Resistance Signaling Pathways Research Articles

Updates on altered signaling pathways in tumor drug resistance

Curing cancer has always presented a formidable clinical challenge. Among the various treatment strategies for combating tumors, the combination of targeted therapy and immunotherapy has recently assumed significant importance. Regrettably, while targeted drugs demonstrate efficiency in the early stages of cancer treatment, patients inevitably develop drug resistance as treatment progresses, ultimately resulting in treatment failure and death. Currently, effective countermeasures against drug-resistant tumor cells remain limited, and research into the mechanisms of drug resistance continues to garner profound interest. The current understanding of drug resistance primarily focuses on two aspects: intrinsic or primary drug resistance, and acquired or secondary drug resistance. Further explanations delve into molecular mechanisms, including acquired gene mutations, epigenetic modifications, the plasticity of cancer stem cells, and the mediation of exosomes. With the advancement of single-cell analysis, our understanding of these potential mechanisms has become more comprehensive. In this review, we initially explore classical signaling pathways related to tumorigenesis and cancer cell progression. Additionally, we summarize recent findings regarding gene mutations and modifications related to drug resistance in cancer. Finally, we discuss the plasticity of cancer stem cells and the latest research on exosome-mediated tumor drug resistance.

Regulation of Nrf2/Keap1 signaling pathway in cancer drug resistance by galectin-1: cellular and molecular implications.

Oxidative stress is characterized by the deregulation of the redox state in the cells, which plays a role in the initiation of various types of cancers. The activity of galectin-1 (Gal-1) depends on the cell redox state and the redox state of the microenvironment. Gal-1 expression has been related to many different tumor types, as it plays important roles in several processes involved in cancer progression, such as apoptosis, cell migration, adhesion, and immune response. The erythroid-2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1) signaling pathway is a crucial mechanism involved in both cell survival and cell defense against oxidative stress. In this review, we delve into the cellular and molecular roles played by Gal-1 in the context of oxidative stress onset in cancer cells, particularly focusing on its involvement in activating the Nrf2/Keap1 signaling pathway. The emerging evidence concerning the anti-apoptotic effect of Gal-1, together with its ability to sustain the activation of the Nrf2 pathway in counteracting oxidative stress, supports the role of Gal-1 in the promotion of tumor cells proliferation, immuno-suppression, and anti-tumor drug resistance, thus highlighting that the inhibition of Gal-1 emerges as a potential strategy for the restraint and regression of tumor progression. Overall, a deeper understanding of the multi-functionality and disease-specific expression profiling of Gal-1 will be crucial for the design and development of novel Gal-1 inhibitors as anticancer agents. Excitingly, although it is still understudied, the ever-growing knowledge of the sophisticated interplay between Gal-1 and Nrf2/Keap1 will enable researchers to gain valuable insights into the underlying causes of carcinogenesis and metastasis.

Exploring the therapeutic mechanisms of Yikang decoction in polycystic ovary syndrome: an integration of GEO datasets, network pharmacology, and molecular dynamics simulations.

The incidence of Polycystic Ovary Syndrome (PCOS) is increasing annually. This study aims to investigate the therapeutic mechanisms of Yikang Decoction (YKD) in the treatment of PCOS through the integration of GEO datasets, network pharmacology, and dynamic simulation. Active ingredients of YKD and their targets were collected from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform. Disease-relevant targets for PCOS were retrieved from several databases, including GeneCards, OMIM, PharmGKB, DrugBank, and GEO. The underlying pathways associated with the overlapping targets between YKD and PCOS were identified using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The mechanisms of interaction between the core targets and components were further explored through molecular docking and molecular dynamics simulations (MD). 139 potential active components and 315 targets of YKD were identified. A topological analysis of the PPI network revealed 10 core targets. These targets primarily participated in the regulation of biological processes, including cell metabolism, apoptosis, and cell proliferation. The pathways associated with treating PCOS encompassed PI3K-Akt signaling pathway, Lipid and atherosclerosis, MAPK signaling pathways, and Endocrine resistance signaling pathways. Moreover, molecular docking and MD have been shown to reveal a good binding capacity between active compounds and screening targets. This study systematically investigates the multi-target mechanisms of YKD in the treatment of PCOS, with preliminary verification provided through molecular docking and MD. The findings offer compelling evidence supporting the efficacy of YKD in treating PCOS.

