This clinical trial evaluated the safety, efficacy, and pharmacokinetics of F14512, a new polyamine-vectorized anticancer drug, in dogs with naturally occurring lymphoma. Twenty-three dogs with stage III-IV lymphoma received intravenous injections of F14512 over three cycles. The trial determined the maximum tolerated dose and found F14512 was well tolerated, with expected hematologic toxicity. F14512 induced an early decrease in tumor cells and achieved a high response rate of 91% among the dogs. Biomarkers like phosphorylation of histone H2AX were studied as potential predictors of drug activity. The trial demonstrated F14512's strong therapeutic efficacy in canine lymphoma and supported further evaluation of F14512
This study examined the expression of HER-2 and LRP in tumor tissue samples from 65 gastric cancer patients and 32 non-gastric cancer patients. Immunohistochemical staining and Western blot analysis were used to test for HER-2 and LRP expression and phosphorylation levels. The results showed that HER-2 and LRP expression in gastric cancer tissues were positively correlated and higher than in non-gastric cancer tissues. HER-2 phosphorylation was highest in gastric cancer patients with lymphatic metastasis. This suggests that LRP may cause primary resistance in gastric cancer and that HER-2 activation may mediate gastric cancer resistance by influencing LRP expression.
Cancer Immunol Res-2015-Manuel-2326-6066.CIR-14-0214 (3)Melanie Lampa
The combination of Salmonella-based therapy targeting indoleamine 2,3-dioxygenase (shIDO-ST) and an enzyme (PEGPH20) that depletes tumor hyaluronan (HA) induced complete regression of aggressive pancreatic tumors in mouse models. Mice treated with this combination showed significant decreases in tumor burden and extended survival, with 30-60% of mice showing no evidence of tumors 8 weeks after treatment. In contrast, other treatment combinations including monotherapies were not effective. The combination of shIDO-ST and PEGPH20 appeared to facilitate entry of bacteria and activated immune cells into otherwise impermeable tumors, highlighting its potential as an effective treatment for pancreatic cancer.
This document discusses the identification of TACC1, NOV, and PTTG1 as potential new biomarkers associated with endocrine therapy resistance in breast cancer. The study used two cell models, MVLN/CL6.7 and VP229/VP267, that were selected for resistance to tamoxifen and acquired cross-resistance to fulvestrant. 26 candidate genes were examined by RTQ-PCR and 8 genes were found overexpressed in breast cancer patient samples that relapsed after tamoxifen treatment. TACC1, NOV, and PTTG1 were identified as independent prognostic markers associated with shorter relapse-free survival. Aberrant mRNA and protein levels of these 3 genes were also observed in
Vitamin D analogs enhance the anticancer activity of 5-fluorouracil in an in ...Enrique Moreno Gonzalez
Active vitamin D analogs that are less toxic than calcitriol can be useful in the combined treatment of patients suffering from colon cancer. In the present study we demonstrate, for the first time in an in vivo model system, the biological effect of combined therapy using 5-fluorouracil (5-FU) along with vitamin D analog PRI-2191 (tacalcitol, 1,24-dihydroxyvitamin D3) or PRI-2205 (5,6-trans-isomer of calcipotriol) on colon cancer.
Scientists screened 18 novel compounds for their ability to kill cancer cells. Several compounds showed potent cytotoxic effects, with IC50 values less than 1 μM on multiple cell lines. Three compounds - TMCOS-3, TMCOS-6, and TMCOS-11 - were found to induce apoptotic cell death through DNA fragmentation and caspase activation. TMCOS-11 was found to specifically inhibit tubulin polymerization and cause cell cycle arrest in the G2/M phase. These findings suggest that some of the compounds may be promising new anti-cancer drugs that work by targeting the microtubule protein tubulin.
1) Prodigiosin, a bacterial metabolite, induces apoptosis in human breast cancer cells. Gene expression profiling found that prodigiosin strongly increased expression of the NAG-1 gene.
2) Experiments showed that prodigiosin triggers accumulation of the tumor suppressor protein p53, but induction of NAG-1 was independent of p53.
3) Prodigiosin causes inhibition of AKT and activation of glycogen synthase kinase-3B (GSK-3B). Induction of NAG-1 and apoptosis correlated with GSK-3B activation. Inhibiting GSK-3B reduced apoptosis, suggesting GSK-3B plays a key role in the proap
This study assessed toxicities of CMF (cyclophosphamide, methotrexate, and 5-fluorouracil) chemotherapy and tamoxifen on rabbit livers and kidneys. Rabbits received either CMF alone, tamoxifen alone, or CMF plus tamoxifen. Liver enzymes and histopathology of livers and kidneys were analyzed. Rabbits that received both CMF and tamoxifen showed the highest liver enzyme levels and most histological changes in the liver including edema, infiltration, and fibrosis. They also exhibited the most kidney mesangial cell proliferation. The study concludes that CMF and tamoxifen chemotherapy can cause toxic effects in the liver and kidney.
Hepatic and serum lipid signatures specific to NASH in mouse modelsFranck Chiappini
This study identified lipid signatures in the liver and serum that are specific to nonalcoholic steatohepatitis (NASH) using mouse models of NAFL and NASH. Mice were fed either a high-fat diet (HFD) to induce NAFL or a methionine choline deficient diet (MCDD) to induce NASH. Comprehensive lipidomic analysis of liver tissues and serum identified 21 lipids in the liver and 14 lipids in the serum that could discriminate mice with NASH from those with NAFL or normal controls. Machine learning techniques were able to characterize lipid signatures specific to NASH in both the liver and serum, opening possibilities for investigating early and non-invasive lipid markers for diagn
This study examined the expression of HER-2 and LRP in tumor tissue samples from 65 gastric cancer patients and 32 non-gastric cancer patients. Immunohistochemical staining and Western blot analysis were used to test for HER-2 and LRP expression and phosphorylation levels. The results showed that HER-2 and LRP expression in gastric cancer tissues were positively correlated and higher than in non-gastric cancer tissues. HER-2 phosphorylation was highest in gastric cancer patients with lymphatic metastasis. This suggests that LRP may cause primary resistance in gastric cancer and that HER-2 activation may mediate gastric cancer resistance by influencing LRP expression.
Cancer Immunol Res-2015-Manuel-2326-6066.CIR-14-0214 (3)Melanie Lampa
The combination of Salmonella-based therapy targeting indoleamine 2,3-dioxygenase (shIDO-ST) and an enzyme (PEGPH20) that depletes tumor hyaluronan (HA) induced complete regression of aggressive pancreatic tumors in mouse models. Mice treated with this combination showed significant decreases in tumor burden and extended survival, with 30-60% of mice showing no evidence of tumors 8 weeks after treatment. In contrast, other treatment combinations including monotherapies were not effective. The combination of shIDO-ST and PEGPH20 appeared to facilitate entry of bacteria and activated immune cells into otherwise impermeable tumors, highlighting its potential as an effective treatment for pancreatic cancer.
This document discusses the identification of TACC1, NOV, and PTTG1 as potential new biomarkers associated with endocrine therapy resistance in breast cancer. The study used two cell models, MVLN/CL6.7 and VP229/VP267, that were selected for resistance to tamoxifen and acquired cross-resistance to fulvestrant. 26 candidate genes were examined by RTQ-PCR and 8 genes were found overexpressed in breast cancer patient samples that relapsed after tamoxifen treatment. TACC1, NOV, and PTTG1 were identified as independent prognostic markers associated with shorter relapse-free survival. Aberrant mRNA and protein levels of these 3 genes were also observed in
Vitamin D analogs enhance the anticancer activity of 5-fluorouracil in an in ...Enrique Moreno Gonzalez
Active vitamin D analogs that are less toxic than calcitriol can be useful in the combined treatment of patients suffering from colon cancer. In the present study we demonstrate, for the first time in an in vivo model system, the biological effect of combined therapy using 5-fluorouracil (5-FU) along with vitamin D analog PRI-2191 (tacalcitol, 1,24-dihydroxyvitamin D3) or PRI-2205 (5,6-trans-isomer of calcipotriol) on colon cancer.
Scientists screened 18 novel compounds for their ability to kill cancer cells. Several compounds showed potent cytotoxic effects, with IC50 values less than 1 μM on multiple cell lines. Three compounds - TMCOS-3, TMCOS-6, and TMCOS-11 - were found to induce apoptotic cell death through DNA fragmentation and caspase activation. TMCOS-11 was found to specifically inhibit tubulin polymerization and cause cell cycle arrest in the G2/M phase. These findings suggest that some of the compounds may be promising new anti-cancer drugs that work by targeting the microtubule protein tubulin.
1) Prodigiosin, a bacterial metabolite, induces apoptosis in human breast cancer cells. Gene expression profiling found that prodigiosin strongly increased expression of the NAG-1 gene.
2) Experiments showed that prodigiosin triggers accumulation of the tumor suppressor protein p53, but induction of NAG-1 was independent of p53.
3) Prodigiosin causes inhibition of AKT and activation of glycogen synthase kinase-3B (GSK-3B). Induction of NAG-1 and apoptosis correlated with GSK-3B activation. Inhibiting GSK-3B reduced apoptosis, suggesting GSK-3B plays a key role in the proap
This study assessed toxicities of CMF (cyclophosphamide, methotrexate, and 5-fluorouracil) chemotherapy and tamoxifen on rabbit livers and kidneys. Rabbits received either CMF alone, tamoxifen alone, or CMF plus tamoxifen. Liver enzymes and histopathology of livers and kidneys were analyzed. Rabbits that received both CMF and tamoxifen showed the highest liver enzyme levels and most histological changes in the liver including edema, infiltration, and fibrosis. They also exhibited the most kidney mesangial cell proliferation. The study concludes that CMF and tamoxifen chemotherapy can cause toxic effects in the liver and kidney.
Hepatic and serum lipid signatures specific to NASH in mouse modelsFranck Chiappini
This study identified lipid signatures in the liver and serum that are specific to nonalcoholic steatohepatitis (NASH) using mouse models of NAFL and NASH. Mice were fed either a high-fat diet (HFD) to induce NAFL or a methionine choline deficient diet (MCDD) to induce NASH. Comprehensive lipidomic analysis of liver tissues and serum identified 21 lipids in the liver and 14 lipids in the serum that could discriminate mice with NASH from those with NAFL or normal controls. Machine learning techniques were able to characterize lipid signatures specific to NASH in both the liver and serum, opening possibilities for investigating early and non-invasive lipid markers for diagn
The document summarizes research on the metastatic spread of breast cancer cells in mice. Key findings include:
- Line 4T1, a metastatic breast cancer cell line, primarily spreads through hematogenous metastasis to the lungs followed later by the liver. Necropsy found lung and liver nodules.
- Line 66cl4 also metastasized to the lungs and liver but spread more through lymph nodes than 4T1.
- The non-metastatic line 67NR was unable to intravasate and spread, as no clonogenic cancer cells were found in distant organs.
This paper reports phase II clinical data on galunisertib, a novel small molecule TGF-beta receptor inhibitor in patients with advanced non-operable hepatocellular carcinoma. This poster was presented at ASCO 2016 in Chicago. Results show activity and mild toxicity.
This study found that colon cancer cells express the chemokine receptor CCR4, which mediates migration of the cells in response to its ligand CCL17 (TARC) through the RhoA/Rho kinase signaling pathway. Quantitative RT-PCR and flow cytometry showed that the colon cancer cell lines HT-29 and AZ-97 expressed CCR4 at both the mRNA and protein levels. Stimulation with CCL17 induced dose-dependent migration of the colon cancer cells, which was inhibited by blocking CCR4 with an antibody or antagonist. CCL17 also increased mRNA levels of RhoA proteins and RhoA activation in the cells. Inhibition of Rho kinase or isoprenylation blocked CCL17-induced cell migration
Vorinostat combined with DNMTi epigenetically controls the proliferation of l...MustafaFathy6
This study evaluated the effects of combining the histone deacetylase inhibitor (HDACi) vorinostat with other chemotherapeutic drugs on lung cancer cells. Vorinostat alone and in combination with carboplatin was most effective at reducing cell viability of A549 lung cancer cells. Global DNA methylation patterns varied depending on the drug combinations, with vorinostat and carboplatin causing hypomethylation and vorinostat and cyclophosphamide resulting in hypermethylation. The results suggest that combining epigenetic and chemotherapeutic drugs may be more effective at controlling lung cancer proliferation than single agents alone. However, more experiments are needed to confirm these findings.
