PDGFs and PDGFRs in canine osteosarcoma: New targets for innovative therapeutic strategies in comparative oncology

Identification of anti-proliferative kinase inhibitors as potential therapeutic agents to treat canine osteosarcoma

“…Interestingly, the PDGFR inhibitor AG1296, which has recently been shown to inhibit AKT phosphorylation in canine OS overexpressing PDGFR (Maniscalco et al, 2013), did not show significant decreased cell viability in our screen. Finally, another study showed that the cMet inhibitor (Liao et al, 2007) PF2362376 could inhibit proliferation of canine OS cells, but this compound was unfortunately not present in our library.…”

“…In particular, several protein kinases are potential interesting drug molecular targets, as these are often overexpressed or dysregulated in cancer cells. Individual kinases have been found to be expressed in canine OS and OS cell lines, including HER2 (Flint et al, 2004), EGFR , cMET (De Maria et al, 2009;Fieten et al, 2009), PDGFR (Fahey et al, 2013;Maniscalco et al, 2013), cKIT (Fahey et al, 2013), and PKC (Hong et al, 2011). This suggests that kinases represent potential targets for canine OS adjuvant therapies.…”

Increased expression of insulin-like growth factor-1 receptor is correlated with worse survival in canine appendicular osteosarcoma

“…Seven primary canine OSA cell lines (Wall, Penny, Dark, Sky, Pedro, Lord and Desmon) (Maniscalco et al, 2013) and normal osteoblasts (OSB1) were cultured in Iscove's standard medium supplemented with 10% fetal bovine serum (FBS), 1% glutamine, 100 μg/mL penicillin and 100 μg/mL streptomycin. NMuMG cells were purchased from American Type Culture Collection (ATCC) and cultured in DMEM medium containing 10% FBS (HyClone), 10 μg/mL insulin (Sigma-Aldrich), and 100 IU/ mL penicillin.…”

“…Total proteins were obtained from primary canine OSA cell lines, OSB1 (De Maria et al, 2009;Maniscalco et al, 2013) and NMuUG cell lines by boiling lysis buffer containing 1% SDS and 0.1 M Tris-HCl (pH 6.8). Total proteins from each sample (40 μg) were separated on an 8% sodium dodecyl sulphate (SDS)-polyacrylamide (PAGE) gel, transferred onto a Hybond-C Extra membrane (Amersham Biosciences) and incubated overnight at 4°C with the primary antibodies (Table 2).…”

“…Furthermore, as in humans, 79% of canine OSA samples overexpress the MET oncogenes; cell motility and invasiveness appear to be MET-dependent since they can be inhibited by small interfering RNAs (siRNAs) that are specific for MET (De Maria et al, 2009). Finally, we have recently reported that PDGF receptors are overexpressed in canine OSA, adding a new potential therapeutic target (Maniscalco et al, 2013).…”