Exploring the potential mechanism of Simiao Yongan decoction in the treatment of diabetic peripheral vascular disease based on network pharmacology and molecular docking technology

The study aims to investigate the potential action targets and molecular mechanisms of Simiao Yongan decoction (SMYAD) in treating diabetic peripheral vascular disease (DPVD) by utilizing network pharmacology analysis and molecular docking technology. The components and targets of SMYAD were screened using the TCMSP database, while DPVD-related genes were obtained from the GeneCards, OMIM, and Disgenet databases. After intersecting the gene sets, a Protein-Protein Interaction (PPI) network was established, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were carried out. The practical chemical components and core targets identified were molecularly docked using AutoDock software. A total of 126 active compounds were screened from which 25 main components included quercetin, rutoside, hesperidin, naringin, and β-sitosterol were determined to be the active components most associated with the core targets. A total of 224 common target genes were obtained. Among them, JUN, AKT1, MAPK3, TP53, STAT3, RELA, MAPK1, FOS, and others are the expected core targets of traditional Chinese medicine. The top-ranked GO enrichment analysis results included 727 biological processes (BP), 153 molecular functions (MF), and 102 cellular components (CC). KEGG pathway enrichment analysis involved mainly 178 signaling pathways, such as cancer signaling pathway, AGE-RAGE signaling pathway, interleukin-17 signaling pathway, tumor necrosis factor signaling pathway, endocrine resistance signaling pathway, cell aging signaling pathway, and so on. The molecular docking results demonstrate that the principal chemical components of SMYAD exhibit considerable potential for binding to the core targets. SMYAD has the potential to treat DPVD through various components, targets, and pathways. Its mechanism of action requires further experimental investigation.

Physiological and transcriptomic analyses of response of walnuts (Juglans regia) to Pantoea agglomerans infection.

Walnut blight is a serious bacterial disease that affects the yield and quality of walnuts. Pantoea agglomerans is one of the main causative agents of walnut blight. However, there have been few studies on the response of walnuts to P. agglomerans infection. In this study, the soluble sugar, photosynthesis, antioxidant enzyme activities, and secondary metabolites were measured, and the transcriptomic analysis was performed to determine the response of walnut tissue cultures to P. agglomerans infection. After pathogen inoculation, the soluble sugar content decreased, and photosynthesis was inhibited. Antioxidant enzyme (superoxide dismutase and peroxidase) activities and secondary metabolites (phenol and flavonoid) contents increased, especially in the early stages of inoculation. Transcriptomic analysis revealed that the phenylpropanoid biosynthesis pathway is induced after infection, and pathogen infection promotes ABA and ethylene signal transduction and inhibits auxin signaling. In addition, SA and JA-related gene expression was altered after inoculation with P. agglomerans, and the FLS- and calcium-mediated disease resistance signaling pathways were activated. Furthermore, our results suggested an involvement of the R-protein RPM-mediated disease resistance pathway in the response of walnuts to bacterial infections. Our findings indicated that phenylpropanoid biosynthesis, hormone signal transduction, and plant-pathogen interaction have key roles in pathogenic inoculation, which provide insights into the molecular mechanisms in the response of walnuts to P. agglomerans infection.

Seed Soaking Using Methanol Kalanchoe pinnata Leaf Extracts Induces Rice Resistance against Bacterial Leaf Blight

Bacterial leaf blight (Xanthomonas oryzae pv. oryzae) is a significant rice disease. Aqueous crude extracts of Kalanchoe pinnata were shown to induce rice resistance against the disease. This study aims at testing the disease-reducing effects of K. pinnata leaf extracts using the liquid–liquid extraction method with three different solvents (dichloromethane, methanol, and water). This serves as a basis to select appropriate extracts for effective disease control. Three concentrations (1, 1.5, and 2%) of each extract were tested using seed soaking. The extracts did not show adverse effects on seed germination and seedling growth. Methanol extracts showed significantly different effects compared to those of the untreated control. The involvement of induced resistance in the disease reduction was shown through activities of the four defense-related and antioxidant enzymes, i.e., peroxidase (POX), catalase (CAT), polyphenol oxidase (PPO), and phenylalanine ammonia lyase (PAL). Using 1% methanol extract, activities of POX and CAT involved in hydrogen peroxide production in rice tissues increased 1–4 days after pathogen inoculation (DAI) and remained at high levels until 6 DAI. Activities of PPO and PAL involved in resistance signaling pathways significantly increased after pathogen inoculation. Activities of the four enzymes generally increased after pathogen inoculation and reached higher levels with extract applications.