Geriatric Care at a Time of Accelerated Aging in the World Population and Eme...science journals
It is a well-known fact that the world population has been aging since mid-20th century. The number of older people aged 60 years and above has more than quadrupled since 1950’s and older people’s share of the world population reached 11.7 percent in 2013.
Poster presentation at the 2016 WORLD GASTROINTESTINAL SYMPOSIUM on tepotinib a selective inhibitor of c-MET by S. Faivre, J.-F. Blanc, P. Merle, A. Fasolo, A. Iacobellis, V. Grando, T. Decaens, J. Trojan, E. Villa, U. Stammberger, R. Bruns, E. Raymond
1Oncology Unit, Beaujon University Hospital, Clichy, France; 2Service d’hépato-gastroentérologie et d’oncologie digestive, Groupe Hospitalier Saint André, Bordeaux, France; 3Service d'Hépato-Gastro-Entérologie, Hôpital de la Croix Rousse, Lyon, France; 4Dipartimento di Oncologia Medica, Ospedale San Raffaele IRCSS, Milan, Italy; 5Servizio di Endoscopia Digestiva, Ospedale Casa Sollievo della Sofferenza IRCCS, San Giovanni Rotondo, Italy; 6Service Hépatologie, Hôpital Jean-Verdier, Bondy, France; 7Service d'hepato-gastro-enterologie, CHU de Grenoble - Hôpital Nord, Grenoble, France; 8Gastrointestinal Oncology, Goethe University Hospital, Frankfurt, Germany; 9Policlinico di Modena, Modena, Italy; 10Merck KGaA, Darmstadt, Germany
This journal article summarizes a study investigating the cytotoxic effects and molecular mechanisms of action of coumarins isolated from Calophyllum brasiliense (mammea A/BA and A/BB) in K562 leukemia cells. The study found that the coumarin mixture induced cytotoxicity in the cancer cells and apoptosis, as shown by TUNEL staining and caspase-3 activation. Genotoxic effects were also observed. Additionally, an in silico analysis found the coumarins complied with criteria for drug-likeness. The results support further development of these natural compounds as potential anticancer agents.
Triple negative breast cancer (TNBC) accounts for 15-25% of breast cancer cases and has a poor prognosis due to a lack of molecular targets. The study characterized expression of the lactate transporter MCT1 and its ancillary protein CD-147 in murine TNBC 4T1 cells. MCT1 and CD-147 were expressed on the cell membrane. L-lactate uptake followed Michaelis-Menten kinetics and was inhibited by the MCT1 inhibitor AR-155858, indicating MCT1's role in uptake. Further studies are needed to investigate MCT1 inhibition as a potential TNBC therapy.
This study tested the hypothesis that depleting intratumoral regulatory T cells (Treg) using Fas ligand (FasL) prior to adoptive T cell therapy could improve the therapeutic efficacy. The researchers found that:
1) Applying FasL intratumorally via protein transfer decreased intratumoral Treg by inducing apoptosis in these cells.
2) Pretreating tumors with intratumoral FasL before infusing tumor-reactive CD8+ T cells enhanced the therapeutic efficacy of adoptive T cell transfer against established tumors, resulting in persistent, systemic antitumor immunity.
3) The effect of intratumoral FasL pretreatment was abolished when Treg were c
1) The document analyzes trends in the development and approval of 147 human monoclonal antibody (mAb) therapeutics that entered clinical studies between 1985-2008.
2) It finds that while only 16 human mAbs entered clinical studies from 1985-1996 due to manufacturing challenges, 131 entered from 1997-2008 due to advances in transgenic mouse and phage display technologies.
3) As of 2008, 7 human mAbs have been approved by the FDA to treat various conditions such as rheumatoid arthritis, cancer, and rare genetic fever disorders.
Preparation and therapeutic evaluation of 188 re thermogelling emulsionSamieh Asadian
This document summarizes a study that evaluated a 188Re-labeled thermosensitive hydrogel emulsion for the treatment of hepatocellular carcinoma in a rat model. The emulsion consisted of Lipiodol encapsulated in micelles formed from a PEG-PLGA-PEG triblock copolymer labeled with 188Re. Biodistribution studies found high uptake of radioactivity in the tumor with low levels in other organs. Rats treated with intratumoral injections of the 188Re-labeled emulsion showed significantly improved survival rates and tumor response compared to untreated controls or injection with 188Re-Lipiodol alone, demonstrating potential for clinical use in hepatoma treatment.
Content Cytotoxicity Studies of Colorectal Carcinoma Cells Using Printed Impe...journalBEEI
Monitoring the effectiveness of drugs on cancer cells is crucial for chemotherapeutics studies. In-vitro cell-based biosensors can be used as an alternative for characteristic studies of cells’ response to drugs. Cell-based sensors provide real-time measurements and require smaller sample volumes compared to conventional T-flask measurement methods. This paper presents a biosensor that detects in real-time, impedance variations of human colon cancer, HCT-116 cells when treated with anti-cancer agent, 5-Fluorouracil (5-FU). Two different extracellular matrix (ECM); polyaniline and gelatin were tested and evaluated in terms of attachment quality. Polyaniline was found to provide the best attachment for HCT-116 cells and was used for cytotoxicity studies. Cytokinetic behavior indicated that 5-FU inhibited HCT-116 cells at IC50 of 6.8 µg/mL. Trypan blue exclusion method for testing cell viability was used to validate the impedance measurements, where the cancer cell concentrations were reduced to ~35% when treated with 2.5 µg/mL, and 50% when treated with 6.8 µg/mL. The results generated by the microfabricated impedance biosensor are comparable to the Trypan blue method since both gave similar cell growth trend. It can be concluded that the impedance biosensor has potential to be used as an alternative method in drug testing applications.
This document describes the development of a phenotypic chemotaxis assay using flow cytometry to quantify the migration of specific cell types from human peripheral blood mononuclear cells (PBMCs) in response to chemokines. The assay was used to measure the migration of CD4+/CLA+ T cells in response to MDC (CCL22) and inhibition by a CCR4 antagonist. Flow cytometry allowed measurement of an EC50 of 1nM for MDC-induced migration and consistency with the antagonist's known potency. This novel assay provides benefits over traditional methods by enabling investigation of rare cell types, accurate cell counts, phenotypic analysis, and use of low sample volumes.
Biochemical and Toxicological Investigations of 5-Fluorouracil, Nimesulide, a...BRNSS Publication Hub
The objective of this study was biochemical and toxicological investigations of 5-fluorouracil (5-FU), nimesulide, and ascorbic acid (Vitamin C) in Wistar rats with hepatocellular carcinoma in. Results showed that DENA increased the level of alpha fetoprotein (AFP), alkaline phosphatase (ALP), serum glutamic oxaloacetate transaminase (SGOT), serum glutamic pyruvate transaminase (SGPT), and total bilirubin which was decreased by the various combinations of 5-FU to normal. On the other hand, DENA resulted in decrease of blood glucose level, DFN decreased more than DF, and DFC showed results similar to DFN, while DFNC led to increased AFP, ALP, SGOT, SGPT, and total bilirubin levels to normal. Histopathological evaluations showed normal architecture of tissues of rat liver in normal group. Lesser damage of hepatocytes and low index of necrosis were in pre- and post-treated group of 5-FU+DF, DFN+DFC groups. DFNC treated group exhibited histological features resembling normal control animals.
Anti- Tumor assay / Screening of Anticancer DrugsPratik Parikh
This document presents information about in vitro and in vivo anti-tumor assays. It discusses several in vitro assays including tetrazolium salt, sulphorhodamine B, and 3H-thymidine uptake assays. It also describes various in vivo models like carcinogen-induced tumors in mice, viral infection models, transplantation models, and genetically engineered mouse models. Specific techniques covered include DMBA-induced skin papillomas in mice and MNU-induced rat mammary gland cancer models. The document concludes by discussing parameters to evaluate anti-tumor effects in an EAC liquid tumor mouse model.
Inhibition of hdac by vorinostat in human lung cancer cellsMustafaFathy6
The document describes a research study aiming to identify the role of Vorinostat, a histone deacetylase (HDAC) inhibitor, in treating lung cancer. The study will culture A519 lung cancer cell lines and treat them with varying concentrations of Vorinostat for 4-5 days. Trypan Blue and MTT tests will then assess cell viability. DNA will be extracted and analyzed using methylation kits to study the drug's effects on methylation. Flow cytometry will examine effects on the cell cycle. The goal is to evaluate Vorinostat's potential as an anti-cancer agent for lung cancer.
The document summarizes the synthesis and evaluation of novel pyridazinone derivatives bearing benzenesulfonamide moieties for their anticancer activity. A series of 8 pyridazinone derivatives (2a-h) were synthesized and 5 (2a, 2b, 2d, 2g, 2h) were evaluated against human cancer cell lines. Compound 2h showed the best results, inhibiting the growth of 34/59 cell lines with GI50 values below 1 μM for several cancer types. Acute toxicity studies in mice found 2h to be well tolerated at 400 mg/kg. Compound 2h was selected for further evaluation based on its promising anticancer activity and favorable toxicity profile.
The document summarizes a study investigating the expression and role of apoptosis-related molecules like TRAIL, FasL, and their receptors in pancreatic adenocarcinoma cell lines. The key findings were:
1) The pancreatic cancer cell lines expressed high levels of apoptosis-inducing ligands and receptors but showed variable susceptibility to TRAIL-induced cell death.
2) Treatment with chemotherapy drugs did not increase their susceptibility to apoptosis, likely due to their differential expression of decoy receptors and inhibitor molecules.
3) This suggests pancreatic cancers develop resistance to immune-mediated apoptosis, allowing immune evasion and tumor progression.
inducing Apoptosis in cancer cell by natural compounds and screening methodssyeddastagir9
This document is a seminar paper on inducing apoptosis in cancer cells by natural compounds. It discusses apoptosis and its importance in cancer treatment. Specifically, it examines two natural compounds - curcumin and piperlongamine - that can induce apoptosis in cancer cells through various pathways like the death receptor pathway, Bcl-2 pathway, and p53 pathway. It describes experiments showing curcumin and piperlongamine's ability to inhibit tumor growth in vivo in nude mice models and influence protein expression in vitro through western blot analysis. The paper concludes that natural compounds providing safe and effective anticancer activity through apoptotic pathways can be promising for cancer therapy.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise causes chemical changes in the brain that may help boost feelings of calmness, happiness and focus.
This document provides an overview, prerequisites, and table of contents for a tutorial on learning the Python programming language from scratch. It is designed for software programmers who want to learn Python. The tutorial will cover Python's history and features, setting up a local environment to run Python, basic syntax like variables, data types, operators, and decision making statements. It will also discuss how to get and install Python, set environment paths, run Python programs, and handle command line arguments. The content is copyrighted and users are prohibited from redistributing it without permission.
This short document promotes creating presentations using Haiku Deck, a tool for making slideshows. It encourages the reader to get started making their own Haiku Deck presentation and sharing it on SlideShare. In a single sentence, it pitches the idea of using Haiku Deck to easily design presentations.
The document summarizes research on the metastatic spread of breast cancer cells in mice. Key findings include:
- Line 4T1, a metastatic breast cancer cell line, primarily spreads through hematogenous metastasis to the lungs followed later by the liver. Necropsy found lung and liver nodules.
- Line 66cl4 also metastasized to the lungs and liver but spread more through lymph nodes than 4T1.
- The non-metastatic line 67NR was unable to intravasate and spread, as no clonogenic cancer cells were found in distant organs.
This paper reports phase II clinical data on galunisertib, a novel small molecule TGF-beta receptor inhibitor in patients with advanced non-operable hepatocellular carcinoma. This poster was presented at ASCO 2016 in Chicago. Results show activity and mild toxicity.
