38th FEBS Congress
Maxim L. Bychkov
Poster for abstract: "DR5-B – DR5-selective mutant variant of cytokine TRAIL overcomes resistance of cancer cells to TRAIL"
Abstract book, p. 595
Targeting cell death receptor ligand systems in cancer treatment by dr muzami...muzamilch5
This is a powerpoint presentation on the topic "Targeting cell death receptor ligand systems in cancer treatment" by Dr Muzamil Ch.
Owner of this Content: Dr. Muzamil Ch
Email: muzamilch2018@gmail.com
Before using this content, you must contact the owner first.
Evaluating the ability of anti-cancer drugs Etoposide and Staurosporine to in...Davient Bala
Cervical cancer is considered one of the most prevalent cancers affecting Singaporean women.
Although many novel chemotherapeutics have been developed recently, little has been done to
determine the efficiency of current anti-cancer agents working in combination. Here, we aimed to
evaluate the apoptosis induction efficiency of Etoposide and Staurosporine in HeLa cells. Cell cultures
were subjected to either 50 μM Etoposide, 10 nM Staurosporine or both for 24 hours prior visualization
under a fluorescence microscope. We found that Etoposide alone had an efficiency of 16.1% while
Staurosporine alone had 18.3%. However, the polytherapy achieved an efficiency of up to 33.6%,
which indicates an additive effect of both drugs to induce apoptosis. Our results demonstrate that
Etoposide and Staurosporine are both capable of inducing apoptosis in HeLa cells. Furthermore, it
reveals the potential of Etoposide-Staurosporine polytherapy to be a potent combinative treatment
option for cervical cancer patients resistant or sensitive to conventional anti-cancer agents.
This document summarizes research on the discovery of a novel class of orally active inhibitors of N-myristoyltransferase (NMT) that are trypanocidal, meaning they can kill the parasite Trypanosoma brucei which causes Human African Trypanosomiasis (HAT). Researchers screened over 63,000 compounds and identified a hit compound (1) that inhibited T. brucei NMT with low micromolar potency and also had activity against the parasite. They then optimized the hit through structure-activity relationship studies, improving potency against the enzyme and parasite as well as developing good oral pharmacokinetics. This led to a lead compound, DDD85646 (
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
Pharmacogenetics involves studying genetic variations that lead to differences in individual drug responses. The goals are to optimize drug therapy and limit toxicity based on a person's genetic profile by choosing the best drug, dose, and duration for that individual. Genetic variations can occur as single nucleotide polymorphisms, which are single base pair differences between individuals. Polymorphisms in genes encoding drug-metabolizing enzymes, such as CYP2D6 and CYP2C9, are clinically significant as they can result in poor, intermediate, extensive, or ultrarapid metabolism of certain drugs. This can impact treatment outcomes and risk of adverse effects.
Three key points:
1. A kinome-centered synthetic lethality screen identified that suppression of the ERBB3 receptor tyrosine kinase sensitizes KRAS mutant lung and colon cancer cells to MEK inhibitors.
2. MEK inhibition results in MYC-dependent transcriptional upregulation of ERBB3, which is responsible for intrinsic drug resistance.
3. Drugs targeting both EGFR and ERBB2, each capable of forming hetero-dimers with ERBB3, can reverse unresponsiveness to MEK inhibition by decreasing inhibitory phosphorylation of the proapoptotic proteins BAD and BIM.
This document describes research to develop new treatments for human African trypanosomiasis (HAT) by optimizing inhibitors of Trypanosoma brucei N-myristoyltransferase (TbNMT). The initial lead compound 1 showed potent activity against TbNMT but poor brain penetration, limiting its efficacy in the later central nervous system stage of HAT. The researchers modified compound 1 to improve blood-brain barrier permeability and selectivity against human NMTs. Key modifications included reducing the compound's polar surface area, replacing the central aromatic ring with a flexible linker, and capping the sulfonamide group. This led to the discovery of compound 40, which demonstrated partial efficacy in a mouse model
Targeting cell death receptor ligand systems in cancer treatment by dr muzami...muzamilch5
This is a powerpoint presentation on the topic "Targeting cell death receptor ligand systems in cancer treatment" by Dr Muzamil Ch.
Owner of this Content: Dr. Muzamil Ch
Email: muzamilch2018@gmail.com
Before using this content, you must contact the owner first.
Evaluating the ability of anti-cancer drugs Etoposide and Staurosporine to in...Davient Bala
Cervical cancer is considered one of the most prevalent cancers affecting Singaporean women.
Although many novel chemotherapeutics have been developed recently, little has been done to
determine the efficiency of current anti-cancer agents working in combination. Here, we aimed to
evaluate the apoptosis induction efficiency of Etoposide and Staurosporine in HeLa cells. Cell cultures
were subjected to either 50 μM Etoposide, 10 nM Staurosporine or both for 24 hours prior visualization
under a fluorescence microscope. We found that Etoposide alone had an efficiency of 16.1% while
Staurosporine alone had 18.3%. However, the polytherapy achieved an efficiency of up to 33.6%,
which indicates an additive effect of both drugs to induce apoptosis. Our results demonstrate that
Etoposide and Staurosporine are both capable of inducing apoptosis in HeLa cells. Furthermore, it
reveals the potential of Etoposide-Staurosporine polytherapy to be a potent combinative treatment
option for cervical cancer patients resistant or sensitive to conventional anti-cancer agents.
This document summarizes research on the discovery of a novel class of orally active inhibitors of N-myristoyltransferase (NMT) that are trypanocidal, meaning they can kill the parasite Trypanosoma brucei which causes Human African Trypanosomiasis (HAT). Researchers screened over 63,000 compounds and identified a hit compound (1) that inhibited T. brucei NMT with low micromolar potency and also had activity against the parasite. They then optimized the hit through structure-activity relationship studies, improving potency against the enzyme and parasite as well as developing good oral pharmacokinetics. This led to a lead compound, DDD85646 (
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
Pharmacogenetics involves studying genetic variations that lead to differences in individual drug responses. The goals are to optimize drug therapy and limit toxicity based on a person's genetic profile by choosing the best drug, dose, and duration for that individual. Genetic variations can occur as single nucleotide polymorphisms, which are single base pair differences between individuals. Polymorphisms in genes encoding drug-metabolizing enzymes, such as CYP2D6 and CYP2C9, are clinically significant as they can result in poor, intermediate, extensive, or ultrarapid metabolism of certain drugs. This can impact treatment outcomes and risk of adverse effects.
Three key points:
1. A kinome-centered synthetic lethality screen identified that suppression of the ERBB3 receptor tyrosine kinase sensitizes KRAS mutant lung and colon cancer cells to MEK inhibitors.
2. MEK inhibition results in MYC-dependent transcriptional upregulation of ERBB3, which is responsible for intrinsic drug resistance.
3. Drugs targeting both EGFR and ERBB2, each capable of forming hetero-dimers with ERBB3, can reverse unresponsiveness to MEK inhibition by decreasing inhibitory phosphorylation of the proapoptotic proteins BAD and BIM.
