MAPK Signal Transduction Pathway Research Articles

Potential Response Patterns of Endogenous Hormones in Cliff Species Opisthopappus taihangensis and Opisthopappus longilobus under Salt Stress.

When plants are exposed to salt stress, endogenous hormones are essential for their responses through biosynthesis and signal transduction pathways. However, the roles of endogenous hormones in two cliff species (Opisthopappus taihangensis and Opisthopappus longilobus (Opisthopappus genus)) in the Taihang Mountains under salt stress have not been investigated to date. Following different time treatments under 500 mM salt concentrations, 239 differentially expressed gene (DEG)-related endogenous hormones were identified that exhibited four change trends, which in Profile 47 were upregulated in both species. The C-DEG genes of AUX, GA, JA, BR, ETH, and ABA endogenous hormones were significantly enriched in Opisthopappus taihangensis (O. taihangensis) and Opisthopappus longilobus (O. longilobus). During the responsive process, mainly AUX, GA, and JA biosynthesis and signal transduction were triggered in the two species. Subsequently, crosstalk further influenced BR, EHT, ABA, and MAPK signal transduction pathways to improve the salt resistance of the two species. Within the protein-protein interactions (PPI), seven proteins exhibited the highest interactions, which primarily involved two downregulated genes (SAUR and GA3ox) and eight upregulated genes (ACX, MFP2, JAZ, BRI1, BAK1, ETR, EIN2, and SNRK2) of the above pathways. The more upregulated expression of ZEP (in the ABA biosynthesis pathway), DELLA (in the GA signaling pathway), ABF (in the ABA signaling pathway), and ERF1 (in the ETH signaling pathway) in O. taihangensis revealed that it had a relatively higher salt resistance than O. longilobus. This revealed that the responsive patterns to salt stress between the two species had both similarities and differences. The results of this investigation shed light on the potential adaptive mechanisms of O. taihangensis and O. longilobus under cliff environments, while laying a foundation for the study of other cliff species in the Taihang Mountains.

CLL Cell-Derived Exosomes Impair Immune Function and Normal Hematopoiesis in CLL

CLL is characterized by immune and hematopoietic abnormalities such as T-cell dysfunction, hypogammaglobulinemia, increased levels of inflammatory cytokines, aberrant monocyte polarity, anemia, neutropenia, and low-grade bone marrow fibrosis. As a result, CLL patients become susceptible to increased rates of infection and/or second neoplasms. Exosomes are nanovesicles that participate in bidirectional communication between neoplastic cells and their microenvironment. Using electron microscopy, we found that CLL cells produce and release CD19 +/CD5 +exosomes, and in agreement with other investigators, we detected high levels of CLL cell-derived exosomes in the plasma of patients with CLL. By using bead-assisted flow cytometry, we found that at least 50% of circulating extracellular vesicles are CLL cell-derived exosomes, identified as CD63 +/CD41a - particles co-expressing CD19 +/CD5 +, whereas only a small proportion of CLL patients' plasma-derived exosomes expressed T-cell or endothelial cell surface markers. Furthermore, we found a positive correlation between levels of CLL cell-derived exosomes in the plasma of patients with CLL and the patients' peripheral white blood cell counts, indicating that CLL cells are the primary source of circulating exosomes. To investigate the effects of CLL cell-derived exosomes, we conducted an analysis of CLL cells and CLL cell-derived exosomes from a cohort of 78 previously untreated patients. To identify the molecular profiles of CLL cell-derived exosomes, we analyzed their RNA transcripts using RNA-seq and qRT-PCR. Our data revealed that CLL cell-derived exosomes expressed significant levels of oncogenic micro-RNAs. Additionally, we identified 1052 differentially expressed genes, and enrichment analysis further unveiled the significant upregulation of gene clusters associated with p53, NF-κB, MAPK, mTOR, and interleukin signal transduction pathways, consistent with their neoplastic cell origin. The distinctive transcriptional profile of CLL cell-derived exosomes led us to explore their effects on normal cell populations. We labeled ultra-centrifuged CLL cell-derived exosomes with fluorescent markers and incubated them with various cell types in culture conditions conducive to their physiological activity. Using confocal microscopy and flow cytometry, we observed significant uptake of CLL cell-derived exosomes by normal monocytes, fibrocytes, B cells, T cells, and CLL cells. Furthermore, incubation with CLL exosomes resulted in notable alterations in normal CD14 + monocyte polarity and subset-specific shifts in the expression of inflammatory cytokines. Remarkably, CLL cell-derived exosomes induced overexpression of the T-cell makers CD5 and CD8 in normal CD19 + cells, suggesting that CLL exosomes may promote immune-evasive reprogramming of normal B cells. Moreover, when normal T cells were incubated with CLL cell-derived exosomes, CyTOF analysis using 46 markers revealed elevated levels of immune checkpoints, including PD1, CTLA4, and LAG3, in both CD4 + and CD8 + subsets, indicating that exosomes derived from CLL cells induce T-cell dysfunction. Given their profound effect on immune cells, we wondered whether CLL exosomes affect hematopoietic cells. We previously found that normal monocyte derived fibrocytes stimulate hematopoietic colony proliferation. In sharp contrast to control fibrocytes, fibrocytes preincubated with CLL exosomes and CLL cell-derived exosomes inhibited normal BFU-E and CFU-GM colony formation, suggesting that CLL exosomes might contribute to CLL patients' anemia or neutropenia. Wondering whether CLL cell-derived exosomes affect the viability of other cell types, we found that they induced a dose-dependent apoptosis of CD19 + B cells, CLL cells but not normal T cells. In summary, CLL cell-derived exosomes exert a profound effect on gene and cell surface protein expression in immune and tissue-remodeling cells, leading to functional modifications, alteration of inflammatory cytokine levels, and suppression of normal hematopoiesis. The impact of CLL exosomes may vary across different cell types; however, as the leukemic cell mass increases, exosome levels escalate, invariably affecting CLL cell environment. The identification of exosomes with distinct molecular signatures holds promising potential for novel therapeutic interventions in future treatment of CLL.

