Screening and Identification of Small Molecule Compounds Perturbing Mitosis Using Time-Dependent Cellular Response Profiles

2010 ◽  
Vol 82 (15) ◽  
pp. 6495-6503 ◽  
Author(s):  
Ning Ke ◽  
Biao Xi ◽  
Peifang Ye ◽  
Wanhong Xu ◽  
Min Zheng ◽  
...  
Keyword(s):  
Small Molecule ◽  
Cellular Response ◽  
Time Dependent ◽  
Screening And Identification ◽  
Response Profiles

Mode of action classification of chemicals using multi-concentration time-dependent cellular response profiles

2014 ◽  
Vol 49 ◽  
pp. 23-35 ◽  
Author(s):  
Zhankun Xi ◽  
Swanand Khare ◽  
Aaron Cheung ◽  
Biao Huang ◽  
Tianhong Pan ◽  
...  
Keyword(s):  
Mode Of Action ◽  
Cellular Response ◽  
Time Dependent ◽  
Action Classification ◽  
Concentration Time ◽  
Response Profiles

D–A–D-type narrow-bandgap small-molecule photovoltaic donors: pre-synthesis virtual screening using density functional theory

2016 ◽  
Vol 18 (22) ◽  
pp. 15054-15059 ◽  
Author(s):  
Yeongrok Gim ◽  
Daekyeom Kim ◽  
Minkyu Kyeong ◽  
Seunghwan Byun ◽  
Yuri Park ◽  
...  
Keyword(s):  
Density Functional Theory ◽  
Virtual Screening ◽  
Small Molecule ◽  
Density Functional ◽  
Density Functional Theory Calculations ◽  
Time Dependent ◽  
Functional Theory ◽  
Narrow Bandgap ◽  
Dependent Density

A new series of D–A–D-type small-molecule photovoltaic donors are designed and screened before synthesis using time-dependent density functional theory calculations.

CRM1/XPO1 Represents a Promising Therapeutic Target for Treatment of Chronic Lymphocytic Leukemia

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 232-232
Author(s):  
Rosa Lapalombella ◽  
Caroline Berglund ◽  
Emilia Mahoney ◽  
Katie Williams ◽  
Shruti Jha ◽  
...  
Keyword(s):  
Small Molecule ◽  
Cytotoxic Effect ◽  
Stromal Cell ◽  
Time Dependent ◽  
Dependent Manner ◽  
Spontaneous Apoptosis ◽  
Stromal Cell Line ◽  
Equity Ownership

Abstract Abstract 232 Exportin 1 (CRM1, XPO1) is a nuclear exporter that promotes the transit of tumor suppressor proteins (TSPs) including p53, I-κB, and FOXO3A out of the nucleus, thereby preventing their activity and contributing to disrupted apoptosis and enhanced proliferation. Recently, whole-genome sequencing in patients with CLL allowed the identification of recurrent mutations in a highly conserved region of CRM1 that can potentially affects its gene function, suggesting a direct role for CRM1 in the pathogenesis of CLL (Puente XS, et al: Nature 75:101, 2011). However the role of CRM1 and the consequences of its mutation in the development of CLL have yet to be explored. CRM1 has been shown to be up-regulated in hematologic and various solid tumors, making it a highly attractive molecular target impacting multiple pro apoptotic pathways. KPT-SINEs are new, potent and irreversible small molecule selective inhibitors of nuclear export developed by Karyopharm that specifically and irreversibly bind to CRM1 and block the function of this protein. CLL is characterized by disrupted apoptosis caused both by co-dependent stromal elements and aberrant activation of several survival-promoting signaling/transcriptional pathways including PI3K/Akt, NF-kB, and p53. Because of the distinct subtypes of CLL and multiple signaling pathways dysregulated, a therapeutic agent targeting a single biological pathway is unlikely to be effective. Thus, pursuit of CRM1 inhibition as a novel strategy aimed to restore multiple death pathways is crucial and has broad implications for many types of patients. Our preliminary work demonstrated CRM1 is over-expressed in CLL cells compared to normal B cells at a protein (3 fold, p<0.005) and mRNA level (2.6 fold p=0.014). Inhibition of CRM1 by KPT-185 induced apoptosis in primary patient CLL cells in a dose and time dependent manner (EC50<500nM) while limited cytotoxicity against normal PBMC and isolated B, NK and T cells was observed (EC50 values >20 μM). Additionally, KPT-185 treatment of NK cells had no effect on their function as measured by ability of NK cells to mediate antibody dependent (ADCC) as wekk as direct cytotoxicity. The effect of KPT-185 on T function is currently under evaluation. Nuclear accumulation of FOXO3, p53 and IkB was also observed in primary CLL cells in a time dependent manner as shown by western blot and confocal microscopy. The evaluation of activated target genes is currently ongoing. Given the importance of microenvironmental stimuli on survival of CLL cells and response to therapy, we evaluated the ability of KPT-185 to induce cytotoxicity of CLL cells in the presence or absence of soluble factors such as CPG, CD40L, BAFF, TNF-α, IL-6, or IL-4, which are known to reduce the spontaneous apoptosis associated with CLL cells. KPT-185 treatment abrogated the protection induced by each of these factors suggesting that KPT-SINEs can disrupt signaling from the microenvironment that lead to in vivo CLL cell survival and potentially drug resistance. Interestingly the cytotoxic effect elicited by KPT-185 was enhanced in CPG activated cells (p=0.02). We also tested the ability of KPT-185 to kill CLL cells under coculture conditions with Hs5 stromal cell line. Coculture of CLL cells alone for 48 hours on the Hs5 stromal cell line resulted in a marked reduction of spontaneous apoptosis suggesting a strong protective effect elicited (P<0.001) by the stromal cells. Interestingly the cytotoxic effect mediated by KPT-185 was enhanced under coculture conditions (p=0.013). KPT-185 was also proven to be effective on murine TCL1+ cells (EC50<500nM) in vitro. The in vivo efficacy of this compound and other structurally related analogs is currently being assessed in an ongoing study in theTCL1 mouse model of CLL. In conclusion CRM1 represents a novel target that has not been adequately explored in CLL. KPT-SINEs are a class of promising therapeutic agents with proven selective in vitro activity in CLL cells providing the rationale for developing small molecule, drug-like CRM1 inhibitors for the treatment of this disease. Disclosures: Sandanayaka: Karyopharm Therapeutics: Employment. Shechter:Karyopharm Therapeutics: Employment. McCauley:Karyopharm Therapeutics: Employment. Shacham:Karyopharm: Equity Ownership. Kauffman:Karyopharm: Equity Ownership.

