scholarly journals Discovery of novel candidates for anti-liposarcoma therapies by medium-scale high-throughput drug screening

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0248140
Author(s):  
Iwona Grad ◽  
Robert Hanes ◽  
Pilar Ayuda-Durán ◽  
Marieke Lydia Kuijjer ◽  
Jorrit M. Enserink ◽  
...  
Keyword(s):  
Cell Lines ◽  
Drug Testing ◽  
Preclinical Development ◽  
Significant Treatment ◽  
Good Efficiency ◽  
Target Drug ◽  
High Throughput Drug Screening ◽  
Advanced Stages ◽  
New Treatment ◽  
Nicotinamide Phosphoribosyl Transferase

Sarcomas are a heterogeneous group of mesenchymal orphan cancers and new treatment alternatives beyond traditional chemotherapeutic regimes are much needed. So far, tumor mutation analysis has not led to significant treatment advances, and we have attempted to bypass this limitation by performing direct drug testing of a library of 353 anti-cancer compounds that are either FDA-approved, in clinical trial, or in advanced stages of preclinical development on a panel of 13 liposarcoma cell lines. We identified and validated six drugs, targeting different mechanisms and with good efficiency across the cell lines: MLN2238 –a proteasome inhibitor, GSK2126458 –a PI3K/mTOR inhibitor, JNJ-26481585 –a histone deacetylase inhibitor, triptolide–a multi-target drug, YM155 –a survivin inhibitor, and APO866 (FK866)–a nicotinamide phosphoribosyl transferase inhibitor. GR50s for those drugs were mostly in the nanomolar range, and in many cases below 10 nM. These drugs had long-lasting effect upon drug withdrawal, limited toxicity to normal cells and good efficacy also against tumor explants. Finally, we identified potential genomic biomarkers of their efficacy. Being approved or in clinical trials, these drugs are promising candidates for liposarcoma treatment.

scholarly journals In Vitro Systematic Drug Testing Reveals Carboplatin, Paclitaxel, and Alpelisib as a Potential Novel Combination Treatment for Adult Granulosa Cell Tumors

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 368
Author(s):  
Joline Roze ◽  
Elena Sendino Garví ◽  
Ellen Stelloo ◽  
Christina Stangl ◽  
Ferdinando Sereno ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Cell Lines ◽  
Combination Treatment ◽  
Drug Testing ◽  
Drug Application ◽  
Hormonal Treatment ◽  
Patient Specific ◽  
Granulosa Cell Tumors ◽  
Potential Synergy

Adult granulosa cell tumors (AGCTs) arise from the estrogen-producing granulosa cells. Treatment of recurrence remains a clinical challenge, as systemic anti-hormonal treatment or chemotherapy is only effective in selected patients. We established a method to rapidly screen for drug responses in vitro using direct patient-derived cell lines in order to optimize treatment selection. The response to 11 monotherapies and 12 combination therapies, including chemotherapeutic, anti-hormonal, and targeted agents, were tested in 12 AGCT-patient-derived cell lines and an AGCT cell line (KGN). Drug screens were performed within 3 weeks after tissue collection by measurement of cell viability 72 h after drug application. The potential synergy of drug combinations was assessed. The human maximum drug plasma concentration (Cmax) and steady state (Css) thresholds obtained from available phase I/II clinical trials were used to predict potential toxicity in patients. Patient-derived AGCT cell lines demonstrated resistance to all monotherapies. All cell lines showed synergistic growth inhibition by combination treatment with carboplatin, paclitaxel, and alpelisib at a concentration needed to obtain 50% cell death (IC50) that are below the maximum achievable concentration in patients (IC50 < Cmax). We show that AGCT cell lines can be rapidly established and used for patient-specific in vitro drug testing, which may guide treatment decisions. Combination treatment with carboplatin, paclitaxel, and alpelisib was consistently effective in AGCT cell lines and should be further studied as a potential effective combination for AGCT treatment in patients.

