Aldo-keto reductase 1C3 mediates chemotherapy resistance in esophageal adenocarcinoma via ROS detoxification

Background: The incidence and mortality of esophageal adenocarcinoma (EAC) are very high, and the prognosis is poor in Europe. During the past years, Aldo-keto reductase 1C 3 (AKR1C3) as an oxidoreductase has attracted much attention in the field of cancer research. Especially in the treatment of tumors, AKR1C3 seems to show its protective effect on tumor therapy in many cancers. However, whether its role in the treatment of EAC is still unknown. Methods: Public datasets were downloaded and applied. Western blot (WB) was used to check the protein level. Immunofluorescence was also used to measure the expression and localization of AKR1C3. Cell lines in SKGT-4 (SKGT-4AKR1C3-KD /SKGT-4AKR1C3-shNT) and OACP4C (OACP4CAKR1C3-KD /OACP4CAKR1C3-shNT), and cell lines in OE33 (OE33AKR1C3-OE /OE33AKR1C3-VEC) and FLO-1 (FLO-1AKR1C3-OE /FLO-1AKR1C3-VEC) were established. Basic cell experiments such as cell proliferation/colony formation/wound healing assay were carried out in this study on those genetically modified EAC cell lines. MTT assay was used to quantify the cells’ viability after anti-cancer drugs treatment. Then cisplatin-induced cell apoptosis was checked by FACS using Annexin V/DAPI. Reactive oxygen species (ROS) of EAC cells were checked by FACS using H2DCFDA. Chromatin-immunoprecipitation (ChIP) was carried out to validate the transcriptional regulation of AKR1C3. SKGT-4NRF2-KD and SKGT-4NRF2-shNT cell lines were established. Then, PCR and WB were used to check its level in SKGT-4NRF2-KD and SKGT-4shNT cell lines. GSH quantification of genetically modified EAC cells was also checked. Moreover, mitochondrial stress test was performed in evaluating oxygen consumption rate (OCR) of EAC cells. Results: AKR1C3 mRNA level in EAC tissues is higher than in the normal squamous esophageal epithelium in the data of GSE26886 and GSE92396 from public database. However, only about half of the 12 patients’ AKR1C3 protein level had elevated in tumor tissue than in matched normal tissue. Although public databases revealed the trend that higher AKR1C3 means poorer survival probabilities, no difference in survival can be observed between the AKR1C3high and AKR1C3low group. Then, we detected its protein level in OE19, OE33, OACP4C, FLO-1, JHesoAD1 and SKGT-4 cell lines. It high-expressed in OE19, OACP4C and SKGT-4 cell lines, and it low-expressed in OE33, JHesoAD1 and FLO-1 cell lines. Immunofluorescence showed the same results with WB. Furthermore, immunofluorescence results also found that AKR1C3 was mainly expressed in the cytoplasm. OE33AKR1C3-OE and FLO-1AKR1C3-OE cell lines were more capable of proliferating/ colony-forming/migrating than OE33AKR1C3-VEC and FLO-1AKR1C3-VEC cell lines. Opposite results were observed in SKGT-4AKR1C3-KD and OACP4CAKR1C3-KD cell lines. Moreover, OE33AKR1C3-OE and FLO-1AKR1C3-OE cells had less apoptosis induced by anti-cancer drugs, while SKGT-4AKR1C3-KD and OACP4CAKR1C3-KD cells showed the opposite results. Furthermore, we found that higher ROS in SKGT-4AKR1C3-KD and OACP4CAKR1C3-KD cells and lower ROS in OE33AKR1C3-OE and FLO-1AKR1C3-OE cells compared with their control cell lines. The nuclear factor erythroid 2-related factor 2 (NRF2) controls redox homeostasis, and it binds with AKR1C3’s promoter and mediates its expression. And their expression was decreased in SKGT-4NRF2-KD cells than in the SKGT-4NRF2-shNT cells. Moreover, the rescue experiment showed that NAC and BSO could make AKR1C3’ effect disappear whether in SKGT-4/OACP4C AKR1C3-KD/shNT cells or OE33/ FLO-1AKR1C3-OE/VEC cells. Furthermore, the phosphorylation of AKT was decreased in SKGT-4AKR1C3-KD and OACP4C AKR1C3-KD cell lines than in SKGT-4AKR1C3-shNT and OACP4CAKR1C3-shNT cell lines. Conversely, it was increased in OE33 AKR1C3-OE and FLO-1 AKR1C3-OE cell lines than OE33 AKR1C3-VEC and FLO-1 AKR1C3-VEC cells lines. AKT inhibitor could inhibit the phosphorylation of AKT but had no effect on AKR1C3 expression. And inhibitor of AKT could diminish the protective effect of the decrease for apoptotic cells caused by the elevated of AKR1C3 in OE33AKR1C3-OE and FLO-1AKR1C3-OE cell lines. Moreover, GSH levels was decreased, and the inhibitor of AKT had the same effect with AKR1C3-KD for its regulation. We found that the OCR is increased in OE33 AKR1C3-OE than in OE33AKR1C3-VEC, while SKGT-4AKR1C3-sh1 and SKGT-4AKR1C3-sh2 showed the opposite results than SKGT-4AKR1C3-shNT. Lipid metabolism substances such as fatty acids, steroids were enriched in AKR1C3high group from public databases Conclusions: AKR1C3 regulates chemotherapy of EAC and may contribute to chemotherapy resistance of tumor cells. The possible mechanism is that the elevated AKR1C3 enhanced its own antioxidant capacity by reducing the ROS levels through AKT/GSH signaling. Tumor cells were exposure in chemotherapy drugs, an abundance of ROS will be produced in tumor cells, while the cells with higher AKR1C3 expression in the cells will show a stronger antioxidant capacity against ROS and thus be more likely to survive. So, this study provides rationale for the design of remedy that targeting AKR1C3 might enhance the anti-tumor effect and prolong survival time of EAC patients

