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

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.

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Functional precision medicine in colorectal cancer based on patient-derived tumoroids and in-vitro sensitivity drug testing.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.e15567 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. e15567-e15567

Author(s):

Lars Henrik Jensen ◽

Anders Kristian Moeller Jakobsen ◽

Birgitte Mayland Havelund ◽

Cecilie Abildgaard ◽

Chris Vagn-Hansen ◽

...

Keyword(s):

Colorectal Cancer ◽

Clinical Trial ◽

Primary Endpoint ◽

Drug Testing ◽

Progression Free Survival ◽

Patient Specific ◽

Precision Oncology ◽

Sensitivity Testing ◽

In Vitro Sensitivity

e15567 Background: Precision oncology based on in-vitro, functional assays has potential advantages compared to the much more common molecular approach, but the clinical benefit is unknown. We here report the results from the largest prospective interventional clinical trial testing the clinical outcome in colorectal cancer patients treated with drugs showing cytotoxic effect in matched patient-derived tumoroids. Methods: This single-center, phase II trial included patients with metastatic colorectal cancer previously exposed to all standard therapies. Specimens from one to three 18-16 G core needle biopsies were manually dissected, enzymatically treated, cultivated, and incubated to form 3D spherical microtumors, i.e. tumoroids. In the assay for in-vitro sensitivity testing, the tumoroids were challenged with single drugs and combinations thereof to determine patient-specific responses. Using tumoroid screening technology (IndiTreat, 2cureX, Copenhagen, Denmark), results were generated by comparing the sensitivity of the individual patient’s tumoroids with a reference panel from other patients. The testing included standard cytostatics and drugs with proven effect in previous early-phase clinical trials, a total of 15 drugs. The primary endpoint was the fraction of patients with progression-free survival (PFS) at two months. Based on placebo arms in randomized last-line trials, a minimal relevant difference of 20% (20% to 40%) was stated. Using Simon's two-stage design, a sample size of 45 patients was calculated with at least 14 PFS at two months (significance 5%, power 90%). Results: Ninety patients were enrolled from 9/2017 to 9/2020. Biopsies from 82 patients were obtained and sent for tumoroid formation of which 44 (54%, 95% CI 42-65) were successful and at least one treatment was suggested. Thirty-four patients initiated treatment according to the response obtained in the drug assays within a median of 51 days from inclusion (IQR 39-63). The primary endpoint, PFS at two months, was met in 17 of 34 patients (50%, 95%CI 32-68). There were no radiological responses. Median PFS was 81 days (95% CI 51-112) and median OS was 189 days (95% CI 103-277). Conclusions: Precision oncology using a functional approach with patient-derived tumoroids and in-vitro drug sensitivity testing seems feasible. The approach is limited by the fraction of patients with successful tumoroid development. The primary endpoint was met, as half of the patients were without progression at two months. Further clinical studies are justified. Clinical trial information: NCT03251612.

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PL3.6 Targeting GSK-3 activity promotes mitotic catastrophe via centrosome destabilisation and enhances the effect of radiotherapy in glioma models

Neuro-Oncology ◽

10.1093/neuonc/noz126.009 ◽

2019 ◽

Vol 21 (Supplement_3) ◽

pp. iii4-iii4

Author(s):

A Bruning-Richardson ◽

H Sanganee ◽

S Barry ◽

D Tams ◽

T Brend ◽

...

Keyword(s):