6-benzylaminopurine causes endothelial dysfunctions to human umbilical vein endothelial cells and exacerbates atorvastatin-induced cerebral hemorrhage in zebrafish.

6-benzylaminopurine (6-BA), a multifunctional plant growth regulator, which is frequently used worldwide to improve qualities of various crops, is an important ingredient in production of "toxic bean sprouts." Although there is no direct evidence of adverse effects, its hazardous effects, as well as joint toxicity with other chemicals, have received particular attention and aroused furious debate between proponents and environmental regulators. By use of human umbilical vein endothelial cells (HUVECs), adverse effects of 6-BA to human-derived cells were first demonstrated in this study. A total of 25-50 mg 6-BA/L inhibited proliferation, migration, and formation of tubular-like structures by 50% in vitro. Results of Western blot analyses revealed that exposure to 6-BA differentially modulated the MAPK signal transduction pathway in HUVECs. Specifically, 6-BA decreased phosphorylation of MEK and ERK, but increased phosphorylation of JNK and P38. In addition, 6-BA exacerbated atorvastatin-induced cerebral hemorrhage via increasing hemorrhagic occurrence by 60% and areas by 4 times in zebrafish larvae. In summary, 6-BA elicited toxicity to the endothelial system of HUVECs and zebrafish. This was due, at least in part, to discoordination of MAPK signaling pathway, which should pose potential risks to the cerebral vascular system.

Dead heart of pith-decayed Scutellariae Radix: a study based on multi-omics

Dead heart is an important trait of pith-decayed Scutellariae Radix. The purpose of this study was to clarify the scientific connotation of the dead heart using multi-omics. Metabolomics and transcriptomics combined with multivariate statistical analysis such as principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were used to systematically compare the differences in chemical composition and gene expression among phloem, outer xylem and near-dead xylem of pith-decayed Scutella-riae Radix. The results revealed significant differences in the contents of flavonoid glycosides and aglycones among the three parts. Compared with phloem and outer xylem, near-dead xylem had markedly lowered content of flavonoid glycosides(including baicalin, norwogonin-7-O-β-D-glucuronide, oroxylin A-7-O-β-D-glucuronide, and wogonoside) while markedly increased content of aglycones(including 3,5,7,2',6'-pentahydroxy dihydroflavone, baicalin, wogonin, and oroxylin A). The differentially expressed genes were mainly concentrated in KEGG pathways such as phenylpropanoid metabolism, flavonoid biosynthesis, ABC transporter, and plant MAPK signal transduction pathway. This study systematically elucidated the material basis of the dead heart of pith-decayed Scutellariae Radix with multiple growing years. Specifically, the content of flavonoid aglycones was significantly increased in the near-dead xylem, and the gene expression of metabolic pathways such as flavonoid glycoside hydrolysis, interxylary cork development and programmed apoptosis was significantly up-regulated. This study provided a theoretical basis for guiding the high-quality production of pith-decayed Scutellariae Radix.

