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The dysregulation of the phosphatidylinositol-3-kinase (PI3K) pathway can lead to uncontrolled cellular growth and tumorigenesis. Targeting PI3K and its downstream substrates has been shown to be effective in preclinical studies and phase III trials with the approval of several PI3K pathway inhibitors by the Food and Drug Administration (FDA) over the past decade. However, the limited clinical efficacy of these inhibitors, intolerable toxicities, and acquired resistances limit the clinical application of PI3K inhibitors. This review discusses the PI3K signaling pathway, alterations in the PI3K pathway causing carcinogenesis, current and novel PI3K pathway inhibitors, adverse effects, resistance mechanisms, challenging issues, and future directions of PI3K pathway inhibitors. [ABSTRACT FROM AUTHOR]

Previous studies have suggested a role of phosphatidylinositol‐3‐kinase gamma (PI3Kγ) in bone remodeling, but the mechanism remains undefined. Here, we explored the contribution of PI3Kγ in the resorption of maxillary bone and dental roots using models of orthodontic tooth movement (OTM), orthodontic‐induced inflammatory root resorption, and rapid maxillary expansion (RME). PI3Kγ‐deficient mice (PI3Kγ−/−), mice with loss of PI3Kγ kinase activity (PI3KγKD/KD) and C57BL/6 mice treated with a PI3Kγ inhibitor (AS605240) and respective controls were used. The maxillary bones of PI3Kγ−/−, PI3KγKD/KD, and C57BL/6 mice treated with AS605240 showed an improvement of bone quality compared to their controls, resulting in reduction of the OTM and RME in all experimental groups. PI3Kγ−/− mice exhibited increased root volume and decreased odontoclasts counts. Consistently, the pharmacological blockade or genetic deletion of PI3K resulted in increased numbers of osteoblasts and reduction in osteoclasts during OTM. There was an augmented expression of Runt‐related transcription factor 2 (Runx2) and alkaline phosphatase (Alp), a reduction of interleukin‐6 (Il‐6), as well as a lack of responsiveness of receptor activator of nuclear factor kappa‐Β (Rank) in PI3Kγ−/− and PI3KγKD/KD mice compared to control mice. The maxillary bones of PI3Kγ−/− animals showed reduced p‐Akt expression. In vitro, bone marrow cells treated with AS605240 and cells from PI3Kγ−/− mice exhibited significant augment of osteoblast mineralization and less osteoclast differentiation. The PI3Kγ/Akt axis is pivotal for bone remodeling by providing negative and positive signals for the differentiation of osteoclasts and osteoblasts, respectively. [ABSTRACT FROM AUTHOR]

Background and objective: Autophagy is the fundamental cell survival machinery that enables cells to respond to metabolic stress by recycling and degrading intracellular components to generate energy and macromolecular precursors. Bone marrowderived mesenchymal stem cells (MSCs), possess the unique ability of self-renewal and development into specialized cells, and long-lived cells with specific metabolic needs. It has been demonstrated that autophagy is essential for MSC differentiation, quiescence, activation, and self-renewal. Thus, the hypothesis that autophagy influences bone marrow-derived MSC post-novodrin-prompted liver dysfunction was investigated in the present study. Methods: Hepatic dysfunction was conducted in a rat model using novodrin (100 mg/kg, subcutaneously), which was divided into two doses for two alternative days, followed by the treatment with 100 μL of intravenous injection of allogeneic MSCs (5 × 106). Results: A month preceding MSC therapy, a marked decline in liver function biomarkers, including alanine aminotransferase and aspartate aminotransferase, was observed, in addition to the significant decrease in oxidative stress biomarker, lipid peroxide. Meanwhile, novodrin significantly elevated the gene expression of cell survival biomarkers, including signal transducer and activator of transcription, phosphatidylinositol-3-kinase, and serine/threonine kinase-1, in addition to the concomitant increase in oncogenic biomarker, phosphatase and tensin homolog, and this was reversed post-MSC implantation. Furthermore, the autophagy biomarkers, including Beclin-1 and X-box binding protein 1, were restored post-MSC implantation. Moreover, the MSCs were labeled using the PKH26 red fluorescent dye, and were traced using a fluorescence microscope for the implant of MSCs on the damaged liver tissue. These were confirmed through histopathological examination. Conclusion: The present study hypothesized that autophagy is essential for stem cell activation, and the differentiation in novodrin-induced liver dysfunction, through the improvement of MSCs, targeting inflamed cells, and self-renewal. [ABSTRACT FROM AUTHOR]

Previous studies have shown that fibroblast growth factor 13 is downregulated in the brain of both Alzheimer’s disease mouse models and patients, and that it plays a vital role in the learning and memory. However, the underlying mechanisms of fibroblast growth factor 13 in Alzheimer’s disease remain unclear. In this study, we established rat models of Alzheimer’s disease by stereotaxic injection of amyloid-β (Aβ1–42)-induced into bilateral hippocampus. We also injected lentivirus containing fibroblast growth factor 13 into bilateral hippocampus to overexpress fibroblast growth factor 13. The expression of fibroblast growth factor 13 was downregulated in the brain of the Alzheimer’s disease model rats. After overexpression of fibroblast growth factor 13, learning and memory abilities of the Alzheimer’s disease model rats were remarkably improved. Fibroblast growth factor 13 overexpression increased brain expression levels of oxidative stress-related markers glutathione, superoxide dismutase, phosphatidylinositol-3-kinase, AKT and glycogen synthase kinase 3β, and anti-apoptotic factor BCL. Furthermore, fibroblast growth factor 13 overexpression decreased the number of apoptotic cells, expression of pro-apoptotic factor BAX, cleaved-caspase 3 and amyloid-β expression, and levels of tau phosphorylation, malondialdehyde, reactive oxygen species and acetylcholinesterase in the brain of Alzheimer’s disease model rats. The changes were reversed by the phosphatidylinositol-3-kinase inhibitor LY294002. These findings suggest that overexpression of fibroblast growth factor 13 improved neuronal damage in a rat model of Alzheimer’s disease through activation of the phosphatidylinositol-3-kinase/AKT/glycogen synthase kinase 3β signaling pathway.

Epilepsy is one of the most common neurological diseases, which is caused by abnormal brain activity. A wide variety of studies have shown the importance of the phosphatidylinositol-3-kinase (PI3K) signaling pathway in epilepsy pathogenesis. Duvelisib (DUV) is a selective inhibitor of PI3K. The present study investigated the anticonvulsant potential of DUV in a rat model of pentylenetetrazole (PTZ)-induced convulsions. Male Wistar rats (200-250 g, 8 weeks old) were injected intraperitoneally (IP) with DUV at different doses of 5 and 10 mg/kg, or vehicle 30 minutes prior to PTZ (70 mg/kg, IP) treatment. Based on Racine's scale, behavioral seizures were assessed. The results showed that pretreatment with DUV prolonged the seizure stages according to the Racine scale, significantly decreased the duration of general tonic-clonic seizure and reduced the number of myoclonic jerks (P <.05). In conclusion, we found that PI3K antagonist DUV significantly reduced PTZ-induced seizures, indicating that DUV exerts an anticonvulsant effect by inhibiting PI3K signaling pathway. [ABSTRACT FROM AUTHOR]

Targeted inhibitors of bromodomain and extraterminal (BET)-bromodomains and phosphatidylinositol-3-kinase (PI3K) signaling demonstrate potent but self-limited antilymphoma activity as single agents in the context of cellular Myelocytomatosis (cMYC) oncogene-dysregulation. However, combined PI3K and BET inhibition imparts synergistic anticancer activity with the potential for more sustained disease responses due to the mutual antagonism of compensatory epigenetic and signaling networks. Here, we describe the mechanistic and therapeutic validation of rationally designed dual PI3K/BET bromodomain inhibitors, built by linkage of established PI3K and BET inhibitor pharmacophores. The lead candidate demonstrates high selectivity, nanomolar range cellular potency, and compelling in vivo efficacy, including curative responses in the aggressive Eµ-Myc lymphoma model. These studies further support the therapeutic strategy of combined PI3K and BET inhibition and provide a potential step-change in approach to orthogonal MYC antagonism using optimized chimeric small-molecule technology [ABSTRACT FROM AUTHOR]