This document describes research to develop new treatments for human African trypanosomiasis (HAT) by optimizing inhibitors of Trypanosoma brucei N-myristoyltransferase (TbNMT). The initial lead compound 1 showed potent activity against TbNMT but poor brain penetration, limiting its efficacy in the later central nervous system stage of HAT. The researchers modified compound 1 to improve blood-brain barrier permeability and selectivity against human NMTs. Key modifications included reducing the compound's polar surface area, replacing the central aromatic ring with a flexible linker, and capping the sulfonamide group. This led to the discovery of compound 40, which demonstrated partial efficacy in a mouse model
Genetic polymorphisms can affect how individuals metabolize and respond to drugs. The document discusses how single nucleotide polymorphisms (SNPs) in genes encoding drug-metabolizing enzymes like the cytochrome P450 system can result in poor, intermediate, normal, extensive, or ultra-rapid metabolizers. This genetic variation impacts the metabolism of many drugs and can influence their effects as well as drug interactions. The cytochrome P450 2C19 enzyme, which is important in metabolizing diazepam, shows polymorphisms that lead to different drug responses and side effects between ethnic populations. Understanding these pharmacogenomic factors is important for optimizing drug therapy.
This document discusses genetic polymorphisms in drug transport proteins and how they can impact drug pharmacokinetics and toxicity. It introduces two major superfamilies of transport proteins - ATP-binding cassette (ABC) transporters and solute carrier (SLC) proteins. Specific ABC transporters discussed include P-glycoprotein (ABCB1), the multidrug resistance proteins ABCC1 and ABCG2. The document also summarizes key SLC transporters and provides examples of important substrates for each. Genetic variations in these transport proteins can significantly influence individual responses to drugs like irinotecan used in cancer chemotherapy.
1) The document reports that 6 out of 16 melanoma tumor samples analyzed acquired expression of EGFR after developing resistance to BRAF or MEK inhibitor drugs.
2) Using a shRNA library targeting chromatin regulators, the researchers found that suppression of SOX10 in melanoma cells leads to activation of TGF-β signaling and upregulation of EGFR and PDGFRB, conferring resistance to BRAF and MEK inhibitors.
3) Expression of EGFR in melanoma cells or treatment with TGF-β results in a slow-growth phenotype with cellular hallmarks of senescence. However, EGFR expression or TGF-β exposure becomes advantageous for proliferation in the presence of BRAF or
This document lists 50 publications by Dr. Gilbert Lefèvre from 1987 to 2008. The publications cover a range of topics including:
- Anti-Müllerian hormone and related monoclonal antibodies (publications 1-3)
- Pharmacokinetic studies of drugs like hirudin, artemether, and rivastigmine (publications 4-10, 23-25, 45-50)
- Drug interaction and safety studies of antimalarial and Alzheimer's drugs (publications 26-28, 30-33, 36, 38, 40, 41, 44)
- Pharmacokinetic studies of antimalarial drugs in specific populations like children and Africans (publications 29, 42, 43)
The document analyzes the tumor transcriptomics profile of a patient with stage IV gallbladder cancer. It identifies the top 200 most significantly activated expression regulators in the patient's tumor using sub-network enrichment analysis software. Key regulators identified include histone deacetylases and DNA methyltransferases. The analysis suggests treatment with the HDAC inhibitor vorinostat and discusses how activated regulators like HDACs, PDCD1, and CTLA4 may be contributing to tumor proliferation and immune evasion. Graphical summaries show pathways enriched with active regulators in the tumor related to cancer hallmarks like histone modification and immune response evasion.
This document discusses epigenetics and epigenetic modifications. It begins by defining the epigenome and how it can change without altering DNA sequence. The main epigenetic modifications discussed are DNA methylation and various histone modifications such as acetylation, methylation, phosphorylation, ubiquitination, and SUMOylation. These modifications can impact gene expression and transcription. The document then examines how epigenetic abnormalities contribute to various diseases like cancer, diabetes, and neurological disorders. Finally, it reviews some FDA-approved epigenetic therapies that target DNA methyltransferases and histone deacetylases, as well as epigenetic drugs currently in clinical trials.
The document discusses anthracycline nano-delivery systems to overcome multiple drug resistance (MDR). It provides an overview of commonly used anthracycline drugs like doxorubicin and challenges with MDR. The review then summarizes over ten types of anthracycline nano-delivery systems that have been developed to overcome MDR, including liposomes, polymeric micelles, nanoparticles, conjugates, and carbon nanotubes. Several of these nano-delivery systems have advanced to clinical trials with the goal of improving cancer treatment by circumventing MDR.
It is my journal club presentation on Synthesis, Docking Studies and Anticancer Activity of New Substituted Pyrimidine and Triazolopyrimidine Glycosides.
I sincerely thank the authors Wael A. El-Sayed, Ashraf M. Mohamed , Hemat S. Khalaf, Dina S. EL-Kady, May Al-Manawaty
Maytansinoid immunoconjugate IMGN901 is cytotoxicEllen Gunn
IMGN901 is a maytansinoid immunoconjugate targeting CD56, which is expressed in 70% of multiple myeloma cases. The study evaluated IMGN901 alone and in combination with lenalidomide and dexamethasone to determine if it can overcome environmental-mediated drug resistance (EM-DR) in multiple myeloma. While adhesion to extracellular matrix reduced IMGN901 potency, it remained cytotoxic with an LC50 of 43 nM. However, only a combination of IMGN901, lenalidomide, and dexamethasone was able to overcome drug resistance arising from direct contact between multiple myeloma and stromal cells. Drug resistance in this condition was associated with upregulation of multi-drug
Dietary Phytochemicals and Histone Modifications in CancerDr. Supratim Ghatak
1. Histone modifications such as acetylation, methylation, and phosphorylation are altered in cancer and can affect gene expression. Certain plant-derived chemicals like resveratrol, curcumin, and anacardic acid can inhibit histone-modifying enzymes and may have anti-cancer effects.
2. Resveratrol, found in grapes, inhibits HDACs and activates SIRT1, inducing apoptosis in cancer cells by downregulating anti-apoptotic proteins. It also reduces heat shock protein Hsp70 levels in leukemia cells.
3. Further studies are needed to understand the effects of resveratrol on other proteins involved in apoptosis signaling pathways and heat shock transcription factors in cancer
1) BRAF(V600E) inhibition through PLX4032 is effective for treating melanoma but not colon cancer, which shows limited response.
2) A genetic screen identified that inhibiting EGFR synergizes with inhibiting BRAF(V600E) in colon cancer cell lines.
3) Mechanistically, inhibiting BRAF(V600E) causes rapid feedback activation of EGFR through inhibition of MEK/ERK and activation of CDC25C, which supports continued proliferation. Inhibiting both BRAF and EGFR prevents this feedback and synergistically inhibits colon cancer cell and tumor growth.
This document compares the efficacy of 5 different FGFR inhibitors - AZD4547, BGJ398, JNJ42756493, PD173074 and TKI258 - at inhibiting FGFR1 signaling and cell proliferation in solid tumor cell lines and leukemia cell lines that overexpress FGFR1. It finds that the pan-FGFR inhibitors AZD4547, BGJ398 and JNJ42756493 more potently suppress FGFR1 phosphorylation and downstream signaling, cause more cell cycle arrest, and have lower GI50 values, indicating they more efficiently inhibit cell growth. It also shows that BGJ398 treatment significantly suppresses leukemia progression in a xenograft model of primary human AML overexpressing FGFR1, supporting
This document describes cardiotoxicity observed in rodents and in rat and human-derived cardiomyocyte cell lines treated with nicotinamide phosphoribosyltransferase inhibitors (NAMPTi). Short-term administration of several NAMPTis to rodents resulted in sudden death and signs of congestive heart failure. Further studies demonstrated that NAMPTi-induced toxicity in rat and human cardiomyocyte cell lines was on-target and likely human-relevant. Co-administration of nicotinic acid partially mitigated toxicity in vitro but not consistently in vivo. Human-derived cardiomyocyte assays were useful for assessing cardiotoxicity of compounds prior to in vivo studies.