An update on Mycobacterium tuberculosis lipoproteins.

Almost 3% of theproteins of Mycobacterium tuberculosis (M. tuberculosis), the main causative agent of human tuberculosis, are lipoproteins. These lipoproteins are characteristic of the mycobacterial cell envelope and participate in many mechanisms involved in the pathogenesis of M. tuberculosis. In this review, the authors provide anupdatedanalysis of M. tuberculosis lipoproteins and categorize them according to their demonstrated or predicted functions, including transport of compounds to and from the cytoplasm, biosynthesis of the mycobacterial cell envelope, defense and resistance mechanisms, enzymatic activities and signaling pathways. In addition, this updated analysis revealed that at least 40% of M.tuberculosis lipoproteins are glycosylated.

Study on resistance of MdMYB gene family in apple

The growth and development of apples are susceptible to environmental stress. The MdMYB gene family is one of the largest gene families in apples. It is widely involved in various resistance signaling pathways in apples and can interact with a variety of genes. Through anthocyanin synthesis, lignin synthesis, epidermal wax synthesis, Na + / H + pump regulation, hormone interaction, etc., the resistance of plants to abiotic and biotic stresses is improved. The MdMYB gene is regulated by multiple factors. It is regulated by epigenetic factors such as methylation and miRNA before transcription. At the transcriptional level, it is regulated by transcription factors such as MdHY5, MdWRKY11 and MdWRKY41. After translation, it is regulated by ubiquitin ligase, which is involved in MdSIZ, MdMEIL and other proteins. This paper reviews the physiological responses and signaling pathways of MdMYB-mediated apple stress resistance and the gene function regulation mechanism of MdMYB, in order to provide new ideas for the study of apple stress resistance.

Bovine oviductal organoids: a multi-omics approach to capture the cellular and extracellular molecular response of the oviduct to heat stress

BackgroundThe mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series of timely and dynamic changes to suitably support early embryogenesis. Climate change-induced heat stress (HS) is one of the largest single stressors compromising reproductive function in humans and farm animals via systemic changes in the redox status of the maternal environment, adversely affecting fertilization and early embryonic development. Oviductal organoids represent a unique 3-dimensional, biomimetic model to study the physiology of the oviduct and its subsequent impact on embryo development under various environmental conditions.ResultsOur study is the first to demonstrate an innovative approach to understanding the cascade of molecular changes sustained by bovine oviductal organoids under HS and the subsequent maternal signals harnessed within their secreted extracellular vesicles (EVs). Transcriptomic analysis of oviductal organoids exposed to HS revealed 2,570 differentially expressed genes (1,222 up‐ and 1,348 downregulated), while EV-coupled miRNome analysis disclosed 18 miRNAs with significant differential expression (12 up- and 6 downregulated) in EVs from thermally stressed organoids compared to EVs released from organoids cultured under thermoneutral conditions. Genes activated in oviductal organoids in response to thermal stress, include: COX1, ACTB, CST6, TPT1, and HSPB1, while miR-1246, miR-148a, miR21-5p, miR-451, and miR-92a represent the top highly abundant EV-coupled miRNAs released in response to HS. Pathway analysis of genes enriched in organoids exposed to thermal stress showed the enrichment of endocrine resistance, cellular senescence, and notch signaling pathways. Similarly, EV-coupled miRNAs released from thermally stressed organoids showed their potential regulation of genes involved in cellular senescence, p53 signaling, and TGF-beta signaling pathways.ConclusionsIn conclusion, the cellular and extracellular response of bovine oviductal organoids to in vitro HS conditions reveal the prospective impact of environmental HS on the physiology of the oviduct and the probable subsequent impacts on oocyte fertilization and early embryo development. Future studies elucidating the potential impact of HS-associated EVs from oviductal organoids on oocyte fertilization and preimplantation embryo development, would justify the use of an organoid model to optimally understand the oviduct-embryo communication under suboptimal environments.

Identification of Potential Mechanisms of Rk1 Combination with Rg5 in the Treatment of Type II Diabetes Mellitus by Integrating Network Pharmacology and Experimental Validation.