This study found that colon cancer cells express the chemokine receptor CCR4, which mediates migration of the cells in response to its ligand CCL17 (TARC) through the RhoA/Rho kinase signaling pathway. Quantitative RT-PCR and flow cytometry showed that the colon cancer cell lines HT-29 and AZ-97 expressed CCR4 at both the mRNA and protein levels. Stimulation with CCL17 induced dose-dependent migration of the colon cancer cells, which was inhibited by blocking CCR4 with an antibody or antagonist. CCL17 also increased mRNA levels of RhoA proteins and RhoA activation in the cells. Inhibition of Rho kinase or isoprenylation blocked CCL17-induced cell migration
Vorinostat combined with DNMTi epigenetically controls the proliferation of l...MustafaFathy6
This study evaluated the effects of combining the histone deacetylase inhibitor (HDACi) vorinostat with other chemotherapeutic drugs on lung cancer cells. Vorinostat alone and in combination with carboplatin was most effective at reducing cell viability of A549 lung cancer cells. Global DNA methylation patterns varied depending on the drug combinations, with vorinostat and carboplatin causing hypomethylation and vorinostat and cyclophosphamide resulting in hypermethylation. The results suggest that combining epigenetic and chemotherapeutic drugs may be more effective at controlling lung cancer proliferation than single agents alone. However, more experiments are needed to confirm these findings.
Geriatric Care at a Time of Accelerated Aging in the World Population and Eme...science journals
It is a well-known fact that the world population has been aging since mid-20th century. The number of older people aged 60 years and above has more than quadrupled since 1950’s and older people’s share of the world population reached 11.7 percent in 2013.
Poster presentation at the 2016 WORLD GASTROINTESTINAL SYMPOSIUM on tepotinib a selective inhibitor of c-MET by S. Faivre, J.-F. Blanc, P. Merle, A. Fasolo, A. Iacobellis, V. Grando, T. Decaens, J. Trojan, E. Villa, U. Stammberger, R. Bruns, E. Raymond
1Oncology Unit, Beaujon University Hospital, Clichy, France; 2Service d’hépato-gastroentérologie et d’oncologie digestive, Groupe Hospitalier Saint André, Bordeaux, France; 3Service d'Hépato-Gastro-Entérologie, Hôpital de la Croix Rousse, Lyon, France; 4Dipartimento di Oncologia Medica, Ospedale San Raffaele IRCSS, Milan, Italy; 5Servizio di Endoscopia Digestiva, Ospedale Casa Sollievo della Sofferenza IRCCS, San Giovanni Rotondo, Italy; 6Service Hépatologie, Hôpital Jean-Verdier, Bondy, France; 7Service d'hepato-gastro-enterologie, CHU de Grenoble - Hôpital Nord, Grenoble, France; 8Gastrointestinal Oncology, Goethe University Hospital, Frankfurt, Germany; 9Policlinico di Modena, Modena, Italy; 10Merck KGaA, Darmstadt, Germany
This journal article summarizes a study investigating the cytotoxic effects and molecular mechanisms of action of coumarins isolated from Calophyllum brasiliense (mammea A/BA and A/BB) in K562 leukemia cells. The study found that the coumarin mixture induced cytotoxicity in the cancer cells and apoptosis, as shown by TUNEL staining and caspase-3 activation. Genotoxic effects were also observed. Additionally, an in silico analysis found the coumarins complied with criteria for drug-likeness. The results support further development of these natural compounds as potential anticancer agents.
Triple negative breast cancer (TNBC) accounts for 15-25% of breast cancer cases and has a poor prognosis due to a lack of molecular targets. The study characterized expression of the lactate transporter MCT1 and its ancillary protein CD-147 in murine TNBC 4T1 cells. MCT1 and CD-147 were expressed on the cell membrane. L-lactate uptake followed Michaelis-Menten kinetics and was inhibited by the MCT1 inhibitor AR-155858, indicating MCT1's role in uptake. Further studies are needed to investigate MCT1 inhibition as a potential TNBC therapy.
This study tested the hypothesis that depleting intratumoral regulatory T cells (Treg) using Fas ligand (FasL) prior to adoptive T cell therapy could improve the therapeutic efficacy. The researchers found that:
1) Applying FasL intratumorally via protein transfer decreased intratumoral Treg by inducing apoptosis in these cells.
2) Pretreating tumors with intratumoral FasL before infusing tumor-reactive CD8+ T cells enhanced the therapeutic efficacy of adoptive T cell transfer against established tumors, resulting in persistent, systemic antitumor immunity.
3) The effect of intratumoral FasL pretreatment was abolished when Treg were c
1) The document analyzes trends in the development and approval of 147 human monoclonal antibody (mAb) therapeutics that entered clinical studies between 1985-2008.
2) It finds that while only 16 human mAbs entered clinical studies from 1985-1996 due to manufacturing challenges, 131 entered from 1997-2008 due to advances in transgenic mouse and phage display technologies.
3) As of 2008, 7 human mAbs have been approved by the FDA to treat various conditions such as rheumatoid arthritis, cancer, and rare genetic fever disorders.
Preparation and therapeutic evaluation of 188 re thermogelling emulsionSamieh Asadian
This document summarizes a study that evaluated a 188Re-labeled thermosensitive hydrogel emulsion for the treatment of hepatocellular carcinoma in a rat model. The emulsion consisted of Lipiodol encapsulated in micelles formed from a PEG-PLGA-PEG triblock copolymer labeled with 188Re. Biodistribution studies found high uptake of radioactivity in the tumor with low levels in other organs. Rats treated with intratumoral injections of the 188Re-labeled emulsion showed significantly improved survival rates and tumor response compared to untreated controls or injection with 188Re-Lipiodol alone, demonstrating potential for clinical use in hepatoma treatment.
Content Cytotoxicity Studies of Colorectal Carcinoma Cells Using Printed Impe...journalBEEI
Monitoring the effectiveness of drugs on cancer cells is crucial for chemotherapeutics studies. In-vitro cell-based biosensors can be used as an alternative for characteristic studies of cells’ response to drugs. Cell-based sensors provide real-time measurements and require smaller sample volumes compared to conventional T-flask measurement methods. This paper presents a biosensor that detects in real-time, impedance variations of human colon cancer, HCT-116 cells when treated with anti-cancer agent, 5-Fluorouracil (5-FU). Two different extracellular matrix (ECM); polyaniline and gelatin were tested and evaluated in terms of attachment quality. Polyaniline was found to provide the best attachment for HCT-116 cells and was used for cytotoxicity studies. Cytokinetic behavior indicated that 5-FU inhibited HCT-116 cells at IC50 of 6.8 µg/mL. Trypan blue exclusion method for testing cell viability was used to validate the impedance measurements, where the cancer cell concentrations were reduced to ~35% when treated with 2.5 µg/mL, and 50% when treated with 6.8 µg/mL. The results generated by the microfabricated impedance biosensor are comparable to the Trypan blue method since both gave similar cell growth trend. It can be concluded that the impedance biosensor has potential to be used as an alternative method in drug testing applications.
This document describes the development of a phenotypic chemotaxis assay using flow cytometry to quantify the migration of specific cell types from human peripheral blood mononuclear cells (PBMCs) in response to chemokines. The assay was used to measure the migration of CD4+/CLA+ T cells in response to MDC (CCL22) and inhibition by a CCR4 antagonist. Flow cytometry allowed measurement of an EC50 of 1nM for MDC-induced migration and consistency with the antagonist's known potency. This novel assay provides benefits over traditional methods by enabling investigation of rare cell types, accurate cell counts, phenotypic analysis, and use of low sample volumes.
Biochemical and Toxicological Investigations of 5-Fluorouracil, Nimesulide, a...BRNSS Publication Hub
The objective of this study was biochemical and toxicological investigations of 5-fluorouracil (5-FU), nimesulide, and ascorbic acid (Vitamin C) in Wistar rats with hepatocellular carcinoma in. Results showed that DENA increased the level of alpha fetoprotein (AFP), alkaline phosphatase (ALP), serum glutamic oxaloacetate transaminase (SGOT), serum glutamic pyruvate transaminase (SGPT), and total bilirubin which was decreased by the various combinations of 5-FU to normal. On the other hand, DENA resulted in decrease of blood glucose level, DFN decreased more than DF, and DFC showed results similar to DFN, while DFNC led to increased AFP, ALP, SGOT, SGPT, and total bilirubin levels to normal. Histopathological evaluations showed normal architecture of tissues of rat liver in normal group. Lesser damage of hepatocytes and low index of necrosis were in pre- and post-treated group of 5-FU+DF, DFN+DFC groups. DFNC treated group exhibited histological features resembling normal control animals.
Anti- Tumor assay / Screening of Anticancer DrugsPratik Parikh
This document presents information about in vitro and in vivo anti-tumor assays. It discusses several in vitro assays including tetrazolium salt, sulphorhodamine B, and 3H-thymidine uptake assays. It also describes various in vivo models like carcinogen-induced tumors in mice, viral infection models, transplantation models, and genetically engineered mouse models. Specific techniques covered include DMBA-induced skin papillomas in mice and MNU-induced rat mammary gland cancer models. The document concludes by discussing parameters to evaluate anti-tumor effects in an EAC liquid tumor mouse model.
Inhibition of hdac by vorinostat in human lung cancer cellsMustafaFathy6
The document describes a research study aiming to identify the role of Vorinostat, a histone deacetylase (HDAC) inhibitor, in treating lung cancer. The study will culture A519 lung cancer cell lines and treat them with varying concentrations of Vorinostat for 4-5 days. Trypan Blue and MTT tests will then assess cell viability. DNA will be extracted and analyzed using methylation kits to study the drug's effects on methylation. Flow cytometry will examine effects on the cell cycle. The goal is to evaluate Vorinostat's potential as an anti-cancer agent for lung cancer.
The document summarizes the synthesis and evaluation of novel pyridazinone derivatives bearing benzenesulfonamide moieties for their anticancer activity. A series of 8 pyridazinone derivatives (2a-h) were synthesized and 5 (2a, 2b, 2d, 2g, 2h) were evaluated against human cancer cell lines. Compound 2h showed the best results, inhibiting the growth of 34/59 cell lines with GI50 values below 1 μM for several cancer types. Acute toxicity studies in mice found 2h to be well tolerated at 400 mg/kg. Compound 2h was selected for further evaluation based on its promising anticancer activity and favorable toxicity profile.
The document summarizes a study investigating the expression and role of apoptosis-related molecules like TRAIL, FasL, and their receptors in pancreatic adenocarcinoma cell lines. The key findings were:
1) The pancreatic cancer cell lines expressed high levels of apoptosis-inducing ligands and receptors but showed variable susceptibility to TRAIL-induced cell death.
2) Treatment with chemotherapy drugs did not increase their susceptibility to apoptosis, likely due to their differential expression of decoy receptors and inhibitor molecules.
3) This suggests pancreatic cancers develop resistance to immune-mediated apoptosis, allowing immune evasion and tumor progression.
inducing Apoptosis in cancer cell by natural compounds and screening methodssyeddastagir9
This document is a seminar paper on inducing apoptosis in cancer cells by natural compounds. It discusses apoptosis and its importance in cancer treatment. Specifically, it examines two natural compounds - curcumin and piperlongamine - that can induce apoptosis in cancer cells through various pathways like the death receptor pathway, Bcl-2 pathway, and p53 pathway. It describes experiments showing curcumin and piperlongamine's ability to inhibit tumor growth in vivo in nude mice models and influence protein expression in vitro through western blot analysis. The paper concludes that natural compounds providing safe and effective anticancer activity through apoptotic pathways can be promising for cancer therapy.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise causes chemical changes in the brain that may help boost feelings of calmness, happiness and focus.
This document provides an overview, prerequisites, and table of contents for a tutorial on learning the Python programming language from scratch. It is designed for software programmers who want to learn Python. The tutorial will cover Python's history and features, setting up a local environment to run Python, basic syntax like variables, data types, operators, and decision making statements. It will also discuss how to get and install Python, set environment paths, run Python programs, and handle command line arguments. The content is copyrighted and users are prohibited from redistributing it without permission.
This short document promotes creating presentations using Haiku Deck, a tool for making slideshows. It encourages the reader to get started making their own Haiku Deck presentation and sharing it on SlideShare. In a single sentence, it pitches the idea of using Haiku Deck to easily design presentations.
Op sales teams wordt vandaag vaak een flinke druk gelegd. Sommigen hebben die druk niet nodig om te presteren. Anderen kunnen de druk niet aan. Voor nog anderen zal geen enkele vorm van druk ooit helpen om hen tot prestaties te brengen.
Waar ligt het verschil?