1) The research tested 25 pyrazoline derivative compounds for their ability to inhibit the growth of the intestinal parasite Entamoeba histolytica.
2) The results showed that series 1a and 1b were the most effective at inhibiting trophozoite growth, with certain compounds performing better than the drug metronidazole.
3) The most promising individual inhibitors were compounds 5 and 14, which showed greater effectiveness than metronidazole at inhibiting growth over time at both tested concentrations.
Conformation of the Human Immunoglobulin G2 Hinge Imparts Superagonistic Prop...World ADC
Monoclonal antibody (mAb) drugs that stimulate antitumor immunity are transforming cancer treatment but require optimization for maximum clinical impact. Here, we show that, unlike other immunoglobulin isotypes, human IgG2 (h2) imparts FcgR-independent agonistic activity to immune-stimulatory mAbs such as anti-CD40, -4-1BB, and -CD28. Activity is provided by a subfraction of h2, h2B, that is structurally constrained due its unique arrangement of hinge region disulfide bonds. Agonistic activity can be transferred from h2 to h1 by swapping their hinge and CH1 domains, and substitution of key hinge and CH1 cysteines generates homogenous h2 variants with distinct agonistic properties. This provides the exciting opportunity to engineer clinical reagents with defined therapeutic activity regardless of FcgR expression levels in the local microenvironment.
The MAT (Monocyte Activation Test) is an in vitro, non-animal alternative to the rabbit pyrogen test. It measures the levels of cytokines like IL-1, IL-6, and TNFα produced by human monocytes when exposed to pyrogenic substances. Elevated cytokine levels indicate the presence of pyrogens, allowing correlation to a product's pyrogen content. The test utilizes cryopreserved human whole blood or isolated PBMCs (peripheral blood mononuclear cells). It involves incubating a test solution with human monocytes and comparing the cytokine response to an endotoxin standard to determine if it meets the defined pyrogen limit.
This document discusses regulation of deoxynucleotide metabolism in cancer and its therapeutic implications. It describes how imbalanced levels of deoxynucleotide triphosphates (dNTPs) can lead to genomic instability and increased cancer risk. Several key points are made: 1) Enzymes like ribonucleotide reductase and SAMHD1 regulate dNTP levels and maintain genomic stability. 2) Mutations or dysregulation of these enzymes can cause elevated dNTP pools and increased mutagenesis, facilitating cancer development. 3) Many cancer therapies target dNTP synthesis pathways to inhibit tumor growth. 4) SAMHD1 specifically acts as a tumor suppressor by maintaining low dNTP levels
CXCR7 is induced by hypoxia and mediates glioma cell migration towards SDF-1a...Enrique Moreno Gonzalez
Glioblastomas, the most common and malignant brain tumors of the central nervous system, exhibit high invasive capacity, which hinders effective therapy. Therefore, intense efforts aimed at improved therapeutics are ongoing to delineate the molecular mechanisms governing glioma cell migration and invasion.
This study describes the structure-based design and evaluation of novel peptide inhibitors of thrombin-induced platelet aggregation. Researchers designed tetrapeptide analogs of a lead thrombin inhibitor by replacing the P3 residue with various unnatural amino acids like D-3,3-di-phenylalanine. All inhibitors competitively inhibited thrombin and improved inhibition of platelet aggregation. Compounds containing D-3,3-di-phenylalanine at P3 completely inhibited thrombin-induced platelet aggregation at concentrations of 300-60 nM. These novel thrombin inhibitors could be used as scaffolds to develop treatments for acute coronary syndrome by inhibiting thrombin-mediated platelet aggregation.
Functional p53 is required for rapid restoration of daunorubicin-induced lesi...Enrique Moreno Gonzalez
This document summarizes a research article that studied the role of p53 in daunorubicin (DNR)-induced lesions in the spleen. The key findings were:
1) DNR treatment caused more rapid cell death and weight loss in the spleens of wild type mice compared to p53-null mice.
2) While wild type mouse spleens recovered normal morphology 8 days after DNR treatment, p53-null mouse spleens still had large necrotic lesions.
3) DNR treatment increased p21 levels in wild type mice but not p53-null mice, indicating p53 is required for p21 induction.
4) The results suggest p53
The document discusses several antibody-drug conjugates (ADCs) that are being studied or developed for the treatment of non-small cell lung cancer (NSCLC). It summarizes clinical trial results of trastuzumab emtansine and trastuzumab deruxtecan for HER2-positive NSCLC, and sacituzumab govitecan and datopotamab deruxtecan for Trop-2 positive NSCLC. It also briefly mentions ADCs in development that target HER3, c-MET, CEACAM5, Axl, PTK7, NaPi2b, and Nectin-4 for lung cancer. The document concludes by reviewing a phase 1 clinical trial of
1) Researchers discovered new lead compounds that inhibit the enzyme N-myristoyltransferase (NMT) in the parasite that causes African sleeping sickness, Trypanosoma brucei. 2) These lead compounds were shown to rapidly kill trypanosomes in vitro and cure infections in mouse models of the disease. 3) The best compound demonstrated good oral bioavailability and cured infections in mice at low doses, comparable to existing drugs. The compound represents a promising new treatment for African sleeping sickness.
Genetic polymorphisms can affect how individuals metabolize and respond to drugs. The document discusses how single nucleotide polymorphisms (SNPs) in genes encoding drug-metabolizing enzymes like the cytochrome P450 system can result in poor, intermediate, normal, extensive, or ultra-rapid metabolizers. This genetic variation impacts the metabolism of many drugs and can influence their effects as well as drug interactions. The cytochrome P450 2C19 enzyme, which is important in metabolizing diazepam, shows polymorphisms that lead to different drug responses and side effects between ethnic populations. Understanding these pharmacogenomic factors is important for optimizing drug therapy.
This document discusses genetic polymorphisms in drug transport proteins and how they can impact drug pharmacokinetics and toxicity. It introduces two major superfamilies of transport proteins - ATP-binding cassette (ABC) transporters and solute carrier (SLC) proteins. Specific ABC transporters discussed include P-glycoprotein (ABCB1), the multidrug resistance proteins ABCC1 and ABCG2. The document also summarizes key SLC transporters and provides examples of important substrates for each. Genetic variations in these transport proteins can significantly influence individual responses to drugs like irinotecan used in cancer chemotherapy.
1) The document reports that 6 out of 16 melanoma tumor samples analyzed acquired expression of EGFR after developing resistance to BRAF or MEK inhibitor drugs.
2) Using a shRNA library targeting chromatin regulators, the researchers found that suppression of SOX10 in melanoma cells leads to activation of TGF-β signaling and upregulation of EGFR and PDGFRB, conferring resistance to BRAF and MEK inhibitors.
