During the reporting period, activities have been implemented in line with the objectives outlined in the Description of the Action, particularly with respect to data generation, technology establishment, and early-stage mechanistic analyses. Key activities included the establishment of multiplex imaging approaches for thick liver sections, as well as the implementation of high-dimensional flow cytometry to comprehensively characterize the tumor microenvironment.
Significant progress has been made in multi-omic data generation and analysis. This includes the analysis of sequencing data from healthy liver and liver metastases, the establishment of spatial transcriptomics pipelines, and the development of data-sharing applications for single-cell RNA sequencing data within the consortium. In parallel, bioinformatic analyses have focused on NK cell biology and their interactions within the tumor microenvironment, including the derivation of cross-tissue NK cell signatures. Experimental work further addressed NK cell heterogeneity through immunophenotyping, cytotoxicity assays, and transcriptomic profiling.
In line with the aims, advanced spatial transcriptomics methodologies have been established, including the development of high-plex panels (e.g. Vizgen MERSCOPE) tailored to characterize liver immune cell populations. Moreover, relevant in vivo models have been successfully implemented, including a syngeneic mouse model of colorectal cancer liver metastases (AKTPF), which recapitulates key molecular and histopathological features of human disease. Complementary analyses on human patient samples (FFPE tissues) using spatial transcriptomics have also been initiated.
Overall, these activities are fully consistent with the planned work programme and provide a strong foundation for the subsequent phases of the project.
