Targeting the “undruggable” cancer driver genes: <i>Ras</i>, <i>myc</i>, and <i>tp53</i>

The term “undruggable” is to describe molecules that are not targetable or at least hard to target pharmacologically. Unfortunately, some targets with potent oncogenic activity fall into this category, and currently little is known about how to solve this problem, which largely hampered drug research on human cancers. Ras, as one of the most common oncogenes, was previously considered “undruggable”, but in recent years, a few small molecules like Sotorasib (AMG-510) have emerged and proved their targeted anti-cancer effects. Further, myc, as one of the most studied oncogenes, and tp53, being the most common tumor suppressor genes, are both considered “undruggable”. Many attempts have been made to target these “undruggable” targets, but little progress has been made yet. This article summarizes the current progress of direct and indirect targeting approaches for ras, myc, two oncogenes, and tp53, a tumor suppressor gene. These are potential therapeutic targets but are considered “undruggable”. We conclude with some emerging research approaches like proteolysis targeting chimeras (PROTACs), cancer vaccines, and artificial intelligence (AI)-based drug discovery, which might provide new cues for cancer intervention. Therefore, this review sets out to clarify the current status of targeted anti-cancer drug research, and the insights gained from this review may be of assistance to learn from experience and find new ideas in developing new chemicals that directly target such “undruggable” molecules.     

<i>In Silico</i> analysis and linking of metabolism-related genes with the immune landscape in head and neck squamous cell carcinoma

Metabolic reprogramming and immunologic suppression are two critical characteristics promoting the progression of head and neck squamous cell carcinoma (HNSCC). The integrative analysis of all the metabolism-related genes (MRGs) in HNSCC is lacking and the interaction between the metabolism and the immune characteristics also requires more exploration to uncover the potential mechanisms. Therefore, this study was designed to establish a prognostic signature based on all the MRGs in HNSCC. Genes of HNSCC samples were available from the TCGA and GEO databases while the MRGs were retrieved from a previous study. Ultimately 4 prognostic MRGs were selected to construct a model possessing robust prognostic value and accuracy in TCGA cohorts. The favorable reproducibility of this model was confirmed in validation cohorts from GEO databases. The risk score calculated by this model was an independent prognostic factor that further classified these HNSCC patients into high-/low-risk groups. GSEA analyses and somatic mutations indicated the low-risk group could activate several anti-tumor pathways and possessed lower TP53 mutation. The results of ESTIMATE, single-sample GSEA, CIBERSORT, and some immune-related molecules analyses suggested the low-risk group exhibited lower metabolic activities and higher immune characteristics. The Spearman correlation test implied most metabolic pathways with tumor-promoting function were negatively correlated with the immune activity, indicating a plausible approach of combining the anti-metabolism and the immunotherapy drugs in the high-risk group to enhance therapeutic effects than applied separately. In conclusion, this prognostic signature linking MRGs with the immune landscape could promote the individualized treatment for HNSCC patients.     

<i>Trypanosoma rangeli:</i> growth in mammalian cells <i>in vitro</i> and action of a repositioned drug (17-AAG) and a natural extract (<i>Artemisia sp</i>. essential oil)

Trypanosoma rangeli and T. cruzi are both parasitic unicellular species that infect humans. Unlike T. cruzi,the causative agent of Chagas disease, T. rangeli is an infective and non-pathogenic parasite for humans, but pathogenicfor vectors from the Rhodnius genus. Because both species can coexist in different hosts and overlap their infectivecycles but very little is known about the infection of T. rangeli in mammalian cells, we decided to characterize both thedevelopment of this parasite in cell culture and the effect of therapeutic agents with potential trypanocidal action onit. We found that T. rangeli exhibits a cycle of infection in Vero cells similar to that for T. cruzi and that the repurposeddrug, 17-AAG, and the natural extract Artemisia sp. essential oil produce a toxic effect on epimastigotes showinga trypanocidal action from the fifth day of culture. Both treatments also affected the infection of trypomastigotesand reduced the capacity of replication of amastigotes of T. rangeli. Since T. cruzi / T. rangeli coinfection cases havebeen reported, the finding of drugs with potential activity against both species could be significant in the future.Furthermore, studies of susceptibility of both species to drugs could also help to know the different mechanisms ofpathogenicity in humans displayed by T. cruzi that are absent in T. rangeli     