3) Expression of EGFR in melanoma cells or treatment with TGF-β results in a slow-growth phenotype with cellular hallmarks of senescence. However, EGFR expression or TGF-β exposure becomes advantageous for proliferation in the presence of BRAF or
This document lists 50 publications by Dr. Gilbert Lefèvre from 1987 to 2008. The publications cover a range of topics including:
- Anti-Müllerian hormone and related monoclonal antibodies (publications 1-3)
- Pharmacokinetic studies of drugs like hirudin, artemether, and rivastigmine (publications 4-10, 23-25, 45-50)
- Drug interaction and safety studies of antimalarial and Alzheimer's drugs (publications 26-28, 30-33, 36, 38, 40, 41, 44)
- Pharmacokinetic studies of antimalarial drugs in specific populations like children and Africans (publications 29, 42, 43)
The document analyzes the tumor transcriptomics profile of a patient with stage IV gallbladder cancer. It identifies the top 200 most significantly activated expression regulators in the patient's tumor using sub-network enrichment analysis software. Key regulators identified include histone deacetylases and DNA methyltransferases. The analysis suggests treatment with the HDAC inhibitor vorinostat and discusses how activated regulators like HDACs, PDCD1, and CTLA4 may be contributing to tumor proliferation and immune evasion. Graphical summaries show pathways enriched with active regulators in the tumor related to cancer hallmarks like histone modification and immune response evasion.
This document discusses epigenetics and epigenetic modifications. It begins by defining the epigenome and how it can change without altering DNA sequence. The main epigenetic modifications discussed are DNA methylation and various histone modifications such as acetylation, methylation, phosphorylation, ubiquitination, and SUMOylation. These modifications can impact gene expression and transcription. The document then examines how epigenetic abnormalities contribute to various diseases like cancer, diabetes, and neurological disorders. Finally, it reviews some FDA-approved epigenetic therapies that target DNA methyltransferases and histone deacetylases, as well as epigenetic drugs currently in clinical trials.
The document discusses anthracycline nano-delivery systems to overcome multiple drug resistance (MDR). It provides an overview of commonly used anthracycline drugs like doxorubicin and challenges with MDR. The review then summarizes over ten types of anthracycline nano-delivery systems that have been developed to overcome MDR, including liposomes, polymeric micelles, nanoparticles, conjugates, and carbon nanotubes. Several of these nano-delivery systems have advanced to clinical trials with the goal of improving cancer treatment by circumventing MDR.
It is my journal club presentation on Synthesis, Docking Studies and Anticancer Activity of New Substituted Pyrimidine and Triazolopyrimidine Glycosides.
I sincerely thank the authors Wael A. El-Sayed, Ashraf M. Mohamed , Hemat S. Khalaf, Dina S. EL-Kady, May Al-Manawaty
Maytansinoid immunoconjugate IMGN901 is cytotoxicEllen Gunn
IMGN901 is a maytansinoid immunoconjugate targeting CD56, which is expressed in 70% of multiple myeloma cases. The study evaluated IMGN901 alone and in combination with lenalidomide and dexamethasone to determine if it can overcome environmental-mediated drug resistance (EM-DR) in multiple myeloma. While adhesion to extracellular matrix reduced IMGN901 potency, it remained cytotoxic with an LC50 of 43 nM. However, only a combination of IMGN901, lenalidomide, and dexamethasone was able to overcome drug resistance arising from direct contact between multiple myeloma and stromal cells. Drug resistance in this condition was associated with upregulation of multi-drug
Dietary Phytochemicals and Histone Modifications in CancerDr. Supratim Ghatak
1. Histone modifications such as acetylation, methylation, and phosphorylation are altered in cancer and can affect gene expression. Certain plant-derived chemicals like resveratrol, curcumin, and anacardic acid can inhibit histone-modifying enzymes and may have anti-cancer effects.
2. Resveratrol, found in grapes, inhibits HDACs and activates SIRT1, inducing apoptosis in cancer cells by downregulating anti-apoptotic proteins. It also reduces heat shock protein Hsp70 levels in leukemia cells.
3. Further studies are needed to understand the effects of resveratrol on other proteins involved in apoptosis signaling pathways and heat shock transcription factors in cancer
1) BRAF(V600E) inhibition through PLX4032 is effective for treating melanoma but not colon cancer, which shows limited response.
2) A genetic screen identified that inhibiting EGFR synergizes with inhibiting BRAF(V600E) in colon cancer cell lines.
3) Mechanistically, inhibiting BRAF(V600E) causes rapid feedback activation of EGFR through inhibition of MEK/ERK and activation of CDC25C, which supports continued proliferation. Inhibiting both BRAF and EGFR prevents this feedback and synergistically inhibits colon cancer cell and tumor growth.
This document compares the efficacy of 5 different FGFR inhibitors - AZD4547, BGJ398, JNJ42756493, PD173074 and TKI258 - at inhibiting FGFR1 signaling and cell proliferation in solid tumor cell lines and leukemia cell lines that overexpress FGFR1. It finds that the pan-FGFR inhibitors AZD4547, BGJ398 and JNJ42756493 more potently suppress FGFR1 phosphorylation and downstream signaling, cause more cell cycle arrest, and have lower GI50 values, indicating they more efficiently inhibit cell growth. It also shows that BGJ398 treatment significantly suppresses leukemia progression in a xenograft model of primary human AML overexpressing FGFR1, supporting
This document describes cardiotoxicity observed in rodents and in rat and human-derived cardiomyocyte cell lines treated with nicotinamide phosphoribosyltransferase inhibitors (NAMPTi). Short-term administration of several NAMPTis to rodents resulted in sudden death and signs of congestive heart failure. Further studies demonstrated that NAMPTi-induced toxicity in rat and human cardiomyocyte cell lines was on-target and likely human-relevant. Co-administration of nicotinic acid partially mitigated toxicity in vitro but not consistently in vivo. Human-derived cardiomyocyte assays were useful for assessing cardiotoxicity of compounds prior to in vivo studies.
1) The research tested 25 pyrazoline derivative compounds for their ability to inhibit the growth of the intestinal parasite Entamoeba histolytica.
2) The results showed that series 1a and 1b were the most effective at inhibiting trophozoite growth, with certain compounds performing better than the drug metronidazole.
3) The most promising individual inhibitors were compounds 5 and 14, which showed greater effectiveness than metronidazole at inhibiting growth over time at both tested concentrations.
Conformation of the Human Immunoglobulin G2 Hinge Imparts Superagonistic Prop...World ADC
Monoclonal antibody (mAb) drugs that stimulate antitumor immunity are transforming cancer treatment but require optimization for maximum clinical impact. Here, we show that, unlike other immunoglobulin isotypes, human IgG2 (h2) imparts FcgR-independent agonistic activity to immune-stimulatory mAbs such as anti-CD40, -4-1BB, and -CD28. Activity is provided by a subfraction of h2, h2B, that is structurally constrained due its unique arrangement of hinge region disulfide bonds. Agonistic activity can be transferred from h2 to h1 by swapping their hinge and CH1 domains, and substitution of key hinge and CH1 cysteines generates homogenous h2 variants with distinct agonistic properties. This provides the exciting opportunity to engineer clinical reagents with defined therapeutic activity regardless of FcgR expression levels in the local microenvironment.