Cell Lines ◽

Cancer Progression ◽

Combination Treatment ◽

Single Agent ◽

Drug Penetration ◽

In Vivo Models ◽

Human Astrocytes ◽

Normal Human

Abstract BACKGROUND Targeting kinases as regulators of cellular processes that drive cancer progression is a promising approach to improve patient outcome in GBM management. The glycogen synthase kinase 3 (GSK-3) plays a role in cancer progression and is known for its pro-proliferative activity in gliomas. The anti-proliferative and cytotoxic effects of the GSK-3 inhibitor AZD2858 were assessed in relevant in vitro and in vivo glioma models to confirm GSK-3 as a suitable target for improved single agent or combination treatments. MATERIAL AND METHODS The immortalised cell line U251 and the patient derived cell lines GBM1 and GBM4 were used in in vitro studies including MTT, clonogenic survival, live cell imaging, immunofluorescence microscopy and flow cytometry to assess the cytotoxic and anti-proliferative effects of AZD2858. Observed anti-proliferative effects were investigated by microarray technology for the identification of target genes with known roles in cell proliferation. Clinical relevance of targeting GSK-3 with the inhibitor either for single agent or combination treatment strategies was determined by subcutaneous and orthotopic in vivo modelling. Whole mount mass spectroscopy was used to confirm drug penetration in orthotopic tumour models. RESULTS AZD2858 was cytotoxic at low micromolar concentrations and at sub-micromolar concentrations (0.01 - 1.0 μM) induced mitotic defects in all cell lines examined. Prolonged mitosis, centrosome disruption/duplication and cytokinetic failure leading to cell death featured prominently among the cell lines concomitant with an observed S-phase arrest. No cytotoxic or anti-proliferative effect was observed in normal human astrocytes. Analysis of the RNA microarray screen of AZD2858 treated glioma cells revealed the dysregulation of mitosis-associated genes including ASPM and PRC1, encoding proteins with known roles in cytokinesis. The anti-proliferative and cytotoxic effect of AZD2858 was also confirmed in both subcutaneous and orthotopic in vivo models. In addition, combination treatment with AZD2858 enhanced clinically relevant radiation doses leading to reduced tumour volume and improved survival in orthotopic in vivo models. CONCLUSION GSK-3 inhibition with the small molecule inhibitor AZD2858 led to cell death in glioma stem cells preventing normal centrosome function and promoting mitotic failure. Normal human astrocytes were not affected by treatment with the inhibitor at submicromolar concentrations. Drug penetration was observed alongside an enhanced effect of clinical radiotherapy doses in vivo. The reported aberrant centrosomal duplication may be a direct consequence of failed cytokinesis suggesting a role of GSK-3 in regulation of mitosis in glioma. GSK-3 is a promising target for combination treatment with radiation in GBM management and plays a role in mitosis-associated events in glioma biology.

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Phenotypic profiling with a living biobank of primary rhabdomyosarcoma unravels disease heterogeneity and AKT sensitivity

Nature Communications ◽

10.1038/s41467-020-18388-7 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Gabriele Manzella ◽

Leonie D. Schreck ◽

Willemijn B. Breunis ◽

Jan Molenaar ◽

Hans Merks ◽

...

Keyword(s):

Cancer Therapy ◽

Tumor Cells ◽

Drug Testing ◽

Specific Treatment ◽

Patient Specific ◽

Molecular Characteristics ◽

Disease Heterogeneity ◽

Molecular Analyses ◽

Functional Profiling

Abstract Cancer therapy is currently shifting from broadly used cytotoxic drugs to patient-specific precision therapies. Druggable driver oncogenes, identified by molecular analyses, are present in only a subset of patients. Functional profiling of primary tumor cells could circumvent these limitations, but suitable platforms are unavailable for most cancer entities. Here, we describe an in vitro drug profiling platform for rhabdomyosarcoma (RMS), using a living biobank composed of twenty RMS patient-derived xenografts (PDX) for high-throughput drug testing. Optimized in vitro conditions preserve phenotypic and molecular characteristics of primary PDX cells and are compatible with propagation of cells directly isolated from patient tumors. Besides a heterogeneous spectrum of responses of largely patient-specific vulnerabilities, profiling with a large drug library reveals a strong sensitivity towards AKT inhibitors in a subgroup of RMS. Overall, our study highlights the feasibility of in vitro drug profiling of primary RMS for patient-specific treatment selection in a co-clinical setting.

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IPI-504, a Novel, Orally Active HSP90 Inhibitor, Prolongs Survival of Mice with BCR-ABL T315I CML and B-ALL.

Blood ◽

10.1182/blood.v108.11.2183.2183 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2183-2183 ◽

Cited By ~ 1

Author(s):

Cong Peng ◽

Julia Brain ◽

Yiguo Hu ◽

Linghong Kong ◽

David Grayzel ◽

...