In this study, we aimed to explore the potential targets and functional mechanisms of Rk1 combined with Rg5 (Rk1+Rg5) against type II diabetes mellitus (T2DM). Network pharmacology and molecular docking were used to predict and verify the targets and signaling pathways of Rk1+Rg5 against T2DM. The results were further confirmed by a db/db mouse model and a model using PA-induced L6 cells. According to network pharmacology, a total of 250 core targets of Rk1+Rg5 towards T2DM were identified; the insulin resistance signaling pathways were enriched by KEGG. Results of molecular docking indicated good binding affinity of Rk1 and Rg5 to Akt1. In vivo and in vitro studies further showed that Rk1+Rg5 is an inhibitor of skeletal muscle insulin resistance. The results showed that Rk1+Rg5 significantly improved the hyperglycemic state of db/db mice, alleviated dyslipidemia, and promoted skeletal muscle glucose uptake. This phenomenon was closely related to the alleviation of the insulin resistance in skeletal muscles. Finally, the combination activated the Akt signaling pathway and promoted GLUT4 translocation to the cell membrane for glucose uptake. Altogether, our findings, for the first time, demonstrate that the combination of Rk1 and Rg5 could be beneficial for anti-T2DM, possibly involving ameliorated insulin resistance.

Tianhuang formula ameliorates non-alcoholic fatty liver diseases in type 2 diabetic mice through CRLS1-ATF3/ChREBP pathway

ObjectiveTianhuang Formula (THF) is a hospital formula summarized by Professor Jiao Guo's 30 years of clinical experience. Some studies have shown that it can alleviate dyslipidemia in the body. The purpose of this study is to confirm whether THF can improve non-alcoholic fatty liver diseases (NAFLD) in type 2 diabetic mice induced by high-fat diet (HFD)/streptomycin (STZ) and to clarify its potential mechanism. MethodsAfter induction of diabetes, mice were administrated with THF (60 ​mg/kg or 120 ​mg/kg) once daily for 10 weeks. Blood glucose (FBG), glucose tolerance, and insulin resistance (IR) were assayed by oral glucose tolerance test (OGTT) and insulin tolerance test (ITT). Blood lipids, alanine transaminase (ALT), and aspartate transaminases (AST) were detected. Serum fasting insulin (INS) and adiponectin (APN) levels were measured using ELISA. Histological changes in liver and pancreatic islets were observed by H&E staining, followed by Oil Red O staining for liver lipid quantification and periodic acid-Schiff (PAS) staining to detect glycogen accumulation. Western blotting detected the levels of fatty cardiolipin synthase 1 (CRLS1), transcription factor activator 3 (ATF3), and carbohydrate-responsive element binding protein (ChREBP) in the liver. The mRNA transcripts of hepatic inflammatory factors, lipogenesis and lipolysis-related genes, and gluconeogenic enzyme-phosphoenolpyruvate carboxykinase (PEPCK), CRLS1, ATF3, and ChREBP mRNA levels were evaluated by RT-qPCR. ResultsTHF restored impaired glucose tolerance and insulin resistance, respectively. There was an improvement in HFD/STZ-induced liver and islet damage, high serum HDL-C and ANP levels, and a significant decrease in FBG, total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), FFA, INS, ALT, and AST, and lipid droplet counts in the T2DM mice treated with THF. CREBBP binding protein ATF3 mediated the insulin resistance signaling pathway, which regulated glucose and lipid metabolism in the liver. THF upregulated CRLS1, and ChREBP downregulated the expression of downstream ATF3 in the liver. RT-qPCR analysis also systemically indicated that THF suppressed the pathway and key regulators related to inflammation, lipid accumulation, and gluconeogenesis. ConclusionOur findings demonstrated that THF ameliorated lipid profile and attenuated liver steatosis in T2DM mice through CRLS1-ATF3/ChREBP pathway activation.

Wnt/β-catenin pathway is a key signaling pathway to trastuzumab resistance in gastric cancer cells