The MAT (Monocyte Activation Test) is an in vitro, non-animal alternative to the rabbit pyrogen test. It measures the levels of cytokines like IL-1, IL-6, and TNFα produced by human monocytes when exposed to pyrogenic substances. Elevated cytokine levels indicate the presence of pyrogens, allowing correlation to a product's pyrogen content. The test utilizes cryopreserved human whole blood or isolated PBMCs (peripheral blood mononuclear cells). It involves incubating a test solution with human monocytes and comparing the cytokine response to an endotoxin standard to determine if it meets the defined pyrogen limit.
This document discusses regulation of deoxynucleotide metabolism in cancer and its therapeutic implications. It describes how imbalanced levels of deoxynucleotide triphosphates (dNTPs) can lead to genomic instability and increased cancer risk. Several key points are made: 1) Enzymes like ribonucleotide reductase and SAMHD1 regulate dNTP levels and maintain genomic stability. 2) Mutations or dysregulation of these enzymes can cause elevated dNTP pools and increased mutagenesis, facilitating cancer development. 3) Many cancer therapies target dNTP synthesis pathways to inhibit tumor growth. 4) SAMHD1 specifically acts as a tumor suppressor by maintaining low dNTP levels
CXCR7 is induced by hypoxia and mediates glioma cell migration towards SDF-1a...Enrique Moreno Gonzalez
Glioblastomas, the most common and malignant brain tumors of the central nervous system, exhibit high invasive capacity, which hinders effective therapy. Therefore, intense efforts aimed at improved therapeutics are ongoing to delineate the molecular mechanisms governing glioma cell migration and invasion.
This study describes the structure-based design and evaluation of novel peptide inhibitors of thrombin-induced platelet aggregation. Researchers designed tetrapeptide analogs of a lead thrombin inhibitor by replacing the P3 residue with various unnatural amino acids like D-3,3-di-phenylalanine. All inhibitors competitively inhibited thrombin and improved inhibition of platelet aggregation. Compounds containing D-3,3-di-phenylalanine at P3 completely inhibited thrombin-induced platelet aggregation at concentrations of 300-60 nM. These novel thrombin inhibitors could be used as scaffolds to develop treatments for acute coronary syndrome by inhibiting thrombin-mediated platelet aggregation.
Functional p53 is required for rapid restoration of daunorubicin-induced lesi...Enrique Moreno Gonzalez
This document summarizes a research article that studied the role of p53 in daunorubicin (DNR)-induced lesions in the spleen. The key findings were:
1) DNR treatment caused more rapid cell death and weight loss in the spleens of wild type mice compared to p53-null mice.
2) While wild type mouse spleens recovered normal morphology 8 days after DNR treatment, p53-null mouse spleens still had large necrotic lesions.
3) DNR treatment increased p21 levels in wild type mice but not p53-null mice, indicating p53 is required for p21 induction.
4) The results suggest p53
The document discusses several antibody-drug conjugates (ADCs) that are being studied or developed for the treatment of non-small cell lung cancer (NSCLC). It summarizes clinical trial results of trastuzumab emtansine and trastuzumab deruxtecan for HER2-positive NSCLC, and sacituzumab govitecan and datopotamab deruxtecan for Trop-2 positive NSCLC. It also briefly mentions ADCs in development that target HER3, c-MET, CEACAM5, Axl, PTK7, NaPi2b, and Nectin-4 for lung cancer. The document concludes by reviewing a phase 1 clinical trial of
1) Researchers discovered new lead compounds that inhibit the enzyme N-myristoyltransferase (NMT) in the parasite that causes African sleeping sickness, Trypanosoma brucei. 2) These lead compounds were shown to rapidly kill trypanosomes in vitro and cure infections in mouse models of the disease. 3) The best compound demonstrated good oral bioavailability and cured infections in mice at low doses, comparable to existing drugs. The compound represents a promising new treatment for African sleeping sickness.
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.
The Radiosensitivity Effect of Hydroxyurea on HT29 Cell Lineiosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Formaldehyde increases the sensitivity of breast and ovarian cancer cells to chemotherapeutic drugs like doxorubicin, cisplatin, and 5-fluorouracil in a BRCA1/2-dependent manner. Experiments showed a synergistic growth inhibition effect when formaldehyde was combined with these drugs at low doses in BRCA1/2 deficient cell lines, but not in BRCA1/2 proficient cell lines. Further experiments indicated this synergistic response was due to increased DNA double-strand breaks and cell death, rather than just growth inhibition, when formaldehyde was combined with doxorubicin. The synergistic, cytotoxic response to formaldehyde combinations was also observed in BRCA1/2 deficient ovarian cancer cell
This document discusses a proposed study to target cancer stem cells in leukemia using tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-based immunotoxins. The objectives are to identify and characterize cancer stem cells from leukemia cell lines and patients, construct recombinant TRAIL-based chimeric proteins to selectively target these cancer stem cells, and test the efficacy and selectivity of the proteins. Cancer stem cells are resistant to chemotherapy and responsible for leukemia recurrence. TRAIL selectively induces apoptosis in cancer cells and has advantages over chemotherapy. The study aims to develop dual receptor targeted immunotoxins linking TRAIL to the interleukin-2 receptor to more effectively treat leukemia by eliminating drug-resistant cancer stem cells.
This document provides an overview of chemotherapy for head and neck cancer. It discusses the cell cycle effects of chemotherapy drugs and classifications including cytotoxic, targeted, and hormonal agents. Specific drugs are explained including mechanisms of action, indications, and side effects. The document also reviews landmark clinical trials establishing concurrent chemoradiotherapy as the standard of care for locally advanced head and neck cancer. Targeted agents like cetuximab are now being used for recurrent/metastatic disease.
Oncodesign aacr 2018 morab-202 a folate receptor alpha-targeted antibody-dr...Florence Fombertasse
Folate receptor alpha (FRA) is a membrane protein with high affinity for binding and transporting folate into cells. Overexpression of FRA may confer a growth advantage to tumors by increasing folate uptake and affecting cell proliferation via alternative cell signaling pathways (1). FRA levels have been found to be elevated in tumors of epithelial origin compared to normal tissue as cancers of the breast (including TNBC (2)), colon, lungs and ovary (3).
In this study, we report the development of MORAb-202, an anti-FRA antibody-drug conjugate (ADC), consisting of a FRA-binding antibody (MORAb-003, farletuzumab) with a cathepsin-cleavable form of eribulin (eribulin mesylate, marketed as Halaven®), a highly potent anti-mitotic agent that induces cell-cycle arrest and cell death by targeting microtubules.*
We first study expression of FRA on a large panel of tumors patient-derived xenograft (PDX) and Cancer Cell Line-derived Xenograft (CDX). Then, we performed in vitro and in vivo anti-proliferation assays and compare antitumor activity of MORAb-202 with free eribulin accordingly to the FRA expression level. FRA expression was found to be determinant in the sensitivity of tumor cells to the cytotoxic effect of the ADC. Moreover, in case of high expression of FRA, MORAb-202 showed a higher antitumor activity compared with free eribulin.
These results suggest that FRA expression could be used as a response-predictive biomarker for this targeted therapy. The ability to identify and treat patients with an effective therapy based on the known expression of the tumor marker is a key point in predictive medicine progress. These findings support the clinical development of MORAb-202 ADC as a novel targeted therapy for patients with FRA-expressing tumors.
The ADC described in this abstract is investigational, as efficacy and safety have not been established. There is no guarantee that this ADC will be available commercially.