Keyword(s):

Cell Lines ◽

Median Survival ◽

Combination Treatment ◽

Lymphoblastic Leukemia ◽

Hsp90 Inhibitor ◽

Leukemic Cells ◽

Prolonged Survival ◽

Hsp90 Inhibition ◽

T315i Mutation

Abstract Development of mutations within the kinase domain is a major drug-resistance mechanism for tyrosine kinase inhibitors (TKIs) in cancer therapy. In CML (chronic myeloid leukemia), a disease driven by the constitutively active BCR-ABL oncoprotein, no available TKIs have been effective in treating patients with the BCR-ABL T315I mutation. Heat shock protein 90 (Hsp90) is a highly conserved, constitutively expressed molecular chaperone that facilitates folding of client proteins like BCR-ABL, and affects the stability of these proteins. Several labs have shown that Hsp90 inhibition in vitro results in the degradation of BCR-ABL T315I and induces potent killing of these cell lines. However, these results have not been demonstrated in animal models for BCR-ABL-induced CML and B-ALL (B-cell acute lymphoblastic leukemia, a disease that does not respond well to TKIs including imatinib and dasatinib). Thus, IPI-504, an orally administered Hsp90 inhibitor, was evaluated in murine models of CML and B-ALL. Treatment of mice with wild type (WT)- or T315I BCR-ABL-induced CML with IPI-504 resulted in BCR-ABL protein degradation and a decrease in circulating BCR-ABL positive cells. In response to treatment with vehicle the median survival time of WT and T315I CML mice is approximately 20 days. While the T315I CML mice were resistant to imatinib with a median survival of 21 days, IPI-504 (50 and 100 mg/kg, PO TIW) demonstrated dose-dependent prolonged survival of these mice by 30 and 70 days, respectively (p<0.001 for both doses). Both imatinib and IPI-504 similarly prolonged survival of mice with BCR-ABL-WT-induced CML. In the T315I CML mice prolonged survival of the IPI-504 treated cohort was associated with decreased peripheral blood BCR-ABL positive leukemia cells during treatment, less splenomegaly and improved pulmonary histopathlogy at necropsy. In CML mice receiving mixed BCR-ABL-WT- or T315I-transduced donor bone marrow cells, Hsp90 inhibition more potently suppressed T315I-expressing leukemia clones relative to the WT clones, consistent with in vitro studies where T315I BCR-ABL was more sensitive to IPI-504 induced degradation in cell lines than WT BCR-ABL. Combination treatment with IPI-504 and imatinib was more effective than either treatment alone in prolonging survival of mice bearing both WT and T315I leukemic cells. IPI-504 also significantly prolonged survival of B-ALL mice bearing the T315I mutation (p<0.001). These results provide a rationale for use of an Hsp90 inhibitor as a novel approach to overcoming resistance to TKIs as well as the potential for first line combination treatment in CML patients. The potential for IPI-504 to eliminate mutant kinases via Hsp90 inhibition provides a new therapeutic strategy for treating BCR-ABL-induced CML, ALL as well as other cancers resistant to treatment with TKIs.

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Vitamin C Enhances PARPi Efficacy for the Treatment of AML

Blood ◽

10.1182/blood-2021-153884 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 1168-1168

Author(s):

John P Brabson ◽

Tiffany Leesang ◽

Byron Fang ◽

Jingjing Wang ◽

Victoria Strippoli ◽

...

Keyword(s):