A novel prognostic gene signature,nomogram and immune landscape based on tanshinone IIA drug targets for hepatocellular carcinoma: Comprehensive bioinformatics analysis and <i>in vitro</i> experiments

Background: Tanshinone IIA, one of the main ingredients of Danshen, is used to treat hepatocellular carcinoma (HCC). However, potential targets of the molecule in the therapy of HCC are unknown. Methods: In this study, we collected the tanshinone IIA targets from public databases for investigation. We screened differentially expressed genes (DEGs) across HCC and normal tissues using mRNA expression profiles from The Cancer Genome Atlas (TCGA). Univariate Cox regression analysis and least absolute shrinkage and selection operator (LASSO) Cox regression models were used to identify and construct the prognostic gene signature. Results: Finally, we discovered common genes across tanshinone IIA targets and HCC DEGs. We reported Fatty acid binding protein-6 (FABP6), Polo-like Kinase 1 (PLK1), deoxythymidylate kinase (DTYMK), Uridine Cytidine Kinase 2 (UCK2), Enhancer of Zeste Homolog 2 (EZH2), and Cytochrome P450 2C9 (CYP2C9) as components of a gene signature. The six-gene signature’s prognostic ability was evaluated using the Kaplan-Meier curve, time-dependent receiver operating characteristic (ROC), multivariate Cox regression analysis, and the nomogram. The mRNA level and protein expression of UCK2 were experimentally validated after treatment with different concentrations of tanshinone IIA in HEPG2 cells. CIBERSORTx, TIMER2.0, and GEPIA2 tools were employed to explore the relationship between the prognostic signature and immune cell infiltration. Conclusion: We established a six-gene signature as a reliable model with significant therapeutic possibility for prognosis and overall survival estimation in HCC patients, which might also benefit medical decision-making for appropriate treatment.     

Gene expression of 49 kDa apyrase,cytoskeletal proteins,ATPase, ADPase and amino acid contents of <i>Pisum sativum</i> (L.) cells germinated in E<i>uryops arabicus</i> (Steud. ex Jaub. & Spach) water extract

The present research reports of quick and marked changes induced by plant extract of Euryops arabicus in thegene expression of 49-kDa apyrases, cytoskeletal proteins, ATPases, ADPase and amount of amino acid of pea (Pisumsativum L. var. Alaska). Pellets of cytoskeletals proteins (27000 xg) were probed with anti-apyrase antibody, biotinylatedanti-rat, actin and alpha and beta-tubulin for Western blotting. ATPase and ADPase activities were determined basedon the hydrolytic efficacy of adenine triphosphate and adenine diphosphate. By 72 hours, the abundance of apyrases,cytoskeletal proteins and amount of amino acid in pellets of 27000 xg of germinated pea seeds in E. arabicus extractswere sharply increased than those sown in distilled water. All the samples exhibited that the stems had more amountfrom apyrases, cytoskeletal proteins, amino acids and ATPase and ADPase activities than primary leaves and primaryroots that were germinated either on E. arabicus water extract or in distilled water. Based on the enzyme’s capability tohydrolyse nucleotide triphosphate and nucleotide diphosphate as well as the direct association between expression of49-kDa apyrase and cytoskeletal proteins, E. arabicus water extract had an important effect on plant germinations.     