Cytotoxicity assessment based on the AUC50 using multi-concentration time-dependent cellular response curves

2013 ◽  
Vol 764 ◽  
pp. 44-52 ◽  
Author(s):  
Tianhong Pan ◽  
Biao Huang ◽  
Weiping Zhang ◽  
Stephan Gabos ◽  
Dorothy Yu Huang ◽  
...  
Keyword(s):  
Cellular Response ◽  
Time Dependent ◽  
Response Curves ◽  
Concentration Time ◽  
Cytotoxicity Assessment

scholarly journals Dichotomy of cellular inhibition by small-molecule inhibitors revealed by single-cell analysis

2016 ◽  
Author(s):  
Robert Vogel ◽  
Amir Erez ◽  
Grégoire Altan-Bonnet
Keyword(s):  
Signal Transduction ◽  
Single Cell ◽  
Small Molecule ◽  
Cellular Response ◽  
Small Molecule Inhibitors ◽  
Single Cell Analysis ◽  
Phenotypic Variability ◽  
Cellular Responses ◽  
Drug Action ◽  
Cell Analysis

AbstractDespite progress in developing small molecule inhibitors, a quantitative understanding of drug action in the physiological context of cells is lacking. Here, we apply single-cell analysis of signal transduction and proliferation to probe cellular responses to small molecule inhibitors. We use the model of cellular activation of T lymphocytes responding to cytokines and antigens. We uncover two distinct modes of drug action, in terms of signaling inhibition: digital inhibition (e.g. when the fraction of activated cells diminishes upon drug treatment, but cells remaining active appear unperturbed), and analog inhibition (e.g. when the fraction of activated cells is unperturbed while their overall activation is diminished). We introduce a computational model of the signaling cascade in order to account for such dichotomy. We test the predictions of our model in terms of the phenotypic variability of cellular responses under drug inhibition. Finally, we demonstrate that the digital/analog dichotomy of cellular response as revealed on short timescales with signal transduction, translates into similar dichotomy on long timescales. Overall, our analysis of drug action at the single cell level illustrates the strength of quantitative approaches to translate the promise of in vitro pharmacology into functionally-relevant cellular settings.

scholarly journals Circadian control of lung inflammation in influenza infection

2018 ◽  
Author(s):  
Shaon Sengupta ◽  
Soon Yew Tang ◽  
Jill Devine ◽  
Soumyashant Nayak ◽  
Shirley Zhang ◽  
...  
Keyword(s):  
Circadian Rhythms ◽  
Cellular Response ◽  
Influenza Infection ◽  
Time Dependent ◽  
Endogenous Rhythms ◽  
Diurnal Variability ◽  
Mortality And Morbidity ◽  
Respiratory Mortality ◽  
Viral Responses ◽  
Host Tolerance

AbstractInfluenza is a leading cause of respiratory mortality and morbidity. While inflammation is necessary for fighting infection, a fine balance of anti-viral defense and host tolerance is necessary for recovery. Circadian rhythms have been known to modulate inflammation. However, the importance of diurnal variability in the timing of influenza infection is not well understood. Here we demonstrate that endogenous rhythms influence the cellular response to infection in bronchoalveolar lavage (BAL), the pulmonary transcriptomic profile and lesional histology. This time dependent variability does not reflect alterations in viral replication. Rather, we found that better time-dependent outcomes were associated with a preponderance of NK and NKT cells and lower proportion of monocytes in the lung. Thus, host tolerance, rather than viral burden underlies the diurnal gating of influenza induced lung injury.Significance statementOur work demonstrates the importance of circadian rhythms in influenza infection --a condition with significant public health implications. Our findings, which establish the role of the circadian rhythms in maintaining the balance between host tolerance pathways and anti-viral responses confers a new framework for evaluating the relevance of circadian influences on immunity.