Vitamin C ◽

Cell Lines ◽

Combination Treatment ◽

Dna Demethylation ◽

Dependent Manner ◽

Publicly Traded ◽

Colony Forming Capacity ◽

Murine Leukemia

Abstract Poly-ADP-ribose polymerase inhibitors (PARPi) are currently in clinical trial to determine their therapeutic efficacy for the treatment of acute myeloid leukemia (AML). We have shown that vitamin C (VitC), an essential micronutrient and co-factor of Ten-Eleven translocation (TET) proteins, enhances AML sensitivity to PARPi, potentially due to an increased dependency on base-excision repair (BER) enzymes needed to remove TET-catalyzed oxidized methylcytosine bases via active DNA demethylation. TET2 is the most frequently mutated TET gene in patients with AML, and vitamin C treatment can mimic genetic restoration of TET2 function, leading to DNA demethylation, differentiation, and leukemia cell death. Whether vitamin C efficacy in combination with PARPi depends on the level of TET2 functional alleles is not yet known and may stratify whether TET2 wild-type or mutant patients should be targeted by vitamin C adjuvant therapy. We have generated primary murine AML-ETO9a+ and MLL-AF9+ leukemia models with Tet2 +/+, Tet2 +/- and Tet2 -/- alleles to determine the Tet2-dependent efficacy of PARPi treatment when combined with vitamin C. Furthermore, we have performed CRISPR gene knockout and drug library screening in human AML cell lines in combination with vitamin C treatment, and tested a panel of 10 AML cell lines with titrating concentrations of PARPi (Olaparib, Talazoparib, Veliparib and Rucaparib) alone or in combination with vitamin C (L-ascorbic acid) mimicking physiological to pharmacological in vivo doses. Primary murine AML cells and human cell lines were assayed for colony-forming capacity, differentiation, cell cycling, viability and effects on DNA methylation, levels of oxidized 5-mC and gene expression upon combination treatment in vitro and in vivo. TET2 mutant PDX and primary murine AMLs treated in vivo with L-ascorbate (4g/kg) and Olaparib (50mg/kg) by daily IP injection were also monitored for disease burden, cellular differentiation and survival. Vitamin C is known to drive the TET-catalyzed iterative oxidation of 5-methylcytosine (5-mC) leading to the formation of 5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC). We show that VitC-PARPi combination treatment causes an accumulation of oxidized 5-mC intermediates in the AML genome that correlates with increased yH2AX formation in mid-S phase and cell cycle stalling. Vitamin C reduces the IC 50 of Olaparib and Talazoparib by greater than 10-fold in human AML cells lines and primary murine leukemia cells, and treatment in combination promotes myeloid differentiation and blocks colony-forming capacity greater than either alone. In both our in vitro and in vivo studies, Tet2 +/- AML cells exhibit increased sensitivity to vitamin C treatment alone or in combination with PARPi compared to either Tet2 +/+ or Tet2 -/- cells, suggesting that patients with TET2 haploinsufficiency, which represents the majority of TET2 mutant cases, could benefit the most from combined treatment. Our findings confirm that vitamin C can act synergistically with PARPi to block AML cell viability, reduce colony-forming capacity, and decrease leukemia burden in PDX and primary murine leukemia models in a TET2 allelic dose-dependent manner. The combinatorial effect works at clinically relevant concentrations of PARPi, and low-pharmacological doses of vitamin C. These studies suggest that vitamin C can be used as a non-toxic therapeutic adjuvant to PARPi therapy for the treatment of AML. Disclosures Neel: Northern Biologics, LTD: Current equity holder in publicly-traded company, Other: Co- Founder; SAB: Other: Co-Founder; Navire Pharma: Consultancy, Current equity holder in publicly-traded company; Jengu Therapeutics: Consultancy, Current equity holder in publicly-traded company, Other: Co-Founder; Arvinas, Inc: Consultancy, Current equity holder in publicly-traded company; Recursion Pharma: Current equity holder in publicly-traded company.

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Carboplatin sensitivity in epithelial ovarian cancer cell lines: The impact of model systems

PLoS ONE ◽

10.1371/journal.pone.0244549 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0244549

Author(s):

Bishnubrata Patra ◽

Muhammad Abdul Lateef ◽

Melica Nourmoussavi Brodeur ◽

Hubert Fleury ◽

Euridice Carmona ◽

...

Keyword(s):

Ovarian Cancer ◽

Epithelial Ovarian Cancer ◽

Cell Line ◽

Cell Lines ◽

Drug Testing ◽

3D Models ◽

Model Systems ◽

Microfluidic Chips ◽

The Impact

Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy in North America, underscoring the need for the development of new therapeutic strategies for the management of this disease. Although many drugs are pre-clinically tested every year, only a few are selected to be evaluated in clinical trials, and only a small number of these are successfully incorporated into standard care. Inaccuracies with the initial in vitro drug testing may be responsible for some of these failures. Drug testing is often performed using 2D monolayer cultures or 3D spheroid models. Here, we investigate the impact that these different in vitro models have on the carboplatin response of four EOC cell lines, and in particular how different 3D models (polydimethylsiloxane-based microfluidic chips and ultra low attachment plates) influence drug sensitivity within the same cell line. Our results show that carboplatin responses were observed in both the 3D spheroid models tested using apoptosis/cell death markers by flow cytometry. Contrary to previously reported observations, these were not associated with a significant decrease in spheroid size. For the majority of the EOC cell lines (3 out of 4) a similar carboplatin response was observed when comparing both spheroid methods. Interestingly, two cell lines classified as resistant to carboplatin in 2D cultures became sensitive in the 3D models, and one sensitive cell line in 2D culture showed resistance in 3D spheroids. Our results highlight the challenges of choosing the appropriate pre-clinical models for drug testing.