Epilepsy results from abnormal brain function and is one of the most common neurological diseases in the world. The phosphatidylinositol-3-kinase (PI3K) signaling pathway has been extensively studied in epilepsy pathogenesis. Idelalisib (IDE) is a selective inhibitor of PI3K. In the present study, IDE was evalu-ated for its potential to reduce convulsions caused by pentylenetetrazole (PTZ). Male Wistar rats (200–250 g, 8 weeks old) were injected intraperitoneally (IP) with IDE at different doses of 40 and 80 mg/kg, or vehicle 30 min prior to PTZ (70 mg/kg, IP) treatment. Racine's scale was used to measure behavioral seizures. The results showed that pretreatment with IDE decreased the seizure stages according to the Racine scale, significantly prolonged the duration of general tonic-clonic seizure (GTCS) and reduced the number of myoclonic jerks (p < 0.05). In conclusion, we found that the PI3K antagonist IDE had a reducing effect on PTZ-induced seizures, suggesting that inhibition of the PI3K signaling pathway by IDE exerts an anticonvulsant effect. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression and clinical significance of phosphatidylinoinosidine-3-kinase (PI3K) and matrix metalloproteinase 9 (MMP-9) in bronchoscopic alveolar lavage fluid of children with severe pneumonia in pediatric intensive care unit (PICU). Methods A total of 103 children with severe pneumonia admitted to PICU of the hospital from January 2021 to June 2022 were enrolled. All patients underwent fiberbronchoscopic alveolar lavage. Real-time fluorescence quantitative polymerase chain reaction was used to determine the expression of PI3K, and enzyme-linked immunosorption was used to determine the expression of MMP-9 in alveolar lavage fluid. The patients were followed up for 21 days after admission to evaluate the prognosis. Logistic regression analysis was used to analyze the factors affecting the prognosis. Pearson correlation was used to analyze the correlation between the expression levels of PI3K and MMP-9 in the bubble lavage solution and the child Critical Case Score (PCIS) and child Organ Dysfunction 2 (PELOD-2) Scores. The values of PI3K and MMP-9 expression levels in alveolar lavage fluid to predict the prognosis were analyzed by receiver operating curve (ROC). Results Among the 103 children, 29 cases had poor prognosis vs 74 cases had good prognosis. Logistic multivariate regression analysis showed that combined shock (OR = 4.104, 95%CI: 1.689 ~ 9.974), PELOD-2 score (OR = 4.764, 95%CI: 1.960 ~ 11.577), PCIS score (OR = 5.068, 95%CI: 2.085 ~ 12.318), PI3K (OR = 3.924, 95%CI: 1.614 ~ 9.536) and MMP-9 (OR = 3.873, 95%CI: 1.594 ~ 9.412) were the prognostic factors of PICU children with severe pneumonia (P < 0.05). Pearson correlation analysis showed that the expression levels of PI3K and MMP-9 in alveolar lavage fluid were negatively correlated with PCIS scores (r = -0.598, P = 0.000; r = -0.537, P = 0.000), PI3K and MMP-9 expression levels in alveolar lavage fluid were positively correlated with PELOD-2 scores (r = 0.546, P = 0.000; r = 0.571, P = 0.000). ROC curve results showed that the sensitivity of PI3K, MMP-9 and their combination to predict the prognosis of PICU children with severe pneumonia was 75.86% (95%CI: 56.08% ~ 88.98%), 72.41% (95%CI: 52.51% ~ 86.55%) and 82.76% (95%CI: 63.51% ~ 93.47%), the specificity was 75.68% (95%CI: 64.07% ~ 84.58%), 79.73% (95%CI: 68.47% ~ 87.85%), 89.19% (95%CI: 79.28% ~ 94.88%), and the AUC was 0.762 (95%CI: 0.666 ~ 0.859), 0.767 (95%CI: 0.670 ~ 0.873), 0.909 (95%CI: 0.848 ~ 0.971). Conclusion The expression of PI3K and MMP-9 in bronchoscopic alveolar lavage fluid was related to the prognosis of children with severe pneumonia in PICU, and the combination in predicting the prognosis was effective, so it has certain clinical value to use them as a sensitive indicator to predict the prognosis of children with severe pneumonia in PICU. [ABSTRACT FROM AUTHOR]

Phosphatidylinositol‐3‐kinases (PI3Ks) are lipid kinases that produce 3‐phosphorylated derivatives of phosphatidylinositol upon activation by various cues. These 3‐phosphorylated lipids bind to various protein effectors to control many cellular functions. Lipid phosphatases such as phosphatase and tensin homolog (PTEN) terminate PI3K‐derived signals and are critical to ensure appropriate signaling outcomes. Many lines of evidence indicate that PI3Ks and PTEN, as well as some specific lipid effectors are highly compartmentalized, either in plasma membrane nanodomains or in endosomal compartments. We examine the evidence for specific recruitment of PI3Ks, PTEN, and other related enzymes to membrane nanodomains and endocytic compartments. We then examine the hypothesis that scaffolding of the sources (PI3Ks), terminators (PTEN), and effectors of these lipid signals with a common plasma membrane nanodomain may achieve highly localized lipid signaling and ensure selective activation of specific effectors. This highlights the importance of spatial regulation of PI3K signaling in various physiological and disease contexts. [ABSTRACT FROM AUTHOR]

Chondrocyte degeneration and extracellular matrix component loss are the primary causes of osteoarthritis (OA). OA can be treated by inhibiting chondrocyte degeneration and increasing extracellular matrix component secretion. Osteopontin (OPN), a multifunctional protein, has gained immense attention with regard to its involvement in OA. This study aimed to explore the therapeutic value and mechanism of action of OPN in OA treatment. Results of the histomorphological analysis revealed a worn-off OA cartilage tissue surface, cartilage matrix layer deterioration, and calcium salt deposition. Compared to that in normal chondrocytes, in OA chondrocytes, the OPN, CD44, and PI3K protein and mRNA expression was upregulated. Further, siOPN, rhOPN, and rhOPN plus LS-C179404 interfered with OA chondrocytes. As verified in mice, OPN directly inhibited the expression level of PI3K in OA chondrocytes by binding with CD44. Morphological analysis of the knee joints demonstrated that OPN effectively inhibited OA progression via the OPN/CD44/PI3K signal axis. In conclusion, OPN activates intracellular PI3K signaling molecules by binding to CD44 on the cell surface to cause downstream cascading effects, thereby delaying chondrocyte degeneration and reducing cartilage matrix component loss; therefore, OPN is a potential therapeutic agent for OA.

Objective: To explore the effect of curcumin on the insulin receptor substrate 1 (IRS1)/phosphatidylinositol-3-kinase (PI3K)/endometrial expression of glucose 4 (GLUT4) signalling pathway and its regulator, phosphatase and tensin homolog (PTEN), in a rat model of polycystic ovarian syndrome (PCOS). Methods: PCOS model was induced by letrozole intragastric administration. Sprague-Dawley rats were randomized into 4 groups according to a random number table: (1) control group; (2) PCOS group, which was subjected to PCOS and received vehicle; (3) curcumin group, which was subjected to PCOS and treated with curcumin (200 mg/kg for 2 weeks); and (4) curcumin+LY294002 group, which was subjected to PCOS, and treated with curcumin and LY294002 (a specific PI3K inhibitor). Serum hormone levels (17 β-estradiol, follicle stimulating hormone, luteinizing hormone, progesterone, and testosterone) were measured by enzyme linked immunosorbent assay, and insulin resistance (IR) was assessed using the homeostasis model assessment of IR. Ovarian tissues were stained with haematoxylin and eosin for pathological and apoptosis examination. Expression levels of key transcriptional regulators and downstream targets, including IRS1, PI3K, protein kinase B (AKT), GLUT4, and PTEN, were measured via reverse transcription polymerase chain reaction and Western blot, respectively. Results: The PCOS group showed impaired ovarian morphology and function. Compared with the PCOS group, curcumin treatment exerted ovarioprotective effects, down-regulated serum testosterone, restored IR, inhibited inflammatory cell infiltration in ovarian tissues, decreased IRS1, PI3K, and AKT expressions, and up-regulated GLUT4 and PTEN expressions in PCOS rats (P<0.05 or P<0.01). In contrast, IRS1, PI3K, AKT, and PTEN expression levels were not significantly different between PCOS and curcumin+LY294002 groups (P>0.05). Conclusion: The beneficial effects of curcumin on PCOS rats included the alteration of serum hormone levels and recovery of morphological ovarian lesions, in which, PTEN, a new target, may play a role in regulating the IRS1/PI3K/GLUT4 pathway. [ABSTRACT FROM AUTHOR]

Papaverine (PPV), a benzylisoquinoline alkaloid, extracted from the Papaverine somniferum plant, is currently in clinical use as a vasodilator. Research has shown that PPV inhibits phosphodiesterase 10A (PDE10A,) resulting in the accumulation of cyclic adenosine 3′, 5′-monophosphate (cAMP) that affects multiple downstream pathways, including phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt), a mammalian target of rapamycin (mTOR) and vascular endothelial growth factor (VEGF). The accumulation of cAMP can further affect mitochondrial metabolism through the activation of protein kinase A (PKA), which activates the mitochondrial complex I. Literature has shown that PPV exerts anti-proliferative affects in several tumorigenic cell lines including adenocarcinoma alveolar cancer (A549) and human hepatoma (HepG-2) cell lines. Cell cycle investigations have shown varying results with the effects dependent on concentration and cell type with data suggesting an increase in cells occupying the sub-G1 phase, which is indicative of cell death. These results suggest that PPV may be a beneficial compound to explore for the use in anticancer studies. More insight into the effects of the compound on cellular and molecular mechanisms is needed. Understanding the effects PPV may exert on tumorigenic cells may better researchers' understanding of phytomedicines and the effects of PPV and PPV-derived compounds in cancer. [ABSTRACT FROM AUTHOR]

