Project Funding Details

Pancreatic adenocarcinoma (PDAC) is one of the deadliest types of cancer. Currently its 5-years survival rate stands around 5-8% and it is projected to be the second cancer-related cause of death by 2030. Most of its mass is composed by stroma, a mixture of fibroblast-like cell, immune cells and ECM.It is now clear that there is a substantial heterogeneity in stromal cell population, mainly formed by the so-called cancer associated fibroblasts (CAFs). So far, four distinct CAFs sub-types have been identified. Myofibroblast CAFs(myCAFs), detected already in the noninvasive state of PDAC; Inflammatory CAFs (iCAFs) observed only in the PDAC state and absent in the noninvasive dysplastic lesions; Antigen-presenting CAF (apCAF) shown to activate CD4+ T cells; Cancer-associated mesenchymal stem cells (CA-MSCs), shown to increase cancer invasion and regulate macrophage polarization. The formation of the PDAC tumour microenvironment (TME) and the characterization of the different cell types that co-exist in its dense stroma has become the main focus of many research teams. The hope is that understanding the complex dynamics of regulating the crosstalk between tumour cells and the surrounding TME will highlight new, more promising, therapeutic targets. The aim of this work is to further characterize the heterogeneous CAFs' population from a genetic and metabolic perspective, in order to be capable of designing more rational stroma-targeted therapies. To this aim, we will extract different CAFs' cells populations from patients' surgical tumour dissections and characterize them using single cell RNAseq.We will evaluate how the ablation and/ or mutation of TP53 modulates the CAFs' sub-types distribution within the PDAC stroma. We will then focus on looking for a metabolic signature.We will evaluate CAFs' mitochondrial respiration, the glycolysis rate, the ATP production and their dependency from the most common substrates (glucose, fatty acids and glutamine). The metabolic analysis will be brought further by measuring the mitochondrial respiration and the glycolysis rate in 3D co-culture spheroid model that recapitulates the interaction of PDAC tumor cells with patients' derived CAFs sub-types.we will take advantage of a well-established PDAC mouse model (KPC-mice) to test the efficacy of a metabolic drug in combination with the existing therapies. We will also use the MKC mouse model, where a proliferative wave has been observed in the splenocytes at a time point where the pancreatic tumour has not appeared yet. In this context we will verify whether some CAFs sub-populations are already located inside the pancreas before, during or after the proliferative wave, thus evaluating their possible role at very early stages of tumour formation. From our work we expect to identify specific metabolic signatures for different CAF sub-types as well as possibile metabolic pathways that can be targeted to implement PDAC therapeutic alternatives. The field of fibroblast heterogeneity is still in its infancy. It will be crucial to further deepen the knowledge about the dynamics and the plasticity of these heterogeneous CAFs' populations to identify specific targetable pathways and, thus, to better dissect PDAC progression.