This document describes a study that conducted an RNA interference screening in breast cancer cells to identify epigenetic factors regulating the mesenchyme to epithelium transition (MET). Researchers designed a siRNA library targeting 729 chromatin modification genes and screened it in the mesenchymal breast cancer cell line MDA-MB-231. They identified 70 candidate genes involved in MET, including known genes like ZEB1, G9a, SMAD5 and SMARCD3, as well as DOT1L which has been implicated in MET. They also identified KAT5 as a novel gene linked to maintaining the mesenchymal phenotype for the first time. The screening approach involved measuring E-cadherin induction and cell
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.
MED12 controls the response to multiple cancer drugs through regulation of TG...Anirudh Prahallad
This study used an RNAi screen to identify MED12, a component of the transcriptional MEDIATOR complex, as a determinant of response to ALK and EGFR inhibitors in lung cancer cell lines. The study found that suppressing MED12 expression resulted in resistance to these targeted therapies. MED12 was found to negatively regulate TGF-β receptor signaling through physical interaction, and its suppression activated TGF-β signaling. Activation of TGF-β signaling led to MEK/ERK pathway activation, which conferred resistance to MEK, BRAF, and other cancer drugs. Loss of MED12 also induced an EMT-like phenotype associated with chemotherapy resistance in other cancer types. Inhibiting TGF-β receptor signaling
1) The study found that the tumor protein p53 regulates the expression of the immune checkpoint protein PDL1 (programmed death ligand 1) via the microRNA miR-34.
2) Delivery of miR-34a in a mouse model of lung cancer using a liposomal formulation (MRX34) reduced PDL1 expression in tumors and increased tumor-infiltrating immune cells.
3) Combining MRX34 with radiation therapy further increased immune cell numbers in tumors and showed potential as a novel cancer therapeutic approach.
Over-expression of APOBEC3B (A3B) has little to no effect on the efficacy of 5-Fluorouracil (5-FU) in killing mammary epithelial MCF10A cells. The study found that transfecting, transducing, or treating MCF10A cells with PMA to induce A3B expression did not significantly impact cell viability when treated with varying concentrations of 5-FU. Repeating the experiments in A3B-high cancer cell lines and investigating the mechanisms of how A3B and 5-FU may complement each other could provide insights for personalized cancer treatment.
This study developed a new mouse model that lacks all isoforms of the Trim24 gene. Mice with complete loss of Trim24 (Trim24-/-) spontaneously developed hepatic lipid accumulation, inflammation, fibrosis, and hepatocellular carcinoma without a high-fat diet or other induction methods. Trim24-/- mice had decreased expression of genes involved in oxidation, lipid metabolism, and increased expression of genes related to endoplasmic reticulum stress and cell cycle pathways. By 6 months of age, Trim24-/- mice exhibited macroscopic white liver lesions composed of steatosis and fibrosis that progressed to hepatocellular carcinoma, recapitulating features of non-alcoholic fatty liver disease in humans
Clinical Development of ADC Drugs Targeting TROP-2.pdfDoriaFang
TROP-2 is expressed in many tumor types, making it an emerging and popular target for ADC development. This article introduces clinical development of ADC drugs targeting TROP-2.
Multidisciplinary approach to the management of leukemias amlmadurai
The document discusses the multidisciplinary approach to managing leukemias like AML and MDS. It presents the case of a 68-year old male patient presenting with fever and fatigue, and details the diagnostic workup showing features consistent with acute myeloid leukemia. The document then reviews classification, prognostic factors, recent treatment trials, and the role of allogeneic stem cell transplantation for AML patients.
TREATMENT OF NON-HIDGKIN'S LYMPHOMA IN ELDERLY PATIENTSspa718
This document summarizes treatment approaches for non-Hodgkin's lymphoma in elderly patients. It discusses palliative options for refractory/relapsed diffuse large B-cell lymphoma such as gemcitabine-based chemotherapy, low-dose oral chemotherapy, and hyperfractionated cyclophosphamide. It also reviews novel anti-CD20 monoclonal antibodies showing efficacy against relapsed/refractory indolent lymphoma, and brentuximab vedotin's mechanism of action and responses seen in relapsed/refractory systemic anaplastic large cell lymphoma. Finally, it provides a high-level overview of the MD Anderson Cancer Center's Department of Lymphoma/Myeloma and its disease-specific
This document summarizes research finding that elevated activity of the Akt protein protects prostate cancer LNCaP cells from apoptosis induced by TRAIL (tumor necrosis factor-related apoptosis-inducing ligand). The researchers found that LNCaP cells have high constitutive Akt activity due to lack of the PTEN lipid phosphatase. Inhibiting PI3-kinase, which activates Akt, sensitized LNCaP cells to TRAIL-induced apoptosis. TRAIL alone activated caspases 8 and XIAP in LNCaP cells but failed to induce full apoptosis. Combining TRAIL with Akt inhibitors allowed cleavage of BID and subsequent mitochondrial apoptosis steps. Akt inhibition of BID cleavage appears to mediate the protective effect
Executive Directors Chat Leveraging AI for Diversity, Equity, and InclusionTechSoup
Let’s explore the intersection of technology and equity in the final session of our DEI series. Discover how AI tools, like ChatGPT, can be used to support and enhance your nonprofit's DEI initiatives. Participants will gain insights into practical AI applications and get tips for leveraging technology to advance their DEI goals.
Strategies for Effective Upskilling is a presentation by Chinwendu Peace in a Your Skill Boost Masterclass organisation by the Excellence Foundation for South Sudan on 08th and 09th June 2024 from 1 PM to 3 PM on each day.
Exploiting Artificial Intelligence for Empowering Researchers and Faculty, In...Dr. Vinod Kumar Kanvaria
Exploiting Artificial Intelligence for Empowering Researchers and Faculty,
International FDP on Fundamentals of Research in Social Sciences
at Integral University, Lucknow, 06.06.2024
By Dr. Vinod Kumar Kanvaria
LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UPRAHUL
This Dissertation explores the particular circumstances of Mirzapur, a region located in the
core of India. Mirzapur, with its varied terrains and abundant biodiversity, offers an optimal
environment for investigating the changes in vegetation cover dynamics. Our study utilizes
advanced technologies such as GIS (Geographic Information Systems) and Remote sensing to
analyze the transformations that have taken place over the course of a decade.
The complex relationship between human activities and the environment has been the focus
of extensive research and worry. As the global community grapples with swift urbanization,
population expansion, and economic progress, the effects on natural ecosystems are becoming
more evident. A crucial element of this impact is the alteration of vegetation cover, which plays a
significant role in maintaining the ecological equilibrium of our planet.Land serves as the foundation for all human activities and provides the necessary materials for
these activities. As the most crucial natural resource, its utilization by humans results in different
'Land uses,' which are determined by both human activities and the physical characteristics of the
land.
The utilization of land is impacted by human needs and environmental factors. In countries
like India, rapid population growth and the emphasis on extensive resource exploitation can lead
to significant land degradation, adversely affecting the region's land cover.
Therefore, human intervention has significantly influenced land use patterns over many
centuries, evolving its structure over time and space. In the present era, these changes have
accelerated due to factors such as agriculture and urbanization. Information regarding land use and
cover is essential for various planning and management tasks related to the Earth's surface,
providing crucial environmental data for scientific, resource management, policy purposes, and
diverse human activities.
