COMBINED ANTICANCER ACTIVITY AND SIRNA DELIVERY BY DENGUE VIRUS CAPSID PROTEIN PEPTIDES PEPR AND PEPM AGAINST CHRONIC MYELOID LEUKEMIA
(Abstract release date: 05/19/16)
EHA Library. Figueiredo I. 06/09/16; 134710; PB1810
Dr. Ines Figueiredo
Contributions
Contributions
Abstract
Abstract: PB1810
Type: Publication Only
Background
Cancer remains a major cause of morbidity and mortality worldwide. Although progress has been made regarding chemotherapeutic agents, new therapies that combine increased selectivity and efficacy with low resistance are still needed. In the search for new anticancer agents, therapies based on biologically active peptides, in particular, antimicrobial peptides (AMPs), have attracted attention for their decreased resistance development and low cytotoxicity. AMPs, which have been essentially studied and developed as potential alternatives for fighting infectious diseases, have been tested as anticancer peptides (ACPs) in cancer therapy either alone or in combination with other conventional drugs. Moreover, the usage of cell-penetrating peptides (CPP) to deliver anticancer drugs, such as siRNA targeting molecular causes of cancer are currently being developed.
Aims
Chronic Myeloid Leukemia (CML) is a myeloproliferative disorder caused by a single genetic mutation, a reciprocal translocation that originates the BCR-ABL gene with constitutive tyrosine kinase activity. Because there is a specific gene associated to this pathology it is an optimum therapeutic target for RNA silencing therapy (siRNA).Thus we developed a siRNA-based therapeutic approach in which the siRNA is delivered by Dengue Virus Capsid Protein-derived peptides, pepM and pepR. These peptides were used with the intention of having have dual role, to deliver siRNA into cells and act as Anticancer molecules by targeting intracellular cancer signaling events.
Methods
Dengue virus-peptides ability to transfect the positive BCR-ABL+ Cell Line (BV-173) was evaluated by confocal microscopy following GFP fluorescence emission after plasmid expression. Anti-BCR-ABL siRNA design was performed using a siRNA design web-tool and the BCR-ABL downregulation kinetics (48h to 168h) after transfection by Dengue virus-peptides was evaluated by RT-PCR. The Anticancer action of either pepR or pepM was assessed by genome-wide analysis microarray and further validated by testing BV 173 cell cycle and cell proliferation analysis by RT-PCR.
Results
siRNA design for BCR-ABL retrieved 148 potential siRNA sequences, which were reduced to 5 BCR-ABL siRNA for in vitro analysis after thorough screening. Positive efficacy of siRNA targeting BCR-ABL was tested using a commercial transfection agent. Significant BCR-ABL gene knockdown were observed for siRNA #3 when delivered by pepM with maximum decrease at 120h. Both pepM and pepR showed downregulation effects on proliferative CML cancer-related signaling pathways having direct impact on BV 173 cell cycle and proliferation at the G2/M phase.
Conclusion
With this work we showed the potential therapeutic technology of combining a drug delivery system (CPP) with anticancer properties to deliver functional siRNA into CML cell model. Acting together, these conjugates significantly decreased BCR-ABL gene expression levels, and perturbed leukemogenic cells homeostasis, revealing a potential scaffold to develop an alternative CML therapy. The development of a selective ACP is still a current challenge. It is not straightforward the prediction and design peptides with antitumor activity and we believe that these results will enrich ACP-structure based prediction.
Session topic: E-poster
Keyword(s): Chronic myeloid leukemia, SiRNA
Type: Publication Only
Background
Cancer remains a major cause of morbidity and mortality worldwide. Although progress has been made regarding chemotherapeutic agents, new therapies that combine increased selectivity and efficacy with low resistance are still needed. In the search for new anticancer agents, therapies based on biologically active peptides, in particular, antimicrobial peptides (AMPs), have attracted attention for their decreased resistance development and low cytotoxicity. AMPs, which have been essentially studied and developed as potential alternatives for fighting infectious diseases, have been tested as anticancer peptides (ACPs) in cancer therapy either alone or in combination with other conventional drugs. Moreover, the usage of cell-penetrating peptides (CPP) to deliver anticancer drugs, such as siRNA targeting molecular causes of cancer are currently being developed.
