ELLIPTICINES DRIVE LEUKAEMIA CELL DEATH VIA MITOCHONDRIAL ROS
(Abstract release date: 05/21/15)
EHA Library. Russell E. 06/12/15; 102696; PB1649
Disclosure(s): University College Cork
Eileen Russell
Contributions
Contributions
Abstract
Abstract: PB1649
Type: Publication Only
Background
Ellipticine is an anticancer agent which induces cell death by multiple mechanisms of action1. Unfortunately, this compound and its analogues have limited solubility and have been associated with several toxic effects which has hampered their clinical development. A panel of novel ellipticine isomers was designed and synthesised with the aim of evaluating their cytotoxic effects2. A preliminary NCI 60-cell screen demonstrated that these compounds display promising anti-tumour activity across a number of different cancer cell types, particularly leukaemia cell lines.
Aims
We examined the effect of these derivatives in detail on the acute myeloid leukaemia (AML) cell line, MV4-11. The most cytotoxic compound, 7-formyl-10-methylisoellipticine was identified from this panel. The mechanism by which this compound induces its cytotoxic effects was investigated.
Methods
Cell cycle was monitored by flow cytometry. Cell number and viability was measured by dye exclusion. Confocal microscopy was used to visualise compound accumulation. Protein levels were evaluated by Western blotting. Reactive oxygen species (ROS) levels were measured by flow cytometry using the ROS probes dihydroethidium and MitoSOX.
Results
Cell cycle analyses revealed that the compounds had a range of distinctive cell cycle effects3. 7-formyl-10-methylisoellipticine showed the most promise with respect to cytotoxic activity. We demonstrated that this compound killed cells by apoptosis which is mediated by an induction of reactive oxygen species (ROS). In a cell, mitochondria and NAPDH oxidase (NOX) are the major sources of ROS. Previous research in our laboratory has shown that NOX derived ROS is critical for cell survival4. ROS inhibitors were employed to reveal the origin of the ROS. Our study suggests that the compound generates ROS in mitochondria which ultimately leads to its cytotoxic effect. Interestingly synergistic effects are seen when 7-formyl-10-methylisoellipticine is used in combination with a NOX inhibitor as well as the clinically used AML chemotherapeutic, daunorubicin.
Summary
This study provides detailed cellular information on the potential use of isoellipticines as chemotherapeutic agents. By probing the mechanism of action of this novel compound class we have uncovered a potential clinical application in the field of adjuvant therapy. We are currently investigating the effects of the compound in a pre-clinical AML mouse model.
Keyword(s): Acute myeloid leukemia, Chemotherapy, Cytotoxicity, Reactive oxygen species
Type: Publication Only
Background
Ellipticine is an anticancer agent which induces cell death by multiple mechanisms of action1. Unfortunately, this compound and its analogues have limited solubility and have been associated with several toxic effects which has hampered their clinical development. A panel of novel ellipticine isomers was designed and synthesised with the aim of evaluating their cytotoxic effects2. A preliminary NCI 60-cell screen demonstrated that these compounds display promising anti-tumour activity across a number of different cancer cell types, particularly leukaemia cell lines.
Aims
We examined the effect of these derivatives in detail on the acute myeloid leukaemia (AML) cell line, MV4-11. The most cytotoxic compound, 7-formyl-10-methylisoellipticine was identified from this panel. The mechanism by which this compound induces its cytotoxic effects was investigated.
Methods
Cell cycle was monitored by flow cytometry. Cell number and viability was measured by dye exclusion. Confocal microscopy was used to visualise compound accumulation. Protein levels were evaluated by Western blotting. Reactive oxygen species (ROS) levels were measured by flow cytometry using the ROS probes dihydroethidium and MitoSOX.
Results
Cell cycle analyses revealed that the compounds had a range of distinctive cell cycle effects3. 7-formyl-10-methylisoellipticine showed the most promise with respect to cytotoxic activity. We demonstrated that this compound killed cells by apoptosis which is mediated by an induction of reactive oxygen species (ROS). In a cell, mitochondria and NAPDH oxidase (NOX) are the major sources of ROS. Previous research in our laboratory has shown that NOX derived ROS is critical for cell survival4. ROS inhibitors were employed to reveal the origin of the ROS. Our study suggests that the compound generates ROS in mitochondria which ultimately leads to its cytotoxic effect. Interestingly synergistic effects are seen when 7-formyl-10-methylisoellipticine is used in combination with a NOX inhibitor as well as the clinically used AML chemotherapeutic, daunorubicin.
Summary
This study provides detailed cellular information on the potential use of isoellipticines as chemotherapeutic agents. By probing the mechanism of action of this novel compound class we have uncovered a potential clinical application in the field of adjuvant therapy. We are currently investigating the effects of the compound in a pre-clinical AML mouse model.
References
1Auclair C (1987) Multimodal action of antitumor agents on DNA: the ellipticine series. Arch Biochem Biophys. Nov 15; 259(1):1-14
2Miller CM, O’Sullivan EC, Devine KJ, McCarthy FO (2012) Synthesis and biological evaluation of novel isoellipticine derivatives and salts. Org Biomol Chem. 10(39):7912-21
3Russell EG, O’Sullivan EC, Miller CM, Stanicka J, McCarthy FO, Cotter TG (2014) Ellipticine derivative induces potent cytostatic effect in acute myeloid leukaemia cells. Investigational New Drugs 32 (6):1113-1122
4Naughton R, Quincey C, Turner SD, Cotter TG (2009) Bcr-Abl-mediated redox regulation of the PI3K/AKT pathway. Leukemia Aug;23(8):1432-40
Keyword(s): Acute myeloid leukemia, Chemotherapy, Cytotoxicity, Reactive oxygen species
Abstract: PB1649
Type: Publication Only
Background
Ellipticine is an anticancer agent which induces cell death by multiple mechanisms of action1. Unfortunately, this compound and its analogues have limited solubility and have been associated with several toxic effects which has hampered their clinical development. A panel of novel ellipticine isomers was designed and synthesised with the aim of evaluating their cytotoxic effects2. A preliminary NCI 60-cell screen demonstrated that these compounds display promising anti-tumour activity across a number of different cancer cell types, particularly leukaemia cell lines.
