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Background: Gastric cancer is a highly heterogeneous disease, presenting a major obstacle to personalized treatment. Effective markers of the immune checkpoint blockade response are needed for precise patient classification. We, therefore, divided patients with gastric cancer according to collagen gene expression to indicate their prognosis and treatment response. Methods: We collected data for 1250 patients with gastric cancer from four cohorts. For the TCGA‐STAD cohort, we used consensus clustering to stratify patients based on expression levels of 44 collagen genes and compared the prognosis and clinical characteristics between collagen subtypes. We then identified distinct transcriptomic and genetic alteration signatures for the subtypes. We analyzed the associations of collagen subtypes with the responses to chemotherapy, immunotherapy, and targeted therapy. We also established a platform‐independent collagen‐subtype predictor. We verified the findings in three validation cohorts (GSE84433, GSE62254, and GSE15459) and compared the collagen subtyping method with other molecular subtyping methods. Results: We identified two subtypes of gastric adenocarcinoma: a high‐expression collagen subtype (CS‐H) and a low‐expression collagen subtype (CS‐L). Collagen subtype was an independent prognostic factor, with better overall survival in the CS‐L subgroup. The inflammatory response, angiogenesis, and phosphoinositide 3‐kinase (PI3K)/Akt pathways were transcriptionally active in the CS‐H subtype, while DNA repair activity was significantly greater in the CS‐L subtype. PIK3CA was frequently amplified in the CS‐H subtype, while PIK3C2A, PIK3C2G, and PIK3R1 were frequently deleted in the CS‐L subtype. CS‐H subtype tumors were more sensitive to fluorouracil, while CS‐L subtype tumors were more sensitive to immune checkpoint blockade. CS‐L subtype was predicted to be more sensitive to HER2‐targeted drugs, and CS‐H subtype was predicted to be more sensitive to vascular endothelial growth factor and PI3K pathway‐targeting drugs. Collagen subtyping also has the potential to be combined with existing molecular subtyping methods for better patient classification. Conclusions: We classified gastric cancers into two subtypes based on collagen gene expression and validated these subtypes in three validation cohorts. The collagen subgroups differed in terms of prognosis, clinical characteristics, transcriptome, and genetic alterations. The subtypes were closely related to patient responses to chemotherapy, immunotherapy, and targeted therapy. [ABSTRACT FROM AUTHOR]

Human astrovirus is a positive-sense, single-stranded RNA virus. Astrovirus infection causes gastrointestinal symptoms and can lead to encephalitis in immunocompromised patients. Positive-strand RNA viruses typically utilize host intracellular membranes to form replication organelles, which are potential antiviral targets. Many of these replication organelles are double-membrane vesicles (DMVs). Here, we show that astrovirus infection leads to an increase in DMV formation through a replication-dependent mechanism that requires some early components of the autophagy machinery. Results indicate that the upstream class III phosphatidylinositol 3-kinase (PI3K) complex, but not LC3 conjugation machinery, is utilized in DMV formation. Both chemical and genetic inhibition of the PI3K complex lead to significant reduction in DMVs, as well as viral replication. Elucidating the role of autophagy machinery in DMV formation during astrovirus infection reveals a potential target for therapeutic intervention for immunocompromised patients. IMPORTANCE These studies provide critical new evidence that astrovirus replication requires formation of double-membrane vesicles, which utilize class III phosphatidylinositol 3-kinase (PI3K), but not LC3 conjugation autophagy machinery, for biogenesis. These results are consistent with replication mechanisms for other positive-sense RNA viruses suggesting that targeting PI3K could be a promising therapeutic option for not only astrovirus, but other positive-sense RNA virus infections.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a devastating disease most often associated with tobacco consumption that induces a field of mutations from which a tumor arises. Identification of ways to prevent the emergence of cancer in high-risk patients is an ultimate goal for combatting all types of cancer, including OSCC. METHODS: Our study employs a mouse model of tongue carcinogenesis induced by tobacco carcinogen mimetic, 4-nitroquinoline 1-oxide (4NQO), to establish tongue dysplasia and OSCC. We use conventional histology, immunohistochemistry, multispectral imaging, mass cytometry, novel cell lines, pharmaceutical inhibition of PI3Kγ, T-cell suppression assays and mouse transplant models in our functional experimentation. RESULTS: In our study, we identify Ly6G+ granulocytes as the most abundant immune cell type in a model of tongue carcinogenesis induced by tobacco carcinogen mimetic 4NQO. Targeting Ly6G+ granulocytes with a pharmacologic inhibitor of PI3Kγ, an isoform of PI3K exclusively expressed by myeloid cells, resulted in reduced tongue dysplasia severity, and reduced rates of OSCC. Importantly, we performed functional assays with the Ly6G+ granulocytes induced in cell line models of 4NQO carcinogenesis to demonstrate that these granulocytes have increased polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) activity against T-cell proliferation and these PMN-MDSCs play a functional role in promoting tumor formation by inhibiting tumor regression in a PI3Kγ-dependent manner. CONCLUSIONS: Overall, our data suggest that recruitment of PMN-MDSCs to sites of dysplasia is critical to immune suppression of CD8 T cells, thereby permitting malignancy, and PI3Kγ inhibitors are one mechanism to reduce PMN-MDSC recruitment, immunosuppression and tumorigenesis in OSCC.

Neurovascular coupling (NVC), a vital physiological process that rapidly and precisely directs localized blood flow to the most active regions of the brain, is accomplished in part by the vast network of cerebral capillaries acting as a sensory web capable of detecting increases in neuronal activity and orchestrating the dilation of upstream parenchymal arterioles. Here, we report a Col4a1 mutant mouse model of cerebral small vessel disease (cSVD) with age-dependent defects in capillary-to-arteriole dilation, functional hyperemia in the brain, and memory. The fundamental defect in aged mutant animals was the depletion of the minor membrane phospholipid phosphatidylinositol 4,5 bisphosphate (PIP2) in brain capillary endothelial cells, leading to the loss of inwardly rectifying K+ (Kir2.1) channel activity. Blocking phosphatidylinositol-3-kinase (PI3K), an enzyme that diminishes the bioavailability of PIP2 by converting it to phosphatidylinositol (3, 4, 5)-trisphosphate (PIP3), restored Kir2.1 channel activity, capillary-to-arteriole dilation, and functional hyperemia. In longitudinal studies, chronic PI3K inhibition also improved the memory function of aged Col4a1 mutant mice. Our data suggest that PI3K inhibition is a viable therapeutic strategy for treating defective NVC and cognitive impairment associated with cSVD.

Abstract Background Insulin resistance (IR) induces hyperinsulinemia, which activates downstream signaling pathways such as the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) pathway, ultimately leading to abnormal proliferation and apoptosis of endometrial cells. This is thought to be a key pathogenic mechanism underlying the development of endometrial polyps (EP). This study aims to investigate the relationship between IR and the development of EP, the expression levels of downstream signaling molecules, including PI3K and AKT, and related laboratory parameters were examined. Methods A total of 100 patients who visited the gynecology outpatient clinic of Zhongda Hospital affiliated with Southeast University from May 2021 to March 2023 and were diagnosed with abnormal endometrial echoes by vaginal ultrasound and underwent hysteroscopic diagnostic curettage were enrolled in this study. General data and relevant hematological indicators were compared, and intraoperative specimens were obtained for pathological examination. Possible factors influencing the development of endometrial polyps were analyzed using Pearson correlation analysis and logistic regression analysis. Results In terms of body mass index, waist circumference, fasting insulin, insulin resistance index, serum total testosterone, and free testosterone index, women of childbearing age in the endometrial polyp group had higher values than those in the non-polyp group, while sex hormone-binding globulin in the endometrial polyp group was lower than that in the non-polyp group, and the differences were statistically significant (P

Background: Insulin resistance (IR) induces hyperinsulinemia, which activates downstream signaling pathways such as the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) pathway, ultimately leading to abnormal proliferation and apoptosis of endometrial cells. This is thought to be a key pathogenic mechanism underlying the development of endometrial polyps (EP). This study aims to investigate the relationship between IR and the development of EP, the expression levels of downstream signaling molecules, including PI3K and AKT, and related laboratory parameters were examined. Methods: A total of 100 patients who visited the gynecology outpatient clinic of Zhongda Hospital affiliated with Southeast University from May 2021 to March 2023 and were diagnosed with abnormal endometrial echoes by vaginal ultrasound and underwent hysteroscopic diagnostic curettage were enrolled in this study. General data and relevant hematological indicators were compared, and intraoperative specimens were obtained for pathological examination. Possible factors influencing the development of endometrial polyps were analyzed using Pearson correlation analysis and logistic regression analysis. Results: In terms of body mass index, waist circumference, fasting insulin, insulin resistance index, serum total testosterone, and free testosterone index, women of childbearing age in the endometrial polyp group had higher values than those in the non-polyp group, while sex hormone-binding globulin in the endometrial polyp group was lower than that in the non-polyp group, and the differences were statistically significant (P < 0.05). The expression scores and mRNA expression levels of PI3K and AKT proteins were higher in the EP group than in the non-EP group (p < 0.05). Pearson correlation analysis showed a positive correlation between HOMA-IR and the expression scores of PI3K and AKT proteins (p < 0.01). Conclusions: Insulin resistance and abnormal activation of the phosphatidylinositol 3-kinase/protein kinase B signaling pathway may be potential pathogenic mechanisms for the development of endometrial polyps. [ABSTRACT FROM AUTHOR]