Reduced urinary opioid levels from pain management patients associated with marijuana use

2019 ◽  
Vol 9 (5) ◽  
pp. 441-447
Author(s):  
Melissa M Goggin ◽  
Breane J Shahriar ◽  
Andy Stead ◽  
Gregory C Janis
Keyword(s):  
Pain Management ◽  
Drug Testing ◽  
Marijuana Use ◽  
Potential Interaction ◽  
Test Results ◽  
Opioid Use ◽  
Target Drug ◽  
Urine Drug ◽  
Urine Drug Test ◽  
Lower Urinary

Aim: Marijuana use has been postulated to modulate opioid use, dependence and withdrawal. Broad target drug testing results provide a unique perspective to identify any potential interaction between marijuana use and opioid use. Materials & methods: Using a dataset of approximately 800,000 urine drug test results collected from pain management patients of a time from of multiple years, creatinine corrected opioid levels were evaluated to determine if the presence of the primary marijuana marker 11-nor-carboxy-tetrahydrocannabinol (THC-COOH) was associated with statistical differences in excreted opioid concentrations. Results & conclusion: For each of the opioids investigated (codeine, morphine, hydrocodone, hydromorphone, oxycodone, oxymorphone, fentanyl and buprenorphine), marijuana use was associated with statistically significant lower urinary opiate levels than in samples without indicators of marijuana use.

scholarly journals Automating Human Induced Pluripotent Stem Cell Culture and Differentiation of iPSC-Derived Retinal Pigment Epithelium for Personalized Drug Testing

2020 ◽  
pp. 247263032097211
Author(s):  
Vincent Truong ◽  
Kevin Viken ◽  
Zhaohui Geng ◽  
Samantha Barkan ◽  
Blake Johnson ◽  
...  
Keyword(s):  
Cell Culture ◽  
Retinal Pigment Epithelium ◽  
Cell Lines ◽  
Drug Screening ◽  
Drug Testing ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Age Related Macular Degeneration ◽  
End To End ◽  
Induced Pluripotent

Derivation and differentiation of human induced pluripotent stem cells (hiPSCs) provide the opportunity to generate medically important cell types from individual patients and patient populations for research and the development of potential cell therapies. This technology allows disease modeling and drug screening to be carried out using diverse population cohorts and with more relevant cell phenotypes than can be accommodated using traditional immortalized cell lines. However, technical complexities in the culture and differentiation of hiPSCs, including lack of scale and standardization and prolonged experimental timelines, limit the adoption of this technology for many large-scale studies, including personalized drug screening. The entry of reproducible end-to-end automated workflows for hiPSC culture and differentiation, demonstrated on commercially available platforms, provides enhanced accessibility of this technology for both research laboratories and commercial pharmaceutical testing. Here we have utilized TECAN Fluent automated cell culture workstations to perform hiPSC culture and differentiation in a reproducible and scalable process to generate patient-derived retinal pigment epithelial cells for downstream use, including drug testing. hiPSCs derived from multiple donors with age-related macular degeneration (AMD) were introduced into our automated workflow, and cell lines were cultured and differentiated into retinal pigment epithelium (RPE). Donor hiPSC-RPE lines were subsequently entered in an automated drug testing workflow to measure mitochondrial function after exposure to “mitoactive” compounds. This work demonstrates scalable, reproducible culture and differentiation of hiPSC lines from individuals on the TECAN Fluent platform and illustrates the potential for end-to-end automation of hiPSC-based personalized drug testing.

Spontaneous keratinocyte cell lines representing early and advanced stages of malignant transformation of the epidermis

2000 ◽  
Vol 9 (2) ◽  
pp. 104-117 ◽  
Author(s):  
C. M. Proby ◽  
K. J. Purdie ◽  
C. J. Sexton ◽  
P. Purkis ◽  
H. A. Navsaria ◽  
...  
Keyword(s):  
Malignant Transformation ◽  
Cell Lines ◽  
Keratinocyte Cell ◽  
Advanced Stages

Comparison of Patients (Pts) Characteristic and Treatment Outcomes in Lymphocyte-Predominant (LPHD) and Classical Hodgkin’s Disease (cHD): A Report from the German Hodgkin Lymphoma Study Group (GHSG).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1309-1309
Author(s):  
Lucia Nogova ◽  
Thorsten Reineke ◽  
Andreas Josting ◽  
Karolin Behringer ◽  
Beate Pfistner ◽  
...  
Keyword(s):  
Treatment Outcome ◽  
Progressive Disease ◽  
Early Stage ◽  
Patient Characteristics ◽  
Late Relapse ◽  
Early Stages ◽  
Comprehensive Picture ◽  
Advanced Stages ◽  
New Treatment ◽  
Lymphoma Study Group

