scholarly journals Studying cellular functions in bipolar disorder: Are there specific predictors of lithium response?

2019 ◽  
Author(s):  
Pradip Paul ◽  
Shruti Iyer ◽  
Ravi Kumar Nadella ◽  
Rashmitha Nayak ◽  
Anirudh S. Chellappa ◽  
...  
Keyword(s):  
Bipolar Disorder ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Lithium Treatment ◽  
Healthy Population ◽  
Outpatient Services ◽  
Mitochondrial Potential ◽  
Lithium Response ◽  
Cellular Phenotypes

ABSTRACTBackgroundLithiumis the first-line mood stabilizer for the treatment of bipolar disorder (BD). In order to interrogate cellular phenotypes related to disease and lithium treatment response, this study used neural precursor cells (NPCs) and lymphoblastoid cell lines (LCLs) from BD patients who are well characterized for clinical lithium response.MethodsBDpatientsdiagnosed according to the DSM-IV criteria; were recruited from the outpatient services of the National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore, India. Clinical lithium response was assessed using the “Alda scale” and “NIMH Retrospective Life chart method”. The controls were ethnically matched healthy subjects with no family history of neuropsychiatric illness. NPCs from two BD patients from the same family who clearly differed in their clinical response to lithium were chosen, and compared with healthy population controls. Whole transcriptome sequencing (RNA-Seq) and analysis were performed, with and withoutin vitrolithium (1mM for 7 days). In addition, mitochondrial membrane potential (MMP), cell viability and cell proliferation parameters were examined. Experiments were also performed in 25 LCLs from BD patients (16 lithium responders and 9 lithium non-responders), and 12 healthy control LCLs, to evaluate them in a system amenable to clinical translation.ResultsRNA-Sequencingand analysis did not reveal differences in NPCs onin vitrolithium treatment. MMP was lower in BD, both in NPCs and LCLs; reversal within vitrolithium happened only in LCLs and was unrelated to lithium response. Cell proliferation was higher in BD compared to controls, and there was no change on lithium addition. Cell viability assays indicated greater cell death in BD; which could only be rescued in LCLs of clinical lithium responders. The latter finding was associated with enhancedBCL2andGSK3Bexpression within vitrolithium.DiscussionOverall, our study findings indicate that there are cellular phenotypes related to the disease (mitochondrial potential, cell proliferation) in NPCs and LCLs. We also observed clinical lithium response related phenotypes (cell viability,BCL2/ GSK3Bexpression) in LCLs. The next step would be to evaluate a larger set of PBMCs from clinical lithium response groups of BD to derive cellular phenotypes related to direct clinical application.

scholarly journals The Olive Leaves Extract Has Anti-Tumor Effects against Neuroblastoma through Inhibition of Cell Proliferation and Induction of Apoptosis

Nutrients ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 2178
Author(s):  
Fabio Morandi ◽  
Veronica Bensa ◽  
Enzo Calarco ◽  
Fabio Pastorino ◽  
Patrizia Perri ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Cell Cycle Progression ◽  
Treatment Strategies ◽  
Annexin V ◽  
Dependent Manner ◽  
Induction Of Apoptosis ◽  
Dose Dependent

Neuroblastoma (NB) is the most common extra-cranial solid tumor of pediatric age. The prognosis for high-risk NB patients remains poor, and new treatment strategies are desirable. The olive leaf extract (OLE) is constituted by phenolic compounds, whose health beneficial effects were reported. Here, the anti-tumor effects of OLE were investigated in vitro on a panel of NB cell lines in terms of (i) reduction of cell viability; (ii) inhibition of cell proliferation through cell cycle arrest; (iii) induction of apoptosis; and (iv) inhibition of cell migration. Furthermore, cytotoxicity experiments, by combining OLE with the chemotherapeutic topotecan, were also performed. OLE reduced the cell viability of NB cells in a time- and dose-dependent manner in 2D and 3D models. NB cells exposed to OLE underwent inhibition of cell proliferation, which was characterized by an arrest of the cell cycle progression in G0/G1 phase and by the accumulation of cells in the sub-G0 phase, which is peculiar of apoptotic death. This was confirmed by a dose-dependent increase of Annexin V+ cells (peculiar of apoptosis) and upregulation of caspases 3 and 7 protein levels. Moreover, OLE inhibited the migration of NB cells. Finally, the anti-tumor efficacy of the chemotherapeutic topotecan, in terms of cell viability reduction, was greatly enhanced by its combination with OLE. In conclusion, OLE has anti-tumor activity against NB by inhibiting cell proliferation and migration and by inducing apoptosis.