BackgroundTrastuzumab is the only approved target agent for the first-line treatment of human epidermal growth factor receptor-2 (HER-2) positive gastric cancer; however, trastuzumab resistance is a major problem in clinical practice. To comprehend the mechanism of trastuzumab resistance, we focused on the Wnt/β-catenin signaling pathway and its influence on the phenotypes and behavior of trastuzumab-resistant gastric cancer cells.MethodsTrastuzumab-resistant NCI-N87R cells were established in vitro from the human gastric cancer cell line NCI-N87 by dose-escalating repeated trastuzumab treatment. We investigated the phenotypes of NCI-N87R cells, including Wnt signaling pathway activity. Gastric cancer organoid cells were incubated with complete medium and Wnt3a-depletion medium, and their resistance to trastuzumab was compared.ResultsNCI-N87R exhibited stemness and epithelial-mesenchymal transition (EMT)-like phenotypes, along with decreased levels of the epithelial marker E-cadherin and increased levels of the mesenchymal markers Vimentin and Snail along with an increased Wnt signaling pathway activity. When gastric cancer cells were incubated in Wnt3a-conditioned medium. Wnt signaling pathway activity and resistance to trastuzumab increased. Gastric cancer patient-derived organoids incubated in Wnt3a-depletion medium were more susceptible to dose-dependent inhibition of cell viability by trastuzumab than those incubated in complete medium.ConclusionsTrastuzumab-resistant gastric cancer cells exhibited EMT-like phenotype, and trastuzumab resistance was promoted by the Wnt/β-catenin signaling pathway. The Wnt/β-catenin pathway is a key signaling pathway for trastuzumab resistance in gastric cancer cells.

AXL receptor tyrosine kinase inhibition improves the anti-tumor effects of CD8+ T cells by inducing CD103+ dendritic cell-mediated T cell priming

AXL is a member of TAM receptor family and has been highlighted as a potential target for cancer treatment. Accumulating evidence has uncovered the critical role of the AXL signaling pathway in tumor growth, metastasis, and resistance against anti-cancer drugs, as well as its association with cancer immune escape. However, the function of AXL as a manipulator of the immune system in the tumor microenvironment (TME) remains unclear. Therefore, in this study, we investigated the impact of AXL on immune cells in the TME of a syngeneic tumor model using AXL knockout (AXL−/−) mice. Compared to AXL wild-type (AXL+/+) mice, tumor growth was significantly suppressed in AXL−/− mice, and an induced population of tumor-infiltrated CD8+ T cells and CD103+ dendritic cells (DCs) was observed. The change of CD8+ T cells and CD103+ DCs was also confirmed in tumor-draining lymph nodes (TdLN). In addition, the clonal expansion of OVA-specific CD8+ T cells was dominant in AXL−/− mice. Finally, anti-PD-1 treatment evidenced synergistic anti-cancer effects in AXL−/− mice. Overall, our data indicate that AXL signaling may inhibit the clonal expansion of tumor-specific CD8+ T cells through the regulation of the migration of CD8+ T cells and DCs in TME. Thus, AXL may be a powerful molecular target to improve anti-cancer effects through single or combined therapy with immune checkpoint inhibitors (ICI).

More than trichomes and acylsugars: the role of jasmonic acid as mediator of aphid resistance in tomato

ABSTRACT This study investigates the impact of priming on the resistance of tomato plants to the potato aphid Macrosiphum euphorbiae, focusing on the role of glandular trichomes. Glandular trichomes are specific hairs that provide protection to tomato plants against herbivorous insects. The experimental priming conducted in this study revealed that prior infestation by Spodoptera littoralis caterpillars increased the plant's resistance against M. euphorbiae, pointing at the jasmonic acid (JA) signaling pathway in regulating this plant-aphid interaction. Glandular trichomes type IV were effective against aphids regardless of the previous infestation. Using JA-deficient tomato (spr2), we observed that M. euphorbiae multiplication increased, while the number of aphids on salicylic -deficient NahG plants was lower than in the wildtype Moneymaker. These findings emphasize the crucial role of the JA signaling pathway in tomato plant resistance to aphids and the importance of glandular trichomes to enhance plant defences against pests.

Current advances in nanoformulations of therapeutic agents targeting tumor microenvironment to overcome drug resistance.