ObjectiveTo investigate the effects of metformin (Met) on the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) pathway and cognitive dysfunction in Alzheimer's disease (AD) rats.MethodsFifty Sprague Dawley rats were selected and divided into the following groups: normal, AD model (AD group), Met low (50 mg/kg), Met medium (100 mg/kg) and Met high (200 mg/kg) according to the random number table method. Each group included 10 rats. With the exception of the normal group, AD models were established by injecting 10 μL streptozotocin (STZ, 3 mg/kg) into the bilateral ventricles. After the model was successfully established, Met at the corresponding dose was administered by gavage. The normal and AD groups were administered the same amount of normal saline by gavage. The medicine was administered once a dayfor 14 days. After the last administration, the Morris water maze test was used to detect the spatial cognitive function of rats. The rats were sacrificed, and the hippocampal tissues were collected. The contents of β-amyloid protein 42 (Aβ42) and phosphorylated tau protein (p-tau) in hippocampus were detected by enzyme-linked immunosorbent assay (ELISA). The pathological changes of neurons in hippocampus were observed by HE staining. The detection of PI3K positive expression was performed by immunohistochemistry. Western blots were used to detect the relative expression levels of PI3K, phosphorylated Akt (p-Akt), insulin receptor substrate-1 (IRS-1), and glycogen synthase kinase-3 (GSK-3) in hippocampus.ResultsCompared with the normal group, there were pathological damages, such as disordered arrangement and decrease of neurons in hippocampus of rats in the AD group. The number of times the platform was crossed, the ratio of swimming distance to the total distance in the original platform quadrant, the ratio of swimming time to the total time, and the protein expressions of PI3K, p-Akt and IRS-1 in hippocampus of the AD group were significantly low (P < 0.05). The average escape latency, the contents of Aβ42 and p-tau, and the protein expression of GSK-3 in hippocampus were high (P < 0.05). Compared with the AD group, the pathological damages of hippocampal tissue in low, medium, and high dose Met rats were alleviated. The number of times the platform was crossed, the ratio of swimming distance to the total distance in the original platform quadrant, the ratio of swimming time to the total time, and the protein expressions of PI3K, p-Akt and IRS-1 in hippocampus of the AD group were significantly high (P < 0.05). The average escape latency, the contents of Aβ42 and p-tau, and the protein expression of GSK-3 in hippocampus were low (P < 0.05). Moreover, the above indexes were dose-dependent in the Met groups.ConclusionMet can activate the PI3K/Akt pathway, decrease the levels of p-tau and Aβ42 in hippocampus, and improve cognitive impairment in AD rats.

Simple Summary: Treatment of glioblastoma is hampered by the activation of compensatory survival mechanisms by malignant cells that lead to drug resistance. Moreover, the blood–brain barrier (BBB) precludes the brain entrance of most drugs. We hypothesized that computer-assisted drug discovery tools would reveal novel multi-targeting drug candidates with BBB-permeant and favorable ADMET properties. We aimed to discover molecules with predicted ability to inhibit the EGFR/PI3Kp110β pathway and to validate their efficacy and safety in biological assays. We used quantitative structure–activity relationship models and structure-based virtual screening, and assessed ADMET properties, to identify BBB-permeant drug candidates. Moreover, we tested their anti-tumor efficacy and BBB safety and permeation in cell models. We found two EGFR, two PI3Kp110β, and, mostly, two dual inhibitors with anti-tumor effects. Among them, one EGFR and two PI3Kp110β inhibitors were able to cross the BBB endothelium without compromising it. These studies revealed novel drug candidates for glioblastoma treatment. The epidermal growth factor receptor (EGFR) is upregulated in glioblastoma, becoming an attractive therapeutic target. However, activation of compensatory pathways generates inputs to downstream PI3Kp110β signaling, leading to anti-EGFR therapeutic resistance. Moreover, the blood–brain barrier (BBB) limits drugs' brain penetration. We aimed to discover EGFR/PI3Kp110β pathway inhibitors for a multi-targeting approach, with favorable ADMET and BBB-permeant properties. We used quantitative structure–activity relationship models and structure-based virtual screening, and assessed ADMET properties, to identify BBB-permeant drug candidates. Predictions were validated in in vitro models of the human BBB and BBB-glioma co-cultures. The results disclosed 27 molecules (18 EGFR, 6 PI3Kp110β, and 3 dual inhibitors) for biological validation, performed in two glioblastoma cell lines (U87MG and U87MG overexpressing EGFR). Six molecules (two EGFR, two PI3Kp110β, and two dual inhibitors) decreased cell viability by 40–99%, with the greatest effect observed for the dual inhibitors. The glioma cytotoxicity was confirmed by analysis of targets' downregulation and increased apoptosis (15–85%). Safety to BBB endothelial cells was confirmed for three of those molecules (one EGFR and two PI3Kp110β inhibitors). These molecules crossed the endothelial monolayer in the BBB in vitro model and in the BBB-glioblastoma co-culture system. These results revealed novel drug candidates for glioblastoma treatment. [ABSTRACT FROM AUTHOR]

ABSTRACT Eosinophilia is associated with various persisting inflammatory diseases and often coincides with chronic fungal infections or fungal allergy as in the case of allergic bronchopulmonary aspergillosis (ABPA). Here, we show that intranasal administration of live Aspergillus fumigatus conidia causes fatal lung damage in eosinophilic interleukin-5 (IL-5)-transgenic mice. To further investigate the activation of eosinophils by A. fumigatus, we established a coculture system of mouse bone marrow-derived eosinophils (BMDE) with different A. fumigatus morphotypes and analyzed the secretion of cytokines, chemokines, and eicosanoids. A. fumigatus-stimulated BMDE upregulated expression of CD11b and downregulated CD62L and CCR3. They further secreted several proinflammatory mediators, including IL-4, IL-13, IL-18, macrophage inflammatory protein-1α (MIP-1α)/CC chemokine ligand 3 (CCL3), MIP-1β/CCL4, and thromboxane. This effect required direct interaction and adherence between eosinophils and A. fumigatus, as A. fumigatus culture supernatants or A. fumigatus mutant strains with impaired adhesion elicited a rather poor eosinophil response. Unexpectedly, canonical Toll-like receptor (TLR) or C-type-lectin receptor (CLR) signaling was largely dispensable, as the absence of MYD88, TRIF, or caspase recruitment domain-containing protein 9 (CARD9) resulted in only minor alterations. However, transcriptome analysis indicated a role for the PI3K-AKT-mTOR pathway in A. fumigatus-induced eosinophil activation. Correspondingly, we could show that phosphatidylinositol 3-kinase (PI3K) inhibitors successfully prevent A. fumigatus-induced eosinophil activation. The PI3K pathway in eosinophils may therefore serve as a potential drug target to interfere with undesired eosinophil activation in fungus-elicited eosinophilic disorders. IMPORTANCE Allergic bronchopulmonary aspergillosis (ABPA) is caused by the fungus Aspergillus fumigatus, afflicts about five million patients globally, and is still a noncurable disease. ABPA is associated with pronounced lung eosinophilia. Activated eosinophils enhance the inflammatory response not only by degranulation of toxic proteins but also by secretion of small effector molecules. Receptors and signaling pathways involved in activation of eosinophils by A. fumigatus are currently unknown. Here, we show that A. fumigatus-elicited activation of eosinophils requires direct cell-cell contact and results in modulation of cell surface markers and rapid secretion of cytokines, chemokines, and lipid mediators. Unexpectedly, this activation occurred independently of canonical Toll-like receptor or C-type lectin receptor signaling. However, transcriptome analysis indicated a role for the PI3K-AKT-mTOR pathway, and PI3K inhibitors successfully prevented A. fumigatus-induced eosinophil activation. The PI3K pathway may therefore serve as a potential drug target to interfere with undesired eosinophil activation in fungus-elicited eosinophilic disorders.