The Novel 2nd Generation Small Molecule MEK Inhibitor, AZD-6244, Induces Cell Death in Lymphoma Cells Lines, Primary Cells, and in a Human Lymphoma Xenograft Model.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 285-285 ◽  
Author(s):  
Savita Bhalla ◽  
Ron Gartenhaus ◽  
Bojie Dai ◽  
Sheila Prachand ◽  
Arindam Mukherjee ◽  
...  
Keyword(s):  
Cell Lines ◽  
Small Molecule ◽  
Tumor Volume ◽  
Xenograft Model ◽  
Time Dependent ◽  
In Vivo Studies ◽  
Primary Cells ◽  
2Nd Generation ◽  
Average Tumor Volume

Abstract Abstract 285 Introduction: MCT-1 is an oncogene that has been shown to induce cell proliferation and activate cell survival pathways and is constitutively expressed in the majority of diffuse large B-cell lymphomas (DLBCL). MCT-1 is phosphorylated by the MEK/ERK kinases, which occurs directly upstream of MCT-1; thus MEK/ERK inhibition blocks the phosphorylation and activation of MCT-1. Potent small-molecule inhibitors of MEK have been developed in pre-clinical and clinical studies that have shown the MEK/ERK pathway can be effectively shut down in a highly selective manner. However, the majority of pre-clinical and clinical information regarding MEK inhibitors to date have emerged mainly from solid tumor studies. The study of anti-MEK therapy is largely unexplored in lymphoma. The benzimidazole AZD-6244 (ARRY-142886) is a novel 2nd generation small molecule MEK antagonist being developed for the treatment of cancer. We investigated the activity of AZD-6244 in DLBCL cell lines, primary cells, and an in vivo xenograft model. Methods: DLBCL cell lines (SUDHL4, SUDHL6, OCI-LY3, and OCI-LY19) and primary cells obtained from a patient with relapsed/refractory transformed DLBCL (absolute lymphocyte count 167.3 K/UL, hemoglobin 9.8 gm/dL, and LDH 1,141 Units/L (high normal 195 Units/L)) were incubated with increasing nanomolar concentrations of AZD-6244 (50-400nM) for 24–72 hours (hr). MTT was calculated and apoptosis was determined by fluorescence-activated cell sorting using AnnexinV-FITC/propidium iodide (AnnV+/PI+) staining. Cleaved caspases and phosphorylated ERK (pERK) and MCT-1 (pMCT-1) were assessed through Western blot studies. In vivo studies were performed with mice bearing subcutaneous tumors of SUDHL6 that were randomly divided into control and AZD-6244 groups. SUDHL6 cells (1.2 × 106) were subcutaneously injected into left and right dorsal flanks of 7-week-old female SCID mice. When the tumor reached the size of approximately 60-163mm3, AZD-6244 was administered intraperitoneally every other day at a dose of 1.0 mg/kg body weight for a total of 3 weeks. Results: Time-dependent cytotoxicity was documented in each of the 4 DLBCL cell lines. IC50 at 48 hours in SUDHL4 and OCI-LY3 was 130nM, 240nM in SUDLH6, and 300nM in OCI-LY19. Dose-dependent apoptosis was also seen in all cell lines. Compared with control at 48 hrs, >50% AnnV+/PI+ was documented in SUDHL-4 at 200nM and with 300-320nM in the three other cell lines (p<0.01). Interestingly, minimal cleavage of caspases 3,8, or 9 were seen. In terms of target inhibition, 100nM of AZD-6244 effectively down-regulated p-ERK in each of the cell lines, while 200-300nM was required for significant down-regulation of pMCT-1. In primary DLBCL cells, dose- and time-dependent apoptosis was also seen (Figure below). At 72 hrs, 100nM of AZD-6244 resulted in >75% AnnV+/PI+ (p<0.001). In terms of the in vivo studies, the average tumor volume of AZD-6244-treated and control SCID mice with standard deviation was calculated as a function of time. From 28 weeks on, mice treated with AZD-6244 had significantly and increasingly less average tumor volume compared with control (p<0.05). Conclusion: Collectively, these data show that the 2nd generation MEK small molecule antagonist AZD-6244 induced significant cell death at nanomolar and clinically achievable concentrations in multiple DLBCL cells lines, primary cells, and in an in vivo xenograft model. In addition, AZD-6244 effectively downregulated the critical pro-growth kinase pERK, and the downstream oncogene, MCT-1. Clinical study of AZD-6244 in DLBCL is warranted, while continued investigation of the role of the MEK/ERK/MCT-1 signaling cascade in lymphogenesis is needed. Disclosures: No relevant conflicts of interest to declare.

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