Aims
Chronic Myeloid Leukemia (CML) is a myeloproliferative disorder caused by a single genetic mutation, a reciprocal translocation that originates the BCR-ABL gene with constitutive tyrosine kinase activity. Because there is a specific gene associated to this pathology it is an optimum therapeutic target for RNA silencing therapy (siRNA).Thus we developed a siRNA-based therapeutic approach in which the siRNA is delivered by Dengue Virus Capsid Protein-derived peptides, pepM and pepR. These peptides were used with the intention of having have dual role, to deliver siRNA into cells and act as Anticancer molecules by targeting intracellular cancer signaling events.
Methods
Dengue virus-peptides ability to transfect the positive BCR-ABL+ Cell Line (BV-173) was evaluated by confocal microscopy following GFP fluorescence emission after plasmid expression. Anti-BCR-ABL siRNA design was performed using a siRNA design web-tool and the BCR-ABL downregulation kinetics (48h to 168h) after transfection by Dengue virus-peptides was evaluated by RT-PCR. The Anticancer action of either pepR or pepM was assessed by genome-wide analysis microarray and further validated by testing BV 173 cell cycle and cell proliferation analysis by RT-PCR.
Results
siRNA design for BCR-ABL retrieved 148 potential siRNA sequences, which were reduced to 5 BCR-ABL siRNA for in vitro analysis after thorough screening. Positive efficacy of siRNA targeting BCR-ABL was tested using a commercial transfection agent. Significant BCR-ABL gene knockdown were observed for siRNA #3 when delivered by pepM with maximum decrease at 120h. Both pepM and pepR showed downregulation effects on proliferative CML cancer-related signaling pathways having direct impact on BV 173 cell cycle and proliferation at the G2/M phase.
Conclusion
With this work we showed the potential therapeutic technology of combining a drug delivery system (CPP) with anticancer properties to deliver functional siRNA into CML cell model. Acting together, these conjugates significantly decreased BCR-ABL gene expression levels, and perturbed leukemogenic cells homeostasis, revealing a potential scaffold to develop an alternative CML therapy. The development of a selective ACP is still a current challenge. It is not straightforward the prediction and design peptides with antitumor activity and we believe that these results will enrich ACP-structure based prediction.
Session topic: E-poster
Keyword(s): Chronic myeloid leukemia, SiRNA
Abstract: PB1810
Type: Publication Only
Background
Cancer remains a major cause of morbidity and mortality worldwide. Although progress has been made regarding chemotherapeutic agents, new therapies that combine increased selectivity and efficacy with low resistance are still needed. In the search for new anticancer agents, therapies based on biologically active peptides, in particular, antimicrobial peptides (AMPs), have attracted attention for their decreased resistance development and low cytotoxicity. AMPs, which have been essentially studied and developed as potential alternatives for fighting infectious diseases, have been tested as anticancer peptides (ACPs) in cancer therapy either alone or in combination with other conventional drugs. Moreover, the usage of cell-penetrating peptides (CPP) to deliver anticancer drugs, such as siRNA targeting molecular causes of cancer are currently being developed.
Aims
Chronic Myeloid Leukemia (CML) is a myeloproliferative disorder caused by a single genetic mutation, a reciprocal translocation that originates the BCR-ABL gene with constitutive tyrosine kinase activity. Because there is a specific gene associated to this pathology it is an optimum therapeutic target for RNA silencing therapy (siRNA).Thus we developed a siRNA-based therapeutic approach in which the siRNA is delivered by Dengue Virus Capsid Protein-derived peptides, pepM and pepR. These peptides were used with the intention of having have dual role, to deliver siRNA into cells and act as Anticancer molecules by targeting intracellular cancer signaling events.
Methods
Dengue virus-peptides ability to transfect the positive BCR-ABL+ Cell Line (BV-173) was evaluated by confocal microscopy following GFP fluorescence emission after plasmid expression. Anti-BCR-ABL siRNA design was performed using a siRNA design web-tool and the BCR-ABL downregulation kinetics (48h to 168h) after transfection by Dengue virus-peptides was evaluated by RT-PCR. The Anticancer action of either pepR or pepM was assessed by genome-wide analysis microarray and further validated by testing BV 173 cell cycle and cell proliferation analysis by RT-PCR.