Introduction: Experimental and clinical studies have shown that exercise training is one of the most effective strategies for reducing the progression of cardiomyopathy and decreasing the incidence of cardiovascular complications and mortality due to myocardial ischemia. This study aimed to investigate the role of high-intensity interval training (HIIT) in the expression of Hand2 and Phosphatidylinositol-3-kinase (PI3K) genes in cardiac ischemia rats. Materials & Methods: In this study, 28 male Wistar rats (200-250 g) were randomly divided into four groups of sham, ischemia, exercise, and exercise-ischemia. Myocardial infarction was performed by the closure of the left anterior descending artery for 30 min. In total, 40-min HIIT (each interval consisted of 4-min very high intensity running with approximately 85% to 90% maximum rate of oxygen consumption [VO2max] followed by 2-min active recovery with approximately 50% to 60% VO2max) were performed three days a week for eight weeks. Ethics code: IR.SSRI.REC.1396.134 Findings: The results showed that the expression level of the Hand2 gene in the ischemia-training group was significantly increased, compared to the sham (P=0.001) and ischemia (P=0.001) groups. Furthermore, the concentration of PI3K in the exercise and exercise-ischemia groups was significantly increased, compared to the sham (P=0.001) and ischemia (P=0.001) groups. However, there was a decrease in the ischemia group, compared to the other groups (P

Background Genetic alterations in ARID1A were detected at a high frequency in cholangiocarcinoma (CCA). Growing evidence indicates that the loss of ARID1A expression leads to activation of the PI3K/AKT pathway and increasing sensitivity of ARID1A-deficient cells for treatment with the PI3K/AKT inhibitor. Therefore, we investigated the association between genetic alterations of ARID1A and the PI3K/AKT pathway and evaluated the effect of AKT inhibition on ARID1A-deficient CCA cells. Methods Alterations of ARID1A, PI3K/AKT pathway-related genes, clinicopathological data and overall survival of 795 CCA patients were retrieved from cBio Cancer Genomics Portal (cBioPortal) databases. The association between genetic alterations and clinical data were analyzed. The effect of the AKT inhibitor (MK-2206) on ARID1A-deficient CCA cell lines and stable ARID1A-knockdown cell lines was investigated. Cell viability, apoptosis, and expression of AKT signaling were analyzed using an MTT assay, flow cytometry, and Western blots, respectively. Results The analysis of a total of 795 CCA samples revealed that ARID1A alterations significantly co-occurred with mutations of EPHA2 (p < 0.001), PIK3CA (p = 0.047), and LAMA1 (p = 0.024). Among the EPHA2 mutant CCA tumors, 82% of EPHA2 mutant tumors co-occurred with ARID1A truncating mutations. CCA tumors with ARID1A and EPHA2 mutations correlated with better survival compared to tumors with ARID1A mutations alone. We detected that 30% of patients with PIK3CA driver missense mutations harbored ARID1A-truncated mutations and 60% of LAMA1-mutated CCA co-occurred with truncating mutations of ARID1A. Interestingly, ARID1A-deficient CCA cell lines and ARID1A-knockdown CCA cells led to increased sensitivity to treatment with MK-2206 compared to the control. Treatment with MK-2206 induced apoptosis in ARID1A-knockdown KKU-213A and HUCCT1 cell lines and decreased the expression of pAKTS473 and mTOR. Conclusion These findings suggest a dependency of ARID1A-deficient CCA tumors with the activation of the PI3K/AKT-pathway, and that they may be more vulnerable to selective AKT pathway inhibitors which can be used therapeutically.

Simple Summary: Failure of chemotherapy is common during the treatment of colon cancer, and there is a compelling need to develop alternative therapeutic approaches against this common malignancy. Metformin, which is an oral hypoglycaemic agent used for treating diabetes mellitus, and vitamin D have shown promising anticancer activities, and both agents boosted the actions of chemotherapy against colon cancer. This study, therefore, measured the potential beneficial effects of adding metformin and/or active vitamin D to the main cytotoxic drug used for treating colon cancer. The results demonstrate that metformin had superior anticancer effects relative to active vitamin D and ameliorated the effects of chemotherapy in animals and in cells. To the best of our knowledge, this study is also the first to report that triple treatment with the drugs of interest showed the best inhibition of cancer progression, which could provide a better therapeutic strategy against colon cancer. Chemoresistance to 5-fluorouracil (5-FU) is common during colorectal cancer (CRC) treatment. This study measured the chemotherapeutic effects of 5-FU, active vitamin D3 (VD3), and/or metformin single/dual/triple regimens as complementary/alternative therapies. Ninety male mice were divided into: negative and positive (PC) controls, and 5-FU, VD3, Met, 5-FU/VD3, 5-FU/Met, VD3/Met, and 5-FU/VD3/Met groups. Treatments lasted four weeks following CRC induction by azoxymethane. Similar regimens were also applied in the SW480 and SW620 CRC cell lines. The PC mice had abundant tumours, markedly elevated proliferation markers (survivin/CCND1) and PI3K/Akt/mTOR, and reduced p21/PTEN/cytochrome C/caspase-3 and apoptosis. All therapies reduced tumour numbers, with 5-FU/VD3/Met being the most efficacious regimen. All protocols decreased cell proliferation markers, inhibited PI3K/Akt/mTOR molecules, and increased proapoptotic molecules with an apoptosis index, and 5-FU/VD3/Met revealed the strongest effects. In vitro, all therapies equally induced G1 phase arrest in SW480 cells, whereas metformin-alone showed maximal SW620 cell numbers in the G0/G1 phase. 5-FU/Met co-therapy also showed the highest apoptotic SW480 cell numbers (13%), whilst 5-FU/VD3/Met disclosed the lowest viable SW620 cell percentages (81%). Moreover, 5-FU/VD3/Met revealed maximal inhibitions of cell cycle inducers (CCND1/CCND3), cell survival (BCL2), and the PI3K/Akt/mTOR molecules alongside the highest expression of cell cycle inhibitors (p21/p27), proapoptotic markers (BAX/cytochrome C/caspase-3), and PTEN in both cell lines. In conclusion, metformin monotherapy was superior to VD3, whereas the 5-FU/Met protocol showed better anticancer effects relative to the other dual therapies. However, the 5-FU/VD3/Met approach displayed the best in vivo and in vitro tumoricidal effects related to cell cycle arrest and apoptosis, justifiably by enhanced modulations of the PI3K/PTEN/Akt/mTOR pathway. [ABSTRACT FROM AUTHOR]

Background. Genetic alterations in ARID1A were detected at a high frequency in cholangiocarcinoma (CCA). Growing evidence indicates that the loss of ARID1A expression leads to activation of the PI3K/AKT pathway and increasing sensitivity of ARID1A-deficient cells for treatment with the PI3K/AKT inhibitor. Therefore, we investigated the association between genetic alterations of ARID1A and the PI3K/AKT pathway and evaluated the effect of AKT inhibition on ARID1A-deficient CCA cells. Methods. Alterations of ARID1A, PI3K/AKT pathway-related genes, clinicopathological data and overall survival of 795 CCA patients were retrieved from cBio Cancer Genomics Portal (cBioPortal) databases. The association between genetic alterations and clinical data were analyzed. The effect of theAKTinhibitor (MK-2206) on ARID1Adeficient CCA cell lines and stable ARID1A-knockdown cell lines was investigated. Cell viability, apoptosis, and expression of AKT signaling were analyzed using anMTTassay, flow cytometry, and Western blots, respectively. Results. The analysis of a total of 795 CCA samples revealed that ARID1A alterations significantly co-occurred with mutations of EPHA2 (p < 0.001), PIK3CA (p D 0.047), and LAMA1 (p D 0.024). Among the EPHA2 mutant CCA tumors, 82% of EPHA2 mutant tumors co-occurred with ARID1A truncating mutations. CCA tumors with ARID1A and EPHA2 mutations correlated with better survival compared to tumors with ARID1A mutations alone. We detected that 30% of patients with PIK3CA driver missense mutations harbored ARID1A-truncated mutations and 60% of LAMA1-mutated CCA co-occurred with truncating mutations of ARID1A. Interestingly, ARID1A-deficient CCA cell lines and ARID1A-knockdown CCA cells led to increased sensitivity to treatment with MK-2206 compared to the control. Treatment with MK- 2206 induced apoptosis in ARID1A-knockdown KKU-213A and HUCCT1 cell lines and decreased the expression of pAKTS473 and mTOR. Conclusion. These findings suggest a dependency of ARID1A-deficient CCA tumors with the activation of the PI3K/AKT-pathway, and that they may be more vulnerable to selective AKT pathway inhibitors which can be used therapeutically. Subjects Bioinformatics, Cell Biology, Genomics, Molecular Biology, Gastroenterology and Hepatology [ABSTRACT FROM AUTHOR]

To determine the clinical benefit of monotherapy with PI3K/AKT/mTOR inhibitors in patients diagnosed with advanced or recurrent ovarian cancer and to investigate the predictive value of current PI3K/AKT/mTOR biomarkers on therapy response. A systematic search was conducted in PubMed, Embase and the Cochrane Library for articles reporting on treatment with PI3K/AKT/mTOR inhibitors in ovarian cancer. The primary endpoint was defined as the clinical benefit rate (CBR), including the proportion of patients with complete (CR) and partial response (PR) and stable disease (SD). Secondary endpoints included the overall response rate (ORR, including CR and PR) and drug-related grade 3 and 4 adverse events. We included 233 patients from 19 studies and observed a pooled CBR of 32% (95% CI 20–44%) and ORR of 3% (95% CI 0–6%) in advanced or recurrent ovarian cancer patients treated with PI3K/AKT/mTOR inhibitors. Subgroup analysis tended to favor the studies who selected patients based on current PI3K/AKT/mTOR biomarker criteria (e.g. genomic alterations or loss of PTEN protein expression), but the difference in CBR was not statistically significant from studies with unselected populations (respectively, CBR of 42% (95% CI 23–62%) and 27% (95% CI 14–42%), P = 0.217). To better reflect true patient benefit, we excluded SD <6 months as a beneficial outcome which resulted in a pooled CBR of 7% (95% CI 2–13%). The overall proportion of patients with drug-related grade 3 and 4 adverse events was 36%. The efficacy of monotherapy with PI3K/AKT/mTOR inhibitors in advanced recurrent ovarian cancer patients is limited to a small subgroup and selection of patients with the use of current biomarkers did not improved the CBR significantly. Given the toxicity profile, we suggest that current treatment with PI3K/AKT/mTOR inhibitors should not be initiated unless in clinical trials. Furthermore, improved biomarkers to measure functional PI3K/AKT/mTOR pathway activity are needed to optimize patient selection. • Monotherapy with PI3K/AKT/mTOR inhibitors resulted in a pooled CBR of 32% and ORR of 3% in ovarian cancer patients. • Exclusion of stable disease for a period below 6 months as a beneficial outcome measure reduced the pooled CBR to 7%. • Drug-related grade 3 and 4 toxicities occurred in 36% (range 0–80%) of the patients. • Current PI3K/AKT/mTOR biomarkers insufficiently predict therapy response indicating the need for improved biomarker assays. • Combined treatments regimes targeting two signaling pathways may provide favorable outcomes over single agent therapy. [ABSTRACT FROM AUTHOR]