scholarly journals SDF-1α/MicroRNA-134 Axis Regulates Nonfunctioning Pituitary Neuroendocrine Tumor Growth via Targeting VEGFA

2020 ◽  
Vol 11 ◽  
Author(s):  
Xiaoyu Wang ◽  
Yuanjian Fang ◽  
Yunxiang Zhou ◽  
Xiaoming Guo ◽  
Ke Xu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Neuroendocrine Tumor ◽  
Tumor Cells ◽  
Tumor Invasion ◽  
Expression Level ◽  
Qrt Pcr ◽  
Proliferation And Invasion

BackgroundNonfunctioning pituitary neuroendocrine tumor (NF-PitNET) is difficult to resect. Except for surgery, there is no effective treatment for NF-PitNET. MicroRNA-134 (miR-134) has been reported to inhibit proliferation and invasion ability of tumor cells. Herein, the mechanism underlying the effect of miR-134 on alleviating NF-PitNET tumor cells growth is explored.MethodsMouse pituitary αT3-1 cells were transfected with miR-134 mimics and inhibitor, followed by treatment with stromal cell-derived factor-1α (SDF-1α) in vitro. MiR-134 expression level: we used quantitative real-time PCR (qRT-PCR) to detect the expression of miR-134. Cell behavior level: cell viability and invasion ability were assessed using a cell counting kit-8 (CCK8) assay and Transwell invasion assay respectively. Cytomolecular level: tumor cell proliferation was evaluated by Ki-67 staining; propidium iodide (PI) staining analyzed the effect of miR-134 on cell cycle arrest; western blot analysis and immunofluorescence staining evaluated tumor migration and invasive ability. Additionally, we collected 27 NF-PitNET tumor specimens and related clinical data. The specimens were subjected to qRT-PCR to obtain the relative miR-134 expression level of each specimen; linear regression analysis was used to analyze the miR-134 expression level in tumor specimens and the age of the NF-PitNET population, gender, tumor invasion, prognosis, and other indicators.ResultsIn vitro experiment, miR-134 was observed to significantly inhibit αT3-1 cells proliferation characterized by inhibited cell viability and expressions of vascular endothelial growth factor A (VEGFA) and cell cycle transition from G1 to S phase (P < 0.01). VEGFA was verified as a target of miR-134. Additionally, miR-134-induced inhibition of αT3-1 cell proliferation and invasion was attenuated by SDF-1α and VEGFA overexpression (P < 0.01). In primary NF-PitNET tumor analysis, miR-134 expression level was negatively correlated with tumor invasion (P = 0.003).ConclusionThe regulation of the SDF-1α/miR-134/VEGFA axis represents a novel mechanism in the pathogenesis of NF-PitNETs and may serve as a potential therapeutic target for the treatment of NF-PitNETs.

scholarly journals The Effect of Parietin Isolated From Rheum ribes L on In Vitro Wound Model Using Human Dermal Fibroblast Cells

2019 ◽  
Vol 18 (1) ◽  
pp. 56-64 ◽  
Author(s):  
Gulsah Gundogdu ◽  
Koksal Gundogdu ◽  
Kemal Alp Nalci ◽  
Alper Kursat Demirkaya ◽  
Seymanur Yılmaz Tascı ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Viability ◽  
Dermal Fibroblast ◽  
Dose Range ◽  
Human Dermal Fibroblast ◽  
Fibroblast Cells ◽  
Wound Model ◽  
Dpph Method ◽  
Pipette Tip