Regulation mechanisms of endocrine disruptors on vasodilation and vasoconstriction: Insights from <i>ex vivo</i> models

Cardiovascular diseases (CVD) are one of the leading causes of death worldwide. The knowledge and understanding of CVD are based on the study of vascular physiology and how the smooth muscle cells and tissues perform their different functions. Exposure to endocrine disruptors (EDCs), such as phytoestrogens, polycyclic aromatic hydrocarbons, flame retardants, plasticizers, pesticides, and cosmetics, is an integral and fundamental part of human exposure. Humans are exposed to EDCs by multiple pathways including air, food, water, and consumer products. However, this exposure can lead to several adverse effects on human health, including on the cardiovascular (CV) system. The negative impact that EDC toxicity has on human CV health is a serious problem that must not be overlooked. In this point of view, we proposed the use of the human umbilical artery as a human model to study the direct effects of EDCs on the vascular level. Several works where these cells were directly exposed to EDC’s were presented to highlight this well-established model as a great strategy to be used. In the future, we emphasize the need to continue to carry out different investigations using HUA to unveil and understand the vascular toxicity of EDCs and improve human CV health.     

KIFC1 overexpression promotes prostate cancer cell survival and proliferation <i>in vitro</i> by clustering of amplified centrosomes via interaction with Centrin 2

Mitotic kinesin KIFC1 plays critical roles in mitosis by regulating the spindle length, pole formation, and known for clustering extra centrosomes in cancer cells. Centrosome clustering is associated with the survival of cancer cells, but this phenomenon remains obscure in prostate cancer (PCa). The present study demonstrated that PCa cells showed centrosome amplification and clustering during interphase and mitosis, respectively. KIFC1 is highly expressed in PCa cells and tumor tissues of prostatic adenocarcinoma (PAC) patients. Up-regulation of KIFC1 facilitated the PCa cell survival in vitro by ensuring bipolar mitosis through clustering the multiple centrosomes, suggesting centrosome clustering could be a leading cause of prostate carcinogenesis. Conversely, the silencing of KIFC1 resulted in normal centrosome number or multipolar mitosis by inhibiting the clustering of amplified centrosomes in PCa cells. Besides, knockdown of KIFC1 by RNAi in PCa cells reduced cancer cell survival, and proliferation. KIFC1 interacted with centrosome structural protein Centrin 2 in clustering of amplified centrosomes in PCa cells to ensure the bipolar mitotic spindle formation. Knockdown of Centrin 2 in PCa cells inhibited the centrosome amplification and clustering. Moreover, up-regulated KIFC1 promotes PCa cell proliferation via progression of cell cycle possibly through aberrant activation of cyclin dependent kinase 1(Cdk1). Therefore, KIFC1 can be a prognostic marker and therapeutic target of PCa for inhibiting the cancer cell proliferation.     

Potential genomic biomarkers of obesity and its comorbidities for phthalates and bisphenol A mixture: <i>In silico</i> toxicogenomic approach

This in silico toxicogenomic study aims to explore the relationship between phthalates and bisphenol A (BPA) co-exposure and obesity, as well as its comorbid conditions, in order to construct a possible set of genomic biomarkers. The Comparative Toxicogenomics Database (CTD; http://ctd.mdibl.org) was used as the main data mining tool, along with GeneMania (https://genemania.org), ToppGene Suite (https://toppgene.cchmc.org) and DisGeNET (http://www.disgenet.org). Among the phthalates, bis(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) were chosen as the most frequently curated phthalates in CTD, which also share similar mechanisms of toxicity. DEHP, DBP and BPA interacted with 84, 90 and 194 obesity-related genes/proteins, involved in 67, 65 and 116 pathways, respectively. Among these, 53 genes/proteins and 42 pathways were common to all three substances. 31 genes/proteins had matching interactions for all three investigated substances, while more than half of these genes/proteins (56.49%) were in co-expression. 7 of the common genes/proteins (6 relevant to humans: CCL2, IL6, LPL, PPARG, SERPINE1, and TNF) were identified in all the investigated obesity comorbidities, while PPARG and LPL were most closely linked to obesity. These genes/proteins could serve as a target for further in vitro and in vivo studies of molecular mechanisms of DEHP, DBP and BPA mixture obesogenic properties. Analysis reported here should be applicable to any mixture of environmental chemicals and any disease present in CTD.     

