Construction of a New Ferroptosis-related Prognosis Model for Survival Prediction in Colorectal Cancer.

AIM: This study was designed to develop a ferroptosis-related gene signature for guiding the prognostic prediction in colorectal cancer (CRC) and to explore the potential in the molecular functions of the gene signature. BACKGROUND: Ferroptosis is mainly characterized by lipid peroxide accumulation on the cell membranes in an iron-dependent manner, resulting in cellular oxidative stress, metabolic disorders, and, ultimately, cell death. This study aimed to develop a prognostic ferroptosis signature in CRC and explore its potential molecular function. OBJECTIVE: The present work was designed to devise a ferroptosis signature for CRC prognosis and explore its potential molecular function. METHODS: Single-cell RNA sequencing data GSE161277 and transcriptome sequencing data GSE17537 and TCGA-CRC from the Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA) databases were downloaded, respectively. Quality control, dimension reduction, clustering, and clustering of single-cell RNA sequencing (scRNA- seq) data were performed using the Seurat package. A total of 259 ferroptosis-correlated genes from the FerrDb database were acquired. The single sample gene set enrichment analysis (ssGSEA) was performed to calculate the scores of genes related to ferroptosis. ESTIMATE was used to calculate immune infiltration. Independent prognostic factors were determined by performing Weighted Gene Co-Expression Network Analysis (WGCNA), univariate and Cox analyses, and Lasso analyses were used to search for independent prognostic factors. RESULTS: From the scRNA-seq (GSE161277) dataset, 22 cell clusters were initially identified, and according to immune cell markers, only 8 types of cells (Follicular B, central memory T cell, Epithelial, Natural killer T cell, Plasma B, M1 macrophage, Fibroblasts, and Mast cell) were finally determined to be related to CRC prognosis. The results of the scRNA-seq analysis showed that the score of ferroptosis-related genes was higher in tumour tissues and in 8 types of cells in tumour samples. In the TCGA dataset, CRC samples were divided into ferroptosis-related high scores, ferroptosis-related median scores, and ferroptosis-related low scores. Immune cell analysis revealed that ferroptosis- related high scores had the highest abundance of immune cells. An 11-gene signature was developed by WGCNA, univariate Cox, and Lasso Cox regression. The prediction ability of the signature was successfully validated in the GSE17537 dataset. A comprehensive nomogram combining the 11 signature genes and clinical parameters could effectively predict the overall survival of CRC patients. CONCLUSIONS: The present molecular signature established based on the 11 ferroptosis-related genes performed well in assessing CRC prognosis. The present discoveries could inspire further research on ferroptosis, providing a new direction for CRC management.

ASB3 promotes hepatocellular carcinoma progression by mediating DR5 ubiquitination in TRAIL resistance.

Ankyrin-repeat proteins with a suppressor of cytokine signaling box (ASB) proteins belong to the E3 ubiquitin ligase family. 18 ASB members have been identified whose biological functions are mostly unexplored. Here, we discovered that ASB3 was essential for hepatocellular carcinoma (HCC) development and high ASB3 expression predicted poor clinical outcomes. ASB3 silencing induced HCC cell growth arrest and apoptosis in vitro and in vivo. Liver-specific deletion of Asb3 gene suppressed diethylnitrosamine (DEN)-induced liver cancer development. Mechanistically, ASB3 interacted with death receptor 5 (DR5), which promoted ubiquitination and degradation of DR5. We further showed that ASB3 knockdown stabilized DR5 and increased the sensitivity of liver cancer cells to the treatment of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in a DR5-dependent manner in cellular and in animal models. In summary, we demonstrated that ASB3 promoted ubiquitination and degradation of DR5 in HCC, suggesting the potential of targeting ASB3 to HCC treatment and overcome TRAIL resistance.

MLN4924 Treatment Diminishes Excessive Lipid Storage in High-Fat Diet-Induced Non-Alcoholic Fatty Liver Disease (NAFLD) by Stimulating Hepatic Mitochondrial Fatty Acid Oxidation and Lipid Metabolites.

MLN4924 is a selective neddylation inhibitor that has shown great potential in treating several cancer and metabolic diseases, including obesity. However, it remains largely unknown whether MLN4924 has similar effect on non-alcoholic liver disease (NAFLD), which is closely associated with metabolic disorders. Here, we investigated the role of MLN4924 in NAFLD treatment and the underlying mechanism of the action using primary hepatocytes stimulated with free fatty acid, as well as high-fat diet (HFD)-induced NAFLD mouse models. We found that MLN4924 can inhibit the accumulation of lipid and reduce the expression of peroxisome proliferator-activated receptor γ (PPARγ), a key player in adipocyte differentiation and function in both in vivo and in vitro models. Moreover, we verified its important role in decreasing the synthesis and accumulation of fat in the liver, thus mitigating the development of NAFLD in the mouse model. The body weight and fat mass in MLN4924-treated animals were significantly reduced compared to the control group, while the metabolic activity, including O2 consumption, CO2 and heat production, also increased in these animals. Importantly, we demonstrated for the first time that MLN4924 can markedly boost mitochondrial fat acid oxidation (FAO) to alter liver lipid metabolism. Finally, we compared the metabolites between MLN4924-treated and untreated Huh7 cells after fatty acid induction using lipidomics methods and techniques. We found induction of several metabolites in the treated cells, including Beta-guanidinopropionic acid (b-GPA) and Fluphenazine, which was in accordance with the increase of FAO and metabolism. Together, our study provided a link between neddylation modification and energy metabolism, as well as evidence for targeting neddylation as an emerging therapeutic approach to tackle NAFLD.

