p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma.

With few curative treatments and a global yearly death rate of over 800,000, hepatocellular carcinoma (HCC) desperately needs new therapies. Although wild-type p53 gene therapy has been shown to be safe in HCC patients, it has not shown enough efficacy to merit approval. This work aims to show how p53 can be re-engineered through fusion to the pro-apoptotic BH3 protein Bcl-2 antagonist of cell death (Bad) to improve anti-HCC activity and potentially lead to a novel HCC therapeutic, p53-Bad*. p53-Bad* is a fusion of p53 and Bad, with two mutations, S112A and S136A. We determined mitochondrial localization of p53-Bad* in liver cancer cell lines with varying p53 mutation statuses via fluorescence microscopy. We defined the apoptotic activity of p53-Bad* in four liver cancer cell lines using flow cytometry. To determine the effects of p53-Bad* in vivo, we generated and analyzed transgenic zebrafish expressing hepatocyte-specific p53-Bad*. p53-Bad* localized to the mitochondria regardless of the p53 mutation status and demonstrated superior apoptotic activity over WT p53 in early, middle, and late apoptosis assays. Tumor burden in zebrafish HCC was reduced by p53-Bad* as measured by the liver-to-body mass ratio and histopathology. p53-Bad* induced significant apoptosis in zebrafish HCC as measured by TUNEL staining but did not induce apoptosis in non-HCC fish. p53-Bad* can induce apoptosis in a panel of liver cancer cell lines with varying p53 mutation statuses and induce apoptosis/reduce HCC tumor burden in vivo in zebrafish. p53-Bad* warrants further investigation as a potential new HCC therapeutic.

Development of mesoporosity in scandia-stabilized zirconia: particle size, solvent, and calcination effects.

We present the mechanisms of formation of mesoporous scandia-stabilized zirconia using a surfactant-assisted process and the effects of solvent and thermal treatments on the resulting particle size of the powders. We determined that cleaning the powders with water resulted in better formation of a mesoporous structure because higher amounts of surfactant were preserved on the powders after washing. Nonetheless, this resulted in agglomerate sizes that were larger. The water-washed powders had particle sizes of >5 µm in the as-synthesized state. Calcination at 450 and 600 °C reduced the particle size to ∼1-2 and 0.5 µm, respectively. Cleaning with ethanol resulted in a mesoporous morphology that was less well-defined compared to the water-washed powders, but the agglomerate size was smaller and had an average size of ∼250 nm that did not vary with calcination temperature. Our analysis showed that surfactant-assisted formation of mesoporous structures can be a compromise between achieving a stable mesoporous architecture and material purity. We contend that removal of the surfactant in many mesoporous materials presented in the literature is not completely achieved, and the presence of these organics has to be considered during subsequent processing of the powders and/or for their use in industrial applications. The issue of material purity in mesoporous materials is one that has not been fully explored. In addition, knowledge of the particle (agglomerate) size is essential for powder handling during a variety of manufacturing techniques. Thus, the use of dynamic light scattering or any other technique that can elucidate particle size is essential if a full characterization of the powders is needed for achieving postprocessing effectiveness.

The Challenges and Prospects of p53-Based Therapies in Ovarian Cancer.

It has been well established that mutations in the tumor suppressor gene, p53, occur readily in a vast majority of cancer tumors, including ovarian cancer. Typically diagnosed in stages three or four, ovarian cancer is the fifth leading cause of death in women, despite accounting for only 2.5% of all female malignancies. The overall 5-year survival rate for ovarian cancer is around 47%; however, this drops to an abysmal 29% for the most common type of ovarian cancer, high-grade serous ovarian carcinoma (HGSOC). HGSOC has upwards of 96% of cases expressing mutations in p53. Therefore, wild-type (WT) p53 and p53-based therapies have been explored as treatment options via a plethora of drug delivery vehicles including nanoparticles, viruses, polymers, and liposomes. However, previous p53 therapeutics have faced many challenges, which have resulted in their limited translational success to date. This review highlights a selection of these historical p53-targeted therapeutics for ovarian cancer, why they failed, and what the future could hold for a new generation of this class of therapies.