Identification of Two Novel Small Compounds that Inhibit Liver Cancer Formation in Zebrafish and Analysis of Their Conjugation to Nanodiamonds to Further Reduce Toxicity

Abstract

Liver cancer, which is ranked fourth in cancer-related mortality worldwide, lacks effective therapeutic treatments. The development of new targeted therapies for liver cancer is urgently needed. The zebrafish is an excellent preclinical model organism for drug screening. Therefore, in a zebrafish model, hundreds of small molecules are screened, and two compounds (LIB1O0078 and LIB1O0144) are identified as the strongest inducers of antiangiogenic effects without side effects. LIB1O0078 exhibits better antiproliferation ability, and LIB1O0144 has better antimigration ability, as shown by xenotransplantation assays. Furthermore, LIB1O0078 and LIB1O0144 exhibit anti-HCC effects after retro-orbital injection into adult Tg (fabp10a:HBx,Src, p53-) triple transgenic fish with obesity and liver cancer. Because of the embryonic toxicity induced by these compounds, they are conjugated with nanodiamonds (ND), which are highly biocompatible function-based carriers. ND-coated small molecules not only reduce the embryonic toxicity and hepatotoxicity of the compounds, also exhibit better anti-HCC effects when administered by oral gavage in adult Tg (fabp10a:HBx,Src, p53-) fish with obesity and liver cancer. This study integrates nanotechnology and biomedical technology, identifies new potential anticancer drugs, and demonstrates the effectiveness of coating them on nanodiamonds in vivo. Facilitated by such a rapid zebrafish screening system, new anticancer drugs can be identified in a personalized and timely manner.

Dengue Virus Envelope Protein Domain III Induces Nlrp3 Inflammasome-Dependent NETosis-Mediated Inflammation in Mice

“…After fixation by 4% paraformaldehyde on coverslips, these neutrophils were then stained with rabbit anti-mouse citrulline Histone H3 antibody (1:1000) and 4',6-diamidino-2-phenylindole (DAPI, 5 μl/ml). A fluorescence microscope (Nikon Eclipse E800; Nikon Taiwan, Taipei, Taiwan) ( 51 ) was used for obtaining the NETosis images. To analyze mitochondria membrane potential, superoxide, and membrane potential levels, mitochondria labeling reagents MitoTracker™ Green FM (Thermo Fisher Scientific, Waltham, MA, USA), MitoSOX™ Red mitochondrial superoxide indicator (Thermo Fisher Scientific), MitoTracker™ Red CMXRos (Thermo Fisher Scientific) were used according to the manufacturer's instructions ( 33 , 52 ).…”

Opportunistic gill infection is associated with TiO2 nanoparticle-induced mortality in zebrafish

“…Zebrafish have been used to study engineered nanomaterials and NPs in various fields, such as biomedical research [38,[56][57][58] and studies on environmental health and safety [59,60]. Most relevant studies have reported that TiO 2 NPs are toxic and induce mortality in zebrafish embryos but are less toxic and cause less mortality in adult zebrafishes; this finding is likely attributable to the fact that these studies have tended to focus only on observations of acute stimulation [18,[61][62][63][64].…”

“…Adult wild zebrafish were used in the experiment and were kept in a semistatic system with charcoal-filtered tap water (pH 7.0-7.4) at 28 ± 0.5˚C as recommended in a previous study PLOS ONE [38], with a 12-h light-12-h dark (12L:12D) photocycle (Classictone incandescent lamp, Philips; Taiwan, without illuminating UV to avoid UV-induced photocatalysis [5,11,12,33,39,40]). The fish were fed newly hatched brine shrimp and pellet food (Zeigler Brothers, Gardners, PA, USA) and were kept in 30-L glass tanks with 20 L of water per tank.…”

Nanodiamond-Induced Thrombocytopenia in Mice Involve P-Selectin-Dependent Nlrp3 Inflammasome-Mediated Platelet Aggregation, Pyroptosis and Apoptosis

“…To prepare the stock solutions of 10 mg/mL TiO 2 (5 and 60 nm; Nanostructured & Amorphous Materials, Katy, TX, USA), 10 mg/mL NDs (5-200 nm; Kay Diamond Products, Boca Raton, FL. USA) ( 56 , 57 ), and red fluorescent NDs (50 nm; brFND-50, nitrogen-vacancy NV centers per particle > 100, FND Biotech, Taipei, Taiwan) ( 58 , 59 ), the nanoparticles (NPs) were dispersed in distilled deionized water under sonication (80 W/L, 46 kHz) for 20 min. Test NP solutions were prepared immediately before use by dilution of the stock solutions with distilled deionized water and sonication (80 W/L, 46 kHz) for 20 min ( 60 ).…”