Crystal structure of Leptospira LSS_01692 reveals a dimeric structure and induces inflammatory responses through Toll-like receptor 2-dependent NF-κB and MAPK signal transduction pathways.

Leptospirosis is a commonly overlooked zoonotic disease that occurs in tropical and subtropical regions. Recent studies have divided the Leptospira spp. into three groups based on virulence, including pathogenic, intermediate, and saprophytic species. Pathogenic species express a protein family with leucine-rich repeat (LRR) domains, which are less expressed or absent in nonpathogenic species, highlighting the importance of this protein family in leptospirosis. However, the role of LRR domain proteins in the pathogenesis of leptospirosis is still unknown and requires further investigation. In this study, the 3D structure of LSS_01692 (rLRR38) was obtained using X-ray crystallography at a resolution of 3.2 Å. The results showed that rLRR38 forms a typical horseshoe structure with 11 α-helices and 11 β-sheets and an antiparallel dimeric structure. The interactions of rLRR38 with extracellular matrix and cell surface receptors were evaluated using ELISA and single-molecule atomic force microscopy. The results showed that rLRR38 interacted with fibronectin, collagen IV, and Toll-like receptor 2 (TLR2). Incubating HK2 cells with rLRR38 induced two downstream inflammation responses (IL-6 and MCP-1) in the TLR2 signal transduction pathway. The TLR2-TLR1 complex showed the most significant upregulation effects under rLRR38 treatment. Inhibitors also significantly inhibited nuclear factor κB and mitogen-activated protein kinases signals transduction under rLRR38 stimulation. In conclusion, rLRR38 was determined to be a novel LRR domain protein in 3D structure and demonstrated as a TLR2-binding protein that induces inflammatory responses. These structural and functional studies provide a deeper understanding of the pathogenesis of leptospirosis.

Flow cytometry-based cell cycle synchronization and transcriptome analysis in cotton (Gossypium arboretum L.)

Cell cycle plays an important role in plant growth and development. Synchronization of cell cycle will help us to elucidate the molecular mechanisms regulating cell cycle and further understand the cell growth and development for various purposes, including crop improvement. However, there are few reports on cell cycle synchronization and its application on transcriptome analysis. In this study, we first developed a cell cycle synchronization technology using flow cytometry technology; then, we deeply sequenced the synchronized cells for studying the gene expression profiling during cell cycle, using cotton as an example, one of the most important fiber and oil crops. Through GO and KEGG enrichment analysis, we obtained some important genes related to cell cycle, including genes associated with hormone and MAPK signal transduction pathways. Our results also showed that tubulin family plays an important role in the cell cycle. This study revealed the differences of gene expression in different cell cycles in cotton for the first time. This study lays a foundation for in-depth understanding of the molecular mechanism and interaction network during cell cycle.

Langerhans Cell Histiocytosis in Children and Adults: Similarities and Differences

Langerhans cell histiocytosis is an uncommon hematological condition affecting children and adults. It is distinguished by neoplastic infiltration of myeloid dendritic cells and an inflammatory response. Any organ can become dysfunctional as a result of tissue destruction and fibrosis. In almost all cases, activating mutations in the MAPK signal transduction pathway have been found. The earlier these mutations appear during myeloid differentiation, the more severe the disease. The skeleton, skin, and pituitary gland are most typically afflicted in children, but the liver, spleen, and hematopoietic system may also be involved, associated with a worse outcome. In adults, manifestations are most commonly observed in the skeleton, lungs, skin, and in the pituitary region. The course of the disease might range from spontaneous remission to persistent relapse. A histological examination is required to get a definitive diagnosis. In youngsters, where therapeutic surgery and radiation are mostly obsolete, medical therapy is the treatment of choice. Chemotherapy is used when special risk lesions are involved or when children have multifocal skeletal involvement and multisystem disease. In adults, however, local treatments are sufficient in unifocal lesions, and supportive therapy with bisphosphonate is suitable for multifocal bone involvement. Systemic therapy is administered only in patients with multisystem disease and treatment-refractory or quickly reoccurring disease activity. Aside from cytostatics, targeted inhibitors of gene mutations are now accessible for this purpose. While most patients survive, chronic recurrent episodes and late sequelae are the main issues.