The tumor microenvironment (TME) plays a pivotal role in cancer development and progression. In this line, revealing the precise mechanisms of the TME and associated signaling pathways of tumor resistance could pave the road for cancer prevention and efficient treatment. The use of nanomedicine could be a step forward in overcoming the barriers in tumor-targeted therapy. Novel delivery systems benefit from enhanced permeability and retention effect, decreasing tumor resistance, reducing tumor hypoxia, and targeting tumor-associated factors, including immune cells, endothelial cells, and fibroblasts. Emerging evidence also indicates the engagement of multiple dysregulated mediators in the TME, such as matrix metalloproteinase, vascular endothelial growth factor, cytokines/chemokines, Wnt/β-catenin, Notch, Hedgehog, and related inflammatory and apoptotic pathways. Hence, investigating novel multitargeted agents using a novel delivery system could be a promising strategy for regulating TME and drug resistance. In recent years, small molecules from natural sources have shown favorable anticancer responses by targeting TME components. Nanoformulations of natural compounds are promising therapeutic agents in simultaneously targeting multiple dysregulated factors and mediators of TME, reducing tumor resistance mechanisms, overcoming interstitial fluid pressure and pericyte coverage, and involvement of basement membrane. The novel nanoformulations employ a vascular normalization strategy, stromal/matrix normalization, and stress alleviation mechanisms to exert higher efficacy and lower side effects. Accordingly, the nanoformulations of anticancer monoclonal antibodies and conventional chemotherapeutic agents also improved their efficacy and lessened the pharmacokinetic limitations. Additionally, the coadministration of nanoformulations of natural compounds along with conventional chemotherapeutic agents, monoclonal antibodies, and nanomedicine-based radiotherapy exhibits encouraging results. This critical review evaluates the current body of knowledge in targeting TME components by nanoformulation-based delivery systems of natural small molecules, monoclonal antibodies, conventional chemotherapeutic agents, and combination therapies in both preclinical and clinical settings. Current challenges, pitfalls, limitations, and future perspectives are also discussed.

Review of Small Molecule Anticancer NF-κB Inhibitors

Cancer is one of the most common diseases in the world. Th e transcription factor NF-κB plays a key role in various physiological processes including immune response, cell proliferation, cell apoptosis and inflammation. Due to participation of NF-kappa B signaling pathways in carcinogenesis, angiogenesis, and tumor resistance to chemo- and radiotherapy, the factor is considered to be the ideal target for pharmacological treatment of cancer. Th e paper presents a literature review of the RSCI, PubMed, Scopus, Web of Science, Chemical Abstracts NCCN databases and other open access data. The carcinogenesis inhibitors were selected from all NF-κB inhibitors found. Inhibitors with antitumor activity of analytical interest include Bruton tyrosine kinase (BTK) inhibitors, cellular inhibitors of apoptosis proteins (c-IAP), proteasome inhibitors, and one inhibitor of NF-κB translocation to the nucleus. Th e authors analyzed products of diff erent development stages with recorded antitumor activity to varying degrees. NF-κB inhibitors are promising drug candidates, but since NF-κB is involved in most biological processes and the broad spectrum of action can cause side eff ects, the selective action of these compounds is to be explored.

Genome-wide identification of glutamate receptor genes in adzuki bean and the roles of these genes in light and rust fungal response

Plant glutamate receptor proteins (GLRs) are involved in plant development, biotic stress, and light-signal transduction. Vigna angularis is a traditional crop with important economic value in China, and the identification of functional genes can facilitate the breeding of stress resistant varieties. Here, we identified the members of the GLR gene family in the adzuki bean genome and investigated gene expression under light and rust fungal (Uromyces vignae) stimuli. Sixteen GLR genes were identified in V. angularis (VaGLRs), and these genes clustered in a single clade (clade III) with two groups. Evolutionary analysis showed that three VaGLRs result from tandem duplications and four result from whole genome/segmental duplications. To understand the regulation of expression of VaGLRs, cis-acting elements were analyzed in the promoter regions of the VaGLRs including cis-acting elements associated with light and stress responsiveness. Expression analysis by qRT-PCR revealed transcripts of eight and 10 VaGLRs in response to light stimuli and rust infection, respectively. For light responsiveness, the expression levels of XP_017430569.1 and XP_017425299.1 were higher under light condition than in darkness, while the expression levels of XP_017406996.1, XP_017425763.1, and XP_017423557.1 gradually recovered during dark treatment. Additionally, the relative expression levels of XP_017413816.1, XP_017436268.1, and XP_017425299.1 were significantly elevated during U. vignae infection in a resistant cultivar compared to the expression levels in a susceptible cultivar. XP_017425299.1 expression was induced both by light and rust infection, suggesting this gene may link light and disease resistance signaling pathways. Our results provide insight into how the VaGLRs contribute to adzuki bean response to light stimulus and pathogen attack. These identified VaGLRs also provide important reference to improve adzuki bean germplasm resources.