Objective: To discuss the regulatory effect of serum and glucocorticoid-induced protein kinase 1 (SGK1) in the early development of fertilized eggs at G1 phase of the mice, and to clarify the related mechanism. Methods: Some female mice aged 4-6 weeks and weighed about 20 g, and several male mice aged over 8 weeks and weighed about 30 g were selected. The female mice were intraperitoneally injected with 10 IU of pregnant mare serum gonadotropin (PMSG), followed by 10 IU of human chorionic gonadotropin (HCG) after 48 h. After HCG injection, the female mice were caged overnight with the male mice at a ratio of 1: 1. The fertilized eggs at G1, S, G2, and M phases were collected at 12-21 h, 21-26 h, 26-28 h, and 28-30 h after injected with HCG, and their cellular morphology at different cell cycles were observed under light microscope. The mouse fertilized eggs at G1 phase after superovulation were collected, the mRNA was synthesized in vitro, and divided into no injection group, Tris-EDTA buffer injection group (TE injection group), and SGK1-mRNA injection group. The SGK1 antibodies were mixed with KSOM culture medium with the concentrations of 1 : 25, 1 : 50, 1 : 100, 1: 200, and 0 to culture the mouse fertilized eggs at G1 phase. Western blotting method was used to detect the expression levels of SGK1 protein in fertilized eggs of the mice in various groups and the dephosphorylation for phosphorylated SGK1-Threonine 256 site tyrosine15 site of cell diusion cyclin 2 (Cdc2) (Cdc2-pTyr15) in the fertilized eggs of the mice in various groups and different concentrations of SGK1 antibody groups and the developmental states of the fertilized eggs in the fertilized eggs of the mice in various groups and different concentrations of SGK1 antibody groups were observed under phase contrast microscope; the expression levels of phosphorylated SGK1-Thr256 (SGK1-pThr256) and Cdc2-pTyr15 proteins in fertilized eggs at different post-HCG injection times were detected by Western blotting method. Results: Compared with no injection and TE injection groups, the expression level of SGK1 protein in the cells in SGK1-mRNA injection group was significantly increased (P<0. 01). 27-28 h after injected with HCG, the phosphorylation signaling of Cdc2-pTyr15 in fertilized eggs of the mice in SGK1-mRNA injection group was gradually disappeared, and there was no phosphorylation signaling 29 h after injected with HCG. At 28-29 h after injected with HCG, the phosphorylation signaling of Cdc2-pTyr15 in fertilized eggs of the mice in no injection and TE injection groups gradually disappeared, completely disappeared at 30 h after injected with HCG. With the increasing of the concentration of SGK1 antibody, the disappearing time of the Cdc2-pTyr15 phosphorylation signaling was increased. At 27 h after injected with HCG, the fertilized eggs of the mice in SGK1-mRNA injection group was initiated cleavage; at 31 h after injected with HCG, nearly all the fertilized eggs turned into G2 phase; at 33 h after injected with HCG, all the fertilized eggs in 0 and 1: 200 SGK1 antibody groups underwent cleavage. However, with the increasing of SGK1 antibody concentration, the cleavage of the fertilized eggs in 1: 25, 1: 50, and 1: 100 SGK1 antibody groups was gradually decreased, particularly at 1: 25 SGK1 antibody group. Compared with no injection and TE injection groups, the death rate of the fertilized eggs of the mice in SGK1-mRNA injection group was significantly decreased at 31 h after injected with HCG (P<0. 05), and the cleavage rate was increased (P<0. 05). With the increasing of the SGK1 antibody concentration, the death rates of the fertilized eggs in different concentrations of SGK1 antibody group were increased (P<0. 05), with the extending of cleavage time was increased, and the cleavage rate of the fertilized eggs was decreased in a dose-dependent manner, and the cleavage rate of fertilized eggs in 1: 25 SGK1 antibody group was the lowest. The expression level of SGK1-pThr256 protein in fertilized eggs of the mice was gradually increased from 27 h after injected with HCG (P<0. 05 or P< 0. 01) in a time-dependent manner; at 28 to 29 h after injected with HCG, the expression levels of Cdc2-Tyr15 protein were gradually decreased (P<0. 05) in a time-dependent manner, and had completely disappeared at 30 h after injected with HCG. Conclusion: Both the over-expression and inhibition of SGK1 can affect the time for the fertilized eggs at G1 phase to entry into M phase, suggesting that SGK1 protein may be one of the regulatory factors in the early development of fertilized eggs at G1 phase of the mice, and it may regulate the development of the fertilized eggs at G1 phase through regulation of Cdc2. [ABSTRACT FROM AUTHOR]

Tumors in dogs and humans share many similar molecular and genetic features, incentivizing a better understanding of canine neoplasms not only for the purpose of treating companion animals, but also to facilitate research of spontaneously developing tumors with similar biologic behavior and treatment approaches in an immunologically competent animal model. Multiple tumor types of both species have similar dysregulation of signal transduction through phosphatidylinositol 3-kinase (PI3K), protein kinase B (PKB; AKT), and mechanistic target of rapamycin (mTOR), collectively known as the PI3K-AKT-mTOR pathway. This review aims to delineate the pertinent aspects of the PI3K-AKT-mTOR signaling pathway in health and in tumor development. It will then present a synopsis of current understanding of PI3K-AKT-mTOR signaling in important canine cancers and advancements in targeted inhibitors of this pathway. [ABSTRACT FROM AUTHOR]

Objective To evaluate the regulatory effect of the adaptor related protein complex 2 subunit μ1 (AP2M1) on proliferation and invasion of diffuse large B-cell lymphoma (DLBCL). Methods Human diffuse large B-cell lymphoma cell line OCI-LY8 was aliquoted into control group, NC-shRNA group, AP2M1-shRNA group, NC-LV group, and AP2M1-LV group. Lipofectamine 2000 was used for cell transfection. Cell proliferation was detected by tetramethylazolium salt (MTT) method, apoptosis was detected by flow cytometry and cell migration and invasion were detected by Transwell assay. The protein expression of AP2M1, epidermal growth factor receptor (EGFR), p-phosphatidylinositol 3 kinase (PI3K), PI3K, p-protein kinase B (Akt) and AKT was detected by Western blot. Results Compared with control group, the relative expression of AP2M1 mRNA and protein in the AP2M1-shRNA group was decreased (P＜0.05). The relative cell viability was increased (P＜0.05). The cell apoptosis rate was decreased (P＜0.05). The counting number of migrating and invading cells was increased (P＜0.05). The relative expression level of EGFR protein and the phosphorylation level of PI3K and AKT were increased(P＜0.05). Compared with Control group, the expression of AP2M1 mRNA and protein relative expression level in AP2M1-LV group was increased (P＜0.05). The relative cell viability was decreased (P＜0.05). The cell apoptosis rate was increased (P＜0.05). The number of migrating and invading cells was decreased(P＜0.05). The relative expression level of EGFR protein and the phosphorylation level of PI3K and AKT were all decreased (P＜0.05). Conclusions The over-expression of AP2M1 partially inhibits the proliferation and invasion of DLBCL cells by inhibiting the EGFR/PI3K/AKT signaling pathway.

Objective To evaluate the regulatory effect of the adaptor related protein complex 2 subunit μ1 (AP2M1) on proliferation and invasion of diffuse large B-cell lymphoma (DLBCL). Methods Human diffuse large B-cell lymphoma cell line OCI-LY8 was aliquoted into control group, NC-shRNA group, AP2M1-shRNA group, NC-LV group, and AP2M1-LV group. Lipofectamine 2000 was used for cell transfection. Cell proliferation was detected by tetramethylazolium salt (MTT) method, apoptosis was detected by flow cytometry and cell migration and invasion were detected by Transwell assay. The protein expression of AP2M1, epidermal growth factor receptor (EGFR), p-phosphatidylinositol 3 kinase (PI3K), PI3K, p-protein kinase B (Akt) and AKT was detected by Western blot. Results Compared with control group, the relative expression of AP2M1 mRNA and protein in the AP2M1-shRNA group was decreased (P＜0.05). The relative cell viability was increased (P＜0.05). The cell apoptosis rate was decreased (P＜0.05). The counting number of migrating and invading cells was increased (P＜0.05). The relative expression level of EGFR protein and the phosphorylation level of PI3K and AKT were increased(P＜0.05). Compared with Control group, the expression of AP2M1 mRNA and protein relative expression level in AP2M1-LV group was increased (P＜0.05). The relative cell viability was decreased (P＜0.05). The cell apoptosis rate was increased (P＜0.05). The number of migrating and invading cells was decreased(P＜0.05). The relative expression level of EGFR protein and the phosphorylation level of PI3K and AKT were all decreased (P＜0.05). Conclusions The over-expression of AP2M1 partially inhibits the proliferation and invasion of DLBCL cells by inhibiting the EGFR/PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

One of the possible candidates for the treatment of diabetic cardiomyopathy is liraglutide, a glucagon-like peptide-1 receptor (GLP1R) agonist. In this study, the impacts of liraglutide on the integrin-linked kinase (ILK)-related PI3K/AKT axis in rats with type 2 diabetes induced via streptozotocin were examined. Twenty-four Wistar albino rats were distributed in four different groups, and a high-fat diet and streptozotocin were used to induce type 2 in two groups. Rats in the untreated control groups were administered 0.9% NaCl solution over a 6-week period, and those in the treatment groups were administered 0.9% NaCl for 3 weeks, followed by subcutaneous injection of liraglutide (150 μg/kg) for an additional 3 weeks. In the liraglutide-treated diabetic group, the heart-to-body weight ratio was significantly reduced, levels of cardiac biomarkers, troponin I and creatine-kinase-MB, were improved; activities of antioxidant enzymes, glutathione peroxidase and superoxide dismutase, were increased; and levels of malondialdehyde were decreased. Western blotting and immunohistochemical studies revealed increased levels of ILK, P-PI3K, P-AKT, and BCL2, as well as those of caspase 3, BAX, and P-PTEN, indicating mitigation of cardiomyocyte apoptosis. Our results show that liraglutide, by targeting GLP1Rs, enhances the expression of proteins in the ILK/PI3K/AKT/PTEN pathway and thereby exerts its cardioprotective effects in rats with DCM. [ABSTRACT FROM AUTHOR]