Objective-Agnostic Enhancement of Molecule Properties via Multi-Stage VAE

Variational autoencoder (VAE) is a popular method for drug discovery and various architectures and pipelines have been proposed to improve its performance. However, VAE approaches are known to suffer from poor manifold recovery when the data lie on a low-dimensional manifold embedded in a higher dimensional ambient space [Dai and Wipf, 2019]. The consequences of it in drug discovery are somewhat under-explored. In this paper, we explore applying a multi-stage VAE approach, that can improve manifold recovery on a synthetic dataset, to the field of drug discovery. We experimentally evaluate our multi-stage VAE approach using the ChEMBL dataset and demonstrate its ability to improve the property statistics of generated molecules substantially from pre-existing methods without incorporating property predictors into the training pipeline. We further fine-tune our models on two curated and much smaller molecule datasets that target different proteins. Our experiments show an increase in the number of active molecules generated by the multi-stage VAE in comparison to their one-stage equivalent. For each of the two tasks, our baselines include methods that use learned property predictors to incorporate target metrics directly into the training objective and we discuss complications that arise with this methodology.Comment: arXiv admin note: text overlap with arXiv:2212.0275

Linking Cancer Stem Cell Plasticity to Therapeutic Resistance-Mechanism and Novel Therapeutic Strategies in Esophageal Cancer

Esophageal cancer (EC) is an aggressive form of cancer, including squamous cell carcinoma (ESCC) and adenocarcinoma (EAC) as two predominant histological subtypes. Accumulating evidence supports the existence of cancer stem cells (CSCs) able to initiate and maintain EAC or ESCC. In this review, we aim to collect the current evidence on CSCs in esophageal cancer, including the biomarkers/characterization strategies of CSCs, heterogeneity of CSCs, and the key signaling pathways (Wnt/β-catenin, Notch, Hedgehog, YAP, JAK/STAT3) in modulating CSCs during esophageal cancer progression. Exploring the molecular mechanisms of therapy resistance in EC highlights DNA damage response (DDR), metabolic reprogramming, epithelial mesenchymal transition (EMT), and the role of the crosstalk of CSCs and their niche in the tumor progression. According to these molecular findings, potential therapeutic implications of targeting esophageal CSCs may provide novel strategies for the clinical management of esophageal cancer

Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional chinese medicine

Species in the ascomycete fungal genus Cordyceps have been proposed to be the teleomorphs of Metarhizium species. The latter have been widely used as insect biocontrol agents. Cordyceps species are highly prized for use in traditional Chinese medicines, but the genes responsible for biosynthesis of bioactive components, insect pathogenicity and the control of sexuality and fruiting have not been determined. Here, we report the genome sequence of the type species Cordyceps militaris. Phylogenomic analysis suggests that different species in the Cordyceps/Metarhizium genera have evolved into insect pathogens independently of each other, and that their similar large secretomes and gene family expansions are due to convergent evolution. However, relative to other fungi, including Metarhizium spp., many protein families are reduced in C. militaris, which suggests a more restricted ecology. Consistent with its long track record of safe usage as a medicine, the Cordyceps genome does not contain genes for known human mycotoxins. We establish that C. militaris is sexually heterothallic but, very unusually, fruiting can occur without an opposite mating-type partner. Transcriptional profiling indicates that fruiting involves induction of the Zn2Cys6-type transcription factors and MAPK pathway; unlike other fungi, however, the PKA pathway is not activated.https://doi.org/10.1186/gb-2011-12-11-r11

The COP9 signalosome complex regulates fungal development and virulence in the wheat scab fungus Fusarium graminearum

