3 T, CD4 T helper, CD8 cytotoxic T, FoxP3 regulatory T, and PD-L1) and
DAPI to assess which analytical approach is best suited to combine morphological and immunohistochemical data into a
cancer score to identify the
cancer area that best matches an independent pathologist's assignment. For each cell, a total of 68 individual cell features were determined, which were used as input for 4 different approaches for computing a
cancer score: a correlation-based selection of individual cell features, a MANOVA-based selection of features, a multilayer perceptron, and a convolutional neural network (a U-net). Accuracy was used to evaluate performance. With a mean accuracy of 75%, the U-net was best capable of identifying the
cancer area. Although individual cell features showed a strong heterogeneity between patients, the spatial representations obtained with the computed
cancer scores delineate HCC well from non-
cancer liver tissues. Future analyses with larger sample sizes will help to improve the model and enable direct, in-depth investigations of prognostic parameters, ultimately enabling
precision medicine.