Transcriptome analysis of gene expression profiles reveals wood formation mechanisms in Chinese fir at different stand ages

Forests are crucial sustainable sources of natural ecosystems and contribute to human welfare. Cunninghamia lanceolata (Chinese fir) is an economically important conifer and occupies the largest area in China that produces global wood resources. Although Chinese fir has high economic value in China, little information is known regarding its mechanisms of wood formation. Therefore, transcriptome analysis was conducted to study the gene expression patterns and associated timber formation mechanisms in Chinese fir at different stand ages. In the present study, a total of 837,156 unigenes were identified in 84 samples from Chinese fir (pith and root) at different stand ages via RNA-Seq. Among them, most of the differentially expressed genes (DEGs) were significantly enrichment in plant hormone signal transduction, flavonoid metabolism pathway, starch and sucrose metabolism, and MAPK signal transduction pathway, which might be associated with the diameter formation in Chinese fir. The DEGs in these pathways were analyzed in Chinese fir and were related to lignin synthesis, cell wall formation and cell wall reinforcement/thickening. These genes might play an important role in regulating timber formation/growth in Chinese fir. In addition, certain transcriptome factors (TFs) related to Chinese fir timber formation were identified, including WRKY33, WRKY22, PYR/PYL, and MYC2. Weighted co-expression network analysis (WGCNA) showed that glucan endo-1,3-beta-d-glucosidase was a hub gene significantly correlated with the growth-related genes in Chinese fir. Sixteen key genes that related to diameter regulation in Chinese fir were verified by qRT-PCR analysis. These key genes might have a fine regulatory role in timber formation in Chinese fir. Our results pave the way for research on the regulatory mechanisms of wood formation, and provide an insight for improving the quality production of Chinese fir.

Picolinafen exposure induces ROS accumulation and calcium depletion, leading to apoptosis in porcine embryonic trophectoderm and uterine luminal epithelial cells during the peri-implantation period

The global use of herbicides accounts for more than 48% of total pesticide usage. Picolinafen is a pyridine carboxylic acid herbicide that is predominantly used to control broadleaf weeds in wheat, barley, corn, and soybeans. Despite its widespread use in agriculture, its toxicity in mammals has rarely been studied. In this study, we first identified the cytotoxic effects of picolinafen on porcine trophectoderm (pTr) and luminal epithelial (pLE) cells, which are involved in the implantation process during early pregnancy. Picolinafen treatment significantly decreased the viability of pTr and pLE cells. Our results demonstrate that picolinafen increased the number of sub-G1 phase cells and early/late apoptosis. In addition, picolinafen disrupted mitochondrial function and resulted in the accumulation of intracellular ROS, leading to a reduction in calcium levels in both the mitochondria and cytoplasm of pTr and pLE cells. Moreover, picolinafen was found to significantly inhibit the migration of pTr. These responses were accompanied by the activation of the MAPK and PI3K signal transduction pathways by picolinafen. Our data suggest that the deleterious effects of picolinafen on the viability and migration of pTr and pLE cells might impair their implantation potential.

Transcriptome analysis reveals key genes involved in the resistance to Cryphonectria parasitica during early disease development in Chinese chestnut

BackgroundChestnut blight, one of the most serious branch diseases in Castanea caused by Cryphonectria parasitica, which has ravaged across American chestnut and most of European chestnut since the early twentieth century. Interestingly, the Chinese chestnut is strongly resistant to chestnut blight, shedding light on restoring the ecological status of Castanea plants severely affected by chestnut blight. To better explore the early defense of Chinese chestnut elicited in response to C. parasitica, the early stage of infection process of C. parasitica was observed and RNA sequencing-based transcriptomic profiling of responses of the chestnut blight-resistant wild resource ‘HBY-1’ at 0, 3 and 9 h after C. parasitica inoculation was performed.ResultsFirst, we found that 9 h was a critical period for Chinese chestnut infected by C. parasitica, which was the basis of further study on transcriptional activation of Chinese chestnut in response to chestnut blight in the early stage. In the transcriptome analysis, a total of 283 differentially expressed genes were identified between T9 h and Mock9 h, and these DEGs were mainly divided into two clusters, one of which was metabolism-related pathways including biosynthesis of secondary metabolites, phenylpropanoid biosynthesis, amino sugar and nucleotide sugar metabolism, and photosynthesis; the other was related to plant-pathogen interaction and MAPK signal transduction. Meanwhile, the two clusters of pathways could be connected through junction among phosphatidylinositol signaling system, phytohormone signaling pathway and α-Linolenic acid metabolism pathway. It is worth noting that genes associated with JA biosynthesis and metabolic pathway were significantly up-regulated, revealing that the entire JA metabolic pathway was activated in Chinese chestnut at the early stage of chestnut blight infection.ConclusionWe identified the important infection nodes of C. parasitica and observed the morphological changes of Chinese chestnut wounds at the early stage of infection. In response to chestnut blight, the plant hormone and MAPK signal transduction pathways, plant-pathogen interaction pathways and metabolism-related pathways were activated at the early stage. JA biosynthesis and metabolic pathway may be particularly involved in the Chinese chestnut resistance to chestnut blight. These results contributes to verifying the key genes involved in the resistance of Chinese chestnut to C. parasitica.