Objective To explore the effect of Huangqi Yanghe Decoction on wound healing of diabetic foot ulcer (DFU) rats based on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/nuclear factor- κB (NF- κB) signal pathway. Methods DFU rat model was constructed, and 48 rats successfully modeled were randomly divided into the model group, the Huangqi Yanghe Decoction low (8.5 g/kg) group, the Huangqi Yanghe Decoction high (17 g/kg) dose group and the Huangqi Yanghe Decoction high dose (17 g/kg)+LY294002 (PI3K/AKT pathway inhibitor, 0.3 mg/kg) group. There were 12 rats in each group. Another 12 rats were selected as the control group. Rats in each group were given corresponding drug intervention for 4 weeks. After the 14th and 28th day-administration, the general state and wound changes of rats were observed, and the wound healing rate was calculated. The fasting blood glucose (FBG) level of rats was measured, and the percutaneous partial pressure of oxygen (TcpO2) of tissue around the wound was detected. Serum levels of vascular endothelial growth factor (VEGF), hypoxia inducible factor-1α (HIF-1α), C-reactive protein (CRP) and interleukin (IL)-6 were determined by enzyme linked immunosorbent assay. Histopathological changes of the wound were observed by hematoxylin-eosin staining. Immunohistochemical staining was used to measure the microvascular density of rat wound tissue. The protein expression levels of PI3K, phosphorylated PI3K (p-PI3K), AKT, phosphorylated AKT (p-AKT), NF-κB p65, phosphorylated NF- κB p65 (p-NF- κB p65) and NF- κB inhibitory protein α (IκB- α) in rat wound tissue were determined by Western blot assay. Results Rats in the control group had smooth hair color, normal diet, drinking water and excretion, more active, wound healing fast, less inflammatory reaction in wound tissue, and there were more new blood vessels. Fibroblasts and collagen matrix were abundant in granulation tissue. In the model group, the fur color of rats was dull and matte, and the activity was reduced. The symptoms of polydipsia, polyphagia and polyuria were appeared in the model group, the wound color was dark, and edema and ulcer appeared in the surrounding tissue, a large number of inflammatory cells infiltrated in the wound tissue, accompanied by tissue necrosis and exudation, fewer neovascularization and fibroblasts were observed. Wound healing rate, TcpO2 in wound surrounding tissue, serum VEGF, HIF-1α, microvascular density, pPI3K, p-AKT and IκB-α protein expression levels in wound tissue were decreased, and FBG, serum CRP, IL-6, p-NF-κB p65 protein expression in wound tissue were increased (P＜0.05). Compared with the model group, the state of rats was gradually improved in the Huangqi Yanghe Decoction low and high dose groups, and the lesion degree of wound tissue was reduced successively, wound healing rate, TcpO2 in wound surrounding tissue, serum VEGF, HIF-1α, microvascular density, p-PI3K, p-AKT and IκB-α protein expression levels in wound tissue were increased in turn (P＜0.05). The FBG, serum CRP, IL-6 and p-NF- κB p65 protein expression in wound tissue were decreased in turn (P＜0.05). LY294002 could partially reverse the therapeutic effect of high-dose Huangqi Yanghe Decoction on DFU rats (P＜0.05). Conclusion Huangqi Yanghe Decoction can regulate PI3K/AKT/NF-κB pathway, inhibit inflammatory response in DFU rats, promote angiogenesis and thus promote wound healing. [ABSTRACT FROM AUTHOR]

This data report, published in Frontiers in Pharmacology, discusses leniolisib, a drug used to treat activated phosphoinositide 3-kinase delta (APDS) syndrome. Leniolisib inhibits the PI3Kδ pathway and has shown positive results in reducing lymphoproliferation and improving immune cell subsets in APDS patients. The report provides information on the development, synthesis, and physicochemical properties of leniolisib. It also mentions that leniolisib is being studied for its potential use in treating other autoimmune diseases associated with overactive PI3Kδ activity. The document also includes references to studies on the role of PI3Kδ in B cell development, the design of inhibitors for the PI3K signaling pathway, and the clinical spectrum and features of APDS. It also provides information on the availability of raw data, author contributions, funding, conflicts of interest, and a disclaimer regarding the views expressed in the article. [Extracted from the article]

Objective To observe the inhibitory effects of baicalin on lipopolysaccharide(LPS)-induced pyroptosis and inflammation of cardiomyocytes and to analyze their mechanism. Methods The rat cardiomyocytes H9C2 were cultivated in vitro and we divided cells in the logarithmic growth phase into the control group, LPS group, and baicalin group. The control group did not receive intervention. In the LPS group, H9C2 cells were stimulated by LPS to construct the cell injury models in vitro. In the baicalin group, cells were induced by LPS and then were added with 10, 20, 40, and 80 µmol/L baicalin, respectively. The cell viability in each group was detected by CCK-8. The concentration of baicalin with the highest cell viability was selected as the optimal concentration for subsequent experiments. Then, H9C2 cells in the logarithmic phase were divided into the control group, LPS group, baicalin group, inhibitor group, baicalin+inhibitor group, and baicalin+activator group. Except the control group without intervention, LPS stimulation was conducted to construct the cell injury models in the other groups; on this basis, the baicalin group was treated with baicalin, while the inhibitor group was treated with phosphatidylinositol 3-kinase(PI3K)/serine protein kinase(AKT) pathway inhibitor LY294002, the baicalin+inhibitor group was treated with baicalin and LY294002, and the baicalin+activator group was treated with baicalin and PI3K/AKT pathway activator insulin-like growth factor 1(IGF-1). The inflammatory cytokines interleukin(IL)-1β and IL-6 in the cell supernatant were detected by ELISA. Western blotting was used to detect the relative expression levels of pyroptosis-related proteins pyroporin D(GSDMD), nucleotide oligomeric domain-like receptor protein 3(NLRP3), Caspase-1, and PI3K/AKT pathway-related proteins PI3K, AKT, phosphorylation(p-) PI3K, and pAKT. Results Compared with the control group, the expression levels of inflammatory cytokines IL-6, IL-1β, pyroptosis-related proteins GSDMD, NLRP3, Caspase-1, p-PI3K and p-AKT increased in the LPS group. Compared with the LPS group, the expression levels of IL-6, IL-1β, GSDMD, NLRP3, Caspase-1, p-PI3K and p-AKT decreased in the baicalin group and inhibitor group. Compared with the baicalin group, the expression levels of IL-6, IL-1β, GSDMD, NLRP3, Caspase-1, p-PI3K and p-AKT decreased in the baicalin + inhibitor group, while the trend was opposite in the baicalin + activator group(all P<0. 05). Conclusion Baicalin could inhibit the inflammatory response and pyroptosis of rat cardiomyocytes in vitro induced by LPS, and the mechanism may be related to inhibiting the activation of PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]

Head and neck squamous cell carcinoma (HNSCC) is commonly associated with tobacco and alcohol consumption that induce a precancerous field, with phosphoinositide 3-kinase (PI3K) signaling being a common driver. However, the preclinical effectiveness of PI3K inhibitors has not necessarily translated to remarkable benefit in HNSCC patients. Thus, we sought to determine how precancerous keratinocytes influence HNSCC proliferation, cancer stem cell (CSC) maintenance, and response to PI3K inhibitors. We used the NOK keratinocyte cell line as a model of preneoplastic keratinocytes because it harbors two frequent genetic events in HNSCC, CDKN2A promoter methylation and TP53 mutation, but does not form tumors. NOK cell coculture or NOK cell-conditioned media promoted HNSCC proliferation, PI3K inhibitor resistance, and CSC phenotypes. SOMAscan-targeted proteomics determined the relative levels of >1300 analytes in the media conditioned by NOK cells and HNSCC cells ± PI3K inhibitor. These results demonstrated that NOK cells release abundant levels of ligands that activate epidermal growth factor receptor (EGFR) and fibroblast growth factor receptor (FGFR), two receptor tyrosine kinases with oncogenic activity. Inhibition of EGFR, but not FGFR, blunted PI3K inhibitor resistance and CSC phenotypes induced by NOK cells. Our results demonstrate that precancerous keratinocytes can directly support neighboring HNSCC by activating EGFR. Importantly, PI3K inhibitor sensitivity was not necessarily a cancer cell-intrinsic property, and the tumor microenvironment impacts therapeutic response and supports CSCs. Additionally, combined inhibition of EGFR with PI3K inhibitor diminished EGFR activation induced by PI3K inhibitor and potently inhibited cancer cell proliferation and CSC maintenance.

Parsaclisib is a potent and highly selective PI3Kδ inhibitor that has shown clinical benefit with monotherapy in a phase 2 study in relapsed or refractory (R/R) follicular lymphoma (FL). CITADEL‐102 (NCT03039114), a phase 1, multicenter study, assessed the efficacy of parsaclisib in combination with obinutuzumab and bendamustine in patients with R/R FL. Patients were ≥18 years of age with histologically confirmed and documented CD20‐positive FL, and R/R to previous rituximab‐containing treatment regimens. Part one (safety run‐in) determined the maximum tolerated dose of parsaclisib in combination with standard dosage regimens of obinutuzumab and bendamustine. Part two (dose expansion) was an open‐label, single‐group design evaluating safety, tolerability (primary endpoint), and efficacy (secondary endpoint) of parsaclisib combination therapy. Twenty‐six patients were enrolled in CITADEL‐102 and all patients received parsaclisib 20 mg once daily for 8 weeks, followed by 20 mg once weekly thereafter, in combination with obinutuzumab and bendamustine. One patient in safety run‐in experienced a dose‐limiting toxicity of grade 4 QT interval prolongation that was considered related to parsaclisib. Eight patients (30.8%) discontinued treatment due to treatment‐emergent adverse events (TEAEs) of colitis (2 [7.7%]), alanine aminotransferase and aspartate aminotransferase increase (both in one patient [3.8%]), neutropenia, thrombocytopenia, QT prolongation, tonsil cancer, and maculopapular rash (each 1 [3.8%]). The most common reported TEAEs were pyrexia (53.8%), neutropenia (50.0%), and diarrhea (46.2%). Twenty‐three patients (88.5%) experienced grade 3 or 4 TEAEs; the most common were neutropenia (34.6%), febrile neutropenia (23.1%), and thrombocytopenia (19.2%). Seventeen patients (65.4%) had a complete response and 3 patients (11.5%) had a partial response, for an objective response rate of 76.9%. Overall, results from CITADEL‐102 suggest that the combination of parsaclisib with obinutuzumab and bendamustine did not result in unexpected safety events, with little evidence of synergistic toxicity, and demonstrated preliminary efficacy in patients with R/R FL who progressed following prior rituximab‐containing regimens. [ABSTRACT FROM AUTHOR]

Graphical AbstractIC50 = 11 nM (PI3Kδ); 244 nM (PI3Kα); 424 nM (PI3Kβ), 2,230 nM (PI3Kγ).