Glioblastoma is one of the most debilitating forms of brain tumour. It accounts for 17% of all the brain tumours and has resulted in 251,329 deaths in 2020 itself. Several studies performed on patient profiling have revealed that many genes tend to be overexpressed or mutated in many GBM patients and hamper the signalling and growth pathways. Additionally, there is a requirement to explore supplementary or alternative treatment that can aid in overcoming the resistance caused by standard care treatment (Temozolomide with radiation). The preliminary analysis on inhibitory potential of certain phytocompounds on the crucial genes involved in GBM has been studied in silico. The network of the genes critical for GBM has been analysed through several plugins of Cytoscape software like BiNGO and MCODE based on factors such as degree, closeness and betweenness. The network analysis gave out a cluster of 97 interactions with 16 interacting proteins. ERBB4 emerged as the seed protein indicating its crucial role in pathways. The BiNGO plugin gave out the gene ontology data describing the functions of the genes of the cluster. Additionally, the properties of ligands were studied by the PharmaGist server and 3D patterns of shared features by all or most input ligands of selected phytocompound were analysed and aligned. Upon docking the seed protein (ERBB4) individually with the best combination of aligned ligands yielded bromelain and EGCG as the compounds with best binding affinity of -9.8 and -9.2 kcal/mol respectively. This study has provided an important lead for further in vitro study to analyse the effect of combination of phytocompounds on the critical signalling pathways of GBM. [ABSTRACT FROM AUTHOR]

The renin–angiotensin system is involved in the regulation of various heart diseases. The present study aimed to determine the effects of angiotensin (Ang)-(3–7) on cardiac remodeling and its downstream signaling pathways in neonatal rat cardiomyocytes (NRCMs) and neonatal rat cardiac fibroblasts (NRCFs). The administration of Ang-(3–7) alleviated isoprenaline (ISO)-induced cardiac hypertrophy and fibrosis of mice. ISO treatment increased the levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and beta-myosin heavy chain (β-MHC) in NRCMs, and reduced the levels of collagen I, collagen III, fibronectin, and alpha-smooth muscle actin (α-SMA) in NRCFs. These changes were inhibited by Ang-(3–7) administration. The levels of protein kinase A (PKA), phosphorylated phosphatidylinositol-3-kinase (p-PI3K), and phosphorylated protein kinase B (p-Akt) were increased in NRCMs and NRCFs treated with ISO. The increase of PKA, but not p-PI3K or p-Akt was attenuated by Ang-(3–7) treatment in NRCMs. The increases of p-PI3K and p-Akt, but not PKA were reversed by Ang-(3–7) treatment in NRCFs. Treatment with cAMP or PKA overexpression reversed the attenuating effects of Ang-(3–7) on ISO-induced hypertrophy of NRCMs. The administration of PI3K inhibitor or Akt inhibitor alleviated ISO-induced fibrosis of NRCFs. These results indicated that Ang-(3–7) could alleviate cardiac remodeling. The administration of Ang-(3–7) attenuated hypertrophy of NRCMs via inhibiting the cAMP/PKA signaling pathway, and alleviated fibrosis of NRCFs via inhibiting PI3K/Akt signaling pathway. [ABSTRACT FROM AUTHOR]

Lupeol exhibits multiple pharmacological activities including, anticancerous, anti-inflammatory, and antioxidant. The aim of this study was to explore the anticancerous activity of lupeol on ovarian cancer cells and examine its mechanism of action. To this end, increasing concentrations of lupeol on cell viability, cell cycle, and apoptosis in Caov-3 cells were evaluated. Lupeol inhibited cell viability, induced G1 phase arrest in cell cycle, increased cell apoptosis, and inhibited the ratio of phospho-Akt/protein kinase B and phospho-mammalian target of rapamycin/mammalian target of rapamycin. In conclusion, these data suggest that lupeol may play a therapeutic role in ovarian cancer. [ABSTRACT FROM AUTHOR]

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Purpose: Everolimus plus exemestane (EVE/EXE) is a registered treatment option for ER-positive, HER2-negative (ER +/HER2-) metastatic breast cancer (MBC), but resistance mechanisms limit efficacy. We aimed to find markers that might help select patients with a higher chance on benefit from EVE/EXE. Methods: Immunohistochemistry (IHC) of PTEN, p-AKT(Thr308), p-AKT(Ser473), p-4EBP1, p-p70S6K, p-S6RP(Ser240/244), p-ERK1/2 and p-S6RP (Ser235/236) was performed on primary tumour tissue and on biopsies immediately taken from ER +/HER2- MBC patients before the start of standard EVE/EXE (Eudract 2013-004120-11). Unsupervised hierarchical clustering was executed to create heatmaps to distinguish subgroups of preferentially activated and less-activated PI3K/MAPK proteins. Uni- and multivariate Cox models were used for associations with PFS. Results: Primary tumour tissue from 145 patients was retrieved. Median PFS was 5.4 months. Patients without (neo)adjuvant therapy (p = 0.03) or bone only disease (p = 0.04) had longer PFS on EVE/EXE. In primary tumours, neither single proteins nor PI3K/MAPK-associated heatmap subgroups were significantly associated with PFS. In 21 patients a non-osseous biopsy obtained before dosing was useful for continuous scoring, which demonstrated upregulation of several proteins as compared to readings in corresponding primary tumour tissues. These comparisons revealed that increased expression of p-4EBP1 was significantly associated with worse PFS (multivariate HR 3.69, p = 0.05). Conclusions: IHC of single proteins or heatmap subgroups of the differentially activated PI3K/MAPK pathways was not able to discriminate patients on EVE/EXE with poor or better PFS. Upregulation of p-4EBP1 in pre-treatment biopsies as compared to levels in primary tumours pointed towards shorter PFS. [ABSTRACT FROM AUTHOR]

Human cancers often related to signal pathway variation, the phosphoinositide-3-kinase (PI3K) pathway is one of it. AKT kinase family is the most common downstream of PI3K pathway. The serum and glucocorticoid kinase 2(SGK2) is similar to AKT as the downstream of PI3K pathway. Up to now, we have few understanding of SGK2 in colon cancer. We determined the 20 colon cancer samples mRNA level. Later, we silence SGK2 in colon cancer cells. We found that SGK2 is up-regulated in colon cancer tissue/cells and have positive correlation with cell migration and invasive potential in human colon cancer cell line CACO2 and HCT116. The expression level of SGK2 have positive correlation with expression of mesenchymal marker N-cadherin and Vimentin and negative correlation with the expression of epithelial marker E-cadherin in CACO2 and HCT116 cells. In short, our research indicate that SGK2 is overexpressed in colon cancer and promotes EMT in colon cancer cells. [ABSTRACT FROM AUTHOR]

Endometrial hyperplasia (EH) is a medical condition that affects many females as it increases their uterine carcinogenic potential. EH results from entangling hormonal imbalance and inflammatory response. The study examined the role of a xanthine oxidase inhibitor, febuxostat, in a rat model of EH. Adult female Wistar albino rats were subjected to estradiol valerate (EV) 2 mg/kg for 10 days to induce EH. Another group was treated concomitantly with febuxostat 10 mg/kg for the same period. The uterine malondialdehyde, reduced glutathione (GSH), and superoxide dismutase (SOD) were assessed by chemical methods. Gene expressions of phosphatidylinositol-3-kinase (PI3K), Akt, and hypoxia-inducible factor 1 alpha were assessed by the quantitative real-time polymerase chain reaction. Moreover, the vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. Histopathology and immunohistochemical techniques were used for the detection of phosphatase and tensin homolog (PTEN). The results revealed that EV administration induced complex EH with focal atypia and loss of PTEN expression by the histological examination. Uteri of the EV group showed a significant drop in GSH content and SOD activity and rise in the expressions of PI3K, Akt, VEGF, and IL-6. Febuxostat administration significantly improved the uterine GSH and SOD levels. It decreased the expressions of PI3K, Akt, VEGF, and IL-6. The endometrium showed a regression of the proliferative epithelium with the restoration of PTEN expression and the absence of the atypical features. In conclusion, febuxostat protected the endometrium against estrogen-induced EH and may be beneficial in the management along with the hormonal therapy. [ABSTRACT FROM AUTHOR]