Determination of the synergistic anti-influenza effect of Huangqin Su tablet and Oseltamivir and investigation of mechanism of the tablet based on gut microbiota and network pharmacology

Huangqin Su (HQS) tablet is mainly composed of baicalein which has been evaluated for its ability to inhibit influenza. The present study aimed to investigate the effect of HQS and oseltamivir phosphate (OS) (single or combination therapy) on influenza-induced acute pneumonia in male and female ICR mice. The regulatory effect of HQS on gut microbiota was also studied by using 16 s rDNA sequencing, and the targets and mechanisms of HQS against influenza were comprehensively analyzed by network pharmacology. Pharmacodynamic results, including lung index and pathological changes, showed that HQS exhibited significant anti-influenza efficacy and could improve the efficacy of low-dose OS (P < 0.05 and P < 0.01, respectively). The results of 16 s rDNA sequencing revealed that HQS modulated the gut microbiota and remarkably enriched the abundance of Lactobacillus. The findings of network pharmacology research suggested that the anti-influenza mechanism of HQS was related to TLRs, MAPK, and other signal transduction pathways. Taken together, this study identified the possibility of the combined use of HQS and OS and demonstrated the role of HQS in modulating the gut microbiota of mice against influenza. Network pharmacology studies also suggested that the anti-influenza effect of HQS was related to TLRs, MAPK, TNF, and other signaling pathways.

Transcriptome analysis reveals the molecular basis of the response to acute hypoxic stress in blood clam Scapharca broughtonii

Hypoxia tolerance and adaptive regulation are important for aquatic animals, especially for species with poor mobility, such as most bivalves. Previous studies have confirmed that the blood clam Scapharca broughtonii has strong hypoxia resistance. However, the molecular mechanism supporting its hypoxic tolerance is still largely limited. To further screen the genes and their potential regulation of hypoxia tolerance, the transcriptome changes of S. broughtonii after acute hypoxic stress were explored by RNA sequencing. In this study, the average value of Q30 is 92.89%, indicating that the quality of sequencing is relatively high. The Unigenes obtained were annotated using four databases, namely Interpo, KEGG, Swisspro and TrEMBL. The annotation rates in these four databases were 71.82%, 75.95%, 92.98%, and 79.26%, respectively. And also, there were 649 DEGs in group B (dissolved oxygen (DO) of 2.5 mg/L) compared with group D (DO of 7.5 mg/L), among which 252 were up-regulated, and 397 were down-regulated. There were 965 DEGs in group A (DO of 0.5 mg/L), 2.5 mg/L, and 7.5 mg/L, compared with group B, among which 530 were up-regulated, and 435 were down-regulated. Meanwhile, there were 2,040 DEGs in group A compared with group D, among which 901 were up-regulated, and 1,139 were down-regulated. The main metabolic-related pathways of KEGG enriched in this study included Insulin secretion, Insulin signaling pathway, MAPK signal transduction pathway, and PPAR signaling pathway. These pathways may be critical metabolic pathways to solve energy demand and rebuild energy balance in S. broughtonii under hypoxic conditions. This study preliminarily clarified the response of S. broughtonii to hypoxia stress on the molecular levels, providing a reference for the following study on the response laws of related genes and pathways under environmental stress of S. broughtonii.

Systematic Pharmacology-Based Strategy to Explore the Mechanism of Bufei Huoxue Capsule in the Treatment of Chronic Obstructive Pulmonary Disease.