Effect of <i>Lycopus lucidus</i> Turcz. supplementation on gut microflora and short chain fatty acid composition in Crj: CD-1 mice

We investigated the diversity and composition of microflora in feces of Lycopus lucidus Turcz.-fed mice. Inaddition, we evaluated the production of major cytokines (Interleukin-6 and -10) which are related to inflammationand fatty acid composition of several tissues. 16S ribosomal DNA sequencing-based microbiome taxonomic profilinganalysis was performed utilizing the EzBioCloud data base. Male mice fed on L. lucidus showed a significantlyreduced number of lactic acid bacteria and coliform in the feces compared with the control group (p < 0.05). 16SrDNA sequencing analysis of fecal samples showed that L. lucidus supplementation decreased the community ofharmful microflora (Enterobacteriaceae including Escherichia coli and Bacteroides sp.) in feces compared with thecontrol group (p < 0.05). There were no significant differences in mRNA expression of cytokine IL-6 and IL-10between the control and L. lucidus fed groups. The fecal fatty acid composition in the L. lucidus group hadpercentages of 4:0, 6:0, 8:0 and 10:0 in the intestine but those short chain fatty acids were not detected in the controlgroup. Our results showed that L. lucidus supplementation influenced gut environment by decreasing harmfulmicroflora and increased the percentages of several short fatty acids.     

An extract of <i>Hypericum perforatum</i> induces wound healing through inhibitions of Ca²⁺ mobilizations,mitochondrial oxidative stress and cell death in epithelial cells: Involvement of TRPM2 channels

The wound is induced by several mechanical and metabolic factors. In the etiology of the wound recovery,excessive oxidative stress, calcium ion (Ca²⁺) influx, and apoptosis have important roles. Ca²⁺-permeable TRPM2 channel is activated by oxidative stress. Protective roles of Hypericum perforatum extract (HP) on the mechanical nerve injury-induced apoptosis and oxidative toxicity through regulation of TRPM2 in the experimental animals wererecently reported. The potential protective roles in HP treatment were evaluated on the TRPM2-mediated cellularoxidative toxicity in the renal epithelium (MPK) cells. The cells were divided into three groups as control, wound,and wound + HP treatment (75 µM for 72 h). Wound diameters were more importantly decreased in the wound+HPgroup than in the wound group. In addition, the results of laser confocal microscopy analyses indicated protectiveroles of HP and TRPM2 antagonists (N-(p-Amylcinnamoyl) anthranilic acid and 2-aminoethyl diphenylborinate)against the wound-induced increase of Ca²⁺ influx and mitochondrial ROS production. The wound-induced increaseof early (annexin V-FITC) apoptosis and late (propidium iodide) apoptosis were also decreased in the cells by the HPtreatment. In conclusion, HP treatment acted protective effects against wound-mediated oxidative cell toxicity andapoptosis through TRPM2 inhibition. These effects may be attributed to their potent antioxidant effect.     

<Emphasis Type="Italic">Development of a Tissue Engineering Scaffold Structure Library for Rapid Prototyping. Part 1: Investigation and Classification</Emphasis>

In tissue engineering (TE), a porous scaffold structure may be required as a template to guide the proliferation, growth and development of cells appropriately in three dimensions. Although TE scaffolds can be created using one of many conventional techniques available, most will suffer from a lack of mechanical strength and/or uniformity in pore distribution and sizes. This study is focused on creating scaffolds using rapid prototyping (RP) techniques. Utilising these novel techniques, a computer-aided design (CAD) of the scaffold structure must first be modelled. The scaffold structure is then fabricated directly from CAD data using a RP system. The objective of this research is to (1) investigate and select various polyhedral shapes suitable for scaffold modelling, (2) classify the selected unit cells, (3) create a parametric library of scaffold structures and (4) verify by building the CAD models using the selective laser sintering process. The first two objectives are covered in Part 1 of this two-part paper. The remaining objectives will be described and discussed in Part 2. ID="A1"Correspondance and offprint requests to: Dr C. K. Chua, School of Mechanical and Production Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798. E-mail: mckchua@ntu.edu.sg