Bij sales profielen is het niet altijd evident om de vinger op de wonde te leggen. En de elementen die veelal fundamenteel aan de basis liggen (persoonlijkheid, motivatoren...) zijn niet makkelijk zichtbaar.
Hebt u er ooit aan gedacht om uw commerciële organisatie een boost te geven?
An employee of the store manager, Darious Judue, has begun having problems showing up late and missing meetings. His performance initially improved but he then came to work smelling of alcohol and wearing the same clothes. When confronted, he claimed personal and family issues. Later he came to work drunk and denied it. The manager met with HR to discuss options according to the discipline process to ensure any firing is legal and fair. The summary ignores addressing the employee's issues led to limited options now.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms.
This document provides strategies for effective persuasion. It discusses how persuasion should produce action, and quotes scripture about having conviction. It recommends choosing words that convey success and eliminating words of doubt. Speak of what you want to experience, not just what you have. Choose positive associations by surrounding yourself with supportive people. Close doors to hindrances and distractions. Keep improving and seeking new opportunities. True possession comes from persevering through challenges with commitment and obedience.
The document discusses a study measuring the effectiveness of digital advertising within videogames. Researchers partnered with game developer EA and advertisers like Samsung to place ads in the game Battlefield 2142. Over 600 gamers were surveyed before and after exposure to see if brand awareness, recall, and perceptions changed. Key findings included that this type of "core" gamer spends significant time playing games instead of traditional media, making them a valuable audience that is hard to reach otherwise. The in-game ads were found to increase awareness for some brands more than others, showing potential for this new advertising medium.
Manish Kumar is seeking a position that offers professional growth and job satisfaction. He has a Bachelor's degree in Mechanical Engineering and over 3 years of experience in aircraft stress analysis. His skills include stress analysis tools like ISAMI, Nastran, and Patran. He has experience analyzing the stress of aircraft structures like the A350 XWB front spar. He also has experience converting 2D models to 3D and designing valves using SolidWorks. His personal traits include being self-reliant, confident, and able to work effectively individually or in a group.
Το Πρόγραμμα Μαθημάτων και το Διαδικτυακό Περιβάλλον Εκπαίδευσης του Έργου Co...eurocy
Αυτή η παρουσίαση αποτελεί μια περίληψη δύο σημαντικών προϊόντων του Έργου Code RED (http://www.codered-project.eu): του ολοκληρωμένου προγράμματος μαθημάτων και της διαδικτυακής πλατφόρμας εκπαίδευσης. Τα δύο αυτά έχουν δημιουργηθεί για να υποστηρίξουν τους εκπαιδευτές στην καθημερινή τους εργασία με το υλικό του Code RED.
This document contains the schedule of meetings and activities for Unisource Inc. on September 9. It includes times for internal meetings on marketing, LinkedIn, and product development. It also lists meeting times on September 11 in Burlingame. The document discusses Unisource's growth, noting it started in 2014 with 200 LinkedIn connections and now has over 400. It achieved 100 new Google searches in 2013. Finally, it includes a brief historical section about the founder starting in 1915.
This short document promotes creating presentations using Haiku Deck, an online presentation tool. It encourages the reader to get started making their own Haiku Deck presentation and sharing it on SlideShare. In a few words, it pitches Haiku Deck as a way to easily make and share online presentations.
This document profiles Danielle, a 20-year-old female student at Northwestern University who loves eating at Chipotle. It provides background on Danielle, including her goals of becoming a director while staying healthy in college. The document discusses how Chipotle helps customers like Danielle by offering custom, affordable, and healthy fast food options. It also includes quotes from Danielle about choosing Chipotle over unhealthier alternatives and how marketing can reach busy college students like her through social media platforms like Facebook and Twitter.
- фишки и дыры гугла в эссейной нише за последние пару лет
- нестандартные методы добычи ссылок
- какие источники работают в нише, какие - не работают
- покажем кейс - сайт, который в топе существует уже много лет и пережил все апдейты. Как создать такой сайт, который не придется продвигать ссылками
Radiotherapy promotes the polarization of tumor-associated macrophages (TAMs) in mice with Lewis lung cancer into anti-tumor M1 macrophages. This is accompanied by increased expression of the long non-coding RNA lincRNA-p21 in the TAMs. TAMs exposed to radiation therapy suppress the viability and invasion of Lewis lung cancer cells in culture. Overexpression of lincRNA-p21 in TAMs enhances their anti-tumor effects, while decreasing lincRNA-p21 reduces the effects of radiation therapy, suggesting lincRNA-p21 plays a key role in the anti-tumor actions of radiotherapy in lung cancer.
Genotyping of 27 Human Papillomavirus Types by Using L1 Consensus PCR Product...Alberto Cuadrado
Amplification of human papillomavirus (HPV) DNA by L1 consensus primer systems (e.g., MY09/11 or
GP51/61) can detect as few as 10 to 100 molecules of HPV targets from a genital sample. However, genotype
determination by dot blot hybridization is laborious and requires at least 27 separate hybridizations for
substantive HPV-type discrimination. A reverse blot method was developed which employs a biotin-labeled
PCR product hybridized to an array of immobilized oligonucleotide probes. By the reverse blot strip analysis,
genotype discrimination of multiple HPV types can be accomplished in a single hybridization and wash cycle.
Twenty-seven HPV probe mixes, two control probe concentrations, and a single reference line were immobilized
to 75- by 6-mm nylon strips. Each individual probe line contained a mixture of two bovine serum albuminconjugated
oligonucleotide probes specific to a unique HPV genotype. The genotype spectrum discriminated on
this strip includes the high-risk, or cancer-associated, HPV genotypes 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 55,
56, 58, 59, 68 (ME180), MM4 (W13B), MM7 (P291), and MM9 (P238A) and the low-risk, or non-cancerassociated,
genotypes 6, 11, 40, 42, 53, 54, 57, 66, and MM8 (P155). In addition, two concentrations of b-globin
probes allowed for assessment of individual specimen adequacy following amplification. We have evaluated the
performance of the strip method relative to that of a previously reported dot blot format (H. M. Bauer et al.,
p. 132–152, in C. S. Herrington and J. O. D. McGee (ed.), Diagnostic Molecular Pathology: a Practical Approach,
(1992), by testing 328 cervical swab samples collected in Digene specimen transport medium (Digene Diagnostics,
Silver Spring, Md.). We show excellent agreement between the two detection formats, with 92%
concordance for HPV positivity (kappa 5 0.78, P < 0.001). Nearly all of the discrepant HPV-positive samples
resulted from weak signals and can be attributed to sampling error from specimens with low concentrations
(<1 copy/ml) of HPV DNA. The primary advantage of the strip-based detection system is the ability to rapidly
genotype HPVs present in genital samples with high sensitivity and specificity, minimizing the likelihood of
misclassification.
Zauderer, M.G., et al. Clinical Cancer Research, 2017.sellasq4
1. This randomized phase II trial evaluated the WT1 peptide vaccine galinpepimut-S combined with GM-CSF and Montanide in patients with malignant pleural mesothelioma after multimodality therapy.
2. The trial randomized 41 patients to galinpepimut-S plus adjuvants or adjuvants alone. The control arm was stopped early due to a futility analysis showing progression within 1 year in over 10 of the first 20 patients.
3. Trends toward improved progression-free survival (10.1 vs 7.4 months) and overall survival (22.8 vs 18.3 months) were observed in the vaccine arm compared to control, but the trial was
Silencing c-Myc translation as a therapeutic strategy through targeting PI3Kd...Mark Lipstein
This document summarizes a study examining the combination of a novel PI3Kδ inhibitor, TGR-1202, with the proteasome inhibitor carfilzomib for treating hematological malignancies. The study found that TGR-1202 synergizes strongly with carfilzomib in lymphoma, leukemia, and myeloma cell lines and primary cells by silencing c-Myc translation. This synergistic effect is driven by TGR-1202's unexpected additional activity of inhibiting CK1ε, which contributes to repressing phosphorylation of 4E-BP1 and lowering c-Myc protein levels. The results suggest that TGR-1202, as a dual PI3Kδ/CK1ε inhibitor, may have
This study examined the potential of using the drug ribavirin to target the oncogene eIF4E in breast cancer cells. eIF4E is overexpressed in over 50% of breast cancers and promotes tumor growth. Ribavirin inhibits eIF4E function and reduced proliferation, clonogenic survival, and levels of eIF4E targets like cyclins in several breast cancer cell lines, with varying sensitivity. Ribavirin treatment was also associated with decreased Akt phosphorylation. Analysis of breast cancer biopsies found elevated eIF4E levels compared to normal tissue, supporting further study of ribavirin as a potential breast cancer therapeutic.
- The document examines the role of plasminogen activator inhibitor 1 (PAI-1) in the recruitment of mast cells (MCs) to glioma tumors.
- It finds that neutralizing PAI-1 attenuates the infiltration of MCs into glioma tumors. It also finds that MCs express the PAI-1 receptor LRP1, and blocking LRP1 also attenuates MC migration.
- Activation of the potential PAI-1/LRP1 axis in MCs by purified PAI-1 promotes increased phosphorylation of STAT3 and subsequent MC exocytosis. This indicates the PAI-1/LRP1 axis influences MC recruitment in glioma tumors.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...JohnJulie1
The IRF family of proteins involves in the tumor progression. However, but the functions of IRF5 in the tumorigenesis are largely unknown. Here, IRF5 was found to be up-regulated in hepatocellular carcinoma (HCC). Interfering with IRF5 inhibited the growth and tumorigenic ability of HCC cells.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...NainaAnon
1. The study found that IRF5 was upregulated in hepatocellular carcinoma (HCC) tissues compared to normal tissues based on mRNA and protein levels.
2. Overexpression of IRF5 promoted the growth and colony formation of HCC cells in vitro, while silencing IRF5 inhibited HCC cell growth and proliferation.
3. TRIM35 was found to interact with and promote the degradation of IRF5. TRIM35 expression was negatively correlated with IRF5 levels in HCC clinical samples.
1) Plumbagin (PL), derived from plants, inhibits prostate tumor development in mice lacking the tumor suppressor PTEN (Pten-KO mice), a model for prostate cancer.
2) PL treatment decreased expression of proteins involved in prostate cancer progression (PKCε, STAT3, AKT) and epithelial-to-mesenchymal transition (EMT).
3) Dietary PL inhibited primary prostate tumor growth and castration-resistant prostate cancer growth in Pten-KO mice, potentially by inhibiting PKCε, STAT3, AKT, and EMT markers.
1. The authors developed a quality-controlled two-color real-time PCR method to reliably test FFPE samples for molecular diagnostics.
2. The method uses internal standards and an internal standard mixture to control for interfering substances in FFPE samples and other experimental variations. It also uses two-color probes and pre-amplification to maximize signal from low RNA samples.
3. The authors validated the method by developing and testing reagents for four genes in a lung cancer diagnostic test. Analytical tests showed the method has acceptable accuracy, precision, and sensitivity. Clinical tests on FFPE lung samples demonstrated 93% diagnostic accuracy, similar to tests on fresh samples.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...daranisaha
The IRF family of proteins involves in the tumor progression. However, but the functions of IRF5 in the tumorigenesis are largely unknown. Here, IRF5 was found to be up-regulated in hepatocellular carcinoma (HCC). Interfering with IRF5 inhibited the growth and tumorigenic ability of HCC cells. When studying the molecular mechanism, it was found that TRIM35 interacted with IRF5, promoting the ubiquitination and degradation of IRF5. In the clinical specimens of HCC, TRIM35 was negatively correlated with the expression of IRF5. These observations reveal the oncogenic function of IRF5 in the progression of HCC, suggesting that IRF5 is a promising target for the therapy of HCC.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...eshaasini
The IRF family of proteins involves in the tumor progression. However, but the functions of IRF5 in the tumorigenesis are largely unknown. Here, IRF5 was found to be up-regulated in hepatocellular carcinoma (HCC). Interfering with IRF5 inhibited the growth and tumorigenic ability of HCC cells. When studying the molecular mechanism, it was found that TRIM35 interacted with IRF5, promoting the ubiquitination and degradation of IRF5. In the clinical specimens of HCC, TRIM35 was negatively correlated with the expression of IRF5. These observations reveal the oncogenic function of IRF5 in the progression of HCC, suggesting that IRF5 is a promising target for the therapy of HCC.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...semualkaira
The IRF family of proteins involves in the tumor progression. However, but the functions of IRF5 in the tumorigenesis are largely unknown. Here, IRF5 was found to be up-regulated in hepatocellular carcinoma (HCC). Interfering with IRF5 inhibited the growth and tumorigenic ability of HCC cells. When studying the molecular mechanism, it was found that TRIM35 interacted with IRF5, promoting the ubiquitination and degradation of IRF5. In the clinical specimens of HCC, TRIM35 was negatively correlated with the expression of IRF5. These observations reveal the oncogenic function of IRF5 in the progression of HCC, suggesting that IRF5 is a promising target for the therapy of HCC.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...semualkaira
The IRF family of proteins involves in the tumor progression. However, but the functions of IRF5 in the tumorigenesis are largely unknown. Here, IRF5 was found to be up-regulated in hepatocellular carcinoma (HCC). Interfering with IRF5 inhibited the growth and tumorigenic ability of HCC cells. When studying the molecular mechanism, it was found that TRIM35 interacted with IRF5, promoting the ubiquitination and degradation of IRF5. In the clinical specimens of HCC, TRIM35 was negatively correlated with the expression of IRF5. These observations reveal the oncogenic function of IRF5 in the progression of HCC, suggesting that IRF5 is a promising target for the therapy of HCC.
