Novel key genes involved in cancer malignancy / A novel approach to address the growing need for advanced lymphatic differentiation protocols

Our group and others have demonstrated that SOX9, a developmental transcription factor that is a master regulator controlling super-enhancer dynamics in stem cell plasticity, exerts a relevant pro-oncogenic role in multiple tumour types by regulating cancer stem cell (CSC) activity. Thus, different genes highly correlated with SOX9 in tumor samples represent outstanding candidates to play a relevant role in CSC activity, tumor malignancy, prognosis and cancer therapy.

We performed computational analyses to identify the top 80 genes most positively correlated with SOX9 expression in GC samples. Among them, we selected a subset of 10 candidates and studied their clinical relevance in multiple cancer types. The expression of 10 candidates displayed prognostic impact in multiple cancer types where they correlated with cancer progression and malignancy. Among them, bioinformatic studies revealed a 4-gene signature (ECT2, TPX2, KIF11 and DIAPH3) associated with poor prognosis, therapy resistance and progenitor population. We selected KIF11 and DIAPH3 and deciphered their role in tumor cell activity finding that both are required for tumor cell survival and gCSC activity in several types of cancer. We identified the molecular mechanisms underlying their activity through OMIC approaches. Of note, the pharmacological inhibition of KIF11 and DIAPH3 severely reduces GC cell viability and self-renewal in vitro, and tumor incidence and growth in vivo.

In summary, we used a novel and unbiased computational approach to identify novel drivers of cancer progression and malignancy. In particular, we identified a novel 4-gene signature related to cancer prognosis and revealed the role of KIF11 and DIAPH3 for tumor stem cell maintenance as well the potential of their inhibition as a promising therapeutic strategy in cancer.

Traditionally, the lymphatic system has been associated with nutrient transport, immunosurveillance, and tissue-fluid homeostasis. However, recent research has highlighted its role in various disease processes, including the spread of solid tumours, obesity, and several neurological disorders. Despite its importance, knowledge about this system has been limited, largely due to the lack of suitable in vitro models. A major obstacle has been the absence of appropriate cells to create these models. To address this, we propose mesenchymal stem cell-derived lymphatic endothelial cells as a clinically relevant cell source. In this project, our goal is to establish a reliable in vitro model to study various lymphatic pathologies. To achieve this, we have developed a novel differentiation protocol that closely mimics the lymphangiogenesis (the formation of new lymphatic vessels) observed in vivo. Our findings not only lay the foundation for creating in vitro systems capable of accurately modelling lymphatic-related pathologies but also present a promising avenue for cellular therapy aimed at regenerating the lymphatic system in vivo.