The COP9 signalosome (Csn) complex is an evolutionarily conserved complex that regulates various important cellular processes. However, the function of the Csn complex in pathogenic fungi remains elusive. Here, the distribution of Csn subunits in the fungal kingdom was surveyed, and their biological functions were systematically characterized in the fungal pathogen Fusarium graminearum, which is among the top 10 plant fungal pathogens. The results obtained from bioinformatic analyses suggested that the F. graminearum Csn complex consisted of seven subunits (Csn1–Csn7) and that Csn5 was the most conserved subunit across the fungi kingdom. Yeast two-hybrid assays demonstrated that the seven Csn subunits formed a complex in F. graminearum. The Csn complex was localized to both the nucleus and cytoplasm and necessary for hyphal growth, asexual and sexual development and stress response. Transcriptome profiling revealed that the Csn complex regulated the transcription abundance of TRI genes necessary for mycotoxin deoxynivalenol (DON) biosynthesis, subsequently regulating DON production to control fungal virulence. Collectively, the roles of the Csn complex in F. graminearum were comprehensively analyzed, providing new insights into the functions of the Csn complex in fungal virulence and suggesting that the complex may be a potential target for combating fungal diseases

Metagenomic Sequencing From Mosquitoes in China Reveals a Variety of Insect and Human Viruses

We collected 8,700 mosquitoes in three sites in China, which belonged to seven species. Their viromes were tested using metagenomic sequencing and bioinformatic analysis. The abundant viral sequences were detected and annotated belonging to more than 50 viral taxonomic families. The results were verified by PCR, followed by phylogenetic analysis. In the present study, we identified partial viral genes of dengue virus (DENV), a novel circovirus (CCV), densovirus (DNV), Japanese encephalitis virus (JEV), and Wuhan mosquito virus (WMV) in mosquitoes. Metagenomic analysis and PCR amplification revealed three DENV sequences, which were as homologous to the NS3 gene of DENV from Singapore isolated in 2005, with at least 91% nucleotide (nt) identity. Seven fragments of JEV encoding structural proteins were identified belonging to genotype I. They all shared high homology with structural protein genes of JEV isolated from Laos in 2009. The production of infectious virus particles of the newly isolated virus YunnanJEV2017-4 increased after passage from the BHK-21 cell line to the Vero cell line. Novel circovirus-related genes were identified and as being related to an unnamed gene of a mosquito circovirus (MCCV) sequence from the USA isolated in 2011, with at least 41% nt identity: this distant relationship suggests that the parent virus might belong to a novel circovirus genus. Additionally, numerous known viruses and some unknown viruses were also detected in mosquitoes from Yunnan province, China, which will be tested for propagation

Metagenomic Analysis of Flaviviridae in Mosquito Viromes Isolated From Yunnan Province in China Reveals Genes From Dengue and Zika Viruses

More than 6,000 mosquitoes of six species from six sites were collected and tested for their virome using metagenomics sequencing and bioinformatic analysis. The identified viral sequences belonged to more than 50 viral families. The results were verified by PCR of selected viruses in all mosquitoes, followed by phylogenetic analysis. In the present study, we identified the partial dengue virus (DENV), Zika virus (ZIKV), and Japanese encephalitis virus (JEV) sequences in mosquitoes. Metagenomic analysis and the PCR amplification revealed three DENV sequences, one of which encodes a partial envelope protein. Two ZIKV sequences both encoding partial nonstructural protein 3 and one JEV sequence encoding the complete envelope protein were identified. There was variability in the viral titers of the newly isolated virus JEV-China/YN2016-1 of different passage viruses. The newly identified Zika virus gene from ZIKV-China/YN2016-1 was an Asian genotype and shared the highest nucleotide sequence identity (97.1%) with a ZIKV sequence from Thailand isolated in 2004. Phylogenetic analysis of ZIKV-China/YN2016-1 and ZIKV-China/YN2016-2 with known Flavivirus genes indicated that ZIKV has propagated in Yunnan province, China

6G Network AI Architecture for Everyone-Centric Customized Services

Mobile communication standards were developed for enhancing transmission and network performance by using more radio resources and improving spectrum and energy efficiency. How to effectively address diverse user requirements and guarantee everyone's Quality of Experience (QoE) remains an open problem. The Sixth Generation (6G) mobile systems will solve this problem by utilizing heterogenous network resources and pervasive intelligence to support everyone-centric customized services anywhere and anytime. In this article, we first coin the concept of Service Requirement Zone (SRZ) on the user side to characterize and visualize the integrated service requirements and preferences of specific tasks of individual users. On the system side, we further introduce the concept of User Satisfaction Ratio (USR) to evaluate the system's overall service ability of satisfying a variety of tasks with different SRZs. Then, we propose a network Artificial Intelligence (AI) architecture with integrated network resources and pervasive AI capabilities for supporting customized services with guaranteed QoEs. Finally, extensive simulations show that the proposed network AI architecture can consistently offer a higher USR performance than the cloud AI and edge AI architectures with respect to different task scheduling algorithms, random service requirements, and dynamic network conditions