IRF5 Promotes the Progression of Hepatocellular Carcinoma and is Regulated by...semualkaira
The IRF family of proteins involves in the tumor progression. However, but the functions of IRF5 in the tumorigenesis are largely unknown. Here, IRF5 was found to be up-regulated in hepatocellular carcinoma (HCC). Interfering with IRF5 inhibited the growth and tumorigenic ability of HCC cells. When studying the molecular mechanism, it was found that TRIM35 interacted with IRF5, promoting the ubiquitination and degradation of IRF5. In the clinical specimens of HCC, TRIM35 was negatively correlated with the expression of IRF5. These observations reveal the oncogenic function of IRF5 in the progression of HCC, suggesting that IRF5 is a promising target for the therapy of HCC.
Inhibition of glutathione by buthionine sulfoximine enhanced the anti-cancer ...Ashujit
Multiple myeloma (MM) is an incurable blood cancer. Melphalan is an alkylating agent given prior to stem cell transplantation to MM patients. Increased glutathione confers resistance to melphalan. This study investigate the effect of inhibition of glutathione by BSO in preclinical models of MM. Pretreatment with BSO enhanced the anti-cancer effect of melphalan in cell lines and animal models. BSO and melphalan combination was well tolerated by animals and enhanced the survival as compared to controls, BSO and melphalan alone. BSO enhanced depth and duration of responses induced by melphalan. In the combination group, majority of treated animals achieved complete response (CR) and more than 20% had maintained CR. Also, the survival of animals was doubled after combination treatment as compared to BSO or melphalan alone. Mechanistic investigation demonstrated that BSO enhanced melphalan induced DNA damage, caspase cleavage and apoptosis. The combination also achieved multi-logs of cells kills in nine human multiple myeloma cell lines and primary MM cells isolated from blood and bone marrows. Interestingly, the effect of BSO and melphalan combination was abolished when cells were treated with N-acetyl cysteine and sodium thiosulfate but not with vitamin C and vitamin E. This observation suggests that effect of BSO is primarily driven by its ability to deplete glutathione and therefore preventing melphalan detoxification. Together, this study provides framework for testing the combination in a Phase I trial.
This clinical trial assessed the safety and immunogenicity of a polyvalent WT1 peptide vaccine in patients with acute myeloid leukemia (AML) in complete remission. Nine evaluable patients received six vaccinations over 12 weeks with four WT1 peptides plus adjuvants. WT1-specific T-cell responses were detected in seven of eight patients by assays such as ELISPOT and tetramer staining. The vaccine was found to be safely administered and induced immune responses against WT1. Further studies are needed to establish the role of vaccination as postremission therapy for AML.
This research article summarizes a study encapsulating curcumin, a natural anti-cancer compound, in polymeric micelles for cancer therapy. The researchers synthesized a diblock copolymer micelle (PNPC) from methoxy polyethylene glycol and oleic acid to encapsulate curcumin. They showed that PNPC had high drug loading efficiency and stability. In vitro, PNPC significantly suppressed the proliferation of breast and liver cancer cell lines. In an animal model of breast cancer, PNPC treatment reduced tumor incidence and size compared to controls. PNPC also increased expression of pro-apoptotic genes and decreased expression of anti-apoptotic and proliferative genes in the tumors. The results suggest PNPC is a
inmunofenotipo para leucemias linfociticas en perros.pdfleroleroero1
This study evaluated 43 dogs diagnosed with chronic lymphocytic leukemia (CLL) to determine if immunophenotype predicts survival time. The dogs were grouped as having B-CLL (CD211), T-CLL (CD31 CD81), or atypical CLL based on immunophenotyping. Survival analysis found that dogs with T-CLL had approximately 3 times and 19 times higher probability of surviving than dogs with B-CLL and atypical CLL, respectively, indicating that immunophenotype predicts survival in canine CLL. Younger age was also associated with shorter survival for dogs with B-CLL, and anemia was linked to poorer prognosis in dogs with T-CLL.
Whiteman et al-1998-international_journal_of_cancerSilvina Verna
This study investigated the relationship between p53 expression and risk factors for cutaneous melanoma using a case-control study design. Tissue samples from 121 melanoma cases were analyzed for p53 expression using immunohistochemistry. Abnormal p53 expression was detected in 22 samples (18%). Risk factors for p53-positive melanoma included an inability to tan and history of non-melanoma skin cancer. Risk factors for p53-negative melanoma included high nevus count and heavy freckling. The results suggest there may be two pathways in melanoma pathogenesis characterized by p53 overexpression related to chronic sun exposure and pigment cell instability.
Whiteman et al-1998-international_journal_of_cancer
Tierny et al
1. Cancer Therapy: Preclinical
Phase I Clinical Pharmacology Study of F14512, a
New Polyamine-Vectorized Anticancer Drug, in
Naturally Occurring Canine Lymphoma
Dominique Tierny1
, Fran¸cois Serres1
, Zacharie Segaoula1,2,3
, Ingrid Bemelmans1
,
Emmanuel Bouchaert1
, Aurelie Petain4
,Viviane Brel5
, Stephane Couffin6
,Thierry Marchal7
,
Laurent Nguyen4
, Xavier Thuru2,3
, Pierre Ferre4
, Nicolas Guilbaud5
, and Bruno Gomes5
Abstract
Purpose: F14512 is a new topoisomerase II inhibitor con-
taining a spermine moiety that facilitates selective uptake by
tumor cells and increases topoisomerase II poisoning. F14512
is currently in a phase I/II clinical trial in patients with acute
myeloid leukemia. The aim of this study was to investigate
F14512 potential in a new clinical indication. Because of the
many similarities between human and dog lymphomas, we
sought to determine the tolerance, efficacy, pharmacokinetic/
pharmacodynamic (PK/PD) relationship of F14512 in this
indication, and potential biomarkers that could be translated
into human trials.
Experimental Design: Twenty-three dogs with stage III–IV
naturally occurring lymphomas were enrolled in the phase I
dose-escalation trial, which consisted of three cycles of F14512
i.v. injections. Endpoints included safety and therapeutic efficacy.
Serial blood samples and tumor biopsies were obtained for
PK/PD and biomarker studies.
Results: Five dose levels were evaluated to determine the
recommended dose. F14512 was well tolerated, with the expected
dose-dependent hematologic toxicity. F14512 induced an early
decrease of tumoral lymph node cells, and a high response rate of
91% (21/23) with 10 complete responses, 11 partial responses, 1
stable disease, and 1 progressive disease. Phosphorylation of
histone H2AX was studied as a potential PD biomarker of F14512.
Conclusions: This trial demonstrated that F14512 can be safely
administered to dogs with lymphoma resulting in strong thera-
peutic efficacy. Additional evaluation of F14512 is needed to
compare its efficacy with standards of care in dogs, and to translate
biomarker and efficacy findings into clinical trials in humans. Clin
Cancer Res; 1–10. Ó2015 AACR.
Introduction
Non-Hodgkin lymphoma is the seventh most common human
systemic malignancy, with an estimated prevalence of 70,000
patients in the United States in 2013 (1). The addition of anti-
CD20 therapy to the multidrug chemotherapy regimen CHOP
(cyclophosphamide–adriamycin–vincristine–prednisolone) great-
ly improved the prognosis of diffuse large B-cell lymphoma
(DLBCL), but nearly one third of patients relapsed, underlining
the considerable possibility for therapeutic improvement (2).
Vectorization is an ingenious way to improve tumor selectivity
of known therapeutic agents, by conjugating them with a chem-
ical entity to target cancer cells more specifically. One possibility is
to exploit a selective transport system, such as the polyamine
transport system, which is overactive in many tumor cells (3). The
anticancer drug candidate F14512 is designed to target cancer cells
through the polyamine transport system. It contains a spermine
chain in place of the C4 glycosidic moiety of etoposide (4). The
positively charged spermine tail contributes to (i) favoring the
selective uptake of the drug by tumor cells via the polyamine
transport system, (ii) increasing DNA binding to reinforce topo-
isomerase II inhibition, (iii) enhancing the water solubility of the
drug. These properties translated into a favorable pharmacologic
profile: F14512 has demonstrated potent in vitro and in vivo
antitumor activities in preclinical studies, and shown to be super-
ior to etoposide, its parent compound (4–11).
With the support of this solid preclinical data, F14512 pro-
gressed to clinical development and a phase I clinical trial was
initiated in refractory/relapsing acute myeloid leukemia (AML).
Promising antileukemic activity was observed at different dose
levels (12) and F14512 is currently in a phase I/II trial in AML in
combination with cytarabine.
Preclinical data showed that lymphoma could be an indication
of interest for F14512 development (4). Dogs may be the most
relevant animal model to study new therapies for this indication.
Actually, naturally occurring lymphomas in dogs are closer to
their human counterparts than any xenograft mice models,
1
Oncovet Clinical Research, SIRIC ONCOLille, Avenue Paul Langevin,
Villeneuve d'Ascq, France. 2
Inserm, UMR-S1172, Jean Pierre Aubert
Research Centre, Lille, France. 3
Universite de Lille, Lille, France. 4
Insti-
tut de Recherche Pierre Fabre, Oncology Pharmacokinetics,Toulouse,
France. 5
Institut de Recherche Pierre Fabre, Experimental Oncology
Research Center, Toulouse, France. 6
Institut de Recherche Pierre
Fabre, Pharmacokinetics, Bel Air de Campans, Castres, France. 7
UPSP
2011-03-101, Interaction Cellules Environnement, Campus Veterinaire
de VetAgro-Sup, Marcy l'Etoile, France.
Note: Supplementary data for this article are available at Clinical Cancer
Research Online (http://clincancerres.aacrjournals.org/).
Corresponding Author: Bruno Gomes, Institut de Recherche Pierre Fabre,
Experimental Oncology Research Center, 3 Avenue Hubert Curien, Toulouse,
France. Current address: iTeos Therapeutics, Rue Auguste Piccard 48, B-6041,
Gosselies, Belgium. E-mail: gomesbruno@live.fr
doi: 10.1158/1078-0432.CCR-14-3174
Ó2015 American Association for Cancer Research.
Clinical
Cancer
Research
www.aacrjournals.org OF1
2. because of their growth over long periods of time in an intact
immune system, interindividual and intratumoral heterogene-
ity, development of recurrent or resistant diseases, and metas-
tasis (13). Clinical presentation, biologic behavior, tumor
genetics, and treatment response are very similar between
canine and human DLBCL (14, 15). Moreover, recent gene
profiling expression studies emphasized the molecular similar-
ities of DLBCL in both species (16, 17). Finally, the relevance of
dogs as a lymphoma model is supported by their use in clinical
trials (18, 19).
Etoposide has been extensively studied in dogs for pharmaco-
kinetics (PK) and toxicologic purposes (20, 21), but little data are
available concerning its potential antitumor efficacy in pet dogs
(22, 23). A retrospective study on 13 dogs with relapsing lym-
phoma treated with etoposide (24) showed that etoposide had a
minimal therapeutic effect. Etoposide phosphate administration
in dogs was associated with hematologic toxicity (20). An acute
pruritic cutaneous reaction was also observed, but it could have
been linked to the vehicle used in this study (24). As etoposide
had poor results in a pet dog lymphoma model, it was of great
interest to evaluate F14512, a vectorized and more soluble form of
etoposide, in this model to validate the proof of concept and the
benefits of this tumor-targeting agent.