Abstract Background Klippel–Trenaunay syndrome (KTS) is a rare slow-flow combined vascular malformation with limb hypertrophy. KTS is thought to lie on the PIK3CA-related overgrowth spectrum, but reports are limited. PIK3CA encodes p110α, a catalytic subunit of phosphatidylinositol 3-kinase (PI3K) that plays an essential role in the PI3K/AKT/mammalian target of rapamycin (mTOR) signaling pathway. We aimed to demonstrate the clinical utility of targeted next-generation sequencing (NGS) in identifying PIK3CA mosaicism in archival formalin-fixed paraffin-embedded (FFPE) tissues from patients with KTS. Results Participants were 9 female and 5 male patients with KTS diagnosed as capillaro-venous malformation (CVM) or capillaro-lymphatico-venous malformation (CLVM). Median age at resection was 14 years (range, 5–57 years). Median archival period before DNA extraction from FFPE tissues was 5.4 years (range, 3–7 years). NGS-based sequencing of PIK3CA achieved an amplicon mean coverage of 119,000x. PIK3CA missense mutations were found in 12 of 14 patients (85.7%; 6/8 CVM and 6/6 CLVM), with 8 patients showing the hotspot variants E542K, E545K, H1047R, and H1047L. The non-hotspot PIK3CA variants C420R, Q546K, and Q546R were identified in 4 patients. Overall, the mean variant allele frequency for identified PIK3CA variants was 6.9% (range, 1.6–17.4%). All patients with geographic capillary malformation, histopathological lymphatic malformation or macrodactyly of the foot had PIK3CA variants. No genotype–phenotype association between hotspot and non-hotspot PIK3CA variants was found. Histologically, the vessels and adipose tissues of the lesions showed phosphorylation of the proteins in the PI3K/AKT/mTOR signaling pathway, including p-AKT, p-mTOR, and p-4EBP1. Conclusions The PI3K/AKT/mTOR pathway in mesenchymal tissues was activated in patients with KTS. Amplicon-based targeted NGS could identify low-level mosaicism from low-input DNA extracted from FFPE tissues, potentially providing a diagnostic option for personalized medicine with inhibitors of the PI3K/AKT/mTOR signaling pathway.

Abstract Background Endometriosis is still a difficult problem for women. The Nuclear Ubiquitous Casein and cyclin-dependent Kinase Substrate 1 (NUCKS1) gene is located on human chromosome 1q32.1. It encodes the NUCKS1 protein, a 27 kDa nuclear DNA binding protein that plays an important role in cell growth and proliferation. NUCKS1 plays an important role in the development of many diseases. However, its role in endometriosis is unclear. Methods Ectopic endometrial tissues and normal tissue specimens were collected, and the expression of NUCKS1, NF-κB and PI3K was detected by RT-qPCR and immunohistochemistry. Inhibition of NUCKS1 in hEM15A cells, study the changes in cell viability, apoptosis, migration and protein expression by CCK8 assay, flow cytometry, wound-healing assay, western blot and ELISA techniques. The comparison of differences between the two groups was implemented using unpaired sample t test or Mann-whitney U test. One-way analysis of variance or Kruskal-wallis test was used for comparisons among the three groups. Results (1) NUCKS1 is highly expressed in endometriosis tissues. (2) Inhibition of NUCKS1 decreases cell viability and capability of migration, and increases apoptosis in endometriosis cells. (3) Expressions of NF-κB and PI3K are increased in endometriosis tissues, and inhibition of NUCKS1 decreases the expression levels of PI3K and NF-κB in endometriosis cells. (4) Inhibition of NUCKS1 decreases the expression of VEGF. Conclusion (1) NUCKS1 is overexpressed in endometriosis, and inhibition of NUCKS1 inhibits cell viability and capability of migration, and increases apoptosis. (2) NUCKS1 promotes the progress of endometriosis through activating PI3K and NF-κB pathways, and VEFG is also involved in this process.

The sterile inflammatory response mediated by Toll-like receptors (TLRs) 4 and 9 is implicated in the massive hepatic damage caused by acetaminophen (APAP)- overdose. There is a crosstalk between TLR-dependent signaling with other intracellular kinases like phosphatidylinositol 3-kinases (PI3Ks). Nevertheless, the detailed role of PI3Kα is still unknown in hepatic sterile inflammation. Accordingly, the effect of the novel PI3Kα inhibitor alpelisib was investigated in the setting of APAP-driven sterile inflammation in the liver. This was examined by pretreating mice with alpelisib (5 and 10 mg/kg, oral) 2 h before APAP (500 mg/kg, i.p.)-intoxication. The results indicated that alpelisib dose-dependently lowered APAP-induced escalation in serum liver function biomarkers and hepatic necroinflammation score. Alpelisib also attenuated APAP-induced rise in cleaved caspase 3 and proliferating cell nuclear antigen (PCNA) in the liver hepatocytes, as indices for apoptosis and proliferation. Mechanistically, inhibition of PI3Kα by alpelisib limited APAP-induced overproduction of the pro-inflammatory tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 in the blood circulation via switching off the activation of several signal transduction proteins, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), signal transducer and activator of transcription-3 (Stat-3), glycogen Synthase Kinase (GSK)-3ß and nuclear factor (NF)-κB. Alpelisib also impaired APAP-instigated immune cell infiltration in the liver via reducing systemic granulocyte/macrophage-colony stimulating factor (GM-CSF) release and reversed APAP-induced abnormalities in the systemic and hepatic levels of the anti-inflammatory IL-10 and IL-22. In conclusion, selective modulation of the PI3Kα activity by alpelisib can hinder the inflammatory response and infiltration of immune cells occurring by APAP-hepatotoxicity. [ABSTRACT FROM AUTHOR]

Background: Klippel–Trenaunay syndrome (KTS) is a rare slow-flow combined vascular malformation with limb hypertrophy. KTS is thought to lie on the PIK3CA-related overgrowth spectrum, but reports are limited. PIK3CA encodes p110α, a catalytic subunit of phosphatidylinositol 3-kinase (PI3K) that plays an essential role in the PI3K/AKT/mammalian target of rapamycin (mTOR) signaling pathway. We aimed to demonstrate the clinical utility of targeted next-generation sequencing (NGS) in identifying PIK3CA mosaicism in archival formalin-fixed paraffin-embedded (FFPE) tissues from patients with KTS. Results: Participants were 9 female and 5 male patients with KTS diagnosed as capillaro-venous malformation (CVM) or capillaro-lymphatico-venous malformation (CLVM). Median age at resection was 14 years (range, 5–57 years). Median archival period before DNA extraction from FFPE tissues was 5.4 years (range, 3–7 years). NGS-based sequencing of PIK3CA achieved an amplicon mean coverage of 119,000x. PIK3CA missense mutations were found in 12 of 14 patients (85.7%; 6/8 CVM and 6/6 CLVM), with 8 patients showing the hotspot variants E542K, E545K, H1047R, and H1047L. The non-hotspot PIK3CA variants C420R, Q546K, and Q546R were identified in 4 patients. Overall, the mean variant allele frequency for identified PIK3CA variants was 6.9% (range, 1.6–17.4%). All patients with geographic capillary malformation, histopathological lymphatic malformation or macrodactyly of the foot had PIK3CA variants. No genotype–phenotype association between hotspot and non-hotspot PIK3CA variants was found. Histologically, the vessels and adipose tissues of the lesions showed phosphorylation of the proteins in the PI3K/AKT/mTOR signaling pathway, including p-AKT, p-mTOR, and p-4EBP1. Conclusions: The PI3K/AKT/mTOR pathway in mesenchymal tissues was activated in patients with KTS. Amplicon-based targeted NGS could identify low-level mosaicism from low-input DNA extracted from FFPE tissues, potentially providing a diagnostic option for personalized medicine with inhibitors of the PI3K/AKT/mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

Background: Endometriosis is still a difficult problem for women. The Nuclear Ubiquitous Casein and cyclin-dependent Kinase Substrate 1 (NUCKS1) gene is located on human chromosome 1q32.1. It encodes the NUCKS1 protein, a 27 kDa nuclear DNA binding protein that plays an important role in cell growth and proliferation. NUCKS1 plays an important role in the development of many diseases. However, its role in endometriosis is unclear. Methods: Ectopic endometrial tissues and normal tissue specimens were collected, and the expression of NUCKS1, NF-κB and PI3K was detected by RT-qPCR and immunohistochemistry. Inhibition of NUCKS1 in hEM15A cells, study the changes in cell viability, apoptosis, migration and protein expression by CCK8 assay, flow cytometry, wound-healing assay, western blot and ELISA techniques. The comparison of differences between the two groups was implemented using unpaired sample t test or Mann-whitney U test. One-way analysis of variance or Kruskal-wallis test was used for comparisons among the three groups. Results: (1) NUCKS1 is highly expressed in endometriosis tissues. (2) Inhibition of NUCKS1 decreases cell viability and capability of migration, and increases apoptosis in endometriosis cells. (3) Expressions of NF-κB and PI3K are increased in endometriosis tissues, and inhibition of NUCKS1 decreases the expression levels of PI3K and NF-κB in endometriosis cells. (4) Inhibition of NUCKS1 decreases the expression of VEGF. Conclusion: (1) NUCKS1 is overexpressed in endometriosis, and inhibition of NUCKS1 inhibits cell viability and capability of migration, and increases apoptosis. (2) NUCKS1 promotes the progress of endometriosis through activating PI3K and NF-κB pathways, and VEFG is also involved in this process. [ABSTRACT FROM AUTHOR]

This study aimed to assess the effects of exogenous hydrogen sulfide (H2S) on abdominal aorta coarctation (AAC) induced myocardial fibrosis (MF) and autophagy in rats. Forty-four Sprague–Dawley rats were randomly divided into control group, AAC group, AAC + H2S group, and H2S control group. After a model of rats with AAC was built surgically, AAC + H2S group and H2S group were injected intraperitoneally with H2S (100 μmol/kg) daily. The rats in the control group and the AAC group were injected with the same amount of PBS. We observed that H2S can improve left ventricular function and the deposition of myocardial collagen fibers, inhibit pyroptosis, down-regulate the expression of P-eif2α in myocardial tissue, and inhibit cell autophagy by activating the phosphatidylinositol 3-kinase (PI3K)/AKT1 signaling pathway (p < 0.05). In addition, angiotensin II (1 μM) H9c2 cardiomyocytes were injured in vitro experiments, and it was also observed that pyroptosis was inhibited after H2S (400 μmol/kg) intervention, the expression of P-eif2α in cardiomyocytes was significantly down-regulated, and the PI3K/AKT1 signaling pathway was activated at the same time. Therefore, increasing the expression of P-eif2α reverses the activation of the PI3K/AKT1 signaling pathway by H2S. In conclusion, these findings suggest that exogenous H2S can ameliorate MF in rats with AAC by inhibiting pyroptosis, and the mechanism may be associated with inhibiting the phosphorylation of eif2α and activating the PI3K/AKT1 signaling pathway to inhibit excessive cell autophagy. [ABSTRACT FROM AUTHOR]