Abstract Introduction: LPHD differs in histological and clinical presentation from cHD. Treatment of LPHD patients (pts) using standard Hodgkin disease (HD) protocols leads to complete remission (CR) in more than 95% of pts. However, differences in terms of relapse rates, survival and freedom from treatment failure (FFTF) between clinical stages of LPHD and cHD pts were suggested by a recent intergroup analysis. To obtain a more comprehensive picture, we reviewed all LPHD-cases registered in the GHSG database and compared patient characteristics and treatment outcome with cHD pts. Patients and methods: We retrospectively analysed 8298 HD pts treated within the GHSG trials (HD4 to HD12): 394 LPHD pts and 7904 cHD pts. From 394 LPHD pts 63% were in early stage, 16% in intermediate and 21% in advanced stage of disease. Of the 7904 cHD pts analysed, 22% were in early, 39% in intermediate and 39% in advanced stages. 9% of LPHD pts had B symptoms compared to 40% in cHD pts. Results: 87.5% LPHD pts reached CR/CRu compared to 81.1% cHD pts. 0.3% LPHD pts developed progressive disease (PD) compared to 3.7% cHD. The relapse rate of LPHD pts was very similar to cHD (6.9%). 0.8% LPHD pts and 3.1% cHD had early relapse, 6.9% LPHD pts and 4.2% cHD pts suffered from late relapse. LPHD pts with late relapse were in intermediate stages (12.5%). In contrast, only 3.9% of cHD pts in intermediate stages had late relapses. There were 2.5% secondary malignancies in LPHD and 3.7% in cHD pts. 4.3% LPHD pts and 8.8% cHD pts died. The FFTF rates for LPHD or cHD pts at a median observation of 41 or 48 months, respectively were as follows: early stages 93% vs. 87%, in intermediate stages 87% vs. 85% and in advanced stages 77% vs. 75%. The OS rates for LPHD or cHD pts were 98% vs. 96% for early stages, 94% vs. 93% for intermediate stages and 91% vs. 87% for advanced stages. The multivariate analysis of prognostic and risk factors will be presented. Conclusion: cHD pts present more frequently with advanced stages and B symptoms compared to LPHD pts. Comparing LPHD and cHD pts we found differences in treatment outcome in respect of CR/CRu, progressive disease and mortality. Surprisingly, there were no differences in terms of relapses. The majority of late relapses in LPHD pts were in intermediate stages. In contrast to previous reports, our data suggest that relapses in LPHD and cHD pts seem to be comparable and not more frequent. Thus new treatment strategy for LPHD pts in intermediate stages should be considered.

Treatment with the Anti-CD20 Antibody (B1) and Irradiation Result in Synergistic Cytotoxicity That Is Dependent on MAPK Activation.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2508-2508
Author(s):  
Andrei Ivanov ◽  
Mark S. Cragg ◽  
Tim M. Illidge
Keyword(s):  
Cell Lines ◽  
Molecular Mechanisms ◽  
Death Receptor ◽  
Clonogenic Survival ◽  
Lymphoma Cell ◽  
Non Hodgkin Lymphoma ◽  
Morphological Studies ◽  
Synergistic Cytotoxicity ◽  
New Treatment ◽  
Anti Cd20

Abstract Radioimmunotherapy using radiolabeled anti-CD20 antibodies (mAb) is an effective new treatment in non-Hodgkin lymphoma with high response rates. However, the molecular mechanisms behind these impressive clinical responses are poorly understood. To elucidate these mechanisms we studied the signaling events evoked in a panel of lymphoma cell lines following treatment with anti-CD20 mAb alone or in combination with irradiation. In all three lymphoma cell-lines tested a synergistic cytotoxic effect was observed when the anti-CD20 mAb B1 was combined with irradiation. The additive effect seen with B1 mAb and radiation was not observed with Rituximab and could be reversed with MEK inhibitors U0126 and PD98059 as well as siRNA targeting MEK1 or 2. Moreover, addition of U0126 reversed the decrease in clonogenic survival triggered by treatment with B1 and irradiation. To further probe the mechanism of this synergistic cell death we used cell lines over-expressing BCL2 or crmA, to block mitochondrial and death receptor pathways, respectively. Although BCL2 and crmA over-expression mediated protection against radiation alone, it had no impact on the increased cytotoxicity induced by B1+irradiation. Morphological studies revealed gross vacuolization of the cytoplasm, yet relatively well preserved nuclei in cells treated with B1+irradiation. Taken together our data indicate that activation of the MAPK cascade is an important factor that contributes to the synergistic effect of anti-CD20 (B1) antibody and irradiation and provides important new insights into how this treatment may work in the clinic.