The aims of this dose-escalation clinical trial were therefore (i)
to assess the potential clinical and hematologic tolerance and to
determine the MTD of F14512 in a population of dogs with
naturally occurring lymphoma, (ii) to describe the PK character-
istics of F14512 in these dogs, and (iii) to identify early signs of
efficacy of this treatment, with an evaluation of the remission rate
after three cycles of therapy. This was a translational research
study, as PK/pharmacodynamic (PD) relationships, biomarker,
and efficacy can be directly translated into ongoing and future
clinical trials of F14512.
Materials and Methods
Chemicals
F14512 was provided by Pierre Fabre Medicament. The design
and synthesis of F14512 have been patented (WO 2005/100363).
Cell culture
Namalwa (Burkitt's lymphoma—ATCC CRL-1432) cells were
grown in RPMI-1640 medium with 10% FCS, 2 mmol/L
L-glutamine, 100 mg/mL penicillin–streptomycin, and 1.25
mg/mL fungizone. Cells were incubated at 37
C in a humidified
atmosphere with 5% CO2 and maintained using standard cell
culture techniques.
Antiproliferative activity
Namalwa cells were seeded in 96-well plates, treated with
increasing concentrations of F14512 and incubated for 72 hours.
Cell viability was then evaluated by dosing the ATP released by
viable cells using the ATPlite assay (PerkinElmer). EC50 values
were determined with a curve-fitting analysis (a nonlinear regres-
sion model with a sigmoidal dose response, variable hill slope
coefficient), performed with the GraphPad Software algorithm.
Cell-cycle analysis
Namalwa cells were seeded in 96-well plates and incubated
with 6 different concentrations ranging from 0.125- to 5-fold the
IC50 value (determined in the ATPlite assay) for 24, 48, and 72
hours. Cells were labeled with the Kit Coulter DNA-Prep Reagents
(Beckman Couter) and analyzed with a Guava PCA-96 flow
cytometer (Merck Millipore). The percentage of cells in each
cell-cycle phase was calculated using M Cycle software.
DNA double-strand breaks detection
Namalwa cells were seeded in 12-well plates and exposed to
increasing concentrations of F14512 for 4, 16, and 24 hours. To
detect DNA double-strand breaks (DSB), cells were fixed and
permeabilized with DNA-prep LPr reagent (Beckman Coulter)
and stained with an Alexa Fluor 647 anti–H2AX-phosphorylated
(ser139) antibody (Biolegend). For the DNA content analysis,
cells were stained with DNA-prep STAIN reagent. Phospho-
H2AX–positive cells were analyzed on a LSR-II cytometer (BD
Biosciences).
Annexin V–PE/Nexin-7AAD staining experiments
Namalwa cells were seeded in 96-well plates and treated with
increasing concentrations of tested compound for 16 hours. The
cells were then trypsinized and stained with the Guava PCA-96
Nexin Kit (Guava Technologies). This kit contained Annexin V
combined with phycoerythrin and 7-amino actinomycin D (7-
AAD) that bound to the intracellular nucleic acid when the
membrane integrity was impaired. Thereafter, samples were ana-
lyzed with a Guava PCA-96 flow cytometer.
Caspases 3/7 activation measurements
Namalwa cells were seeded in 96-well plates, treated with
different concentrations of F14512 and incubated for 16 hours.
Caspases 3/7 enzyme activities were measured using the Caspase-
Glo 3/7 assay (Promega Corp.), in accordance with the manu-
facturer's instructions.
Fine-needle aspirates
Fine-needle aspirates were performed by a clinical trial and
veterinary care assistant using 21-gauge needles. Cells were col-
lected from three different lymph nodes on which an average of
three punctures had been performed. Aspirates were then flushed
in a 4 mL vial containing PBS (adapted from refs. 25 and 26). The
needle was redirected several times during aspiration to avoid cell
Translational Relevance
Despite the approval over the past 10 years of several
targeted therapeutic drugs, it is important to continue to test
cytotoxic agents that still play a major role in high-grade
lymphoma therapy. Drugs that are specifically vectorized to
cancer cells, such as the new polyamine-vectorized drug
F14512, should offer reinforced activity to tackle tumor cells
while sparing normal cells, resulting in an improved thera-
peutic index and/or reducing unwanted toxicities. Although
traditional preclinical mice models often fail to accurately
predict the antitumor activity and toxicity of new therapies,
comparative oncology has revealed that spontaneous lympho-
mas in dogs share clinical, biologic, genetic, and therapeutic
similarities with their human counterparts. This study showed
the therapeutic relevance of the vectorized drug F14512 in a
canine lymphoma model and has potential translational
relevance for both the ongoing clinical development of
F14512 and the treatment of lymphomas in humans.
Tierny et al.
Clin Cancer Res; 2015 Clinical Cancer ResearchOF2
3. collection from the same area. Subsequent sample collections
were carried out on the same lymph nodes used for the initial
diagnosis and evaluation (as described by Williams and collea-
gues; ref. 27). A cell count was performed twice on each sample
using Nexcelom Cellometer Auto T4 (Ozyme Biosciences) and
Covalab cell chambers.
Ex vivo P-H2AX cytometry analysis
Phospho-H2AX levels were quantified using the flow-cytome-
try technique adapted from Huang and colleagues (28). Experi-
ments were performed on fine-needle aspirate samples. Cells were
fixed in a 2% formalin solution and permeabilized in a 70% cold
ethanol solution then washed and rehydrated in PBS, 4% FCS and
0.1% Triton X-100. Nuclear H2AX staining was performed using a
rabbit anti canine-gamma-H2AX antibody (NB100-384; Novus
biologicals) and a secondary antibody (Cy5-goat anti-rabbit IgG
A10931; Invitrogen Life Technologies).
Dog selection
This study was conducted by Oncovet Clinical Research (OCR)
as part of a collaborative research project between OCR and Pierre
Fabre Medicament. As the drug provider, Pierre Fabre Medica-
ment also provided technical and scientific assistance. The trial
was available for all dogs that (i) had new or previously diagnosed
multicentric lymphoma; (ii) had a measurable disease at study
entry (allowing diagnosis and staging) with no restriction on the
stage of the disease; (iii) had not responded to standard therapies
(including chemotherapy and/or glucocorticoids) or whose own-
ers had declined standard therapies; (iv) had no anticancer
treatment in the month before entry to the study; (v) had no
significant biochemical abnormality or cytopenia, which preclud-
ed the use of cytotoxic drugs; (vi) had no concurrent serious
systemic disorder incompatible with this study; (vii) had a life
expectancy of at least 9 weeks, according to veterinary opinion.
Initial staging was performed on all dogs using a standardized
protocol, with a histologic confirmation of diagnosis. The dogs
were staged at the time of diagnosis based on the World Health
Organization (WHO) classification: five-stage criteria for canine
lymphoma and lymph node size were assessed using published
recommendations (29). Staging tests included a complete blood
count, chemistry panel, two-view chest X-rays, an abdominal
ultrasound, and a bone marrow aspirate.
This study was approved by the OCR Ethical Committee. Dogs
were only enrolled after the obtention of a signed informed
consent from the owner and were able to withdraw from the
study at any time.
Pathology
Biopsy specimens from enlarged lymph nodes were fixed in
10% neutral-buffered formalin for 48 hours and embedded in
paraffin wax. Four micrometer-thick sections were stained with
hematoxylin and eosin. Immunophenotyping was performed
using antibodies targeting human antigens but cross-reacting with
the equivalent canine antigens. An antibody targeting CD-3 was
used as a pan-T marker (monoclonal mouse anti-human F7.2.38;
Dako) and an antibody targeting CD-20 was used as a pan-B
marker (rabbit anti-human RB-9013-P; Thermoscientific). One
null cell neoplasm (negative for both CD20 and CD3) was also
evaluated for PAX5 (clone 24; Cell Mark), and BLA36 (clone A27-
42; Biogenex) expression (two antigens expressed by B-cell neo-
plasms). The original diagnosis for each case was made by one
pathologist (I. Bemelmans). A second pathologist worked inde-
pendently as an expert reviewer (T. Marchal). The classification of
the 23 cases was based on cellular morphology and immuno-
phenotypes according to WHO criteria.
Assessment of hematologic and coagulation status
Blood was collected using jugular or cephalic venipuncture.
Blood samples were placed into EDTA vacutainer glass tubes and
heparinized blood tubes. Analyses were performed immediately.
The baseline assessment included the measurement of the chem-
istry profile, including an ionized calcium assessment (Catalyst
biochemistry analyzer, IDEXX Laboratories Inc.), a complete
blood count, including platelet concentration (Procyte Hematol-
ogy analyzer, IDEXX Laboratories Inc.), and D-dimers concentra-
tion using a turbidometric immunoassay (Nyco Card Reader II,
NYCOMED).
Treatment schedule
All the dogs involved in the study followed the same protocol
over a period of 6 weeks. This consisted of a 3-hour F14512 i.v.
infusion once daily for 3 consecutive days, repeated every 2 weeks
(days 1–3, days 15–17, and days 22–24). The first dose level of
0.05 mg/kg and the schedule of administration were chosen
following previous preclinical studies performed in beagle dogs
and to be consistent with ongoing and future clinical trials in
humans.
Side effect assessment and medical intervention
Toxicities were graded according to the Veterinary Cooperative
Oncology Group criteria for adverse events (30). The toxicity
grade assigned to each dog was based on the nadir neutrophil
or platelet count, the highest documented biochemical enzyme
noted and the highest grade of gastrointestinal or constitutional
symptom/toxicity noted. A total of 16 interim visits were planned
in the protocol (on days 1, 3, 4, 7, 9, 11, 15, 17, 18, 22, 25, 29, 31,
32, 36, and 38). At each follow-up visit, owners were questioned
about signs of adverse clinical effects, daily water intake, appetite,
urination, vomiting, stool consistency/frequency, energy level,
mood, and exercise tolerance. A complete blood count was
performed at each visit. All dogs were monitored for a mini-
mum of 8 weeks. Dose-limiting toxicity (DLT) was defined as
prolonged (48 hours) asymptomatic grade 4 neutropenia,
febrile neutropenia, grade 4 anorexia, grade 4 vomiting, grade
4 diarrhea, or death. In all cases, the maximal toxicity grade
recorded during all follow-up examinations was used for the
determination of the MTD. Gastrointestinal adverse events
were treated with symptomatic treatments. A prophylactic
broad spectrum antibiotherapy was administrated in the case
of severe asymptomatic neutropenia (grades 3 and 4). Dogs
with febrile neutropenia were hospitalized and treated with
intravenous fluids and antibiotics.
Response assessment and follow-up
The response to the drug was evaluated at each treatment
session by an oncologist in accordance with previously published
criteria (31). The remission status was assessed on the basis of
physical examination and mandatory lymph node cytology of the
remaining enlarged lymph nodes. Two weeks after the end of the
protocol (day 45), all dogs underwent complete end-staging,
similar to initial staging. If requested, a complementary CHOP-
based chemotherapy protocol was proposed to the dog's owner.
F14512 Dose-Escalation Trial in Spontaneous Canine Lymphoma
www.aacrjournals.org Clin Cancer Res; 2015 OF3
4. F14512 analysis in plasma
The PK of F14512 and its active metabolite F16490 were
evaluated throughout the treatment. Six blood samples were
drawn on day 1 of cycle 1 (before dosing and 1.5, 3, 3.5, 5, and
6 hours after the start of the 3-hour infusion). Several samples
were also obtained during and after cycles 2 and 3. Plasma
concentrations of F14512 and F16490 were quantified using a
validated LC/MS-MS method with a lower limit of quantification
(LLOQ) of 0.25 ng/mL.
Statistical analysis
Data are expressed either as mean Æ SD or as percentages. The
primary endpoint of this study was to determine the clinical
response rate (stable disease, complete or partial remission) on
day 45. The selected level of significance was set at P 0.05.