The phosphatidylinositol‐3‐kinase (PI3K) and mitogen‐activated protein kinase (MAPK) pathways are frequently activated in breast cancer. We recently demonstrated the importance of analyzing multiple proteins as read‐out for pathway activation in ER+/HER2− breast cancer, since single proteins are known to provide insufficient information. Here, we determined pathway activation in other primary breast cancer intrinsic subtypes derived from postmenopausal patients. Tumor blocks were recollected, and immunohistochemistry was performed using antibodies against PTEN, p‐AKT(Thr308), p‐AKT(Ser473), p‐p70S6K, p‐4EBP1, p‐S6RP(Ser235/236) and p‐ERK1/2, followed by unsupervised hierarchical clustering. In 32 ER+/HER2+, 37 ER−/HER2+ and 74 triple‐negative breast cancer patients, subgroups were identified with preferentially activated (A) and preferentially not activated (N) proteins. These subgroups likely reflect tumors with differences in biological behavior as well as treatment outcome. [ABSTRACT FROM AUTHOR]

Purpose: Oestrogen receptor β is believed to exert a cardioprotective effect against ischaemic injury. Nonetheless, the mechanism underlying its protective action remains to be fully elucidated. Recently, increased attention has been focused on Notch1 signalling for ameliorating cardiac ischaemic injury. Here, we hypothesised that oestrogen receptor β activation attenuates myocardial infarction (MI)-induced cardiac damage by modulating the Notch1 signalling pathway. Methods: Male C57BL/6 mice were used to establish an MI model through the ligation of the anterior descending branch of the left coronary artery. Two chemical drugs, 2,3-Bis(4-hydroxyphenyl)-propionitrile (DPN) and N-[N-(3,5-difluorophenacetyl)-l-alanyl]-s-phenylglycine t-butyl ester (DAPT), a specific inhibitor of Notch1 signalling) were administered via intraperitoneal injection to change oestrogen receptor β and Notch1 activities. Immunohistochemistry, western blot analysis, enzyme-linked immunosorbent assay (Elisa) assessment and echocardiography were used in this study to analyse cardiac oxidative stress, apoptosis, infraction volume, fibrosis and cardiac function. Results: DPN-mediated oestrogen receptor β activation effectively protected cardiomyocytes from MI-induced oxidative damage and apoptosis. Furthermore, oestrogen receptor β activation reduced the infarct size and lowered the levels of myocardial enzymes in the serum, thereby leading to greater overall cardiac function improvement. Ischaemic injury–induced myocardial fibrosis was attenuated by oestrogen receptor β activation. Nevertheless, all of these cardioprotective effects of oestrogen receptor β activation were almost abrogated by DAPT administration, i.e. DAPT attenuated the anti-oxidative and anti-apoptotic effects and the decrease in infarct and fibrotic areas and reversed cardiac functional recovery. The levels of phospho-phosphatidylinositol-3-kinase (PI3K) and phospho-protein kinase B (Akt) were increased after DPN administration, and this change was reversed after DAPT was administered. Conclusions: All of these new findings indicate that oestrogen receptor β activation is effective in ameliorating MI-induced cardiac dysfunction by enhancing Notch1 signalling and that PI3K/Akt signalling is the downstream mediator. [ABSTRACT FROM AUTHOR]

[Display omitted] • Tiron exerted a protective effect against AA- induced colitis in rats. • Tiron attenuated oxidative stress (↑SOD, GSH, TAC and ↓MDA). • Tiron significantly lowered TGF-β-1 that triggered downregulation of PEGFR. • Tiron significantly lowered PI3K, AKT, NF-κB, IL-6 and IFN-γ levels. • Tiron elevated the levels of GLP-1 and TFF-3. Ulcerative colitis (UC), an ongoing inflammatory disorder of the colon, is marked by persistent mucosal surface irritation extending from the rectum to the near-proximal colon. Tiron is a synthetic analogue of vitamin E which is known to have antioxidant and anti-inflammatory effects in various animal models, so the goal of this study was to find out whether Tiron had any preventive impacts on UC inflicted by acetic acid (A.A) exposure in rats. Tiron (235 and 470 mg/kg) was administered intra-peritoneally for 2 weeks, and A.A (2 ml, 3 % v/v) was injected intra-rectally to cause colitis. Colon tissues and blood samples were then collected for measurement of various inflammatory and oxidative stress biomarkers. Tiron administration significantly diminished lactate dehydrogenase (LDH), C-reactive protein (CRP), colon weight, and the weight/length ratio of the colon as compared to A.A-injected rats. Additionally, Tiron attenuated oxidative stress biomarkers. Tiron also enforced the levels of Glucagon-like peptide-1 (GLP-1) and trefoil factor-3 (TFF-3), while it greatly lowered the expression of nuclear factor kappa B (NF-κB), interleukin-6 (IL-6), interferon-γ (IFN-γ), and transforming growth factor-1(TGF-β1), phosphorylated epidermal growth factor receptor (P-EGFR), phosphatidylinositol-3-kinase (PI3K) and protein kinase B (AKT) expression in colonic cellular structures. Furthermore, colonic histopathologic damages, revealed by hematoxylin and eosin (H&E) and Alcian Blue stain, were significantly decreased upon Tiron administration. Tiron prevented A.A-induced colitis in rats via modulating inflammatory pathway TGF-β1/P-EGFR/PI3K/AKT/NF-κB, alongside managing the oxidant/antioxidant equilibrium, and boosting the reliability of the intestinal barrier. [ABSTRACT FROM AUTHOR]

Epidermolysis bullosa acquisita (EBA) is a rare, but prototypical, organ-specific autoimmune disease, characterized and caused by autoantibodies against type VII collagen (COL7). Mucocutaneous inflammation, blistering, and scarring are the clinical hallmarks of the disease. Treatment of EBA is difficult and mainly relies on general immunosuppression. Hence, novel treatment options are urgently needed. The phosphatidylinositol-3-kinase (PI3K) pathway is a putative target for the treatment of inflammatory diseases, including EBA. We recently discovered LAS191954, an orally available, selective PI3Kδ inhibitor. PI3Kδ has been shown to be involved in B cell and neutrophil cellular functions. Both cell types critically contribute to EBA pathogenesis, rendering LAS191954 a potential drug candidate for EBA treatment. We, here, demonstrate that LAS191954, when administered chronically, dose-dependently improved the clinical phenotype of mice harboring widespread skin lesions secondary to immunization-induced EBA. Direct comparison with high-dose corticosteroid treatment indicated superiority of LAS191954. Interestingly, levels of circulating autoantibodies were unaltered in all groups, indicating a mode of action independent of the inhibition of B cell function. In line with this, LAS191954 also hindered disease progression in antibody transfer-induced EBA, where disease develops dependent on myeloid, but independent of B cells. We further show that, in vitro, LAS191954 dose-dependently impaired activation of human myeloid cells by relevant disease stimuli. Specifically, immune complex-mediated and C5a-mediated ROS release were inhibited in a PI3Kδ-dependent manner. Accordingly, LAS191954 also modulated the dermal–epidermal separation induced in vitro by co-incubation of immune complexes with polymorph nuclear cells, thus pointing to an important role of PI3Kδ in EBA effector functions. Altogether, these results suggest a new potential mechanism for the treatment of EBA and potentially also other autoimmune bullous diseases.

Objective: To observe neuroprotective effects of Ca 2+ /calmodulin-dependent kinase Ⅱ (CaMK Ⅱ)γ and CaMkII δ against acute neuronal ischemic reperfusion injury in mice and explore the underlying mechanism., Methods: Primary cultures of brain neurons isolated from fetal mice (gestational age of 18 days) were transfected with two specific siRNAs (si-CAMK2G and si-CAMK2D) or a control sequence (si-NT). After the transfection, the cells were exposed to oxygen-glucose deprivation/reperfusion (OGD/R) conditions for 1 h followed by routine culture. The expressions of phosphatidylinositol-3-kinase/extracellular signal-regulated kinase (PI3K/Akt/Erk) signaling pathway components in the neurons were detected using immunoblotting. The expressions of the PI3K/Akt/Erk signaling pathway proteins were also detected in the brain tissues of mice receiving middle cerebral artery occlusion (MCAO) or sham operation., Results: The neuronal cells transfected with siCAMK2G showed significantly lower survival rates than those with si-NT transfection at 12, 24, 48, and 72 h after OGD/R ( P < 0.01), and si-CAMK2G transfection inhibited OGD/R-induced upregulation of CaMKⅡγ expression. Compared to si-NT, transfection with si-CAMK2G and si-CAMK2D both significantly inhibited the expressions of PI3K/Akt/Erk signaling pathway components ( P < 0.01). In the mouse models of MCAO, the expressions of CaMKⅡδ and CaMKⅡγ were significantly increased in the brain, where activation of the PI3K/Akt/Erk signaling pathway was detected. The expression levels of CaMKⅡδ, CaMKⅡγ, Erk, phosphorylated Erk, Akt, and phosphorylated Akt were all significantly higher in MCAO mice than in the sham-operated mice at 24, 48, 72, and 96 h after reperfusion ( P < 0.05)., Conclusion: The neuroprotective effects of CaMKⅡδ and CaMKⅡγ against acute neuronal ischemic reperfusion injury are mediated probably by the PI3K/Akt/Erk pathway.