Parietin is one of the well-known anthraquinone compounds that can be extracted from Rheum ribes L. In this study, we aimed to investigate the effects of parietin isolated from Rheum ribes L on an in vitro wound model using human dermal fibroblast cells and compare its effectiveness against zinc. The antioxidant effect of parietin was determined by using the 1,1-diphenyl-2-picrylhydrazine (DPPH) method. Human dermal fibroblast cells were cultured in proculture medium and were kept until 100% confluence was achieved. The wound model was created by using a pipette tip. After that, different concentrations of parietin and zinc (final concentrations in the well to be 5-250 µM and 25-200 µM, respectively) were added into the medium. The proliferation-inducing effect on cell viability was determined by using MTT assay. Images of cells were taken at 0, 12, and 24 hours. According to the DPPH method, parietin exhibited have antioxidant activity. According to the MTT results, parietin exhibited significant proliferation-inducing effect on cell viability in a dose range of 5 to 10 M, and zinc showed significant proliferation-inducing effect on cell viability at dose 50 µM ( P < .05). In addition, the image of cell proliferation was also shown at the same doses at 24 hours. In this study, we claim that parietin induces cell proliferation at low doses in cases of dermal fibroblast loss. In conclusion, parietin as an alternative to zinc in wound healing could be used by clinicians in the future with more extensive studies.

The effect of pH on cell viability, cell migration, cell proliferation, wound closure, and wound reepithelialization: In vitro and in vivo study

2017 ◽  
Vol 25 (2) ◽  
pp. 260-269 ◽  
Author(s):  
Carla R. Kruse ◽  
Mansher Singh ◽  
Stefan Targosinski ◽  
Indranil Sinha ◽  
Jens A. Sørensen ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Migration ◽  
Cell Viability ◽  
Wound Closure ◽  
In Vivo Study ◽  
Effect Of Ph

scholarly journals Microglia induce neurogenesis by stimulating PI3K/AKT intracellular signaling in vitro

2019 ◽  
Author(s):  
Kristi Lorenzen ◽  
Nicholas W. Mathy ◽  
Erin R. Whiteford ◽  
Alex Eischeid ◽  
Jing Chen ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Viability ◽  
Intracellular Signaling ◽  
Cortical Cell ◽  
Model System ◽  
Microglial Cells ◽  
Cortical Cells ◽  
Akt Phosphorylation ◽  
Cortical Neurogenesis

Abstract Background: Emerging evidence suggests that microglia can support neuronal survival, synapse development, and neurogenesis in classic neurogenic niches. Little is known about the ability of microglia to regulate the cortical environment and stimulate cortical neurogenesis outside classic neurogenic niches. We used an in vitro co-culture model system to investigate the hypothesis that microglia respond to soluble signals from cortical cells, particularly following injury, to alter the cortical environment and promote cortical cell proliferation, differentiation, and maintain cortical cell survival via activation of specific cortical intracellular signaling pathways. Results: Analyses of cell proliferation, apoptosis, protein expression, and intracellular signaling pathway activation were performed on uninjured and injured cortical cells in co-culture with EOC2 microglia. EOC2 microglia in co-culture enhanced cortical cell viability and proliferation of uninjured and injured cortical cells. Co-culture of injured cortical cells with EOC2 microglial cells significantly reduced cortical cell apoptosis. Microglial co-culture significantly increased Nestin+ and a-internexin+ cells within and outside of the injury site. NeuN+ cells increased in injured cortical cultures with microglia. Multiplex ELISA assays showed decreased levels of inflammatory cytokines in conditioned media from injured cortical cell and microglial co-culture. RTPCR analysis of microglial mRNA was performed. EOC2 microglial co-culture environment increased AKT phosphorylation in cortical cells particularly following injury. Inhibition of AKT phosphorylation in cortical cells blocked the microglial-enhanced cortical cell viability and expression of neurogenic markers in vitro. Conclusion: The in vitro model system presented here allows for assessment of the effect of microglial-derived soluble signals, independent of cell-cell contact, on cortical cell viability, proliferation, and stages of differentiation during homeostasis or following injury. These data suggest that microglia downregulate inflammatory cytokine production following activation by acute cortical injury to enhance proliferation of new cells capable of neurogenesis. Inhibition of AKT signaling in cortical cells blocks the enhanced proliferation and expression of neurogenic markers in cortical cells. Increasing our understanding of the mechanisms that drive cortical neurogenesis stimulated by microglial cells during homeostasis and following injury will provide insight into the potential mechanisms of neuroprotective role of immune activity in the central nervous system (CNS).