Accurate understanding of land use and cover is imperative for the development planning
of any area. Consequently, a wide range of professionals, including earth system scientists, land
and water managers, and urban planners, are interested in obtaining data on land use and cover
changes, conversion trends, and other related patterns. The spatial dimensions of land use and
cover support policymakers and scientists in making well-informed decisions, as alterations in
these patterns indicate shifts in economic and social conditions. Monitoring such changes with the
help of Advanced technologies like Remote Sensing and Geographic Information Systems is
crucial for coordinated efforts across different administrative levels. Advanced technologies like
Remote Sensing and Geographic Information Systems
9
Changes in vegetation cover refer to variations in the distribution, composition, and overall
structure of plant communities across different temporal and spatial scales. These changes can
occur natural.
How to Add Chatter in the odoo 17 ERP ModuleCeline George
In Odoo, the chatter is like a chat tool that helps you work together on records. You can leave notes and track things, making it easier to talk with your team and partners. Inside chatter, all communication history, activity, and changes will be displayed.
Walmart Business+ and Spark Good for Nonprofits.pdfTechSoup
"Learn about all the ways Walmart supports nonprofit organizations.
You will hear from Liz Willett, the Head of Nonprofits, and hear about what Walmart is doing to help nonprofits, including Walmart Business and Spark Good. Walmart Business+ is a new offer for nonprofits that offers discounts and also streamlines nonprofits order and expense tracking, saving time and money.
The webinar may also give some examples on how nonprofits can best leverage Walmart Business+.
The event will cover the following::
Walmart Business + (https://business.walmart.com/plus) is a new shopping experience for nonprofits, schools, and local business customers that connects an exclusive online shopping experience to stores. Benefits include free delivery and shipping, a 'Spend Analytics” feature, special discounts, deals and tax-exempt shopping.
Special TechSoup offer for a free 180 days membership, and up to $150 in discounts on eligible orders.
Spark Good (walmart.com/sparkgood) is a charitable platform that enables nonprofits to receive donations directly from customers and associates.
Answers about how you can do more with Walmart!"
This slide is special for master students (MIBS & MIFB) in UUM. Also useful for readers who are interested in the topic of contemporary Islamic banking.
DR5-B – DR5-selective mutant variant of cytokine TRAIL overcomes resistance of cancer cells to TRAIL
1. HCT 116
Agent: EC 50, ng/ml Death*, %
Control
TRAIL 0,82±0,23 38,85±3,31
DR5-B 1,17±0,14 67,38±1,98
Taxol
(5 nM)
TRAIL 0,47±0,29 57,29±4,43
DR5-B 0,48±0,12 88,49±3,89
Bortezomib
(1 nM)
TRAIL 1,07±0,13 97,50±1,27
DR5-B 0,01±0,02 97,95±1,27
Doxorubicin
(100 nM)
TRAIL 0,56±0,25 43,00±2,21
DR5-B 1,05±0,10 93,03±3,06
Jurkat
Agent: EC 50, ng/ml Death*, %
Control
TRAIL 0,30 ± 0,06 45,14±0,40
DR5-B 0,08 ± 0,03 67,57±3,28
Taxol
(0,1 nM)
TRAIL 0,1 ± 0,05 67,41±3,07
DR5-B 0,07 ± 0,02 88,17±6,17
Bortezomib
(0,1 nM)
TRAIL 0,07 ± 0,05 55,87±3,26
DR5-B 0,05 ± 0,02 76,47±4,12
Doxorubicin
(0,1 nM)
TRAIL 0,1±0,05 63,49±2,04
DR5-B 0,09±0,03 75,46±3,47
U 937
Agent: EC 50, ng/ml Death*, %
Control
TRAIL 0,81±0,03 19,14±4,08
DR5-B 0,08±0,02 39,57±4,34
Taxol
(1 nM)
TRAIL 0,07±0,01 54,96±4,25
DR5-B 0,068±0,03 69,54±3,00
Bortezomib
(1 nM)
TRAIL 0,08±0,01 66,03±1,67
DR5-B 0,1±0,06 95,47±6,74
Doxorubicin
(100 nM)
TRAIL 0,1±0,02 60,45±5,05
DR5-B 0,072±0,05 96,65±0,89
A 549
Agent: EC 50, ng/ml Death*, %
Control
TRAIL 1,03 ± 0,08 16,41±1,05
DR5-B 0,12 ± 0,03 35,55±3,22
Taxol
(100 nM)
TRAIL 1,00 ± 0,07 44,37±3,34
DR5-B 0,09 ± 0,04 74,02±3,19
Bortezomib
(50 nM)
TRAIL 0,90 ± 0,07 62,12±3,42
DR5-B 0,09 ± 0,03 88,52±5,01
Doxorubicin
(100 nM)
TRAIL 0,073±0,03 42,22±3,19
DR5-B 0,070±0,04 65,91±3,35
HT 29
Agent: EC 50, ng/ml Death*, %
Control
TRAIL 0,72±0,02 5,43±0,06
DR5-B 0,54±0,22 40,64±0,09
Taxol
(1 nM)
TRAIL 0,09±0,06 37,62±2,54
DR5-B 0,08±0,05 82,12±1,34
Bortezomib
(1 nM)
TRAIL 0,10±0,09 45,31±3,35
DR5-B 0,10±0,05 82,01±4,28
Doxorubicin
(100 nM)
TRAIL 0,07±0,02 46,51±1,70
DR5-B 0,08±0,05 75,27±4,08
MCF-7
Agent: EC 50, ng/ml Death*, %
Control
TRAIL 1,16±0,07 11,02±0,40
DR5-B 0,45±0,12 53,52±2,02
Taxol
(100 nM)
TRAIL 0,078±0,01 52,33±4,03
DR5-B 0,08±0,05 83,28±4,52
Bortezomib
(10 nM)
TRAIL 0,08±0,02 38,23±3,09
DR5-B 0,095±0,03 96,62±4,23
Doxorubicin
(100 nM)
TRAIL 0,96±0,02 39,18±3,03
DR5-B 0,5±0,05 69,63±3,33
0
5
10
15
20
25
HCT116 U937 Jurkat A549 HT29 MCF-7
FluorescenceG-mean
DR5
DR4
DcR1
DcR2
-5
5
15
25
35
45
55
0 0,1 0,5 1 5 10 50 100 5001000
Celldeath,%
TRAIL / DR5-B, ng/ml
HT 29 (colorectal carcinoma)
TRAIL
DR5-B
0
10
20
30
40
50
60
70
80
90
0 0,1 0,5 1 5 10 50 100 5001000
Celldeath,%
TRAIL / DR5-B, ng/ml
HCT116 (colorectal carcinoma)
TRAIL
DR5-B
0
10
20
30
40
50
60
70
0 0,1 0,5 1 5 10 50 100 5001000
Celldeath,%
TRAIL / DR5-B, ng/ml
Jurkat (T cell leukemia)
TRAIL
DR5-B
0
10
20
30
40
50
0 0,1 0,5 1 5 10 50 100 5001000
Celldeath,%
TRAIL / DR5-B, ng/ml
A 549 (lung carcinoma)
TRAIL
DR5-B
0
10
20
30
40
50
60
70
80
0 0,1 0,5 1 5 10 50 100 5001000
Celldeath,%
TRAIL / DR5-B, ng/ml
MCF-7 (breast adenocarcinoma)
TRAIL
DR5-B
DR5-B – DR5-selective mutant variant of cytokine TRAIL
overcomes resistance of cancer cells to TRAIL.