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Application of Patient-Specific iPSCs for Modelling and Treatment of X-Linked Cardiomyopathies

International Journal of Molecular Sciences ◽

10.3390/ijms22158132 ◽

2021 ◽

Vol 22 (15) ◽

pp. 8132

Author(s):

Jennifer Zhang ◽

Oscar Hou-In Chou ◽

Yiu-Lam Tse ◽

Kwong-Man Ng ◽

Hung-Fat Tse

Keyword(s):

Drug Testing ◽

Control Cell ◽

Induced Pluripotent Stem Cell ◽

Patient Specific ◽

Mortality And Morbidity ◽

Danon Disease ◽

Disease Phenotypes ◽

Induced Pluripotent ◽

And Control

Inherited cardiomyopathies are among the major causes of heart failure and associated with significant mortality and morbidity. Currently, over 70 genes have been linked to the etiology of various forms of cardiomyopathy, some of which are X-linked. Due to the lack of appropriate cell and animal models, it has been difficult to model these X-linked cardiomyopathies. With the advancement of induced pluripotent stem cell (iPSC) technology, the ability to generate iPSC lines from patients with X-linked cardiomyopathy has facilitated in vitro modelling and drug testing for the condition. Nonetheless, due to the mosaicism of the X-chromosome inactivation, disease phenotypes of X-linked cardiomyopathy in heterozygous females are also usually more heterogeneous, with a broad spectrum of presentation. Recent advancements in iPSC procedures have enabled the isolation of cells with different lyonisation to generate isogenic disease and control cell lines. In this review, we will summarise the current strategies and examples of using an iPSC-based model to study different types of X-linked cardiomyopathy. The potential application of isogenic iPSC lines derived from a female patient with heterozygous Danon disease and drug screening will be demonstrated by our preliminary data. The limitations of an iPSC-derived cardiomyocyte-based platform will also be addressed.

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Bringing an Old Drug to a New Treatment Strategy in Treating FLT-ITD AML - Combination of Homoharringtonine and Sorafenib

Blood ◽

10.1182/blood.v124.21.3610.3610 ◽

2014 ◽

Vol 124 (21) ◽

pp. 3610-3610 ◽

Cited By ~ 1

Author(s):

Eric S.K. Ho ◽

Stephen Sze Yuen Lam ◽

Cheuk Him Man ◽

Chae Yin Cher ◽

Yok Lam Kwong ◽

...

Keyword(s):