Objective: To discuss the potential mechanism of Panax quinquefolium triolsaponins (PQTS) in the occurrence and development of ischemic stroke by using network pharmacology and molecular docking technique. Methods: The potential targets of PQTS acing on IS were obtained through Swiss Target Prediction Database, Encyclopedia of Traditional Chinese Medicine (ETCM) Database, SEA Search Server Database, DisGeNET Database, and so on; the protein-protein interaction (PPI) network diagram of the key potential targets was established by STRING Database and Cytoscape 3. 9. 1 software; the core tagets of PQTS acting on IS were got by topology network analysis; Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis were used to analyze the potential targets through DAVID online analysis website; the PQTStarget-signaling pathway network was constructed by Cytoscape 3. 9. 1 software and the topology network analysis was used to obtain the potential main active compositions; AutoDock Vina software was used to verify the molecular docking between the active ingredients and core targets. Results : There were 122 potential targets of PQTS acting on IS; the GO function enrichment analysis was mainly included the regulation of apoptosis, intracellular signal transduction process, and regulations of extracellular substances by cells; the KEGG function analysis included the interleukins signaling pathways, phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signaling pathway and phosphatidylinositol 3 kinase-protein kinase B-mammalian target of rapamycin (PI3K-Akt-mTOR) signaling pathway. The molecular docking analysis results showed that pseudo-ginsenoside F11, 20 (S)-protopanaxatriol, ginsenoside Rg1, ginsenoside Rh1, pseudo-ginsenoside RT5, and ginsenoside Re could form the stable conformations with signal transducer and activator of transcription 3 (STAT3), phosphatidylinositol 3-kinases catalytic suburit α (PIK3CA), epidermal growth factor receptor (EGFR), and mitogen-activated protein kinase 14 (MAPK14) with lower binding energy. Conclusion: The protective effect of PQTS on IS may be related to the STAT3, PIK3CA, EGFR, MAPK14, and PI3K/Akt signaling pathways. [ABSTRACT FROM AUTHOR]

Precision medicine in oncology leverages clinical observations of exceptional response. Toward an understanding of the molecular features that define this response, we applied an integrated, multiplatform analysis of RNA profiles derived from clinically annotated glioblastoma samples. This analysis suggested that specimens from exceptional responders are characterized by decreased accumulation of microglia/macrophages in the glioblastoma microenvironment. Glioblastoma-associated microglia/macrophages secreted interleukin 11 (IL11) to activate STAT3-MYC signaling in glioblastoma cells. This signaling induced stem cell states that confer enhanced tumorigenicity and resistance to the standard-of-care chemotherapy, temozolomide (TMZ). Targeting a myeloid cell restricted an isoform of phosphoinositide-3-kinase, phosphoinositide-3-kinase gamma isoform (PI3Kγ), by pharmacologic inhibition or genetic inactivation disrupted this signaling axis by reducing microglia/macrophage-associated IL11 secretion in the tumor microenvironment. Mirroring the clinical outcomes of exceptional responders, PI3Kγ inhibition synergistically enhanced the anti-neoplastic effects of TMZ in orthotopic murine glioblastoma models. Moreover, inhibition or genetic inactivation of PI3Kγ in murine glioblastoma models recapitulated expression profiles observed in clinical specimens isolated from exceptional responders. Our results suggest key contributions from tumor-associated microglia/macrophages in exceptional responses and highlight the translational potential for PI3Kγ inhibition as a glioblastoma therapy.

Autophagy is a cellular process that degrades cytoplasmic cargo by engulfing it in a double-membrane vesicle, known as the autophagosome, and delivering it to the lysosome. The ATG12-5-16L1 complex is responsible for conjugating members of the ubiquitin-like ATG8 protein family to phosphatidylethanolamine in the growing autophagosomal membrane, known as the phagophore. ATG12-5-16L1 is recruited to the phagophore by a subset of the phosphatidylinositol 3-phosphate-binding seven-bladedß -propeller WIPI proteins. We determined the crystal structure of WIPI2d in complex with the WIPI2 interacting region (W2IR) of ATG16L1 comprising residues 207-230 at 1.85 Å resolution. The structure shows that the ATG16L1 W2IR adopts an alpha helical conformation and binds in an electropositive and hydrophobic groove between WIPI2 ß-propeller blades 2 and 3. Mutation of residues at the interface reduces or blocks the recruitment of ATG12-5-16 L1 and the conjugation of the ATG8 protein LC3B to synthetic membranes. Interface mutants show a decrease in starvation-induced autophagy. Comparisons across the four human WIPIs suggest that WIPI1 and 2 belong to a W2IR-binding subclass responsible for localizing ATG12-5-16 L1 and driving ATG8 lipidation, whilst WIPI3 and 4 belong to a second W34IR-binding subclass responsible for localizing ATG2, and so directing lipid supply to the nascent phagophore. The structure provides a framework for understanding the regulatory node connecting two central events in autophagy initiation, the action of the autophagic PI 3-kinase complex on the one hand and ATG8 lipidation on the other.

ObjectiveAIBP (apolipoprotein A-I binding protein) is an effective and selective regulator of lipid rafts modulating many metabolic pathways originating from the rafts, including inflammation. The mechanism of action was suggested to involve stimulation by AIBP of cholesterol efflux, depleting rafts of cholesterol, which is essential for lipid raft integrity. Here we describe a different mechanism contributing to the regulation of lipid rafts by AIBP. Approach and Results: We demonstrate that modulation of rafts by AIBP may not exclusively depend on the rate of cholesterol efflux or presence of the key regulator of the efflux, ABCA1 (ATP-binding cassette transporter A-I). AIBP interacted with phosphatidylinositol 3-phosphate, which was associated with increased abundance and activation of Cdc42 and rearrangement of the actin cytoskeleton. Cytoskeleton rearrangement was accompanied with reduction of the abundance of lipid rafts, without significant changes in the lipid composition of the rafts. The interaction of AIBP with phosphatidylinositol 3-phosphate was blocked by AIBP substrate, NADPH (nicotinamide adenine dinucleotide phosphate), and both NADPH and silencing of Cdc42 interfered with the ability of AIBP to regulate lipid rafts and cholesterol efflux.ConclusionsOur findings indicate that an underlying mechanism of regulation of lipid rafts by AIBP involves PIP-dependent rearrangement of the cytoskeleton.

One of the possible candidates for the treatment of diabetic cardiomyopathy is liraglutide, a glucagon-like peptide-1 receptor (GLP1R) agonist. In this study, the impacts of liraglutide on the integrin-linked kinase (ILK)-related PI3K/AKT axis in rats with type 2 diabetes induced via streptozotocin were examined. Twenty-four Wistar albino rats were distributed in four different groups, and a high-fat diet and streptozotocin were used to induce type 2 in two groups. Rats in the untreated control groups were administered 0.9% NaCl solution over a 6-week period, and those in the treatment groups were administered 0.9% NaCl for 3 weeks, followed by subcutaneous injection of liraglutide (150 μg/kg) for an additional 3 weeks. In the liraglutide-treated diabetic group, the heart-to-body weight ratio was significantly reduced, levels of cardiac biomarkers, troponin I and creatine-kinase-MB, were improved; activities of antioxidant enzymes, glutathione peroxidase and superoxide dismutase, were increased; and levels of malondialdehyde were decreased. Western blotting and immunohistochemical studies revealed increased levels of ILK, P-PI3K, P-AKT, and BCL2, as well as those of caspase 3, BAX, and P-PTEN, indicating mitigation of cardiomyocyte apoptosis. Our results show that liraglutide, by targeting GLP1Rs, enhances the expression of proteins in the ILK/PI3K/AKT/PTEN pathway and thereby exerts its cardioprotective effects in rats with DCM.

The sterile inflammatory response mediated by Toll-like receptors (TLRs) 4 and 9 is implicated in the massive hepatic damage caused by acetaminophen (APAP)-overdose. There is a crosstalk between TLR-dependent signaling with other intracellular kinases like phosphatidylinositol 3-kinases (PI3Ks). Nevertheless, the detailed role of PI3Kα is still unknown in hepatic sterile inflammation. Accordingly, the effect of the novel PI3Kα inhibitor alpelisib was investigated in the setting of APAP-driven sterile inflammation in the liver. This was examined by pretreating mice with alpelisib (5 and 10 mg/kg, oral) 2 h before APAP (500 mg/kg, i.p.)-intoxication. The results indicated that alpelisib dose-dependently lowered APAP-induced escalation in serum liver function biomarkers and hepatic necroinflammation score. Alpelisib also attenuated APAP-induced rise in cleaved caspase 3 and proliferating cell nuclear antigen (PCNA) in the liver hepatocytes, as indices for apoptosis and proliferation. Mechanistically, inhibition of PI3Kα by alpelisib limited APAP-induced overproduction of the pro-inflammatory tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in the blood circulation via switching off the activation of several signal transduction proteins, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), signal transducer and activator of transcription-3 (Stat-3), glycogen Synthase Kinase (GSK)-3β and nuclear factor (NF)-κB. Alpelisib also impaired APAP-instigated immune cell infiltration in the liver via reducing systemic granulocyte/macrophage-colony stimulating factor (GM-CSF) release and reversed APAP-induced abnormalities in the systemic and hepatic levels of the anti-inflammatory IL-10 and IL-22. In conclusion, selective modulation of the PI3Kα activity by alpelisib can hinder the inflammatory response and infiltration of immune cells occurring by APAP-hepatotoxicity.

