Liposomal delivery of hydrophobic RAMBAs provides good bioavailability and significant enhancement of retinoic acid signalling in neuroblastoma tumour cells

Retinoid treatment is employed during residual disease treatment in neuroblastoma, where the aim is to induce neural differentiation or death in tumour cells. However, although therapeutically effective, retinoids have only modest benefits and suffer from poor pharmacokinetic properties. In vivo, retinoids induce CYP26 enzyme production in the liver, enhancing their own rapid metabolic clearance, while retinoid resistance in tumour cells themselves is considered to be due in part to increased CYP26 production. Retinoic acid metabolism blocking agents (RAMBAs), which inhibit CYP26 enzymes, can improve retinoic acid pharmacokinetics in pre-clinical neuroblastoma models. Here we demonstrate that in cultured neuroblastoma tumour cells, RAMBAs enhance retinoic acid action as seen by morphological differentiation, AKT signalling and suppression of MYCN protein. Although active as retinoid enhancers, these RAMBAs are highly hydrophobic and their effective delivery in humans will be very challenging. Here we demonstrate that such RAMBAs can be loaded efficiently into cationic liposomal particles, where the RAMBAs achieve good bioavailability and activity in cultured tumour cells. This demonstrates the efficacy of RAMBAs in enhancing retinoid signaling in neuroblastoma cells and shows for the first time that liposomal delivery of hydrophobic RAMBAs is a viable approach, providing novel opportunities for their delivery and application in humans

The liposomal delivery of hydrophobic oxidovanadium complexes imparts highly effective cytotoxicity and differentiating capacity in neuroblastoma tumour cells

Oxidovanadium complexes with organic ligands are well known to have cytotoxic or differentiating capabilities against a range of cancer cell types. Their limited use in clinical testing though has resulted largely from uncertainties about the long-term toxicities of such complexes, due in part to the speciation to vanadate ions in the circulation. We hypothesised that more highly stable complexes, delivered using liposomes, may provide improved opportunities for oxidovanadium applications against cancer. In this study we sourced specifically hydrophobic forms of oxidovanadium complexes with the explicit aim of demonstrating liposomal encapsulation, bioavailability in cultured neuroblastoma cells, and effective cytotoxic or differentiating activity. Our data show that four ethanol-solubilised complexes with amine bisphenol, aminoalcohol bisphenol or salan ligands are equally or more effective than a previously used complex bis(maltolato)oxovanadium(V) in neuroblastoma cell lines. Moreover, we show that one of these complexes can be stably incorporated into cationic liposomes where it retains very good bioavailability, apparently low speciation and enhanced efficacy compared to ethanol delivery. This study provides the first proof-of-concept that stable, hydrophobic oxidovanadium complexes retain excellent cellular activity when delivered effectively to cancer cells with nanotechnology. This offers the improved prospect of applying oxidovanadium-based drugs in vivo with increased stability and reduced off-target toxicity.</p

Pharmacological development of target-specific delocalized lipophilic cation-functionalized carboranes for cancer therapy

PURPOSE: Tumor cell heterogeneity and microenvironment represent major hindering factors in the clinical setting toward achieving the desired selectivity and specificity to malignant tissues for molecularly targeted cancer therapeutics. In this study, the cellular and molecular evaluation of several delocalized lipophilic cation (DLC)-functionalized carborane compounds as innovative anticancer agents is presented. METHODS: The anticancer potential assessment of the DLC-carboranes was performed in established normal (MRC-5, Vero), cancer (U-87 MG, HSC-3) and primary glioblastoma cancer stem (EGFRpos, EGFRneg) cultures. Moreover, the molecular mechanism of action underlying their pharmacological response is also analyzed. RESULTS: The pharmacological anticancer profile of DLC-functionalized carboranes is characterized by: a) a marked in vitro selectivity, due to lower concentration range needed (ca. 10 fold) to exert their cell growth-arrest effect on U-87 MG and HSC-3, as compared with that on MRC-5 and Vero; b) a similar selective growth inhibition behavior towards EGFRpos and EGFRneg cultures (>10 fold difference in potency) without, however, the activation of apoptosis in cultures; c) notably, in marked contrast to cancer cells, normal cells are capable of recapitulating their full proliferation potential following exposure to DLC-carboranes; and, d) such pharmacological effects of DLC-carboranes has been unveiled to be elicited at the molecular level through activation of the p53/p21 axis. CONCLUSIONS: Overall, the data presented in this work indicates the potential of the DLC-functionalized carboranes to act as new selective anticancer therapeutics that may be used autonomously or in therapies involving radiation with thermal neutrons. Importantly, such bifunctional capacity may be beneficial in cancer therapy

Exploring the therapeutic potential of protein tyrosine phosphatase inhibition in neuroblastoma

Neuroblastoma accounts for 15% of paediatric cancer deaths and there is an urgent need for improved therapeutic strategies. Phosphotyrosine signalling, regulated by the opposing actions of protein tyrosine kinases and protein tyrosine phosphatases (PTPs), is critical for virtually all aspects of cell behaviour, and is commonly perturbed in cancer. We have previously shown that pan-inhibition of PTPs using oxidovanadium induces cytotoxicity in a panel of neuroblastoma cell lines. We therefore hypothesise that there exist specific PTPs that promote tumour cell survival, and that their specific or pan-inhibition may be beneficial for the treatment of neuroblastoma. Whilst promising preclinical data using vanadium-derived compounds in in vitro and in vivo models of cancer has been reported, clinical trials have been prevented in part due to concerns surrounding off-target tissue toxicity. I have taken several approaches to harness the cytotoxic properties of oxidovanadium, and PTP inhibition, with the aim to develop new therapeutic strategies for neuroblastoma. The tumour-promoting roles of specific PTPs were investigated using loss-of-function approaches including RNAi and CRISPR/Cas9 gene knockout. The dual specificity phosphatase CDC14B was identified as a potential candidate, although further validation studies need to be considered for this enzyme to be taken forward as a potential therapeutic target. In a parallel study, the first genome-wide transcriptomic analysis in neuroblastoma cells treated with oxidovanadium has revealed a potential role for cAMP signalling in bismaltolato oxidovanadium (BMOV)-induced cytotoxicity. This pathway and others that are affected by oxidovanadium may be a source of useful therapeutic targets for neuroblastoma in the future. Finally, I have shown for the first time that hydrophobic oxidovanadium can be packaged into liposomes and maintains its cytotoxicity when delivered to neuroblastoma cells. This presents a novel opportunity to deliver vanadium with potentially fewer safety concerns, whilst retaining its broad activity and high levels of anti-cancer efficacy