Bone Marrow ◽

Cell Lines ◽

Combination Treatment ◽

Protein Translation ◽

Hematopoietic Stem ◽

Clinical Prognosis ◽

Protein Levels ◽

Downstream Effectors ◽

Flt3 Inhibitors

Abstract Acute myeloid leukemia (AML) is a heterogeneous group of diseases characterized by an abnormal increase of myeloblasts in bone marrow (BM) and peripheral blood (PB). Conventional approach of treatment with standard induction chemotherapy and allogeneic hematopoietic stem cell transplantation (HSCT) has reached an impasse with a cure rate of only 30%. A gain-of-function internal tandem duplication (ITD) of fms-like tyrosine kinase 3 (FLT3) was found in 30% of AML and was associated with an inferior treatment response and clinical prognosis. Despite much interests in FLT3 inhibitors in clinical trials, the response was at best transient, limiting their clinical application. Homoharringtonine (HHT) is a natural alkaloid widely used in Mainland China for the treatment of AML. It is a protein translation inhibitor and affects primarily proteins with short half-lives, including many of the downstream effectors of FLT3 signaling. In this study, we evaluated if HHT can be used in combination with sorafenib in the treatment of FLT3-ITD+ AML and examined the mechanistic basis of their synergism. The anti-leukemia effects of drugs on AML cell lines with or without FLT3-ITD mutation were evaluated by a high throughput PrestoBlue® fluorometric assay (a measure of viable cell number) after 3-day culture. Synergism between drugs in combination treatment was evaluated based on Excess Over Bliss Additivism (EOBA). The drug effects on leukemia initiating cells (LIC) activity were examined by xenotransplantation using NOD/SCID/IL2Rg-/- NSG mice and engraftment was enumerated by the presence of human CD45+/mouse CD45.1- cells 6 weeks after transplantation. The effects of HHT and FLT3 inhibitors on FLT3 signaling were examined by Western Blot and Phospho-flow analysis by flow cytometry. HHT exhibited more potent growth inhibitory effect on FLT3-ITD+ AML cell lines, MV4-11 and MOLM-13 (IC50: 3.65 and 3.67 nM) than other AML cell lines (IC50: 7.7 - 32.3 nM). Combination of HHT and sorafenib (H+S) showed pronounced synergism in growth inhibition based on EOBA (29.7% ± 5%) at 3.65 nM (HHT) and 3.85 nM (S). H+S also induced significant increase in apoptosis in MV4-11 based on Annexin V+ population (H+S: 21.3% ± 2.3%; HHT: 11.7% ± 1.9%; S: 8.4% ± 1.0%). Synergism with HHT could also be demonstrated with other FLT3 inhibitors, quizartinib (EOBA: 16% ± 8%) and ponatinib (EOBA: 24% ± 6%) on MV4-11 cell lines. There was no synergism between sorafenib and conventional chemotherapy as exemplified by cytarabine. H+S in vitro significantly reduced engraftment of MV4-11 cells (Vehicle: 65.0% ± 9.7%; H+S: 21.8% ± 8.7%, p<0.01, n=6). HHT alone also inhibited leukemia growth of primary AML samples (IC50: 38.4 ± 7.2 nM, n=88), that was below the peak plasma concentration of HHT (66 nM) in patients receiving treatment. Synergism between H+S was also seen in primary AML samples. Mechanistically, HHT treatment for 6 hours reduced total FLT3 and p-FLT3 protein levels in MV4-11 and MOLM-13. Protein levels of downstream effectors of FLT3 pathway including total Stat5, pStat5, pStat3, pErk were also reduced. A phase II clinical trial of sorafenib (200-400 mg twice daily continuous) and HHT (1.5 mg/m2 for 7 days every 28 days for the first cycle and for 4 days in subsequent cycles, until leukemia progression) combination treatment in patients with chemo-refractory FLT3-ITD+ AML has begun in Hong Kong since January 2014. Bone marrow was performed before and on day 21 after treatment and repeated every two cycles thereafter. Five patients have been treated, including two patients who were primarily refractory to sorafenib monotherapy. Complete remission (CR) was achieved in 1 patient and CR with incomplete hematological recovery (CRi) in four others after 1 cycle. The FLT3-ITD allelic burdens before treatment and at CR/CRi were 77.7 ± 9.6% and 20.0 ± 9.6% (p=0.007). As of 5th August 2014, the two primary sorafenib refractory patients had leukemia progression at 65 and 95 days after treatment. The other three patients remained in remission at 113, 134 and 150 days after treatment. In conclusion, HHT and sorafenib demonstrated significant synergistic effect in suppressing the growth of FLT3-ITD+ AML cells both in vitro and in vivo. It provides a promising strategy in improving treatment outcome of FLT3-ITD+ AML patients. Disclosures No relevant conflicts of interest to declare.

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Combination Treatment With Trabectedin and Irinotecan or Topotecan Has Synergistic Effects Against Ovarian Clear Cell Carcinoma Cells

International Journal of Gynecological Cancer ◽

10.1097/igc.0000000000000143 ◽

2014 ◽

Vol 24 (5) ◽

pp. 829-837 ◽

Cited By ~ 4

Author(s):

Mahiru Kawano ◽

Seiji Mabuchi ◽

Toshiko Kishimoto ◽

Takeshi Hisamatsu ◽

Yuri Matsumoto ◽

...