scholarly journals OS8.7 Targeting Pi3k-Akt-mTOR and MAPKinase pathways in aggressive meningiomas: in vitro study

2019 ◽  
Vol 21 (Supplement_3) ◽  
pp. iii17-iii17
Author(s):  
G Mondielli ◽  
F Darriet ◽  
C Roche ◽  
C Lisbonis ◽  
A Querdray ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Mtor Inhibitor ◽  
Primary Cell ◽  
Preclinical Model ◽  
Maximal Effect ◽  
Mtor Inhibition ◽  
Mek Kinase

Abstract BACKGROUND Recurrent and aggressive meningiomas remain an unmet medical need in neuro-oncology. In our preclinical model on meningioma primary cell culture, the combination of the mTOR inhibitor, everolimus and the somatostatin analog, octreotide decreased cell viability and proliferation without inducing apoptosis. The interest of the combined treatment everolimus plus octreotide were clinically validated by the CEVOREM clinical trial demonstrating a decrease in growth rate of meningiomas in most treated patients. Nevertheless, everolimus induced an increase in Akt activity in vitro, which probably limited everolimus efficiency. We hypothesized that targeting Pi3K could prevent this positive feedback on Akt phosphorylation, induced by mTOR inhibition. The involvement of MAPKinase pathway in meningioma tumorigenesis was recently demonstrated. Our aim was to decipher the effect of the Pi3k α inhibitor Alpelisib (BYL719) and the specific inhibitor of MEK1,2 Mekinist (Trametinib) alone or in combination, in comparison to the mTor inhibitor everolimus on human meningioma primary cell cultures. MATERIAL AND METHODS The impact of the drugs was studied on 40 meningiomas, well characterized at clinical, histological and molecular level. The cell viability, cell proliferation and apoptosis were analyzed under drugs. RESULTS BYL719 induced a dose dependent decrease in cell viability (maximal effect -90%, IC50 10-6M) in all tested meningiomas (n=30). This effect was stronger than those of everolimus (maximal effect -50%, n=24, IC50 10-8M). 22 tumors were sensitive to Trametinib (maximal effect <-50%, IC50 10–5 M), 14 were partially sensitive (maximal effect between -20 and -40% IC50 10–5 M) and 3 were resistant. An apoptotic effect was observed under BYL719 in 6/18 tested tumors whereas no apoptosis was observed under Trametinib and Everolimus. Combination of BYL719 and Trametinib induced a significant stronger decrease in cell viability than each drug alone. Correlation analysis between these functional results, the tumor genomic profile and the activation of ERK/MEK kinase pathway is ongoing. CONCLUSION PI3K-Akt-mTOR and ERK/MEK kinase pathways constitute relevant targets in aggressive meningioma therapy. In our preclinical model, previously validated by CEVOREM clinical trial, co-targeting Pi3k-Akt-mTOR and MAPkinase pathways improved cell proliferation inhibition in comparison to the target of each pathway alone. BYL719 induced apoptosis which was not achieved by everolimus. These results support the ongoing clinical trial ALTREM combining Alpelisib and Trametinib on patients harboring aggressive recurrent meningiomas.