Background: Korean Red Ginseng has been used as a traditional oriental medicine to treat illness and to promote health for several thousand years in Eastern Asia. It is widely accepted that ginseng saponins, ginsenosides, are the major active ingredients responsible for Korean Red Ginseng’s therapeutic activity against many kinds of illness. Although the crude saponin fraction (CSF) displayed antiplatelet activity, the molecular mechanism of its action remains to be elucidated. Methods: The platelet aggregation was induced by collagen, the ligand of integrin αIIβI and glycoprotein VI. The crude saponin’s effects on granule secretion [e.g., calcium ion mobilization and adenosine triphosphate (ATP) release] were determined. The activation of mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated protein kinase 1/2 (ERK1/2), c-Jun N-terminal kinases (JNKs), and p38 MAPK, and phosphoinositide 3-kinase (PI3K)/Akt was analyzed by immunoblotting. In addition, the activation of integrin αIIbβIII was examined by fluorocytometry. Results: CSF strongly inhibited collagen-induced platelet aggregation and ATP release in a concentration-dependent manner. It also markedly suppressed [Ca2+]i mobilization in collagen-stimulated platelets. Immunoblotting assay revealed that CSF significantly suppressed ERK1/2, p38, JNK, PI3K, Akt, and mitogen-activated protein kinase kinase 1/2 phosphorylation. In addition, our fraction strongly inhibited the fibrinogen binding to integrin αIIbβ3. Conclusion: Our present data suggest that CSF may have a strong antiplatelet property and it can be considered as a candidate with therapeutic potential for the treatment of cardiovascular disorders involving abnormal platelet function.

Cervical cancer is one of the most common malignancies that seriously threatens women's health. Krüppel-like factors (KLFs) have been reported to be associated with the progression of cervical cancer. The role of KLF1 in cervical cancer, which still remains unclear, was investigated in the present study. The expression of KLF1 was detected in different cervical cell lines by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. Cell proliferation, metastasis and invasion were respectively detected by Cell Counting Kit-8, wound healing and transwell assays. Associated factor expression was also detected by RT-qPCR and western blotting. In addition, the phosphorylation levels of phosphatidylinositol-3-kinase (PI3K) and protein kinase B (Akt) were determined by western blot analysis. The results revealed that KLF1 expression was promoted in SiHa, Caski and C4-1 cervical cancer cells. However, KLF1 knockdown suppressed cell proliferation, metastasis and invasion in SiHa cervical cancer cells. KLF1 knockdown also inhibited the expressions of Ki67, metastasis-associated antigen 1 and matrix metalloproteinase (MMP)-2. KLF1 knockdown promoted the expressions of nonmetastatic clone 23 type 1 and tissue inhibitor of metalloproteinase-2, and the expression of MMP-9 was promoted slightly as well. In addition, KLF1 knockdown inhibited the PI3K/Akt signaling pathway. Hence, it was concluded that KLF1 promoted metastasis and invasion via the PI3K/Akt signaling pathway in cervical cancer cells. [ABSTRACT FROM AUTHOR]

Hepatic ischemia reperfusion (I/R) injury is very common in liver transplantation and major liver surgeries and may cause liver failure or even death. Docosahexaenoic acid (DHA) has displayed activities in reducing oxidative stress and inflammatory reaction in many disorders. In the present study, we investigated the protective effects of DHA against I/R-induced injury and the underlying mechanisms. Here, we show that DHA protected hepatic I/R injury by reducing aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and decreasing the oxidative stress in liver tissues. The viability of Buffalo rat liver (BRL) cells was reduced by hypoxia/restoration (H/R) but restored by DHA. DHA significantly downregulated the expression of pyroptosis-related proteins including NLR pyrin domain containing 3 (NLRP3), apoptotic speck-like protein containing CARD (ASC) and cleaved caspase-1 and reduced the secretion of pro-inflammatory cytokines. The above results were supported by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. However, incubation with LY294002, a specific inhibitor of phosphatidylinositol-3-kinase (PI3K), abolished the effects of DHA, since it increased the expression of cleaved caspase-1 and the production of inflammatory cytokines. The present results have demonstrated that DHA ameliorated I/R-induced injury by inhibiting pyroptosis of hepatocytes induced in liver I/R injury in vivo and in vitro through the PI3K/Akt pathway, providing a potential therapeutic option to prevent liver injury by I/R. [ABSTRACT FROM AUTHOR]

The present study investigated the effect and underling mechanisms of aclidinium bromide, a novel, inhaled long‑acting muscarinic antagonist, on the development of gastric cancer. Human gastric cancer MKN‑28 cells, as a model in vitro, were treated with aclidinium bromide and dimethyl sulfoxide. Cell Counting Kit‑8 assay, transwell assay and flow cytometry were used to assess cell proliferation, invasion/migration and apoptosis, respectively. In addition, western blotting was performed to determine the relative expression of proteins associated with apoptosis and the phosphatidylinositol‑3‑kinase (PI3K) signaling pathway. Optical density values of MKN‑28 cells were decreased in a time‑ and dose‑dependent manner in the aclidinium bromide treated group. Matrigel invasion analysis demonstrated the number of invasive cells were significantly decreased in the aclidinium bromide‑treated group when compared with the control group. Furthermore, aclidinium bromide led to the marked reduction of the number of MKN‑28 cells passing though the microwells of the transwell chamber. The expression levels of the anti‑apoptotic protein B‑cell lymphoma 2 (Bcl‑2) decreased, and the expression of pro‑apoptotic proteins active Caspase3 and Bcl‑2‑associated X protein increased concurrently following aclidinium bromide stimulation using western blotting. The phosphorylation of protein kinase B and mechanistic target of rapamycin were significantly inhibited in MKN‑28 cells treated with aclidinium bromide; and the activity of the downstream proteins such as p70S6K and Cyclin D1 were also significantly decreased. In conclusion, aclidinium bromide could inhibit gastric cancer cell proliferation and metastasis, which may be associated with the enhancement of apoptosis induced by the PI3K signaling pathway. [ABSTRACT FROM AUTHOR]

Epidermolysis bullosa acquisita (EBA) is a rare, but prototypical, organ-specific autoimmune disease, characterized and caused by autoantibodies against type VII collagen (COL7). Mucocutaneous inflammation, blistering, and scarring are the clinical hallmarks of the disease. Treatment of EBA is difficult and mainly relies on general immunosuppression. Hence, novel treatment options are urgently needed. The phosphatidylinositol-3-kinase (PI3K) pathway is a putative target for the treatment of inflammatory diseases, including EBA. We recently discovered LAS191954, an orally available, selective PI3Kδ inhibitor. PI3Kδ has been shown to be involved in B cell and neutrophil cellular functions. Both cell types critically contribute to EBA pathogenesis, rendering LAS191954 a potential drug candidate for EBA treatment. We, here, demonstrate that LAS191954, when administered chronically, dose-dependently improved the clinical phenotype of mice harboring widespread skin lesions secondary to immunization-induced EBA. Direct comparison with high-dose corticosteroid treatment indicated superiority of LAS191954. Interestingly, levels of circulating autoantibodies were unaltered in all groups, indicating a mode of action independent of the inhibition of B cell function. In line with this, LAS191954 also hindered disease progression in antibody transfer-induced EBA, where disease develops dependent on myeloid, but independent of B cells. We further show that, in vitro, LAS191954 dose-dependently impaired activation of human myeloid cells by relevant disease stimuli. Specifically, immune complex-mediated and C5a-mediated ROS release were inhibited in a PI3Kδ-dependent manner. Accordingly, LAS191954 also modulated the dermal–epidermal separation induced in vitro by co-incubation of immune complexes with polymorph nuclear cells, thus pointing to an important role of PI3Kδ in EBA effector functions. Altogether, these results suggest a new potential mechanism for the treatment of EBA and potentially also other autoimmune bullous diseases. [ABSTRACT FROM AUTHOR]

The phosphatidylinositol-3-kinase (PI3K)/Akt pathway is important for multiple stages of herpesvirus replication including virus entry, replication, latency, and reactivation. Recently, patients with gain-of-function mutations in the p110δ-catalytic subunit of PI3K or in the p85-regulatory subunit of PI3K have been reported. These patients have constitutively active PI3K with hyperactivation of Akt. They present with lymphoproliferation and often have infections, particularly recurrent respiratory infections and/or severe virus infections. The most frequent virus infections are due to Epstein–Barr virus (EBV) and cytomegalovirus (CMV); patients often present with persistent EBV and/or CMV viremia, EBV lymphoproliferative disease, or CMV lymphadenitis. No patients have been reported with CMV pneumonia, colitis, or retinitis. Other herpesvirus infections have included herpes simplex pneumonia, recurrent zoster, and varicella after vaccination with the varicella vaccine. Additional viral infections have included adenovirus viremia, severe warts, and extensive Molluscum contagiosum virus infection. The increased susceptibility to virus infections in these patients is likely due to a reduced number of long-lived memory CD8 T cells and an increased number of terminally differentiated effector CD8 T cells.