Background: N-linoleyltyrosine (NITyr), one of the anandamide analogs, exerts activity via the endocannabinoid receptors (CB1 and CB2), which showed antitumor effects in various tumors. Therefore, we speculated that NITyr might show anti-non-small cell lung cancer (NSCLC) effects via the CB1 or CB2 receptor. The purpose of the investigation was to reveal the anti-tumor ability of NITyr on A549 cells and its mechanisms. Methods: The viability of A549 cells was measured by MTT assay, and the cell cycle and apoptosis were both examined by flow cytometry; in addition, cell migration was tested by wound healing assay. Apoptosis-related markers were measured by immunofluorescence. The downstream signaling pathways (PI3K, ERK, and JNK) of CB1 or CB2 were examined through Western blotting. The expressions of CB1 and CB2 were detected by immunofluorescence. Finally, the AutoDock software was used to validate the binding affinity between the targets, such as CB1 and CB2, with NITyr. Results: We found that NITyr inhibited cell viability, hindered the cell cycle, resulted in apoptosis, and inhibited migration. The CB1 inhibitor, AM251, and the CB2 inhibitor, AM630, weakened the aforementioned phenomenon. The immunofluorescence assay suggested that NITyr upregulated the expression of CB1 and CB2. Western blot analysis indicated that NITyr upregulated the expression of p-ERK, downregulated the expression of p-PI3K, and did not affect p-JNK expression. In conclusion, NITyr showed a role in inhibiting NSCLC through the activation of CB1 and CB2 receptors involved in PI3K and ERK pathways. [ABSTRACT FROM AUTHOR]

Objective To explore the effect of Taohe Chengqi Decoction on phosphatidylinositol 3-kinase (PI3K)/serine/threonine protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway in ovarian tissue of a rat model of polycystic ovary syndrome (PCOS). Methods Ninety-six female SD rats were randomly divided into the control group, the PCOS group, the metformin group, the Taohe Chengqi Decoction low, medium and high dose groups, with 16 rats in each group. Except for the control group, the PCOS model was constructed by subcutaneous injection of dehydroepiandrosterone in the other groups. After successful model building, the metformin group was given 0.1 g/kg metformin by gavage, and the Taohe Chengqi Decoction low, medium and high dose groups were given 1.89 g/kg, 3.78 g/kg and 7.56 g/kg Taohe Chengqi Decoction by gavage, respectively. The control group and the PCOS group were given the same amount of normal saline by gavage, 1 time/d for 28 days. The fasting blood glucose (FPG) level was measured by blood glucose meter. Enzyme linked immunosorbent assay was used to detect serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), testosterone (T) and fasting insulin (FINS). Insulin resistance index (HOMA-IR) was calculated. Hematoxylin eosin staining was used to observe the histopathology of rat ovary. The mRNA and protein expression of PI3K/AKT/mTOR pathway in rat ovarian tissue were detected by fluorescence quantitative PCR and Western blot assay. Results Compared with the control group, the volume of follicles increased, contained follicular fluid, and the cell space increased, corpus luteum decreased, and the cell layer of theca proliferated, serum FSH, E2, mRNA and protein expression of PI3K/AKT/mTOR pathway in ovarian tissue were decreased, and levels of LH, T, FPG, FINS and HOMA-IR were increased in the PCOS group (P＜0.05). In the metformin group and the Taohe Chengqi Decoction low-dose, medium-dose and high-dose groups, the volume of follicles decreased, the number of corpus luteum increased, and the proliferation of alveolar cell layer decreased. Serum levels of FSH, E2, mRNA and protein expression of PI3K/AKT/mTOR pathway in ovarian tissue were increased, and levels of LH, T, FPG, FINS and HOMA-IR were decreased (P＜0.05). Changes of above indexes were dose-dependent in the low, medium and high dose groups of Taohe Chengqi Decoction. There were significant differences in changes of above indexes between the Taohe Chengqi Decoction low, medium-dose groups and the metformin group. There was no significant difference between the high-dose of Taohe Chengqi Decoction group and the metformin group. Conclusion Taohe Chengqi Decoction can restore normal levels of sex hormones and inhibit insulin resistance in PCOS model rats, which may be related to its promotion of PI3K/AKT/mTOR signal pathway activation. [ABSTRACT FROM AUTHOR]

Background: It has been shown that glycolytic protein ENO1 (alpha-enolase) contributes to the pathogenesis of pulmonary hypertension through acting smooth muscle cells; however, the roles of ENO1-caused endothelial and mitochondrial dysfunctions in Group 3 pulmonary hypertension remain unexplored. Methods: PCR array and RNA sequencing were used to screen and decipher the differential gene expression by hypoxia-treated human pulmonary artery endothelial cells. Techniques of small-interfering RNA, specific inhibitor and plasmids carrying gene of ENO1, interventions with specific inhibitor and AAV-ENO1 delivery were employed to explore the role of ENO1 in hypoxic pulmonary hypertension in vitro and in vivo, respectively. Assays for cell proliferation, angiogenesis, and adhesion were employed to analyze cell behaviors, while seahorse analysis was used to measure mitochondrial function of human pulmonary artery endothelial cells. Results: PCR array data showed that ENO1 expression increased in human pulmonary artery endothelial cells exposed to hypoxia, as well as in lung tissues from patients with chronic obstructive lung disease-associated pulmonary hypertension and murine model of hypoxic pulmonary hypertension. Inhibition of ENO1 restored the hypoxia-induced endothelial dysfunction, including excessive proliferation, angiogenesis, and adhesion, while overexpression of ENO1 promotes these disorders of human pulmonary artery endothelial cells. RNA-seq showed that ENO1 targets mitochondrion-related genes and PI3K-Akt signaling pathway, which were validated in vitro and in vivo. Mice treated with ENO1 inhibitor exhibited ameliorated pulmonary hypertension and improved right ventricular failure induced by hypoxia. A reversal effect was observed in mice exposed to hypoxia and inhaled adeno-associated virus overexpressing ENO1. Conclusions: These results indicate that hypoxic pulmonary hypertension is associated with an increased level of ENO1 and that targeting ENO1 might reduce experimental hypoxic pulmonary hypertension by improving endothelial and mitochondrial dysfunction via PI3K-Akt-mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

This review explores the role of G protein-coupled estrogen receptor (GPER) and its interaction with the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinase (ERK), phosphatidylinositol 3-kinase (PI3K)/AKT, and hedgehog (Hh) signaling pathways in prostate cancer (PCa) progression. PCa is a significant health problem globally, and GPER has been shown to play a crucial role in the regulation of cell proliferation and survival in PCa cells. The MAPK/ERK pathway, activated by GPER, is involved in cell growth and differentiation, while the PI3K/AKT pathway influences cell cycle progression and survival. The Hh pathway is important for prostate development and has been implicated in the formation and progression of PCa. However, the specific contributions of GLI1 and GLI2 factors and their correlation with Gleason scores and GPER expression in PCa are not yet fully understood. Understanding the interactions between GPER and these signaling pathways provides valuable insights for the development of targeted therapies for PCa. Further research is needed to explore the potential of GLI1 and GLI2 factors and their relationship with GPER expression in PCa progression and metastasis. [ABSTRACT FROM AUTHOR]

Objective To explore the regulatory role of PI3K/Akt signaling pathway in the hypertrophic scar. Methods The rabbit ear hypertrophic scar model was established and randomly divided into four groups, including normal rabbit ear skin group (A), hypertrophic scar model group (B), DMSO-stimulated hypertrophic scar model group (C), and PI3K-specific inhibitor (LY294002) stimulated model group (D). Morphological changes of rabbit ear scar were observed by histopathology. The expression of phosphorylation of Akt[S473] and Akt[T308] was examined by immunohistochemistry and western blot. Immunofluorescence double staining was performed to assess the co-localization of p-Akt[S473] and p-Akt[T308]. The TUNEL assay was used to detect the fibroblasts' apoptosis in skin tissues. Results Scar thickness and fibroblast count were significantly higher in the D group than that in the B and C groups, but the difference was not statistically significant between the B group and the C group. The results of immunohistochemistry and western blot assay confirmed that the expression of p-Akt[T308] and p-Akt[S473] were significantly higher in the B group than in the A group, but significantly lower in the D group than in the C group. The expression of p-Akt[T308] and p-Akt[S473] was significantly enhanced in the B group compared with the C group. Immunofluorescence double staining showed that p-Akt[T308] and p-Akt[S473] were mainly co-localized in the cytoplasm in scar tissue, and their co-expression was lower in the D group than in the B group. TUNEL assay results confirmed that the apoptosis level of fibroblast was significantly downregulated in skin tissue in the B group compared with the A group, and the apoptosis level of fibroblast was higher in the D group than in the C (P < 0.01). Conclusion Blocking the PI3K/Akt signaling pathway can accelerate fibroblast apoptosis by inhibiting Akt phosphorylation, and then alleviate hypertrophic scar formation. [ABSTRACT FROM AUTHOR]

Objective To observe the effect of curcumin on proliferation and invasion of the gastric cancer MGC-803 cells in vitro and in vivo, and to discuss the mechanism of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian rapamycin target protein (mTOR) signaling pathway. Methods The gastric cancer MGC-803 cells were divided into control group, low dose (10 μmol·L－1) of curcumin group, middle dose (20 μmol·L－1) of curcumin group，and high dose (40 μmol·L－1) of curcumin group. The proliferation abilities of the MGC-803 cells in various groups were detected by CCK-8 method；the invasion abilities of the MGC-803 cells in various groups were detected by Transwell method；the apoptotic rates of the MGC-803 cells were detected by flow cytometry. The MGC-803 cells were used to establish the gastric cancer transplanted tumor model and the mice were treated with low (0.5 mg·kg－1), middle (1.0 mg·kg－1)，and high (2.0 mg·kg－1) doses of curcumin, and the inhibitory effect of curcumin on the tumor growth of the mice in various groups was observed. TUNEL method was used to detect the apoptosis of the tumor cells in transplanted tumor tissue of the mice in various groups；the expression levels of PI3K-Akt-mTOR signaling pathway proteins in transplacted tumor tissue of the mice in various groups were detected by Western blotting ththod. Results Compared with control group, the proliferation abilities and number of invasion cells in different doses of curcumin groups were significantly decreased (P<0.05), and the apoptotic rates were significantly increased (P<0.05)； the cell proliferation ability and the number of invasion cells in low dose of curcumin group were significantly higher than those in high dose of curcumin group (P<0.05), and the apoptotic rate was significantly lower than that in high dose of curcumin group (P<0.05). Compared with control group, the volumes of transplanted tumor of the mice in different doses of curcumin groups were significantly decreased (P<0.05). The apoptotic rate of the cells in transplanted tumor tissue of the mice in control group was low, and the apoptotic rate of the cells in transplated tumor tissue of the mice in middle dose of curcumin group was significantly higher than that in low dose of curcumin group (P<0.05), while the apoptotic rate of the cells in transplated tumor tissue of the mice in high dose of curcumin group was significantly higher than that in middle dose of curcumin group (P<0.05). Compared with control group, the proteins expression levels of PI3K, phosphorylated Akt (p-Akt) and phosphorylated mTOR (p-mTOR) in transplanted tumor tissue of the mice in different doses of curcumin groups were significantly decreased (P<0.05)；the expression levels of PI3K, p-Akt, and p-mTOR proteins in transplanted tumor tissue of the mice in low dose of curcumin group were significantly lower than those in middle dose of curcumin group (P<0.05)； the expression levels of PI3K, p-Akt, and p-mTOR proteins in transplanted tumor tissue of the mice in high dose of curcumin group were significantly lower than those in middle dose of curcumin group (P<0.05). Conclusion Curcumin can effectively inhibit the proliferation of tumor, decrease the invasion ability, and increase the apoptotic rate of the tumor cells；its mechanism may be related to down-regulation of expressions of PI3K/Akt/mTOR signaling pathway proteins. [ABSTRACT FROM AUTHOR]