Antihyperlipidemic mechanisms of a formula containing Curcuma xanthorrhiza, Sechium edule, and Syzigium polyanthum: In silico and in vitro studies

Herbal medicines are multi-component and can exhibit synergistic effects in the treatment of diseases. Sechium edule, Syzigium polyanthum, and Curcuma xanthorrhiza have been used in traditional medicine to reduce serum lipid levels. However, the molecular mechanism was not described clearly, especially as a mixture. Thus, we performed a network pharmacology study combined with molecular docking to find a rational explanation regarding the molecular mechanisms of this antihyperlipidemic formula. According to the network pharmacology study, we predicted that this extract mixture would act as an antihyperlipidemic agent by modulating several pathways including insulin resistance, endocrine resistance, and AMP-activated protein kinase (AMPK) signaling pathway. Based on the topology parameters, we identified 6 significant targets that play an important role in reducing lipid serum levels: HMG-CoA reductase (HMGCR), peroxisome proliferator-activated receptor alpha (PPARA), RAC-alpha serine/threonine-protein kinase (AKT1), epidermal growth factor receptor (EGFR), matrix metalloproteinase-9 (MMP9), and tumor necrosis factor-alpha (TNF). Meanwhile, 8 compounds: β-sitosterol, bisdesmethoxycurcumin, cucurbitacin D, cucurbitacin E, myricetin, phloretin, quercitrin, and rutin were the compounds with a high degree, indicating that these compounds have a multitarget effect. Our consensus docking study revealed that HMGCR was the only protein targeted by all potential compounds, and rutin was the compound with the best consensus docking score for almost all targets. The in vitro study revealed that the extract combination could inhibit HMGCR with an IC50 value of 74.26 µg/mL, indicating that HMGCR inhibition is one of its antihyperlipidemic mechanisms.

Expression Analysis of Trihelix Transcription Factor Family in Strawberries and Functional Characterization of FvTrihelix6

The Trihelix is a plant-specific transcription factor family and has critical roles in plant growth and development and stress resistance. There is less information about Trihelix transcription factor genes and their potential functions in strawberries (Fragaria vesca). In the present study, we performed a detailed bioinformatics analysis of the Trihelix family in strawberries including physicochemical properties, chromosomal location, exon–intron distribution, domain arrangement, and subcellular localization. Thirty Trihelix family members were identified and divided into five subfamilies. The expression of FvTrihelix genes in different tissues/organs, i.e., root, stolon, leaf, flower, and fruit, was measured in strawberries after infection with Colletotrichum. gloeosporioides and foliar applications of salicylic acid (SA) and jasmonic acid (JA). Most of the genes showed differential expression responses following C. gloeosporioides infection and hormone treatments (SA and JA), suggesting critical roles in disease resistance and hormonal signaling pathways. As anticipated, the ectopic expression of FvTrihelix6 in Arabidopsis thaliana increased resistance against Colletotrichum. higginsianum infection. FvTrihelix6 protein was localized in the nucleus. We surmise that FvTrihelix6 enhances resistance against pathogens through the SA and JA signaling pathways. This study provides novel insights into the strawberry Trihelix transcription factor genes and new candidates for disease-resistance breeding of strawberries.

HvMPK4 phosphorylates HvWRKY1 to enhance its suppression of barley immunity to powdery mildew fungus

Mitogen-activated protein kinase (MAPK) cascades play important roles in disease resistance in model plant species. However, the functions of MAPK signaling pathways in crop disease resistance are largely unknown. Here we report the function of HvMKK1-HvMPK4-HvWRKY1 module in barley immune system. HvMPK4 is identified to play a negative role in barley immune response against Bgh, as virus-induced gene silencing of HvMPK4 results in enhanced disease resistance whilst stably overexpressing HvMPK4 leads to super-susceptibility to Bgh infection. Furthermore, the barley MAPK kinase HvMKK1 is found to specifically interact with HvMPK4, and the activated HvMKK1DD variant specifically phosphorylates HvMPK4 in vitro. Moreover, the transcription factor HvWRKY1 is identified to be a downstream target of HvMPK4 and phosphorylated by HvMPK4 in vitro in the presence of HvMKK1DD. Phosphorylation assay coupled with mutagenesis analyses identifies S122, T284, and S347 in HvWRKY1 as the major residues phosphorylated by HvMPK4. HvWRKY1 is phosphorylated in barley at the early stages of Bgh infection, which enhances its suppression on barley immunity likely due to enhanced DNA-binding and transcriptional repression activity. Our data suggest that the HvMKK1-HvMPK4 kinase pair acts upstream of HvWRKY1 to negatively regulate barley immunity against powdery mildew.