Characterization Of FLT3/ITD Resistant Cells and Identification Of Novel Therapeutic Targets Utilizing High Throughput Drug Screening

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2522-2522
Author(s):  
Katherine Tarlock ◽  
C. Anthony Blau ◽  
Timothy Martins ◽  
Soheil Meshinchi
Keyword(s):  
Cell Lines ◽  
High Throughput ◽  
Drug Screening ◽  
Therapeutic Options ◽  
High Throughput Drug Screening ◽  
Mutational Profiling ◽  
Flt3 Inhibitors ◽  
Resistant Cells

Abstract The overall survival (OS) of pediatric acute myeloid leukemia (AML) exceeds 60%, however for high risk (HR) patients, including high allelic ratio FLT3/ITD+, survival remains poor. FLT3/ITD is one of the first genomic alterations in AML to be exploited for therapeutic benefit as it has greater sensitivity to the pro-apoptotic effects of FLT3-inhibitors. Children’s Oncology Group (COG) phase III AML trial AAML1031 is investigating the role of sorafenib in combination with chemotherapy in HR FLT3/ITD+ patients. In vitro and in vivo studies indicate that resistance to FLT3-inhibitors can develop through varying mechanisms including up-regulation of FLT3 receptor, acquisition of secondary mutations, or activation of alternate survival mechanisms leading to apoptotic escape. For FLT3/ITD+ patients who relapse despite treatment with FLT3-inhibitors, there are often no therapeutic options and survival is very poor. In evaluation of therapeutic options for those who relapse on sorafenib, we developed an in vitro resistance model using the FLT3/ITD+ cell line MV4-11. Resistance was induced thru long-term exposure to incrementally increasing doses of sorafenib. Two distinct cell lines with resistance at 10 and 100 fold above the IC50 of naïve MV4-11 were generated for experimental evaluation. Genotypic and phenotypic characterization of the resistant cells was conducted by multidimensional flow cytometry (MDF), conventional karyotyping, and mutational profiling. MDF revealed an overall similar immunophenotype, however the resistant cells were significantly more homogeneous for expression of HLA-DR and had significantly higher CD11b expression compared to their naïve counterparts. CD135 expression was minimally increased in the resistant cells. In comparison of the karyotypes, the resistant cells were a more homogenous population with emergence of one dominant clone and disappearance of a number of pre-existing sub-clones. Mutational profiling by Sanger sequencing revealed a novel N841Y mutation in activation loop, an area implicated in TKI-resistance. Using a high throughput drug screening assay, we explored sensitivity profiles of the naïve and resistant MV4-11 cells to 163 oncology agents, including 45 FDA approved and 118 investigational agents that target a number of key pathways regulating cell growth, differentiation, and survival. The naïve MV4-11s had a sorafenib IC50 of 1.3 nM (published 1-5nM) and resistant cells had IC50 of approximately 2-log folds above the naïve, which was consistent to what we had seen in our lab-based validations. We initially assessed whether resistance to sorafenib induces cross-resistance to other TKIs. Agents in the panel with previously demonstrated efficacy for FLT3/ITD included quizartinib (AC-220), tandutinib, ponatinib, sunitinib, and midostaurin, and in all cases sorafenib-resistant cells were also more resistant to these agents. We then examined whether we could identify agents with efficacy in the resistant cells. We identified 5 novel agents to which the resistant cells retained sensitivity. Two bcl-2 inhibitors tested maintained sensitivity in the resistant cells with IC50s in the 20-100nM range. In addition, YM-155, a survivin inhibitor, also maintained sensitivity in the resistant cells with IC50s of approximately 25-50nM across the cell lines. Survivin over-expression is associated with AML stem progenitor cells and decreased OS in adults, and transcription regulation has been linked to the FLT3/STAT5 pathway. Two CRM inhibitors, a novel class of agents which inhibit nuclear export to restore tumor suppressor function, also maintained sensitivity in the resistant cell lines with an approximate 3-fold increase in IC50 from 12nM in the naïve to 32-40nM in the resistant cells. Experience with the use of directed therapy to target specific somatic events has provided evidence that leukemic evolution can continue under this selection pressure and therapeutic options for patients with emergent disease is often insufficient. Using the high throughput drug assay in a FLT3/ITD+ cell line as an in vitro model for sorafenib-resistant FLT3/ITD patients, we identified classes of targeted agents that maintain sensitivity in resistant cells. Further validation of the targets in specimens from those with resistance to such TKIs can inform on the class of agents that can be used to treat or prevent refractory disease FLT3/ITD+ patients. Disclosures: No relevant conflicts of interest to declare.

