Combination of phenethyl isothiocyanate and dasatinib inhibits hepatocellular carcinoma metastatic potential through FAK/STAT3/Cadherin signalling and reduction of VEGF secretion

Cancerous cells are characterised by their ability to invade, metastasise and induce angiogenesis. To achieve this, tumour cells use various molecules that can be targeted to reverse these processes. Dasatinib, a potent Src inhibitor, has shown promising results in treating hepatocellular carcinoma (HCC) in vitro and in vivo. However, its effectiveness is limited by FAK activation. Isothiocyanates (ITCs), on the other hand, are phytochemicals with broad anticancer activity and FAK inhibition capabilities. This study evaluated the synergistic effects of dasatinib and phenethyl isothiocyanate (PEITC) on HCC. The combination was tested using various assays, including MTT, adhesion, scratch wound healing, Boyden chamber, chorioallantoic membrane (CAM), and yolk sac mem-brane (YSM) assays, to evaluate the effect of the drug combination on HCC metastatic potential and angiogenesis in vitro and in vivo. The results showed that the combination inhibited adhesion, migration, and invasion of HepG2 cells and reduced xenograft volume in the CAM assay. Addi-tionally, the combination reduced angiogenesis in vitro, reducing the growth of new vessels in the tube formation assay. The inhibition of FAK/STAT3 signalling led to increased E-cadherin ex-pression and reduced VEGF secretion, thus reducing HCC metastatic potential. Therefore, com-bination of PEITC and dasatinib could be a potential therapeutic strategy for the treatment of HCC

The role of Scara5 in adipogenesis and breast cancer pathogenesis

Affecting millions of people around the world, osteoporosis and bone fragility represent a serious health concern and one of the major threats for elderly people, since the risk of fracture tends to increase with ageing. Similarly, metabolic disorders such as diabetes and obesity stand as major public health problems worldwide. Both bone loss-related diseases and metabolic disorders impact on mesenchymal stromal cells (MSCs) differentiation fate, with adipogenesis favoured at the expense of osteogenesis and resulting in increased adiposity and decreased bone mass. A plethora of factors have been identified as crucial for MSCs lineage differentiation; scavenger receptor class A member 5 (Scara5) recently emerged as a regulator of adipogenesis in vitro. Among its many functions, Scara5 is a ferritin receptor and is reportedly downregulated in human breast cancer (BrCa) cell lines. Studies conducted on Scara5 -knock out (KO) mice revealed an increased bone mass and a reduced fat deposition. However, bone/fat-related pathologies (i.e. osteoporosis, diabetes) were not influenced by the genetic deletion. In line with these results, Scara5-KO bone marrow MSCs (BMSCs) showed a propensity to become osteoblasts and a reduced ability to differentiate into adipocytes. Mechanistic studies proved that Scara5 deficiency impinged on BMSCs mitochondrial activity during adipogenic differentiation, as well as on murine kidney iron levels. The reduced iron content and adipose tissue deposition also impacted on BrCa development; indeed, Scara5-KO mice developed smaller tumours upon cancer cell injection into the mammary fat pad, as a result of an iron homeostasis perturbation and the consequent reduced fat deposition. This thesis presents Scara5 as a central regulator of iron homeostasis, MSCs adipogenesis, and breast cancer pathogenesis. Although a reduction in Scara5 has been associated with health problems (i.e. cancer), these findings propose that targeting Scara5 may be useful for developing new therapies to counteract obesity and BrCa pathogenesis

Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells Both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

Mesenchymal stem cells (MSCs) have been recently introduced in veterinary medicine as a potential therapeutic tool for several pathologies. The large-scale in vitro expansion needed to ensure the preparation of a suitable number of MSCs for clinical application usually requires the use of xenogeneic supplements like the fetal bovine serum (FBS). The substitution of FBS with species-specific supplements would improve the safety of implanted cells, reducing the risk of undesired immune responses following cell therapy. We have evaluated the effectiveness of canine adipose tissue-derived stromal vascular fraction (SVF) and MSCs (ADMSCs) expansion in the presence of canine blood-derived supplements. Cells were cultured on traditional plastic surface and inside a 3D environment derived from the jellification of different blood-derived products, i.e., platelet-poor plasma (PPP), platelet-rich plasma (PRP), or platelet lysate (PL). PPP, PRP, and PL can contribute to canine ADMSCs in vitro expansion. Both allogeneic and autologous PPP and PL can replace FBS for ADMSCs culture on a plastic surface, exhibiting either a similar (PPP) or a more effective (PL) stimulus to cell replication. Furthermore, the 3D environment based on homospecific blood-derived products polymerization provides a strong stimulus to ADMSCs replication, producing a higher number of cells in comparison to the plastic surface environment. Allogeneic or autologous blood products behave similarly. The work suggests that canine ADMSCs can be expanded in the absence of xenogeneic supplements, thus increasing the safety of cellular preparations. Furthermore, the 3D fibrin-based matrices could represent a simple, readily available environments for effective in vitro expansion of ADMSCs using allogeneic or autologous blood-products