Results
siRNA design for BCR-ABL retrieved 148 potential siRNA sequences, which were reduced to 5 BCR-ABL siRNA for in vitro analysis after thorough screening. Positive efficacy of siRNA targeting BCR-ABL was tested using a commercial transfection agent. Significant BCR-ABL gene knockdown were observed for siRNA #3 when delivered by pepM with maximum decrease at 120h. Both pepM and pepR showed downregulation effects on proliferative CML cancer-related signaling pathways having direct impact on BV 173 cell cycle and proliferation at the G2/M phase.
Conclusion
With this work we showed the potential therapeutic technology of combining a drug delivery system (CPP) with anticancer properties to deliver functional siRNA into CML cell model. Acting together, these conjugates significantly decreased BCR-ABL gene expression levels, and perturbed leukemogenic cells homeostasis, revealing a potential scaffold to develop an alternative CML therapy. The development of a selective ACP is still a current challenge. It is not straightforward the prediction and design peptides with antitumor activity and we believe that these results will enrich ACP-structure based prediction.
Session topic: E-poster
Keyword(s): Chronic myeloid leukemia, SiRNA
Type: Publication Only
Background
Cancer remains a major cause of morbidity and mortality worldwide. Although progress has been made regarding chemotherapeutic agents, new therapies that combine increased selectivity and efficacy with low resistance are still needed. In the search for new anticancer agents, therapies based on biologically active peptides, in particular, antimicrobial peptides (AMPs), have attracted attention for their decreased resistance development and low cytotoxicity. AMPs, which have been essentially studied and developed as potential alternatives for fighting infectious diseases, have been tested as anticancer peptides (ACPs) in cancer therapy either alone or in combination with other conventional drugs. Moreover, the usage of cell-penetrating peptides (CPP) to deliver anticancer drugs, such as siRNA targeting molecular causes of cancer are currently being developed.
Aims
Chronic Myeloid Leukemia (CML) is a myeloproliferative disorder caused by a single genetic mutation, a reciprocal translocation that originates the BCR-ABL gene with constitutive tyrosine kinase activity. Because there is a specific gene associated to this pathology it is an optimum therapeutic target for RNA silencing therapy (siRNA).Thus we developed a siRNA-based therapeutic approach in which the siRNA is delivered by Dengue Virus Capsid Protein-derived peptides, pepM and pepR. These peptides were used with the intention of having have dual role, to deliver siRNA into cells and act as Anticancer molecules by targeting intracellular cancer signaling events.
Methods
Dengue virus-peptides ability to transfect the positive BCR-ABL+ Cell Line (BV-173) was evaluated by confocal microscopy following GFP fluorescence emission after plasmid expression. Anti-BCR-ABL siRNA design was performed using a siRNA design web-tool and the BCR-ABL downregulation kinetics (48h to 168h) after transfection by Dengue virus-peptides was evaluated by RT-PCR. The Anticancer action of either pepR or pepM was assessed by genome-wide analysis microarray and further validated by testing BV 173 cell cycle and cell proliferation analysis by RT-PCR.
Results
siRNA design for BCR-ABL retrieved 148 potential siRNA sequences, which were reduced to 5 BCR-ABL siRNA for in vitro analysis after thorough screening. Positive efficacy of siRNA targeting BCR-ABL was tested using a commercial transfection agent. Significant BCR-ABL gene knockdown were observed for siRNA #3 when delivered by pepM with maximum decrease at 120h. Both pepM and pepR showed downregulation effects on proliferative CML cancer-related signaling pathways having direct impact on BV 173 cell cycle and proliferation at the G2/M phase.
Conclusion
With this work we showed the potential therapeutic technology of combining a drug delivery system (CPP) with anticancer properties to deliver functional siRNA into CML cell model. Acting together, these conjugates significantly decreased BCR-ABL gene expression levels, and perturbed leukemogenic cells homeostasis, revealing a potential scaffold to develop an alternative CML therapy. The development of a selective ACP is still a current challenge. It is not straightforward the prediction and design peptides with antitumor activity and we believe that these results will enrich ACP-structure based prediction.
Session topic: E-poster
Keyword(s): Chronic myeloid leukemia, SiRNA
{{ help_message }}
{{filter}}