To explore the effects and mechanisms of Bufei Huoxue Capsule (BHC) on chronic obstructive pulmonary disease (COPD) based on network pharmacology. The effective components and related targets of BHC were collected by searching TCMSP, HERB, and ETCM databases, after which the related targets of COPD were obtained on GeneCards and OMIM databases. The common targets were imported into the STRING database and Cytoscape database to construct a target interaction network and screen core targets. Next, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed on the Metascape platform. According to the prediction results of network pharmacology, the action mechanism was further examined in an animal model of COPD. The pathological changes of lung tissue were observed by HE staining; goblet cells and mucus secretion in lung tissue were observed by AB-PAS staining, airway collagen deposition was observed by Masson staining, and the expression of NE, TGF-β1, P-EGFR/EGFR, P-ERK1/2/ERK1/2, P-JNK/JNK, and P-P38/P38MAPK protein was detected by Western blot analysis. A total of 379 targets related to BHC and 7391 targets related to COPD were obtained, including 313 potential targets of BHC in treating chronic obstructive pulmonary disease, with JUN, AKT1, TNF, IL6, EGFR, MAPK1, and MAPK14 as the core targets. Through enrichment analysis, BHC may interfere with COPD by regulating the MAPK signal pathway, HIF-1 signal pathway, NF-κB signal pathway, cAMP signal pathway, cGMP-PKG signal pathway, and so on. Animal experiments showed that the BHC could reduce airway inflammatory cell infiltration, inhibit airway epithelial goblet cell proliferation, reduce mucus secretion, and improve small airway collagen fiber deposition in COPD model rats. Besides, BHC could downregulate the protein expression of NE, TGF-β1, P-EGFR, P-ERK1/2, and P-P38MAPK. BHC can reduce airway inflammation, inhibit mucus hypersecretion, and improve airway remodeling by regulating the MAPK signal transduction pathway.

De novo transcriptome assembly and comprehensive assessment provide insight into fruiting body formation of Sparassis latifolia

The genes associated with fruiting body formation of Sparasis latifolia are valuable for improving mushroom breeding. To investigate this process, 4.8 × 108 RNA-Seq reads were acquired from three stages: hyphal knot (SM), primordium (SP), and primordium differentiation (SPD). The de novo assembly generated a total of 48,549 unigenes, of which 71.53% (34,728) unigenes could be annotated by at least one of the KEGG (Kyoto Encyclopedia of Genes and Genomes), GO (Gene Ontology), and KOG (Eukaryotic Orthologous Group) databases. KEGG and KOG analyses respectively mapped 32,765 unigenes to 202 pathways and 19,408 unigenes to 25 categories. KEGG pathway enrichment analysis of DEGs (differentially expressed genes) indicated primordium initiation was significantly related to 66 pathways, such as “Ribosome”, “metabolism of xenobiotics by cytochrome P450”, and “glutathione metabolism” (among others). The MAPK and mTOR signal transduction pathways underwent significant adjustments during the SM to SP transition. Further, our research revealed the PI3K-Akt signaling pathway related to cell proliferation could play crucial functions during the development of SP and SPD. These findings provide crucial candidate genes and pathways related to primordium differentiation and development in S. latifolia, and advances our knowledge about mushroom morphogenesis.

Abstract 2643: Design and discovery of novel oral INV-88 macrophage migration inhibitory factor (MIF) inhibitors: Potent anti-tumor activity in vitro and in vivo

Abstract Background: MIF is a pleiotropic cytokine critically implicated in tumorigenesis. Tumor cell- and immune effector cell-derived MIF are localized to exosomal, extracellular, cytoplasmic and intranuclear compartments. MIF activates MAPK and PI3K signal transduction, inhibits apoptosis, promotes angiogenesis, elicits premetastatic niche formation and resolves DNA replication stress; contributing to unperturbed cancer cell growth and metastasis. MIF overexpression in several cancers is a negative prognostic indicator associated with poor overall patient survival. Methods &amp; Results: Active novel INV-88 MIF inhibitors were discovered using structure-based drug design against critical MIF pharmacophoric binding sites guided by high-resolution X-ray crystallography (Brookhaven) of INV-88 compound:hu-MIF protein bound complexes. Following an iterative medicinal chemistry campaign; stringent biochemical, pharmacological and ADMET funnel screening identified promising INV-88 lead candidates with potent antitumor activities. Strong MIF binding by INV-88 compounds was demonstrated using thermal shift and fluorescence polarization assays. Profound in vitro antitumor pharmacological properties of INV-88 were established on several murine &amp; human tumor cell lines (HCT116, MeWo, DU145, 4T1, B16-F10, CT26). INV-88 inhibitors demonstrated potent anti-proliferative effects down to nM IC50 potencies and impeded 3D spheroid growth &amp; viability. Disruption of MIF:CD74/CD44-mediated MAPK and PI3K signal transduction pathways by INV-88 compounds blocked ERK and AKT phosphorylation leading to decreased pERK1/2 and pAKT levels. INV-88 inhibitors increased Caspase-3/7, p53 and Bax but decreased Bcl-2 levels. INV-88 lead compounds inhibited tumor MIF protein production in 2D &amp; 3D cell culture systems with concomitant pan-inhibition of tumorigenic proinflammatory cytokine (TNF-α, IL-1β, IL-6, and IL-8) expression by stromal cells. Optimal ADMET properties of INV-88 compounds enabled oral dosing of select INV-88 clinical candidate compounds in mouse syngeneic B16-F10 melanoma and CT26 colon carcinoma models. Mice were dosed daily with INV-88 or vehicle per oral gavage starting in BALB/c and in C57/BL6 mice upon demonstrating palpable tumors after s.c. implantation of CT26 and B16-F10 tumor cells, respectively. Tumor growth inhibition was observed with rates of 71% on Day 24 (CT26) and 75% on Day 20 (B16-F10). INV-88 treatment significantly inhibited CT26 [p&amp;lt;0.0001] and B16-F10 tumor growth [p&amp;lt;0.0001] versus controls. INV-88 was well-tolerated by tumor-bearing mice which exhibited normal body condition scores and body weight. Conclusion: The data presented highlight the potential therapeutic utility of pharmacological MIF inhibition with oral INV-88 new chemical entities as a novel immunotherapeutic treatment modality for cancer. Citation Format: Anderson Gaweco, Vincent Chung, Jefferson Tilley, William Windsor, Terry Stouch, John Ellingboe, Gregg Crichlow, Philipp Sabanas, Christian Sebesta. Design and discovery of novel oral INV-88 macrophage migration inhibitory factor (MIF) inhibitors: Potent anti-tumor activity in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2643.