179 EFFECTS OF AN ENDOCANNABINOID, ANANDAMIDE, ON BOVINE BLASTOCYST DEVELOPMENT IN VITRO

2006 ◽  
Vol 18 (2) ◽  
pp. 197
Author(s):  
M. Y. Turco ◽  
K. Matsukawa ◽  
P. Loi ◽  
G. Ptak
Keyword(s):  
Cell Proliferation ◽  
Cell Viability ◽  
Protein Localization ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Santa Cruz ◽  
Negative Effects ◽  
Blastocyst Development ◽  
Development In Vitro

Cannabinoids cause many adverse effects on reproductive functions including fetal loss and pregnancy failure (Paria et al. 1995 PNAS 92, 9460–9464). N-arachidonylethanolamine (anandamide), an endogenous cannabinomimetic lipid derivative, binds with high affinity to brain type and spleen type cannabinoid receptors (CB1-R and CB2-R) and mimics most of the effects of Δ9-tetrahydrocannabinol [(−)THC], a psychoactive derivative of marijuana. In this study, we investigated the effects of anandamide on ovine blastocyst development in vitro. In vitro-matured oocytes were chemically activated and cultured to the blastocyst stage in our standard media (Ptak et al. 1999 Biol. Reprod. 61, 1568–1574). The development rate of blastocysts was 41%. Day 7 blastocysts were exposed to 28 nM anandamide with or without 20 nM SR141716A (an antagonist of CB1-R) for 48 h. In Experiment 1, we examined the CB1-R protein localization on blastocysts by anti CB1-R antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). In Experiment 2, to investigate the possible effect of anandamide on blastocyst development, we used the cell viability assay by propidium iodide, a cell proliferation assay (5-bromo-1′-deoxyuridine incorporation), and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay (Qbiogene, Inc., Carlsbad, CA, USA). Data were analyzed by one-way ANOVA. CB1-R signals were detected on ovine blastocysts, and these signals were effectively improved in the anandamide co-cultured group. The results of Experiment 2 are summarized in Table 1. Our results demonstrate that CB1-R was expressed in the ovine blastocyst, and anandamide exerts negative effects on in vitro blastocyst development by inhibiting cell proliferation and increasing apoptotic rate, but not cell viability. Furthermore, SR141716A can effectively block the negative effects of anandamide. Table 1. Effect of anandamide (AEA) on sheep blastocyst development assessed by viability, cell proliferation, and TUNEL staining assays

scholarly journals Combining schizophrenia and depression polygenic risk scores improves the genetic prediction of lithium response in bipolar disorder patients

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Klaus Oliver Schubert ◽  
Anbupalam Thalamuthu ◽  
Azmeraw T. Amare ◽  
Joseph Frank ◽  
Fabian Streit ◽  
...  
Keyword(s):  
Bipolar Disorder ◽  
Treatment Response ◽  
Molecular Mechanisms ◽  
Lithium Treatment ◽  
Meta Analysis ◽  
Risk Scores ◽  
Poor Responders ◽  
Genetic Trait ◽  
Polygenic Risk ◽  
Lithium Response

AbstractLithium is the gold standard therapy for Bipolar Disorder (BD) but its effectiveness differs widely between individuals. The molecular mechanisms underlying treatment response heterogeneity are not well understood, and personalized treatment in BD remains elusive. Genetic analyses of the lithium treatment response phenotype may generate novel molecular insights into lithium’s therapeutic mechanisms and lead to testable hypotheses to improve BD management and outcomes. We used fixed effect meta-analysis techniques to develop meta-analytic polygenic risk scores (MET-PRS) from combinations of highly correlated psychiatric traits, namely schizophrenia (SCZ), major depression (MD) and bipolar disorder (BD). We compared the effects of cross-disorder MET-PRS and single genetic trait PRS on lithium response. For the PRS analyses, we included clinical data on lithium treatment response and genetic information for n = 2283 BD cases from the International Consortium on Lithium Genetics (ConLi+Gen; www.ConLiGen.org). Higher SCZ and MD PRSs were associated with poorer lithium treatment response whereas BD-PRS had no association with treatment outcome. The combined MET2-PRS comprising of SCZ and MD variants (MET2-PRS) and a model using SCZ and MD-PRS sequentially improved response prediction, compared to single-disorder PRS or to a combined score using all three traits (MET3-PRS). Patients in the highest decile for MET2-PRS loading had 2.5 times higher odds of being classified as poor responders than patients with the lowest decile MET2-PRS scores. An exploratory functional pathway analysis of top MET2-PRS variants was conducted. Findings may inform the development of future testing strategies for personalized lithium prescribing in BD.