The cytokine oncostatin M (OSM) is regarded as a critical mediator in various inflammatory responses. While the gaseous signaling molecule hydrogen sulfide (H2S) plays a role in a variety of pathophysiological conditions, such as hypertension, inflammatory pain, osteoarthritis, ischemic stroke, oxidative stress, retinal degeneration, and inflammatory responses, the underlying mechanism of H2S action on OSM expression in neutrophils needs to be clarified. In this work, we studied how H2S reduces OSM expression in neutrophil-like differentiated (d)HL-60 cells. To evaluate the effects of H2S, sodium hydrosulfide (NaHS, a donor that produces H2S), ELISA, real-time PCR (qPCR), immunoblotting, and immunofluorescence staining were utilized. Although exposure to granulocyte–macrophage colony-stimulating factor (GM-CSF) resulted in upregulated levels of production and mRNA expression of OSM, these upregulated levels were reduced by pretreatment with NaHS in dHL-60 cells. Similarly, the same pretreatment lowered phosphorylated levels of phosphatidylinositol 3-kinase, Akt, and nuclear factor-kB that had been elevated by stimulation with GM-CSF. Overall, our results indicated that H2S could be a therapeutic agent for inflammatory disorders via suppression of OSM. [ABSTRACT FROM AUTHOR]

Background: N-linoleyltyrosine (NITyr), one of the anandamide analogs, exerts activity via the endocannabinoid receptors (CB1 and CB2), which showed anti-tumor effects in various tumors. Therefore, we speculated that NITyr might show anti-non-small cell lung cancer (NSCLC) effects via the CB1 or CB2 receptor. The purpose of the investigation was to reveal the anti-tumor ability of NITyr on A549 cells and its mechanisms.Methods: The viability of A549 cells was measured by MTT assay, and the cell cycle and apoptosis were both examined by flow cytometry; in addition, cell migration was tested by wound healing assay. Apoptosis-related markers were measured by immunofluorescence. The downstream signaling pathways (PI3K, ERK, and JNK) of CB1 or CB2 were examined through Western blotting. The expressions of CB1 and CB2 were detected by immunofluorescence. Finally, the AutoDock software was used to validate the binding affinity between the targets, such as CB1 and CB2, with NITyr.Results: We found that NITyr inhibited cell viability, hindered the cell cycle, resulted in apoptosis, and inhibited migration. The CB1 inhibitor, AM251, and the CB2 inhibitor, AM630, weakened the aforementioned phenomenon. The immunofluorescence assay suggested that NITyr upregulated the expression of CB1 and CB2. Western blot analysis indicated that NITyr upregulated the expression of p-ERK, downregulated the expression of p-PI3K, and did not affect p-JNK expression. In conclusion, NITyr showed a role in inhibiting NSCLC through the activation of CB1 and CB2 receptors involved in PI3K and ERK pathways.

PurposePhosphatidylinositol 3-kinase (PI3K) signaling is highly active in glioblastomas. We assessed pharmacokinetics, pharmacodynamics, and efficacy of the pan-PI3K inhibitor buparlisib in patients with recurrent glioblastoma with PI3K pathway activation.MethodsThis study was a multicenter, open-label, multi-arm, phase II trial in patients with PI3K pathway-activated glioblastoma at first or second recurrence. In cohort 1, patients scheduled for re-operation after progression received buparlisib for 7 to 13 days before surgery to evaluate brain penetration and modulation of the PI3K pathway in resected tumor tissue. In cohort 2, patients not eligible for re-operation received buparlisib until progression or unacceptable toxicity. Once daily oral buparlisib 100 mg was administered on a continuous 28-day schedule. Primary end points were PI3K pathway inhibition in tumor tissue and buparlisib pharmacokinetics in cohort 1 and 6-month progression-free survival (PFS6) in cohort 2.ResultsSixty-five patients were treated (cohort 1, n = 15; cohort 2, n = 50). In cohort 1, reduction of phosphorylated AKTS473 immunohistochemistry score was achieved in six (42.8%) of 14 patients, but effects on phosphoribosomal protein S6S235/236 and proliferation were not significant. Tumor-to-plasma drug level was 1.0. In cohort 2, four (8%) of 50 patients reached 6-month PFS6, and the median PFS was 1.7 months (95% CI, 1.4 to 1.8 months). The most common grade 3 or greater adverse events related to treatment were lipase elevation (n = 7 [10.8%]), fatigue (n = 4 [6.2%]), hyperglycemia (n = 3 [4.6%]), and elevated ALT (n = 3 [4.6%]).ConclusionBuparlisib had minimal single-agent efficacy in patients with PI3K-activated recurrent glioblastoma. Although buparlisib achieved significant brain penetration, the lack of clinical efficacy was explained by incomplete blockade of the PI3K pathway in tumor tissue. Integrative results suggest that additional study of PI3K inhibitors that achieve more-complete pathway inhibition may still be warranted.

Abstract Background Ovarian cancer is the most lethal gynecologic disease and is one of the most commonly diagnosed cancers among women worldwide. The phosphatidylinositol 3-kinase (PI3K) family plays an important regulatory role in various cancer signaling pathways, including those involved in ovarian cancer development; however, its exact function remains to be fully understood. We conducted this study to understand the role of P13K in the molecular mechanisms underlying ovarian cancer development. Methods To determine the differential gene expression of phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), a regulatory subunit of PI3K, in normal, tumor, and metastatic ovary tissues, TNM plotter analysis was performed. The microarray dataset GSE53759 was downloaded from Gene Expression Omnibus. ROC plotter analysis was conducted to understand the potential of PIK3R3 as a predictive marker for effectiveness of therapy in ovarian cancer. muTarget was used to identify mutations that alter PIK3R3 expression in ovarian cancer. To determine the interacting partners for PIK3R3 in ovarian tissues, the interactome-atlas tool was used. The Kyoto encyclopedia of genes and genomes (KEGG) analysis was conducted to identify the pathways in which these interacting partners were primarily enriched. Results PIK3R3 was overexpressed in ovarian and metastatic tumors. Elevated PIK3R3 levels were observed in ovarian cancer stem cells, wherein inhibiting PIK3R3 expression significantly reduced the size of ovarian cancer spheroids. Treatment of ovarian cancer stem cells with PF-04691502 (10 μM), an inhibitor of both PI3K and mTOR kinases, also reduced the size of spheroids and the level of OCT4. PIK3R3 was highly expressed in ovarian cancer with several somatic mutations and was predicted better outcomes in patients undergoing Avastin® chemotherapy using bioinformatic tool. Protein interaction analysis showed that PIK3R3 interacts with 157 genes, including GRB2, EGFR, ERBB3, PTK2, HCK, IGF1R, YES1, and PIK3CA, in the ovary. KEGG enrichment analysis revealed that the interacting partners of PIK3R3 are involved in the ErbB signaling pathway, proteoglycans in cancer, FoxO, prolactin, chemokine, and insulin signaling pathways. Conclusions PIK3R3 plays a pivotal role in ovarian cancer development and is therefore a potential candidate for developing novel therapeutic approaches.

Tomasz J&eogon;drzejewski,1 Justyna Soboci&nacute;ska,1 Ma&lstrok;gorzata Pawlikowska,1 Artur Dzialuk,2 Sylwia Wrotek1 1Department of Immunology, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University, Torun, 87-100, Poland; 2Department of Genetics, Faculty of Biological Sciences, Kazimierz Wielki University, Bydgoszcz, 85-090, PolandCorrespondence: Tomasz J&eogon;drzejewski, Department of Immunology, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University, 1 Lwowska Str., Torun, 87-100, Poland, Tel +48 56 611 25 18, Email tomaszj@umk.pl Artur Dzialuk, Department of Genetics, Faculty of Biological Sciences, Kazimierz Wielki University, 10 Powsta&nacute;ców Wielkopolskich Ave., Bydgoszcz, 85-090, Poland, Tel +48 52 34 19 281, Email dzialuk@ukw.edu.plPurpose: Extract from the fungus Coriolus versicolor (CV) is classified as an immunological response modifier. Previously, we have shown that this extract induces interleukin 6 (IL-6)-related extension of lipopolysaccharide (LPS)-induced fever. This study investigated the effect of CV extract on the production of pro-inflammatory cytokines and the expression of components of signal transduction pathways leading to the secretion of cytokines from RAW 264.7 macrophages stimulated with different doses of LPS.Methods: RAW 264.7 cells were stimulated with CV extract alone or co-treated with CV extract and LPS. The level of IL-6 and tumour necrosis factor α (TNF-α) in the culture media was measured using ELISA. Protein expression of Toll-like receptor (TLR) 4, phosphorylated IκB (p-IκB), CD14 glycoprotein and phospho-phosphatidylinositol 3-kinase (p-PI3K) was evaluated using Western blot. The effects of TLR4, nuclear factor κB (NF-κB) and p-PI3K on cytokine secretion were estimated using inhibitors: TAK-242, JSH-23 and LY294002.Results: CV extract itself stimulates the secretion of IL-6 and TNF-α and increases the expression of TLR4, p-IκB and p-PI3K. The presence of CV extract during the treatment of cells with lower concentrations of LPS (10 and 100 ng/mL) increases the cytokine production. Co-stimulation of cells with CV extract and LPS at a higher dose (500 ng/mL) decreases the secretion of cytokines. This effect is related to the changes in the expression of TLR4, CD14 glycoprotein, p-IκB and p-PI3K.Conclusion: This is the first report showing that the CV extract-induced production of cytokines is mediated by the PI3K signalling pathway. This extract acts antagonistically or additively with LPS on the production of IL-6 and TNF-α, depending on the LPS concentration. Our results are helpful for illustrating the mechanisms for the immunostimulatory effect of CV extract in inflammatory processes.Keywords: Coriolus versicolor, pro-inflammatory cytokines, Toll-like receptor 4, phosphorylated IκB, phosphatidylinositol 3-kinase, CD14 glycoprotein