A photo-thermal nanocomposite capable of relieving inflammatory response to compete multidrug-resistant pseudomonas aeruginosa infection

Lung infection associated with drug-resistant P. aeruginosa and their wide spread have become a serious problem to public health. Effective antibacterial strategy without antibiotics must be developed for treatment of these infections. To combat pneumonia infected by multidrug-resistant P. aeruginosa (MRPA), a novel antibacterial nanocomoposite of AuNRs@PBArPEGMA was prepared. AuNRs@PBArPEGMA could bind with P. aeruginosa through the interaction betweeen PBArPEGMA and bacteria proved by alizarin red assay. The nanocomposite could relieve inflammtory response in MRPA-infected macrophages by downregulating MAPK and NFκB signal-transduction pathways. AuNRs@PBArPEGMA showed well photo-thermal conversion capacity leading to the temperature increasing from 20 to 80 °C under near-infrared (NIR) irradiation for 5 min. The photo-thermal capacity endowed the nanocomposites efficient antibacterial activity against MRPA. Simultaneously, AuNRs@PBArPEGMA with NIR irradiation could kill biofilm bacteria while traditional antibiotics were non-effetive. AuNRs@PBArPEGMA with NIR irradiation could treat MRPA-related lung infection. The nanocomposite with NIR reduced the inflammatory cytokines of TNF-α, IL-1β, and IL-6 by downregulating the MAPK signal pathway in bronchoalveolar lavage fluids. The number of P. aeruginosa and expression of NF-κB p65 in lung also decreased. Above all, our study provides a non-antibiotic therapeutic strategy in clinical application and demonstrated that AuNRs@PBArPEGMA with NIR irradiation showed a potential treatment for drug-resistant P. aeruginosa infection in Lung.

Physiological and Molecular Responses of Zoysia japonica to Rust Infection.

Zoysiagrass (Zoysia japonica) is a popular turfgrass species and is widely used for sport turf and urban landscape. Zoysiagrass is often infected by Puccinia zoysiae, which causes a loss in turf quality. The physiological and molecular mechanisms of rust resistance are poorly understood in this species. In this study, the rust-resistant and susceptible lines of zoysiagrass were inoculated with P. zoysiae, and alterations of leaf cell structure, physiological indicators and transcriptomic response were investigated at the various stages of inoculation. After inoculation, the cell membranes, nucleus, mitochondria, and chloroplast were all impaired, followed by abnormal physiological metabolism. The damage occurred earlier and more severely in the susceptible line. Changes in electrolyte leakage and chlorophyll content varied with the genotype and the inoculation stages. The transcriptome analysis showed that plant hormones, MAPK signal transduction pathway, photosynthesis and energy generation pathways were significantly enriched in the early response, in both the resistant and susceptible lines. The results provided insights into the physiological and molecular mechanisms of rust disease resistance and would benefit the breeding of rust-resistant varieties in zoysiagrass and related turfgrass species.

Hyperactivation of MAPK Induces Tamoxifen Resistance in SPRED2-Deficient ERα-Positive Breast Cancer.