Aims
We examined the effect of these derivatives in detail on the acute myeloid leukaemia (AML) cell line, MV4-11. The most cytotoxic compound, 7-formyl-10-methylisoellipticine was identified from this panel. The mechanism by which this compound induces its cytotoxic effects was investigated.
Methods
Cell cycle was monitored by flow cytometry. Cell number and viability was measured by dye exclusion. Confocal microscopy was used to visualise compound accumulation. Protein levels were evaluated by Western blotting. Reactive oxygen species (ROS) levels were measured by flow cytometry using the ROS probes dihydroethidium and MitoSOX.
Results
Cell cycle analyses revealed that the compounds had a range of distinctive cell cycle effects3. 7-formyl-10-methylisoellipticine showed the most promise with respect to cytotoxic activity. We demonstrated that this compound killed cells by apoptosis which is mediated by an induction of reactive oxygen species (ROS). In a cell, mitochondria and NAPDH oxidase (NOX) are the major sources of ROS. Previous research in our laboratory has shown that NOX derived ROS is critical for cell survival4. ROS inhibitors were employed to reveal the origin of the ROS. Our study suggests that the compound generates ROS in mitochondria which ultimately leads to its cytotoxic effect. Interestingly synergistic effects are seen when 7-formyl-10-methylisoellipticine is used in combination with a NOX inhibitor as well as the clinically used AML chemotherapeutic, daunorubicin.
Summary
This study provides detailed cellular information on the potential use of isoellipticines as chemotherapeutic agents. By probing the mechanism of action of this novel compound class we have uncovered a potential clinical application in the field of adjuvant therapy. We are currently investigating the effects of the compound in a pre-clinical AML mouse model.
Keyword(s): Acute myeloid leukemia, Chemotherapy, Cytotoxicity, Reactive oxygen species
Type: Publication Only
Background
Ellipticine is an anticancer agent which induces cell death by multiple mechanisms of action1. Unfortunately, this compound and its analogues have limited solubility and have been associated with several toxic effects which has hampered their clinical development. A panel of novel ellipticine isomers was designed and synthesised with the aim of evaluating their cytotoxic effects2. A preliminary NCI 60-cell screen demonstrated that these compounds display promising anti-tumour activity across a number of different cancer cell types, particularly leukaemia cell lines.
Aims
We examined the effect of these derivatives in detail on the acute myeloid leukaemia (AML) cell line, MV4-11. The most cytotoxic compound, 7-formyl-10-methylisoellipticine was identified from this panel. The mechanism by which this compound induces its cytotoxic effects was investigated.
Methods
Cell cycle was monitored by flow cytometry. Cell number and viability was measured by dye exclusion. Confocal microscopy was used to visualise compound accumulation. Protein levels were evaluated by Western blotting. Reactive oxygen species (ROS) levels were measured by flow cytometry using the ROS probes dihydroethidium and MitoSOX.
Results
Cell cycle analyses revealed that the compounds had a range of distinctive cell cycle effects3. 7-formyl-10-methylisoellipticine showed the most promise with respect to cytotoxic activity. We demonstrated that this compound killed cells by apoptosis which is mediated by an induction of reactive oxygen species (ROS). In a cell, mitochondria and NAPDH oxidase (NOX) are the major sources of ROS. Previous research in our laboratory has shown that NOX derived ROS is critical for cell survival4. ROS inhibitors were employed to reveal the origin of the ROS. Our study suggests that the compound generates ROS in mitochondria which ultimately leads to its cytotoxic effect. Interestingly synergistic effects are seen when 7-formyl-10-methylisoellipticine is used in combination with a NOX inhibitor as well as the clinically used AML chemotherapeutic, daunorubicin.
Summary
This study provides detailed cellular information on the potential use of isoellipticines as chemotherapeutic agents. By probing the mechanism of action of this novel compound class we have uncovered a potential clinical application in the field of adjuvant therapy. We are currently investigating the effects of the compound in a pre-clinical AML mouse model.
References
1Auclair C (1987) Multimodal action of antitumor agents on DNA: the ellipticine series. Arch Biochem Biophys. Nov 15; 259(1):1-14
2Miller CM, O’Sullivan EC, Devine KJ, McCarthy FO (2012) Synthesis and biological evaluation of novel isoellipticine derivatives and salts. Org Biomol Chem. 10(39):7912-21
3Russell EG, O’Sullivan EC, Miller CM, Stanicka J, McCarthy FO, Cotter TG (2014) Ellipticine derivative induces potent cytostatic effect in acute myeloid leukaemia cells. Investigational New Drugs 32 (6):1113-1122
4Naughton R, Quincey C, Turner SD, Cotter TG (2009) Bcr-Abl-mediated redox regulation of the PI3K/AKT pathway. Leukemia Aug;23(8):1432-40
Keyword(s): Acute myeloid leukemia, Chemotherapy, Cytotoxicity, Reactive oxygen species
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