Conclusions:
1.Recombinant preparation of DR5-selective TRAIL mutant variant DR5-B induces
apoptosis of TRAIL sensitive cells more effectively than wild type TRAIL.
2.Utilization of DR5-selective TRAIL mutant variant DR5-B can overcome the
resistance of TRAIL-insensitive colon, breast and lung cancer cells.
3.Chemotherapeutic agents (bortezomib, taxol, and doxorubicin) greatly increase
the efficacy of TRAIL and DR5-B on different TRAIL-sensitive and TRAIL-
insensitive cancer cell lines.
4.TRAIL and DR5-B preparations are non-toxic to normal cells even in
concentrations of 15 µg/ml.
Introduction
Cytokine TRAIL (Apo2L) induces apoptosis by binding to death receptors
DR4 and DR5 in broad spectrum of cancer cells while sparing normal cells (1), so
recombinant TRAIL preparations are considered as potential anti-cancer drugs (2).
However, many tumor cell lines are resistant to TRAIL, despite of DR4 and DR5
expression, partially due to the competition between death and decoy receptors
(membrane-bound DcR1, DcR2 and soluble OPG) for binding to TRAIL (3).
Recently, unique DR5-selective mutant variant of TRAIL (DR5-B) was generated in
our laboratory. DR5-B practically doesn't interact with decoy receptors and DR4
(4).
The aim of this work was to investigate the cytotoxic activity of recombinant
preparations of wild type TRAIL and its DR5-selective mutant variant DR5-B on
TRAIL-sensitive and resistant cancer cell lines, and also to study cytotoxicity of
these preparations on normal human cell lines.
Methods
Surface expression of TRAIL receptors was examined by flow cytometry (Becton
Dickinson), cytotoxicity of recombinant TRAIL and DR5-B preparations was studied
using MTT assay. Apoptotic cell death was confirmed by fluorescent microscopy
(Hoerst 33342 cell nuclei staining).
Thus, DR5 selective mutant variant of TRAIL DR5-B can be used as a powerful tool in therapy of wild type TRAIL resistant cancers.
Maxim L. Bychkov*, Marine E. Gasparian**, Dmitry A. Dolgikh*, Mikhail P. Kirpichnikov* (Moscow, Russia)
TRAIL
variants
KD,10-9 M
DR5 DR4 DcR1 DcR2 OPG
Wild type 0.51 ± 0.028 0.46 ± 0.014 1.09 ± 0.069 0.36 ± 0.009 0.99 ± 0.017
DR5-B 0.71 ± 0.013 NB NB 101 ± 7.0
143 ± 13*
221 ± 19*
Table. 1. Dissociation constants of TRAIL and DR5-B, measured by SPR (acc. to 4).
* Determined by equilibrium plots, NB – not binding
References:
1. Ashkenazi A., Rai R.C., Fong S. et al. (1999). Safety and antitumor
activity of recombinant soluble Apo2 ligand. J. Clin. Invest.; 104: 155
– 162.
2. Micheau O., Shirley S., Dufour F. (2013). Death receptors as targets
in cancer. Br J Pharmacol. 2013 May 3 . [Epub ahead of print].
3. Wajant H, Gerspach J, Pfizenmaier K. 2011. Engineering death
receptor ligands for cancer therapy. Cancer Lett. 2011 Jan 13. [Epub
ahead of print].
4. Gasparian M.E., Chernyak B.V., Dolgikh D.A. et al. (2009).
Generation of new TRAIL mutants DR5A and DR5B with improved
selectivity to death receptor 5. Apoptosis; 14: 778 – 787.
Table 2. Effective concentrations of TRAIL and
DR5-B on Jurkat, U 937 and HCT 116 cells alone
and in co-incubation with chemotherapeutic
agents (24-hour incubation).
Table 3. Effective concentrations of TRAIL and
DR5-B on MCF-7, HT 29 and A 549 cells alone
and in co-incubation with chemotherapeutic
agents (24-hour incubation).
Fig. 5. Viability of MCF-7, HT 29 and
A 549 cell lines after 24-hour
incubation with TRAIL and DR5-B.
Fig. 4. Viability of Jurkat, U 937 and
HCT 116 cell lines after 24-hour
incubation with TRAIL and DR5-B.
Fig. 1. Expression of TRAIL membrane receptors on surface of different cancer cell lines.
0
1
2
3
4
5
6
7
HUVEC HFF
FluorescenceG-mean
DR5
DR4
DcR1
DcR2
0
20
40
60
80
100
120
0
0,1
1
10
100
500
1000
2500
5000
7500
10000
15000
Cellviability,%
TRAIL/DR5-B, ng/ml
Human Foreskin Fibroblasts
TRAIL
DR5-B
0
20
40
60
80
100
120
0
0,1
1
10
100
500
1000
2500
5000
7500
10000
15000
Cellviability,%
TRAIL/DR5-B, ng/ml
HUVEC (Endothelial cells)
TRAIL
DR5-B
Fig. 3. Viability of normal fibroblasts and
endothelium cells after 24-hour incubation
with TRAIL and DR5-B.
Fig. 2. Expression of TRAIL membrane receptors
on surface of normal cell lines.
0
10
20
30
40
50
60
0 0,1 0,5 1 5 10 50 100 5001000
Celldeath,%
TRAIL / DR5-B, ng/ml
U 937 (monoblastic leukemia)
TRAIL
DR5-B
Results
DR5-B induced apoptosis 1.3-10-fold more effectively than wild type TRAIL either
in TRAIL-sensitive or in TRAIL-resistant cell lines which express all TRAIL membrane
receptors. Combinational treatment of cancer cell lines with chemotherapeutic agents
such as doxorubicin, taxol and bortezomib (1-100nM) and TRAIL or DR5-B preparations
resulted in more than 90 % apoptosis of Jurkat, HCT116 and U937 cells. Chemotherapeutic
agents enhanced DR5-induced apoptosis of TRAIL-resistant cell lines A549, HT29 and MCF-
7 much more than wild type TRAIL-induced cell death (maximal cell death 65-98 % and 35-
60 % for DR5-B and wild type TRAIL respectively). Both preparations of DR5-B and wild
type TRAIL were nontoxic to normal fibroblasts and endothelial cells (HUVEC) expressing
all TRAIL receptors.
These results demonstrate that DR5-selective TRAIL mutant variant DR5-B can
be used for therapy of TRAIL-resistant cancers.
1. TRAIL receptors are expressed either on TRAIL-sensitive or on TRAIL-resistant cell
lines.
3. DR5-B is 3-4 fold more active than wild type TRAIL on different cancer cell lines alone, as well as in combination with
chemotherapeutic agents (bortezomib, taxol and doxorubicin).
2. TRAIL and DR5-B preparations
are non-toxic to normal cells,
expressing all membrane TRAIL
receptors.
*Lomonosov Moscow State University. Biological faculty, department of bioengineering. 119234, Moscow, Russia. e-mail: maksim.bychkov@gmail.com
**Federal State Institution of science Institute of bioorganic chemistry RAS. Laboratory of protein engineering. 117997, Moscow, Russia. +7 (929) 910 86 26
*Death after 24-hour incubation with 100 ng/ml of TRAIL or DR5-B *Death after 24-hour incubation with 100 ng/ml of TRAIL or DR5-B