Results
F14512 effects in lymphoma cell lines
We previously found that F14512 (see structure in Supplemen-
tary Fig. S1A) displayed strong efficacy in a human lymphoma cell
line, both in vitro and in vivo in a mouse model (4, 8). In this study,
wefurtheranalyzed the effect ofF14512 on the Burkitt's lymphoma
cell line Namalwa. This drug induced a dose-dependent growth
inhibition after a 72-hour treatment (Supplementary Fig.S1B) with
a calculated EC50 value of 46 nmol/L (95% confidence interval;
35–61 nmol/L). We observed that Namalwa cells treated with
F14512 were blocked in the G2–M phase in a dose-dependent
manner (55% and 62% after a 48-hour and 72-hour treatment,
respectively, at 2 EC50; Supplementary Fig. S1C). The induction of
apoptosis was first evaluated via the conventional annexin V–PE/
7-AAD staining procedure. F14512 showed no effect on Namalwa
cells after a 16-hour treatment at doses up to 10 mmol/L (Supple-
mentary Fig. S1D). These results were confirmed by measuring
caspases 3 and 7 activities after exposure to the drug for 16 hours.
F14512 only induced a dose-dependent increase of caspases-3/7
activity at high doses (above 3 mmol/L; Supplementary Fig. S1E),
underlining that F14512 did not behave as a proapoptotic agent.
A key step of the mechanism of action of F14512 is the
topoisomerase II-dependent generation of DNA damages. Inhi-
bition of topoisomerase II leads to the generation of cleavable
complexes and DSB of DNA that are characterized by phosphor-
ylation of histone H2AX on Ser-139. P-H2AX is used as a reporter
of DNA damage.
We then evaluated the impact of F14512 on DNA damage by
measuring the phosphorylation of histone H2AX on Ser-139 in
Namalwa cells. F14512 induced DNA-damage in a dose- and
time-dependent manner (Supplementary Fig. S2). P-H2AX
increased as early as 4 hours after F14512 incubation and the
percentage of stained cells stabilized after a 16-hour incubation
period (55% and 67% of P-H2AX–positive cells at the dose
corresponding to the antiproliferative EC50 and 2 mmol/L, respec-
tively). The presence of Ser-139–phosphorylated H2AX was also
explored as an in vivo PD biomarker of F14512.
P-H2AX is a PD biomarker of F14512 in dogs
As a proof of concept of F14512-targeting tumors in dogs with
naturally occurring lymphoma, we looked at P-H2AX induction
in tumoral lymph node fine-needle aspirates. Four lymphoma-
bearing dogs received a single F14512 i.v. injection at a low dose
of 0.05 mg/kg, and serial fine-needle aspirates were performed. A
P-H2AX induction was observed by flow cytometry as early as 2
hours (not shown) after the end of the F14512 infusion and
increased at 4 hours (Fig. 1). P-H2AX induction was heteroge-
neous among the 4 patients but an increase was observed in all
dogs. These PD data further supported the clinical evaluation and
dose-escalation trial of F14512 in lymphoma-bearing dogs. These
4 dogs were subsequently included in the first cohort of the trial.
Epidemiologic characteristics and staging of dogs with
spontaneous lymphoma
Twenty-three dogs with naturally occurring lymphomas were
enrolled in the dose-escalation trial consisting of three cycles of
F14512 i.v. injection between November 2013 and March 2014.
The epidemiologic characteristics of these patients are summa-
rized in Table 1. There were 14 female and 9 male dogs. The mean
age of all 23 dogs was 8.0 Æ 2.6 years. The mean weight of all dogs
was 29.5 Æ 17 kg. Seventeen different canine breeds known as
being predisposed to lymphoma were represented (32). The
majority of cases (15/23) were classified as DLBCLs, 12 as cen-
troblastic (DLBCL-CB) and 3 as immunoblastic (DLBCL-IB).
Three other B-cell lymphoma cases were identified as late-stage
marginal zone lymphomas in transition into DLBCL-IB. The other
subtypes were: two peripheral T-cell lymphomas (2/23), includ-
ing one pleomorphic small lymphoma and one pleomorphic
mixed lymphoma (according to the updated Kiel Malignant
Lymphoma Classification); and a T-cell lymphoblastic lympho-
ma (1/23). The lymphoma subtype could not be determined in
two cases due to the lack of biopsy material, but the diagnosis of
high-grade lymphoma was based on the cytologic examination.
Two dogs had previously received long-term corticosteroid mono-
therapy before the study, with partial responses and rapid
relapses. Four dogs had previously received various CHOP-based
chemotherapy treatments with a complete response and relapse
before inclusion.
Pharmacokinetics
A total of five cohorts were successively initiated, with an
initial dosage of 0.050, 0.060, 0.070, 0.085, and 0.075 mg/kg
(cohorts 1 to 5). Although the cohort size per dose level was
small and the dose range explored was narrow, F14512 and
F16490 plasma AUC increased overall with the dose level (Fig.
2A). Figure 2B and C represents plasma concentrations versus
time of F14512 and its metabolite F16490 on day 1 for all dogs
treated at 0.075 mg/kg (recommended dose). F14512 plasma
concentrations were in the range of the IC50 value estimated in
the Namalwa model (46 nmol/L ¼ 29 ng/mL) for approxi-
mately 2 to 3 hours in most dogs. Interestingly, the patient with
the lowest plasma concentration was the only one that did not
have a clinical response to treatment at this dose level. Else-
where, the AUC of the active metabolite F16490 and of F14512
was proportional and, on average, the F16490 AUC represented
23% of the F14512 AUC.
Pharmacodynamics
Lymph node tumor cells and blood cell numeration were
monitored early during this clinical trial to determine whether
F14512 therapy was associated with any biologic effect. Serial
fine-needle aspirates were performed in tumoral lymph nodes in
the hours following the first injection of F14512 to look for PD
markers of F14512. The total cell number in these serial aspirates
was evaluated and normalized to the aspirate volume. A rapid and
Tierny et al.
Clin Cancer Res; 2015 Clinical Cancer ResearchOF4
5. dramatic decrease in the number of cells was observed (Fig. 3A) as
early as 2 hours after the beginning of the F14512 infusion. By the
end of the first cycle of the F14512 therapy, the number of cells
was dramatically reduced. Such a potent effect on tumoral cells
made the analysis of P-H2AX induction impossible in all patients.
Indeed, F14512 induced a strong decrease in viable cells, gener-
ating a lot of necrotic cells and debris (Supplementary Fig. S3),
and thus rendering any reliable P-H2AX evaluation using flow
cytometry impossible. This significant decrease in total lymph
node cell numbers was noticed at all dose levels (not shown) and
was clearly a PD marker of the F14512 efficacy.
Blood cell counts could be both a marker of efficacy and
toxicity. As expected, a decrease in the white blood cell count,
including neutrophils, lymphocytes, and monocytes, was
observed after each cycle of F14512 therapy, with a nadir on day
nine. A dose–effect relationship was observed between the dose of
F14512 and the number of neutrophils (Fig. 3B). This decrease
was reversible and baseline levels were recovered by the beginning
of the next cycle. The evaluation of the decrease in white blood
cells as well as the duration of the decrease correlated with the
dose of F14512 and allowed to determine DLTs.
Toxicities
All 23 dogs were evaluated for tolerance. Signs of toxicity
included neutropenia, anemia, thrombocytopenia, and digestive
disorders (diarrhea and vomiting). Toxicities are reported
in Table 2. Gastrointestinal adverse events (diarrhea for 6 dogs
and vomiting for 2 dogs) were mild in 6 dogs (grade 1) and
moderate in 1 dog (grade 2). They were not dose related (no
gastrointestinal toxicity observed with the highest dosage) and the
side effects resolved rapidly with the use of symptomatic treat-
ments. Severe adverse events (grade 3 and above) were limited to
neutropenia, thrombocytopenia, and anemia, and were all revers-
ible. Grade 4 neutropenia was observed at all dosages, but was
short (less than 48 hours) and clinically well tolerated for the first
three dose levels. Cohorts 1 and 2 were extended to confirm the
short duration and reversibility of the neutropenia at these dose
levels. Grade 4 neutropenia lasting more than 48 hours was
observed in 2 dogs from cohort 4, and was considered as a DLT.
A fifth cohort with an intermediate dosage (0.075 mg/kg) was,
therefore, constituted and determined as the recommended dose.
There were no treatment-related deaths, but 1 dog died due to the
progress of the disease on day 27.
Figure 1.
P-H2AX is a PD biomarker of F14512 in dogs. In vivo analysis of P-H2AX staining in fine-needle aspirates of tumor lymph nodes from 4 dogs (A–D) treated
with 0.05 mg/kg F14512. Isotype control (blank), before treatment (shaded) and 4 hours after the end of F14512 infusion (black).
F14512 Dose-Escalation Trial in Spontaneous Canine Lymphoma
www.aacrjournals.org Clin Cancer Res; 2015 OF5
6. Clinical outcome
Stable or progressive disease was observed in 2 dogs, both
having been treated before entering the study (Table 3). One of
these dogs, treated in cohort 1, died on day 27 with progressive
disease, whereas the other dog, treated in cohort 5, died on day
128 (after showing a stable disease on day 45). Ten dogs achieved
a complete regression, whereas 11 dogs experienced a partial
remission (Table 3). Pretreatment and inclusion in one of the
first three cohorts were associated with a tendency toward a lower
chance of achieving a complete remission, although the difference
was not statistically significant. Twenty dogs received additional
treatments following F14512 (consisting of various chemother-
apy protocols).
Discussion
Naturally occurring canine tumors represent a useful and
valuable model for deciphering many aspects of human cancers
(33). As part of the broader field of comparative oncology,
translational drug development studies in dogs with spontaneous
cancers have been used to define doses and schedules for thera-
peutic agents through rigorous PK–PD endpoints, often involving
serial biopsies of tumor tissue and the collection of biologic
materials before and after exposure to novel therapeutics (34,
35). Comparative oncology has been focusing on the study of
homologies, differences, and translational relevance of various
cancers, including lymphomas (36), osteosarcomas (37, 38), soft
tissue sarcomas (39), urinary bladder cancer (40), mammary
cancers (41), and others.
In this dose-escalating study, we have shown that the response
rate (complete and partial responses) of F14512 in treated canine
patients with lymphoma is 91%. Direct comparison of median
survival time or time to relapse with published data with other
chemotherapy agents is difficult because of the heterogeneity of
our population. Moreover, for obvious ethical reasons, some
patients were treated with another type of chemotherapy after
the F14512 study. Nevertheless, we can compare the clinical
response rate with F14512 with the reported response rate with
doxorubicin, which is commonly recognized as the most efficient
single agent for the treatment of canine high-grade non-Hodgkin
lymphoma. In a previous study (42), the response rate of dogs
treated with doxorubicin as a first line agent was 74%, which can
be compared with the result observed in our study (91%).
Therefore, our results are promising and emphasize the potential
of F14512, which could be investigated further either alone or in
combination with other agents. Because the treatment schedule
only included three cycles of F14512 injections, additional cycles
could be considered in future studies to potentially increase
clinical efficacy. As the main toxicity of F14512 is febrile neutro-
penia, a combination with conventional chemotherapy should be
chosen rationally to avoid major toxicities. In human lympho-
mas, the CHOP-based chemotherapy is associated with anti-
CD20 antibodies. It should be noted that some laboratories are
now working on therapeutic canine anti-CD20 antibodies (43).