Proteomic analysis of extracellular vesicles from medullospheres reveals a role for iron in the cancer progression of medulloblastoma

2019 ◽  
pp. 1-12
Author(s):  
Maja Larsen ◽  
Matthias Kuhlmann Kuhlmann ◽  
Michael Hvam ◽  
Kenneth Howard
Keyword(s):  
Cell Lines ◽  
Cancer Progression ◽  
Early Stage ◽  
Cell Communication ◽  
Treatment Strategies ◽  
Iron Chelators ◽  
High Morbidity ◽  
New Treatment ◽  
Stem Cell Populations

Background: Medulloblastoma (MB) is the most common malignant childhood brain tumor with the propensity todisseminate at an early stage, and is associated with high morbidity. New treatment strategies are needed toimprove cure rates and to reduce life-long cognitive and functional deficits associated with current therapies.Extracellular Vesicles (EVs) are important players in cell-to-cell communication in health and diseases. A clearerunderstanding of cell-to-cell communication in tumors can be achieved by studying EV secretion inmedullospheres. This can reveal subtle modifications induced by the passage from adherent to non-adherentgrowth, as spheres may account for the adaptation of tumor cells to the mutated environment.Methods: Formation of medullospheres from MB cell lines stabilized in adherent conditions was obtained throughculture conditioning based on low attachment flasks and specialized medium. EVs collected by ultracentrifugation,in adherent conditions and as spheres, were subjected to electron microscopy, NanoSight measurements andproteomics.Results: Interestingly, iron carrier proteins were only found in EVs shed by CSC-enriched tumor cell population ofspheres. We used iron chelators when culturing MB cell lines as spheres. Iron chelators induced a decrease innumber/size of spheres and in stem cell populations able to initiate in vitro spheres formation.Conclusions: This work suggests a not yet identified role of iron metabolism in MB progression and invasion andopens the possibility to use chelators as adjuvants in anti-tumoral chemotherapy.

Preclinical development of small-molecule CRM1 inhibitors as novel therapy for the treatment of colorectal cancer (CRC).

2011 ◽  
Vol 29 (4_suppl) ◽  
pp. 430-430 ◽  
Author(s):  
S. Shacham ◽  
M. Kauffman ◽  
V. Sandanayaka ◽  
G. Draetta ◽  
S. Shechter ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Cell Lines ◽  
Small Molecule ◽  
Nuclear Export ◽  
Cysteine Proteases ◽  
Preclinical Development ◽  
Normal Cells ◽  
Regulatory Pathways ◽  
Multiple Tumor ◽  
Hct 116

430 Background: CRM1 (XPO1) is a key nuclear export protein which controls the location of multiple tumor suppressor (TSP) and growth regulatory (GRP) proteins including p53, PI3K/AKT, Wnt/ß-catenin and NF-kB. Forced nuclear expression of TSP and GRP by CRM1 inhibition can lead to apoptosis in cancer cells while sparing normal cells. Methods: Novel small-moleculeCRM1 inhibitors were synthesized and nuclear distribution studies were performed in cells transfected with HIV-rev GFP proteins. Cell proliferation studies were performed in 16 CRC cell lines: LS-123, SW-626, Colo-201, Colo-205, Colo-320DM, Colo-320HSR, Lovo, DLD-1, HCT-15, WiDi, LS-174T, LS-180, SW-620, C2BBe1, HCT-8, HCT-116, and in human peripheral leukocytes (PBMC). Cellular distribution and apoptosis assays were performed on HCT-116. Antitumor activity is assessed in human HCT-116 xenografts in scid-mice. Results: The lead CRM1 inhibitor, KPT-0127, blocks CRM1 mediated nuclear export of HIV-Rev-GFP, FOXO, and p53 with an IC50 of ∼300 nM. KPT-0127 is cytotoxic to various CRC cell lines with EC50s of 0.07-1.1 μM; in 9 CRC lines EC50s were < 0.3 mM. In contrast, normal cell lines and PBMCs had EC50 > 5-20 μM. In HCT- 116 cells, KPT-0127 induces cell cycle arrest at both G1/S and G2/M checkpoints and dose dependently increases nuclear p53, followed by an increase in caspase 3. KTP-0127 10μM shows no significant effect on 37 proteins including several cysteine proteases. In mice, KPT-0127 given by SC injection of 30-100 mg/kg leads to serum levels exceeding the effective CRM1 inhibitory concentration for at least 4 hours and is well tolerated. KPT-0127 given SC to mice bearing HCT-116 colon xenografts results in dose-dependent antitumor activity. Conclusions: The novel small- molecule CRM1 inhibitor KTP-0127 kills CRC lines with multiple TSP, GRP, and oncogenic abnormalities, including p53 mutations/deletions and PTEN deficiency/AKT activation, while sparing normal cells. This likely reflects the ability of CRM1 inhibition to affect multiple critical and non-redundant regulatory pathways. These results support the development of CRM1 inhibitors for the treatment of CRC. IND-enabling CMC and toxicology work are in preparation. [Table: see text]

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