Phosphatidylinositol 3-Kinase (PI3K) is a key regulator of cell motility during chemotaxis and plays an important role in relaying and amplifying the shallow gradient of chemoattractant signals to ultimately mediate rearrangements of the actin cytoskeleton. To determine whether PI3K plays a similar role in electrotaxis as in chemotaxis, we examined directional cell migration in response to an electric field (EF) and unexpectedly found that the role of PI3K in regulating cell motility differs depending on the state of Dictyostelium cells. Contrary to chemotaxis experiments using aggregation-competent cells, in the cell migration assay, we used a recently developed method for electrotaxis using 3-h starved cells. Wild-type cells starved for 3 h showed increased motility in the presence of LY294002, a PI3K inhibitor, whereas aggregation-competent cells showed slightly decreased motility, indicating the effect of LY294002 on cell motility differ depending on the state of the cells. Consistent with these results, pi3k null cells in the vegetative state exhibited increased motility similar to that in the presence of LY294002 , compared to wild-type cells. These findings were confirmed through random migration experiments. These results suggest that PI3Ks play a suppressive role in regulating cell motility of vegetative Dictyostelium cells and that the suppressive effect is reversed on inhibition or lack of PI3Ks, leading to high motility. • PI3K is a key regulator of cell motility in directional cell migration. • Roles of PI3K in cell motility differ depending on the state of Dictyostelium cells. • PI3K plays an inhibitory role in regulating cell motility of vegetative cells. • Whereas, it plays a stimulatory role in cell motility of aggregation-competent cells. [ABSTRACT FROM AUTHOR]

Objective To explore whether miR-9 can inhibit the growth of hypertrophic scar fibroblasts by targeting PI3K. Methods From September 2020 to September 2021, 12 healthy adult New Zealand long-eared rabbits were randomly divided into control group (6 rabbits) and scar group (6 rabbits). The histological morphology of both groups was detected by HE staining. The mRNA expression of miR-9 and PI3K in tissues was detected by real-time fluorescent quantitative PCR, and the expression of miR-9 and PI3K was analyzed by Pearson's correlation coefficient. The human hypertrophic scar fibroblasts (HSFb) were divided into psiCHECK-WT-PI3K+miR-9 control group, psiCHECK-WT-PI3K+miR-9 mimic group, psiCHECK-MUT-PI3K+miR-9 control group and psiCHECK-MUT-PI3K+ miR-9 mimic group. The corresponding sequences were transfected respectively. The expression of luciferase and renilla luciferase were detected by luciferase reporter gene assay at 48 h after transfection. The HSFb was divided into miR-9 control group and miR-9 mimic group. At 0, 12, 24, 36 and 48 h after transfection, the cell proliferation was detected by MTT. At 24 h after treatment, the cell apoptosis was detected by Annexin V-FITC/PI kit, and the expression of PI3K, caspase 3 and Bcl-2 protein was detected by protein immunoblotting. The number of samples in cell experiment was 3. The data were analyzed by t-test and Pearson's correlation coefficient. Results In HE staining, the excessive proliferation of fibroblasts and inflammatory cell infiltration were found in scar tissue. The model was successfully constructed. Compared with the control group, the miR-9 expression was significantly down-regulated (t=11.020, P<0.05) and PI3K expression was significantly up-regulated in the scar group (t=10.840, P<0.05). Pearson's correlation analysis showed that the expression of miR-9 and PI3K was negatively correlated in all tissues. At 48 h after transfection, the PI3K activity in the psiCHECK-WT-PI3K + miR-9 mimic group was significantly lower than that in the psiCHECK-WT-PI3K+miR-9 control group (t=8.648, P=0.001). The PI3K activity in the psiCHECK-MUT-PI3K+miR-9 control group was similar to that in the psiCHECKMUT-PI3K+miR-9 mimic group (t=2.053, P=0.109). At 12, 24, 36 and 48 h after transfection, the proliferation ability of miR-9 mimic group was significantly lower than that of the miR-9 control group (P<0.05). At 24 h after transfection, the early apoptotic rate of HSFb in the miR-9 mimic group was higher than that in the miR-9 control group (t=11.210, P=0.001). MiR-9 can down regulate the expression of PI3K, inhibit the expression of Bcl-2 and up regulate the content of caspase 3. Conclusion MiR-9 is down-regulated in the hypertrophic scar tissue, which can inhibit the growth of HSFb by targeting PI3K. [ABSTRACT FROM AUTHOR]

Indoleamine 2,3‐dioxygenase 1 (IDO1) catalyzes the initial rate‐limiting step in the degradation of the essential amino acid tryptophan along the kynurenine pathway. When discovered more than 50 years ago, IDO1 was thought to be an effector molecule capable of mediating a survival strategy based on the deprivation of bacteria and tumor cells of the essential amino acid tryptophan. Since 1998, when tryptophan catabolism was discovered to be crucially involved in the maintenance of maternal T‐cell tolerance, IDO1 has become the focus of several laboratories around the world. Indeed, IDO1 is now considered as an authentic immune regulator not only in pregnancy, but also in autoimmune diseases, chronic inflammation, and tumor immunity. However, in the last years, a bulk of new information—including structural, biological, and functional evidence—on IDO1 has come to light. For instance, we now know that IDO1 has a peculiar conformational plasticity and, in addition to a complex and highly regulated catalytic activity, is capable of performing a nonenzymic function that reprograms the expression profile of immune cells toward a highly immunoregulatory phenotype. With this state‐of‐the‐art review, we aimed at gathering the most recent information obtained for this eclectic protein as well as at highlighting the major unresolved questions. [ABSTRACT FROM AUTHOR]

Chronic relapsing inflammatory bowel disease is strongly linked to an increased risk of colitis-associated cancer (CAC). One of the well-known inflammatory carcinogenesis pathways, phosphatidylinositol 3-kinase (PI3K), was identified to be a crucial mechanism in long-standing ulcerative colitis (UC). The goal of this study was to identify somatic variants in the cytokine-induced PI3K-related genes in UC, colorectal cancer (CRC) and CAC. Thirty biopsies (n = 8 long-standing UC, n = 11 CRC, n = 8 paired normal colorectal mucosa and n = 3 CAC) were subjected to targeted sequencing on 13 PI3K-related genes using Illumina sequencing and the SureSelectXT Target Enrichment System. The Genome Analysis Toolkit was used to analyze variants, while ANNOVAR was employed to detect annotations. There were 5116 intronic, 355 exonic, 172 untranslated region (UTR) and 59 noncoding intronic variations detected across all samples. Apart from a very small number of frameshifts, the distribution of missense and synonymous variants was almost equal. We discovered changed levels of IL23R, IL12Rß1, IL12Rß2, TYK2, JAK2 and OSMR in more than 50% of the samples. The IL23R variant in the UTR region, rs10889677, was identified to be a possible variant that might potentially connect CAC with UC and CRC. Additional secondary structure prediction using RNAfold revealed that mutant structures were more unstable than wildtype structures. Further functional research on the potential variants is, therefore, highly recommended since it may provide insight on the relationship between inflammation and cancer risk in the cytokine-induced PI3K pathway. [ABSTRACT FROM AUTHOR]

Glutamate-induced neural toxicity in autophagic neuron death is partially mediated by increased oxidative stress. Therefore, reducing oxidative stress in the brain is critical for treating or preventing neurodegenerative diseases. Selaginella tamariscina is a traditional medicinal plant for treating gastrointestinal bleeding, hematuria, leucorrhea, inflammation, chronic hepatitis, gout, and hyperuricemia. We investigate the inhibitory effects of Selaginella tamariscina ethanol extract (STE) on neurotoxicity and autophagic cell death in glutamate-exposed HT22 mouse hippocampal cells. STE significantly increased cell viability and mitochondrial membrane potential and decreased the expression of reactive oxygen species, lactate dehydrogenase release, and cell apoptosis in glutamate-exposed HT22 cells. In addition, while glutamate induced the excessive activation of mitophagy, STE attenuated glutamate-induced light chain (LC) 3 II and Beclin-1 expression and increased p62 expression. Furthermore, STE strongly enhanced the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) phosphorylation activation. STE strongly inhibited glutamate-induced autophagy by activating the PI3K/Akt/mTOR signaling pathway. In contrast, the addition of LY294002, a PI3K/Akt inhibitor, remarkably suppressed cell viability and p-Akt and p62 expression, while markedly increasing the expression of LC3 II and Beclin-1. Our findings indicate that autophagy inhibition by activating PI3K/Akt/mTOR phosphorylation levels could be responsible for the neuroprotective effects of STE on glutamate neuronal damage. [ABSTRACT FROM AUTHOR]

We developed a general approach for investigation of how cellular processes become adapted for specific cell types during differentiation. Previous studies reported substantial differences in the morphology and dynamics of clathrin-mediated endocytosis (CME) sites. However, associating specific CME properties with distinct differentiated cell types and determining how these properties are developmentally specified during differentiation have been elusive. Using genome-edited human embryonic stem cells, and isogenic fibroblasts and neuronal progenitor cells derived from them, we established by live-cell imaging and platinum replica transmission electron microscopy that CME site dynamics and ultrastructure on the plasma membrane are precisely reprogrammed during differentiation. Expression levels for the endocytic adaptor protein AP2μ2 were found to underlie dramatic changes in CME dynamics and structure. Additionally, CME dependency on actin assembly and phosphoinositide-3 kinase activity are distinct for each cell type. Collectively, our results demonstrate that key CME properties are reprogrammed during differentiation at least in part through AP2μ2 expression regulation.