Apart from rituximab, no other targeted therapy has emerged and
been developed so far for DLBCL, and the canine patient as a
cancer model may accelerate research. Ibrutinib, an irreversible
BTK inhibitor, benefited from its evaluation in canine lymphomas
(44) and is now proven to be efficacious in humans (45): The
canine lymphoma model enabled the authors to demonstrate the
full-target occupancy at various dosages, and to achieve some
objective clinical responses (44). Second-generation BTK inhibi-
tors are now being evaluated in dogs (46) in proof-of-concept
studies along with their development pathway, whereas clinical
Table 1. Epidemiologic and clinical characteristics at diagnosis of the whole study population composed of 23 dogs with na€ve and relapsing lymphoma
Epidemiologic characteristics Population studied
Sex
Male 39.1% (9/23)
Female 60.9% (14/23)
Age, y, mean Æ SD (range) 8.0 Æ 2.6 (4.0–15.0)
Body weight, kg, mean Æ SD (range) 29.5 Æ 17 (4.3–54.0)
Clinical characteristics
Breed
Rottweiler 13.0% (3/23)
Cavalier King Charles spaniel, Bernese mountain dog, Jack Russel terrier, Labrador Retriever 8.7% (2/23 each 4 breeds)
Yorkshire terrier, Cocker spaniel, West highland white terrier, Golden Retriever, Bull terrier, French
Bouledogue, Dogue de Bordeaux, Weimaraner, Mastiff, Greyhound, German Shepherd, cross-breed
4.3% (1/23 each 12 breeds)
Clinical stage
Stage 3 17.4% (4/23)
Stage 4 82.6% (19/23)
Clinical substage
a 52.1% (12/23)
b 47.9% (11/23)
Hypercalcemia 8.7% (2/23)
Pretreatment
Corticosteroid 8.7% (2/23)
Chemotherapy 17.4% (4/23)
Lymphoma subtype
B cell
Diffuse large B-cell lymphoma 65.2% (15/23)
Marginal zone lymphoma 13.0% (3/23)
T cell
Peripheral T cell 8.7% (2/23)
T-cell lymphoblastic 4.3% (1/23)
Unclassified 8.7% (2/23)
Tierny et al.
Clin Cancer Res; 2015 Clinical Cancer ResearchOF6
7. studies of combination of ibrutinib with polychemotherapy are
ongoing in DLBCL in humans. Thus, we believe that a combina-
tion of F14512 with targeted therapies such as anti-CD20 mAb or
BTK inhibitors should be considered in the future.
As a vectorized form of etoposide, F14512 was shown to be
more potent than etoposide in vitro (4) and to be superior to
etoposide in terms of efficacy and therapeutic window in xeno-
graft mice models (8, 9). F14512 also revealed a convincing
antitumor activity in naturally occurring lymphomas in dogs,
with 22 of 23 patients displaying a clinical response (either stable
disease, partial, or complete remission). These clinical results in
dogs clearly appear to be superior to those described for etoposide
(24). Etoposide was also shown to be poorly active in a retro-
spective study on 13 animals, with only 2 dogs displaying a
Figure 2.
F14512 PK data in dogs. A, descriptive statistics of F14512 and F14512 main
metabolite (F16490; AUClast by dose level). B and C, F14512 and
F16490 plasma concentrations versus time for all dogs treated at the
recommended dose level of 0.075 mg/kg, after the first infusion (cycle 1,
day 1). F14512 infusions started at t ¼ 0 hours and lasted between
3.25 and 3.63 hours ($).
Figure 3.
Tumoral and hematologic PD markers of F14512. A, F14512 induces a strong
and early decrease of tumoral lymph node cell number in dogs. Fine-needle
aspirates were performed in tumoral lymph nodes all along the first cycle
of F14512 administration. F14512 infusions were initiated at t ¼ 0 hours and
lasted 3 hours at day 1 (0–3 hours), day 2 (24–27 hours), and day 3
(48–51 hours). The graph shows the total cell count per milliliter of fine-needle
aspirates, from all dogs treated at all dose levels (results, mean þ SD; data
were analyzed using ANOVA followed by PLSD Fisher post hoc; Ã
, P 0.05;
ÃÃ
, P 0.01; and ÃÃÃ
, P 0.001, n ¼ 23 dogs. B, F14512 induces a dose-
dependent decrease in circulating neutrophils in treated dogs. The graph
shows the dose–effect relationship between the dose level of F14512 and the
number of neutrophils (median absolute neutrophil count vs. time by dose
level in mg/kg). F14512 was administered on days 1, 2, and 3.
F14512 Dose-Escalation Trial in Spontaneous Canine Lymphoma
www.aacrjournals.org Clin Cancer Res; 2015 OF7
8. clinical response. Moreover, as F14512 was shown to be a poor
substrate of the efflux protein PgP (unpublished data), it would be
of great interest to look for F14512 efficacy in relapsing and/or
refractory chemoresistant dog lymphomas who have a poor prog-
nosis (47). The canine multidrug resistance associated protein has
been molecularly identified (48). In thepresent study, 4 dogs hada
previous polychemotherapy course. Even if their PgP/Mdr status
was not known, a clinical response was observed in 3 of them.
Therefore, this F14512 phase I study warrants further evaluation of
F14512 in relapsing and/or chemoresistant lymphoma cases.
Because of the unequal distribution of dogs depending on the
lymphomasubtype, it was not possible to see any relation between
the clinical response and the lymphoma subtype.
In this study, we looked at F14512-induced DNA damages and
demonstrated that P-H2AX signaling (49) could be used as a PD
biomarker. We took the opportunity to design a preliminary study
in 4 dogs treated with a single injection of low-dose F14512 and
unraveled an early in vivo P-H2AX induction as a PD response.
Unfortunately, we could not correlate P-H2AX induction with the
F14512 AUC or the clinical response owing to the rapid onset of
lymphoma cell death induced by F14512. We were not able to
recover enough living cells from all treated dogs to perform a
relevant flow-cytometry study of P-H2AX. We did not expect such
a tumoral cell death (as early as a couple of hours after initiating
F14512 infusion). A preferential and rapid uptake of F14512 by
tumor cells, with an added cytotoxic effect brought by F16490
metabolite, may explain this strong tumoral cytotoxicity. We
previously demonstrated the preferential uptake of F14512 by
tumor cells (4, 5), and as a consequence plasma levels of F14512
and F16490 may only partially reflect the concentrations reached
in tumors. F16490 alone displays cytotoxic activity with an EC50
of 74 nmol/L in the Namalwa cell line. This rapid cell death
induction was observed in all dogs, even in the 2 dogs that did not
display a clinical response later. This latter observation confirms
the strong cytotoxicity of F14512 in dog lymphoma and supports
further evaluation either in combination and/or with the addition
of other cycles of treatment to reinforce and extend its efficacy.
Careful monitoring of patients may be required in human clinical
trials for the early detection of tumor lysis syndrome that may
occur.
An interesting point about the study we performed on dogs is
that there is an ongoing clinical assessment of F14512 in humans,
so we are able to compare and translate the clinical data and
findings from both species, even if the clinical tumor indications
are not the same. The first-in-man multicenter phase I trial on
F14512 as a single drug was conducted in adult patients with
relapsed or refractory AML (12, 50). Patients received a daily i.v.
infusion for 5 consecutive days every 2 to 6 weeks depending on
the leukemia response as well as the recovery of sufficient hema-
topoiesis and the resolution of toxicities. The main toxicity was
myelosuppression, which was dose dependent and reversible. The
MTD reached was 44 mg/m2
and the recommended dose was
determined to be 39 mg/m2
. Antileukemic activity was observed
at different dose levels with 10% complete responses, 8% com-
plete responses with incomplete recovery and 3 patients who
experienced hematologic improvements. As with humans, we
Table 2. Toxicity observed in the whole study population composed of 23 dogs with na€ve and relapsing lymphoma
Hematologic toxicity grade (number of dogs)
Gastrointestinal toxicity
grade (number of dogs)
Cohort (dose) Number of dogs Hemoglobin HCT Platelet Neutrophils Diarrhea Vomiting
Cohort 1 (0.05 mg/m2
) 6 Grade 2 (1) Grade 2 (1) Grade 3 (1) Grade 4 (3)a
Grade 1 (1) Grade 1 (1)
Grade 1 (4) Grade 1 (4) Grade 2 (3) Grade 3 (1)
Cohort 2 (0.06 mg/m2
) 4 Grade 4 (1) Grade 4 (1) Grade 2 (2) Grade 4 (2)a
Grade 1 (2) Grade 1 (1)
Grade 2 (1) Grade 3 (1) Grade 1 (1) Grade 3 (1)
Grade 1 (1) Grade 2 (1)
Cohort 3 (0.07 mg/m2
) 3 Grade 3 (1) Grade 3 (1) Grade 2 (1) Grade 4 (1)a
Grade 2 (1) 0
Grade 1 (2) Grade 1 (2) Grade 3 (1)
Cohort 4 (0.085 mg/m2
) 4 Grade 2 (1) Grade 2 (2) 0 Grade 4 (2)b
0 0
Grade 1 (3) Grade 1 (2) Grade 4 (1)a
Grade 2 (1)
Cohort 5 (0.075 mg/m2
) 6 Grade 2 (3) Grade 2 (4) Grade 4 (1) Grade 4 (1)b
Grade 1 (2) 0
Grade 1 (3) Grade 1 (2) Grade 3 (1) Grade 4 (2)a
Grade 2 (2) Grade 3 (1)
a
Asymptomatic neutropenia lasting less than 48 hours.
b
Febrile neutropenia lasting more than 48 hours.
Table 3. Clinical response of the whole study population composed of 23 dogs with na€ve and relapsing lymphoma
Clinical response at day 45
Population Progressive disease Stable disease Partial response Complete response
Whole population 4.4% (1/23) 4.4% (1/23) 47.8% (11/23) 43.5% (10/23)
Pretreatment
Yes (n ¼ 6) 16.7% (1/6) 16.7% (1/6) 66.6% (4/6) 0
No (n ¼ 17) 0 0 58.8% (10/17) 41.2% (7/17)
Cohort
Cohort 1: 0.050 mg/kg (n ¼ 6) 16.7% (1/6) 0 50.0% (3/6) 33.3% (2/6)
Cohort 2: 0.060 mg/kg (n ¼ 4) 0 0 75.0% (3/4) 25.0% (1/4)
Cohort 3: 0.070 mg/kg (n ¼ 3) 0 0 66.6% (2/3) 33.3% (1/3)
Cohort 4: 0.085 mg/kg (n ¼ 4) 0 0 50.0% (2/4) 50.0% (2/4)
Cohort 5: 0.075 mg/kg (n ¼ 6) 0 16.7% (1/6) 16.7% (1/6) 66.6% (4/6)
Tierny et al.
Clin Cancer Res; 2015 Clinical Cancer ResearchOF8
9. also observed clinical responses in dogs at all dose levels, but
relapses were more frequently noticed at the first dose levels (even
if not statistically significant). When we looked at PD markers of
F14512 in dogs, the tumor lymph node cell counts and the
circulating blood cells counts (Fig. 3) revealed a favorable ther-
apeutic window: A strong decrease of tumor cells was observed at
all doses whereas the hematologic adverse events were increased
only at higher doses. Interestingly, the tolerance profile was very
similar between humans and dogs, with the hematotoxicity being
the major adverse event observed. A phase II trial of F14512 in
combination with cytosine arabinoside is currently ongoing in
patients with AML.
In conclusion, our results provide a strong evidence of the
clinical efficacy of the new vectorized drug F14512 in a pet dog
model of lymphoma. The translational value of canine lympho-
ma warrants further studies of F14512 and strongly supports its
clinical development.
Disclosure of Potential Conflicts of Interest
No potential conflicts of interest were disclosed.
Authors' Contributions
Conception and design: D. Tierny, F. Serres, A. Petain, N. Guilbaud, B. Gomes
Development of methodology: Z. Segaoula, I. Bemelmans, E. Bouchaert
Acquisition of data (provided animals, acquired and managed patients,
provided facilities, etc.): D. Tierny, F. Serres, V. Brel, T. Marchal
Analysis and interpretation of data (e.g., statistical analysis, biostatistics,
computational analysis): D. Tierny, Z. Segaoula, A. Petain, V. Brel, S. Couffin,
L. Nguyen, X. Thuru, B. Gomes
Writing, review, and/or revision of the manuscript: D. Tierny, F. Serres, V. Brel,
S. Couffin, P. Ferre, N. Guilbaud, B. Gomes
Administrative, technical, or material support (i.e., reporting or organizing
data, constructing databases): Z. Segaoula, I. Bemelmans, E. Bouchaert
Study supervision: D. Tierny, B. Gomes
Acknowledgments
The authors thank Marie Bosredon for her kind assistance in the F14512
batch management and the flow core facility of BICeL-IFR114 for the technical
assistance.
Grant Support
This work was supported by grant from BPI-France, through the CaninCa
project.
The costs of publication of this article were defrayed in part by the
payment of page charges. This article must therefore be hereby marked
advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate
this fact.
Received December 8, 2014; revised June 13, 2015; accepted July 4, 2015;
published OnlineFirst July 13, 2015.
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