ONX0912, a selective oral proteasome inhibitor, triggering mitochondrial apoptosis and mitophagy in liver cancer.

Proteasome inhibitors represent effective anti-tumor drugs. ONX0912 is a novel oral proteasome inhibitor that selectively targets the chymotrypsin-like activity of 20S proteasome subunits ß5 and LMP7 （Low molecular mass polypeptide-7）. It has been shown to be effective in hematologic malignancies. However, its anti-tumor effect in solid tumors remains unclear. Here, we discovered that ONX0912 suppressed the expansion of liver cancer cells. ONX0912 treatment led to an increased level of mitochondrial membrane potential collapse and mitochondrial ROS in tumor cells in a concentration- and exposure time-dependent manner, indicating ONX0912 triggers apoptosis through the intrinsic mitochondrial pathway. ONX0912 also induced mitophagy by activating Parkin/Pink pathway. Silencing mitophagy receptor protein, p62, aggravated the ONX0912-mediated apoptosis, which implied a new mechanism for the conversion between autophagy and apoptosis. Furthermore, we found that the ONX0912 target protein, LMP7 was overexpressed in liver cancer tissues compared to their adjacent tissues and increased level of LMP7 predicted worse clinical characteristics and poorer prognosis. In conclusion, we demonstrated that ONX0912 suppressed liver cancer cell expansion by inducing apoptosis and mitophagy. Our data also revealed ONX0912 as a potential clinical therapeutic drug for liver cancer therapy, and inhibition of mitophagy may sensitize the anti-tumor effect of ONX0912.

Comparative proteomic analysis of Aurelia coerulea for its locomotion system molecular structure-function inference.

BACKGROUND: Rhythmic contraction and autonomous movement play a key role in the predation, production and displacement of jellyfish. METHODS: Four independent body parts of the jellyfish Aurelia coerulea, including Bell, Tentacle, Oral arm and Gastric pouch were extracted and have been carried out a compared proteomics by liquid chromatography-mass spectrometry/mass-spectrometry (LC-MS/MS). ResultsA total of 13,429 peptides and 1916 proteins with molecular weights in the range of 10.6-980.9 kDa were identified, where 1916, 1562, 1474 and 1441 proteins were matched in the Gastric pouch, Tentacle, Oral arm and Bell, respectively. Gene Ontology (GO) analysis showed that translation, cytoplasma and ATP binding occupy the top differential terms of the three subdomains Biological process, Cellular Component and Molecular Function. A total of 326 pathways were successfully mapped that are mainly associated with intracellular synthesis, metabolism as well as intracellular functions. Moreover, a total of 27 contractile machinery associated proteins including 22 myosin, 3 actin and 2 tropomyosin were identified. CONCLUSIONS: Our results provide a composition profile in the four independent body parts of the jellyfish A. coerulea, of which the identified muscular proteins will greatly help in the understanding of the structural and functional relationship, as well as their operating mechanisms in the jellyfish locomotion system. SIGNIFICANCE: Omics studies have gained a new overall insight into the function of gene and protein networks during the development of motor systems in both bilateral and radial symmetrical animals. A compared proteomics using the label-free method of nano-LC-MS/MS has been performed through the four independent body parts of the moon jellyfish A. coerulea, including Bell, Tentacle, Oral arm and Gastric pouch. In addition to conventional bioinformatics analyses such as GO and KEGG, we have scanned the locomotion-related components, aligned their sequences, simulated three dimensional structures as well as did the molecular phylogenetic analyses. Our investigation provides a composition profile in the four independent body parts of the jellyfish A. coerulea, of which the identified muscular proteins will greatly help in the understanding of the structural and functional relationship, as well as their operating mechanisms in the jellyfish locomotion system.

De novo transcriptomic analysis of the venomous glands from the scorpion Heterometrus spinifer revealed unique and extremely high diversity of the venom peptides.

Scorpion, as an ancient species, has been widely used on dozens of human diseases in traditional Chinese Medicine. Although the scorpion venom from the Buthidae family with the potent toxicity attracts more interests, toxins from the non-Buthidae family draw great attention as well because of its abundance and complexity even without harm to mammals. Moreover, several toxic components of scorpion venom have been identified as valuable scaffolds for the drug design and development. Using the Next Generation Sequencing (NGS) technique, here we reported the transcriptome of the venomous glands of Heterometrus spinifer, a non-Buthidae scorpion that only a few toxic and complete components have been identified known-to-date. The total mRNA extracted from the venomous glands of H. spinifer was subjected to illumina sequencing with a strategy of de novo assembly, and a total of 54 189 transcripts were unigenes from a total of 88 311 600 determined reads. We annotated 18 567 (34.26%) unigenes from NR database, 12 258 (22.62%) from SWISSPROT database, 11 161 (20.60%) from GO database, 10 159 (18.75%) from COG database and 5059 (9.34%) from KEGG database, respectively. 2843 unigenes were further selected against the toxin-related sub-database of SWISSPROT. After removing the redundancy, 13 common toxin-related subfamilies with 62 unigenes were manually confirmed, including 8 K-toxins, 1 calcin, 3 Imperatoxin I-like, 2 La1-like, 1 scorpin-like, 3 antimicrobial peptides, two types of protease inhibitors such as 8 Kunitz-type protease inhibitors and 3 Ascaris-type protease inhibitors, and 33 proteases including 16 serine proteinases, 7 phospholipases, 5 metalloproteases, 3 hyaluronidases and 2 phosphatases. Our report is the first transcriptomic analyses of venomous glands from the scorpion H. spinifer, serving as a public information platform for the development of novel bio-therapeutics.