Simple SummaryTamoxifen has been used for more than 40 years to treat breast tumors that are dependent on the hormone estrogen for their growth. However, resistance and recurrence of the tumors during the course of the treatment are common. Understanding the mechanisms that drive tamoxifen resistance and discovering new biomarkers for early detection are keys for designing appropriate personalized therapies. Here, we show that low levels of SPRED2 may be useful as a novel biomarker of tamoxifen resistance. We found that SPRED2 deficiency causes a hyperactivation of the mitogen-activated protein kinases (MAPKs) ERK1/ERK2, which in turn enhances estrogen signaling and diminishes the toxic effects of tamoxifen on breast cancer cells. Treatment with the ERK1/2 inhibitor, ulixertinib, could restore their sensitivity to tamoxifen. Therefore, we propose that patients with estrogen-dependent breast cancer characterized by low expression levels of SPRED2 may be candidates for a combination therapy with tamoxifen and ulixertinib.Breast cancer is the number one cause of cancer-related mortality in women worldwide. Most breast tumors depend on the expression of the estrogen receptor α (ERα) for their growth. For this reason, targeting ERα with antagonists such as tamoxifen is the therapy of choice for most patients. Although initially responsive to tamoxifen, about 40% of the patients will develop resistance and ultimately a recurrence of the disease. Thus, finding new biomarkers and therapeutic approaches to treatment-resistant tumors is of high significance. SPRED2, an inhibitor of the MAPK signal transduction pathway, has been found to be downregulated in various cancers. In the present study, we found that SPRED2 is downregulated in a large proportion of breast-cancer patients. Moreover, the knockdown of SPRED2 significantly increases cell proliferation and leads to tamoxifen resistance of breast-cancer cells that are initially tamoxifen-sensitive. We found that resistance occurs through increased activation of the MAPKs ERK1/ERK2, which enhances the transcriptional activity of ERα. Treatment of SPRED2-deficient breast cancer cells with a combination of the ERK 1/2 inhibitor ulixertinib and 4-hydroxytamoxifen (4-OHT) can inhibit cell growth and proliferation and overcome the induced tamoxifen resistance. Taken together, these results indicate that SPRED2 may also be a tumor suppressor for breast cancer and that it is a key regulator of cellular sensitivity to 4-OHT.

Front Cover: VF‐4 and DR‐8 Derived from Salted Egg White Inhibit Inflammatory Activity via NF‐κB/PI3K‐Akt/MAPK Signal Transduction Pathways in HT‐29 Cells Induced by TNF‐α

Mol. Nutr. Food Res. 2022, 66, 202100682 DOI: 10.1002/mnfr.202100682 The picture includes the duck egg, the molecular structure of the two peptides, and the enlarged view of the human colon. It represents that the two peptides VF-4 and DR-8 isolated from salted egg white play an anti-inflammatory role in human colon cells. This is reported by Yan Zhao, Yonggang Tu, and co-workers in article number 2100682.

Machine learning assisted analysis of breast cancer gene expression profiles reveals novel potential prognostic biomarkers for triple-negative breast cancer

Tumor heterogeneity and the unclear metastasis mechanisms are the leading cause for the unavailability of effective targeted therapy for Triple-negative breast cancer (TNBC), a breast cancer (BrCa) subtype characterized by high mortality and high frequency of distant metastasis cases. The identification of prognostic biomarker can improve prognosis and personalized treatment regimes. Herein, we collected gene expression datasets representing TNBC and Non-TNBC BrCa. From the complete dataset, a subset reflecting solely known cancer driver genes was also constructed. Recursive Feature Elimination (RFE) was employed to identify top 20, 25, 30, 35, 40, 45, and 50 gene signatures that differentiate TNBC from the other BrCa subtypes. Five machine learning algorithms were employed on these selected features and on the basis of model performance evaluation, it was found that for the complete and driver dataset, XGBoost performs the best for a subset of 25 and 20 genes, respectively. Out of these 45 genes from the two datasets, 34 genes were found to be differentially regulated. The Kaplan-Meier (KM) analysis for Distant Metastasis Free Survival (DMFS) of these 34 differentially regulated genes revealed four genes, out of which two are novel that could be potential prognostic genes (POU2AF1 and S100B). Finally, interactome and pathway enrichment analyses were carried out to investigate the functional role of the identified potential prognostic genes in TNBC. These genes are associated with MAPK, PI3-AkT, Wnt, TGF-β, and other signal transduction pathways, pivotal in metastasis cascade. These gene signatures can provide novel molecular-level insights into metastasis.