Phosphatidylinositol-3-kinase p110δ (PI3Kδ) inhibition by Idelalisib (CAL-101) in hematological malignancies directly induces apoptosis in cancer cells and disrupts immunological tolerance by depleting regulatory T cells. Yet, little is known about the direct impact of PI3Kδ blockade on effector T cells from CAL-101 therapy. Herein, we demonstrate a direct effect of p110δ inactivation via CAL-101 on murine and human CD8+ T cells that promotes a strong undifferentiated phenotype (elevated CD62L/CCR7, CD127, and Tcf7). These CAL-101 T cells also persisted longer after transfer into tumor bearing mice in both the murine syngeneic and human xenograft mouse models. The less differentiated phenotype and improved engraftment of CAL-101 T cells resulted in stronger antitumor immunity compared to traditionally expanded CD8+ T cells in both tumor models. Thus, this report describes a novel direct enhancement of CD8+ T cells by a p110δ inhibitor that leads to markedly improved tumor regression. This finding has significant implications to improve outcomes from next generation cancer immunotherapies.

Curcumin as one kind of phenolic pigment extracted from dietary spice turmeric, has various pharmacological activity, which including anti-inflammatory, antioxidation, anticoagulation, hypolipidemic, anti-atherosclerotic, antineoplastic and antimutagenicity. Inducing apoptosis of tumor cells is one of the mechanisms of anti-tumor as well as anti-tumor-angiogenesis. The mechanisms of anti-angiogenesis include effecting the proliferation and apoptosis of vascular endothelial cell; inhibiting the expression of angiogenesis-promoting factor, such as inhibiting vascular endothelial growth factor(VEGF)and its receptor to inhibit the formation of angiogenesis of tumor; reducing the activity of matrix metalloproteinases(MMPs)and acting through the phosphatidylinositol-3-kinase(PI3K)signaling pathway, the mechanism of which is to down-regulating the PI3K/AKT and MAPK pathways in order to down-regulating VEGF, Ang-1,Ang-2, VEGFR-2 and etc. Further research for effects of curcumin for mechanism of anti-tumor-angiogenesis will lay the foundation of more effective therapy on anti-tumor-angiogenesis, which is useful for prevention and treatment of cancer.

The role of phospholipase D as a positive modulator of glucose uptake activation by insulin in the neocortex of 3-month-old rats were studied by using specific inhibitors of signaling pathways of insulin (wortmannin, LY294002, halopemide, C6-ceramide). It was found that in the rat neocortex inhibitors decreased the insulin-induced accumulation of phosphatidylethanol from 163% in the control to 115% under the influence of wortmannin and 112% under the action of LY294002. Inhibition of insulin stimulated phospholipase D by phosphatidylinositol-3-kinase inhibitors (wortmannin and LY294002) indicated downstream regulation of phospholipase D by phosphatidylinositol-3-kinase. The suppression of this phospholipase by a specific inhibitor haloperimide is accompanied by a decrease in the amount of labeled glucose in the neocortical tissue from 154% in the control to 111% under the influence of the inhibitor. Inhibition of insulin-induced glucose uptake by specific inhibitor of phospholipase D halopemide points to the key role of the enzyme in the regulation of glucose uptake in the rat neocortex. In addition, phospholipase D-dependent insulin signaling and glucose uptake in the rat cerebral cortex are highly sensitive to intracellular ceramide content. The results obtained in this study provide that modulation of PLD activity, as well as ceramides content in the cells, can be a useful tool for manipulating the nerve tissue sensitivity to insulin action. [ABSTRACT FROM AUTHOR]

The phosphatidylinositol-3-kinase (PI3K)/Akt pathway is important for multiple stages of herpesvirus replication including virus entry, replication, latency, and reactivation. Recently, patients with gain-of-function mutations in the p110δ-catalytic subunit of PI3K or in the p85-regulatory subunit of PI3K have been reported. These patients have constitutively active PI3K with hyperactivation of Akt. They present with lymphoproliferation and often have infections, particularly recurrent respiratory infections and/or severe virus infections. The most frequent virus infections are due to Epstein-Barr virus (EBV) and cytomegalovirus (CMV); patients often present with persistent EBV and/or CMV viremia, EBV lymphoproliferative disease, or CMV lymphadenitis. No patients have been reported with CMV pneumonia, colitis, or retinitis. Other herpesvirus infections have included herpes simplex pneumonia, recurrent zoster, and varicella after vaccination with the varicella vaccine. Additional viral infections have included adenovirus viremia, severe warts, and extensive Molluscum contagiosum virus infection. The increased susceptibility to virus infections in these patients is likely due to a reduced number of long-lived memory CD8 T cells and an increased number of terminally differentiated effector CD8 T cells. [ABSTRACT FROM AUTHOR]

The phytohormone abscisic acid (ABA) exists in animal tissues particularly in the brain. However, the neurophysiological effects of ABA have not yet been fully determined. Signaling molecules such as PKC and PI3K have been implicated in anxiety-like behavior as well as learning and memory processes. Recently, it has been demonstrated that PKC and PI3K signaling pathways participate in some biological effects of ABA. This study was designed to evaluate the effects of central injection of ABA on spatial learning and memory and anxiety-like behavior and determine its possible signaling mechanisms. Spatial learning and memory and anxiety-like behaviors were determined using Morris water maze (MWM) and plus maze tests, respectively. ABA alone or in accompanied with selective inhibitors of PKC (chelerythrine) and PI3K (LY2940029) was injected bilaterally into the cerebral lateral ventricles. The results indicated that ABA (10 μg/rat) significantly improved rats’ performance in MWM which was blocked by PKC or PI3K inhibitors. In addition, ABA showed anti-anxiety effect in plus maze test, which was not observed in PKC or PI3K inhibitor-treated rats. Overall, the results indicated that ABA has positive effect on spatial learning and memory performance and elicits anti-anxiety effects which are performed, at least in part, through PI3K/PKC signaling pathway. [ABSTRACT FROM AUTHOR]

Neural stem/progenitor cells (NSPCs) reside not only in the developing brain, but also in the adult brain within specialized microenvironments that regulate their function. In vitro and in vivo studies have revealed strong regulatory links between hypoxic/ischemic insults and activation of NSCPs. However, the underlying mechanisms of this activation remain unclear. In this study, we found that cell proliferation is promoted by hypoxia, and accompanied by increasing expression of miR-21 in cultured rat NSPCs. Moreover, qRT-PCR analysis indicated that expression of miR-21 increases in a time-dependent manner. 5-Bromo-2-deoxyUridine (BrdU) staining and flow cytometry showed that overexpression of miR-21 further promoted proliferation of NSPCs in the presence of hypoxia. Knocking down of miR-21 partially abolished the proliferative effect of hypoxia treatment on cell proliferation. Western blot demonstrated that overexpression of miR-21 enhanced expression of cyclin D1, while knock down of miR-21 suppressed cyclin D1 expression under hypoxic conditions. Furthermore, overexpression of miR-21 also increased levels of p-AKT. These results demonstrate that miR-21 plays a role in regulating the proliferation of cultured rat NSPCs undergoing hypoxia, and the activation of the PI-3-K signaling pathway might be one of the underlying mechanisms. These findings prompt a molecular study investigating potential mechanisms for stem cell treatment of cerebral ischemia. [ABSTRACT FROM AUTHOR]

Phosphatidylinositol-3-kinase p110δ (PI3Kδ) inhibition by Idelalisib (CAL-101) in hematological malignancies directly induces apoptosis in cancer cells and disrupts immunological tolerance by depleting regulatory T cells. Yet, little is known about the direct impact of PI3Kδ blockade on effector T cells from CAL-101 therapy. Herein, we demonstrate a direct effect of p110δ inactivation via CAL-101 on murine and human CD8+ T cells that promotes a strong undifferentiated phenotype (elevated CD62L/CCR7, CD127, and Tcf7). These CAL-101 T cells also persisted longer after transfer into tumor bearing mice in both the murine syngeneic and human xenograft mouse models. The less differentiated phenotype and improved engraftment of CAL-101 T cells resulted in stronger antitumor immunity compared to traditionally expanded CD8+ T cells in both tumor models. Thus, this report describes a novel direct enhancement of CD8+ T cells by a p110δ inhibitor that leads to markedly improved tumor regression. This finding has significant implications to improve outcomes from next generation cancer immunotherapies. [ABSTRACT FROM AUTHOR]