Keyword(s):

Cell Carcinoma ◽

Cell Lines ◽

Combination Treatment ◽

Clear Cell ◽

Clear Cell Carcinoma ◽

Synergistic Effects ◽

Chemotherapeutic Agents ◽

Ovarian Clear Cell Carcinoma ◽

Combination Treatments

ObjectivesThe objective of this study was to investigate the chemotherapeutic agents that produce the strongest synergistic effects when combined with trabectedin against ovarian clear cell carcinoma (CCC), which is regarded as an aggressive chemoresistant histological subtype.MethodsUsing 4 human CCC cell lines (RMG1, RMG2, KOC7C, and HAC2), the cytotoxicities of trabectedin, SN-38, topotecan, doxorubicin, cisplatin, and paclitaxel as single agents were first assessed using the MTS assay. Then, the cytotoxicities of combination treatments involving trabectedin and 1 of the other 4 agents were evaluated by isobologram analysis to examine whether these combinations displayed synergistic, additive, or antagonistic effects. The antitumor activities of the combination treatments were also examined using cisplatin-resistant and paclitaxel-resistant CCC sublines, which were derived from the parental CCC cells by continuously exposing them to cisplatin or paclitaxel. Finally, we determined the effect of everolimus on the antitumor efficacy of trabectedin-based combination chemotherapy.ResultsConcurrent exposure to trabectedin and SN-38 or topotecan resulted in synergistic interactions in all 4 CCC cell lines. Among the tested combinations, trabectedin plus SN-38 was the most effective cytotoxic regimen. The combination of trabectedin plus SN-38 also had strong synergistic effects on both the cisplatin-resistant and paclitaxel-resistant CCC cell lines. Treatment with everolimus significantly enhanced the antitumor activity of trabectedin plus SN-38 or topotecan.ConclusionsCombination treatment with trabectedin and SN-38 displays the greatest cytotoxic effect against ovarian CCC. Our in vitro study provides the rationale for future clinical trials of trabectedin plus irinotecan with or without everolimus in patients with ovarian CCC in both the front-line chemotherapy setting and as a second-line treatment of recurrent CCC that had previously been treated with cisplatin or paclitaxel.

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RUNX3 Promotes the Tumorigenic Phenotype in KGN, a Human Granulosa Cell Tumor-Derived Cell Line

International Journal of Molecular Sciences ◽

10.3390/ijms20143471 ◽

2019 ◽

Vol 20 (14) ◽

pp. 3471 ◽

Cited By ~ 1

Author(s):

Huachen Chen ◽

Powel Crosley ◽

Abul K. Azad ◽

Nidhi Gupta ◽

Nisha Gokul ◽

...

Keyword(s):

Cell Line ◽

Granulosa Cell ◽

Cell Lines ◽

Granulosa Cell Tumor ◽

Tumor Formation ◽

Dominant Negative ◽

Cell Cycle Regulators ◽

Cyclin D2

Granulosa cell tumors of the ovary (GCT) are the predominant type of ovarian sex cord/stromal tumor. Although prognosis is generally favorable, the outcome for advanced and recurrent GCT is poor. A better understanding of the molecular pathogenesis of GCT is critical to developing effective therapeutic strategies. Here we have examined the potential role of the runt-related transcription factor RUNX3. There are only two GCT cell lines available. While RUNX3 is silenced in the GCT cell line KGN cells, it is highly expressed in another GCT cell line, COV434 cells. Re-expression of RUNX3 promotes proliferation, anchorage-independent growth, and motility in KGN cells in vitro and tumor formation in mice in vivo. Furthermore, expression of a dominant negative form of RUNX3 decreases proliferation of COV434 cells. To address a potential mechanism of action, we examined expression of cyclin D2 and the CDK inhibitor p27Kip1, two cell cycle regulators known to be critical determinants of GCT cell proliferation. We found that RUNX3 upregulates the expression of cyclin D2 at the mRNA and protein level, and decreases the level of the p27Kip1 protein, but not p27Kip1 mRNA. In conclusion, we demonstrate that RUNX proteins are expressed in GCT cell lines and human GCT specimens, albeit at variable levels, and RUNX3 may play an oncogenic role in a subset of GCTs.

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