scholarly journals In Vitro and in Silico Evaluation of Bikaverin as a Potent Inhibitor of Human Protein Kinase CK2

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1380 ◽  
Author(s):  
Samer Haidar ◽  
Dagmar Aichele ◽  
Robin Birus ◽  
Janine Hielscher ◽  
Tuomo Laitinen ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Protein Kinase ◽  
Cell Viability ◽  
Binding Site ◽  
Protein Kinase Ck2 ◽  
Potent Inhibitor ◽  
Cell Permeability ◽  
Atp Binding Site ◽  
Kinase Ck2

Protein kinase CK2 is an emerging target for therapeutic intervention in human diseases, particularly in cancer. Inhibitors of this enzyme are currently in clinical trials, indicating the druggability of human CK2. By virtual screening of the ZINC database, we found that the natural compound bikaverin can fit well in the ATP binding site of the target enzyme CK2. By further in vitro evaluation using CK2 holoenzyme, bikaverin turned to be a potent inhibitor with an IC50 value of 1.24 µM. In this work, the cell permeability of bikaverin was determined using a Caco-2 cell permeability assay as a prerequisite for cellular evaluation and the compound turned out to be cell permeable with a Papp- value of 4.46 × 10−6 cm/s. Bikaverin was tested for its effect on cell viability using a MTT assay and cell proliferation using an EdU assay in different cancer cell lines (MCF7, A427 and A431 cells). Cell viability and cell proliferation were reduced dramatically after treatment with 10 µM bikaverin for 24 h. Additionally the IncuCyte® live-cell imaging system was applied for monitoring the cytotoxicity of bikaverin in the three tested cancer cell lines. Finally, molecular dynamic studies were performed to clarify the ligand binding mode of bikaverin at the ATP binding site of CK2 and to identify the amino acids involved.

scholarly journals Starting lithium prophylaxis earlyv.late in bipolar disorder

2014 ◽  
Vol 205 (3) ◽  
pp. 214-220 ◽  
Author(s):  
Lars Vedel Kessing ◽  
Eleni Vradi ◽  
Per Kragh Andersen
Keyword(s):  
Bipolar Disorder ◽  
Psychiatric Hospital ◽  
Lithium Treatment ◽  
Preventive Treatment ◽  
Hazard Ratio ◽  
Mixed Episode ◽  
Start Up ◽  
Late Intervention ◽  
Lithium Response ◽  
First Contact

BackgroundNo study has investigated when preventive treatment with lithium should be initiated in bipolar disorder.AimsTo compare response rates among patients with bipolar disorder starting treatment with lithium earlyv.late.MethodNationwide registers were used to identify all patients with a diagnosis of bipolar disorder in psychiatric hospital settings who were prescribed lithium during the period 1995–2012 in Denmark (n= 4714). Lithium responders were defined as patients who, following a stabilisation lithium start-up period of 6 months, continued lithium monotherapy without being admitted to hospital. Earlyv.late intervention was defined in two ways: (a) start of lithium following first contact; and (b) start of lithium following a diagnosis of a single manic/mixed episode.ResultsRegardless of the definition used, patients who started lithium early had significantly decreased rates of nonresponse to lithium compared with the rate for patients starting lithium later (adjusted analyses: firstv.later contact:P<0.0001; hazard ratio (HR) = 0.87, 95% CI 0.76–0.91; single manic/mixed episodev.bipolar disorder:P<0.0001; HR = 0.75, 95% CI 0.67–0.84).ConclusionsStarting lithium treatment early following first psychiatric contact or a single manic/mixed episode is associated with increased probability of lithium response.

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