DEFINING NEW THERAPEUTIC AGENTS THAT TARGET THE ONCOGENIC TRANSLATION PROGRAM IN MULTIPLE MYELOMA
(Abstract release date: 05/19/16)
EHA Library. Manier S. 06/11/16; 135205; S449
Dr. Salomon Manier
Contributions
Contributions
Abstract
Abstract: S449
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:00 - 12:15
Location: Auditorium 2
Background
Despite significant therapeutic advances, Multiple Myeloma (MM) remains an incurable hematological malignancy. Therefore, there is a need for the development of new therapeutic options with novel mechanisms of action that have not been previously explored in MM.
Aims
To identify new potential therapies in MM, we screened a library of small molecules against lymphoid cell lines and defined the mode of action for the most active class of hits that we found. These compounds act as potent, highly selective inhibitors of translation initiation and therefore inhibit the oncogenic translation program of MM.
Methods
A chemically diverse small-molecule drug screen of over 3,000 compounds from the Boston University Center for Molecular Discovery was performed using H929 and NAMALWA cell lines. Follow-up validation and structure-activity relationship (SAR) studies were performed in several cell lines including H929, MM1S, MM1R, U266, RPMI and OPM2. RNA sequencing was performed using the NEBNext kit and Illumina HiSeq 2500. Quantitative proteomic analysis was performed by Tandem Mass Tag (TMT) with mass spectrometry (MS). For animal studies, SCID mice were injected with 5x106 MM1S luc-GFP cells and followed by blood cell count, body weight, bioluminescence imaging (BLI) and survival.
Results
A primary screen of over 3,000 small-molecules identified 45 compounds with potent activity on H929 and NAMALWA. Further validation of these hits in 6 cell lines highlighted 3 compounds of the rocaglate class as by far the most effective. A follow-up SAR screen including 40 rocaglate derivatives identified a compound called CMLD10509 with an IC50 below 10nM against several MM cell lines, but no cytotoxicity on human PBMCs, suggesting a useful therapeutic window for this agent in MM. To define the mode of action for CMLD10509 against MM, we performed RNA-seq on drug- vs. DMSO-treated cell lines. By GSEA analysis, the most enriched pathways were regulators of transcriptional activation and translation inhibition. To validate these findings we queried our compound’s signature against the LINCS Cloud database - a large catalog of gene-expression profiles collected from human cells exposed to chemical and genetic perturbations - and identified a positive correlation between our drug signature and translation inhibitors as well as KD of ribosome subunits. To better define the consequences of CMLD10509-induced translation inhibition, we performed a quantitative proteomic MS experiment. We identified 7312 proteins of which 54 were significantly down-regulated by CMLD10509 exposure (p<0.05 and FC>2), among them key oncoproteins in MM such as MYC, MDM2, CCND1, MCL1 and MAF (Fig. 1). Strikingly, down-regulated proteins were strongly associated with KEGG cancer pathways, representing the oncogenic translation program in MM. Correlation between TMT and RNA-seq (confirmed by immunoblot and qRT-PCR) validated a purely translational mechanism as responsible for their depletion in MM cells. Importantly, increased expression of these same rocaglate-sensitive genes was significantly enriched in MM patients in several datasets. Finally, CMLD10509 was both active and well tolerated in a xenograft model of MM (n=10 per group), with significantly lower BLI counts (p<0.001) and prolonged survival (median OS 35 vs. 47 days, p<0.001).
Conclusion
From a large-scale small molecule screen, we identified CMLD10509 as a potent compound, which selectively inhibits the translation initiation and impairs an oncogenic translation program supporting MM cells that includes MYC, MDM2, CCND1 and MCL1. As proof of concept for the potential of CMLD10509 as a novel therapeutic strategy, we found it to be highly active and well tolerated in a mouse model of MM.
Session topic: New biological markers in MM
Keyword(s): MYC, Myeloma, Targeted therapy
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:00 - 12:15
Location: Auditorium 2
Background
Despite significant therapeutic advances, Multiple Myeloma (MM) remains an incurable hematological malignancy. Therefore, there is a need for the development of new therapeutic options with novel mechanisms of action that have not been previously explored in MM.
Aims
To identify new potential therapies in MM, we screened a library of small molecules against lymphoid cell lines and defined the mode of action for the most active class of hits that we found. These compounds act as potent, highly selective inhibitors of translation initiation and therefore inhibit the oncogenic translation program of MM.
Methods
A chemically diverse small-molecule drug screen of over 3,000 compounds from the Boston University Center for Molecular Discovery was performed using H929 and NAMALWA cell lines. Follow-up validation and structure-activity relationship (SAR) studies were performed in several cell lines including H929, MM1S, MM1R, U266, RPMI and OPM2. RNA sequencing was performed using the NEBNext kit and Illumina HiSeq 2500. Quantitative proteomic analysis was performed by Tandem Mass Tag (TMT) with mass spectrometry (MS). For animal studies, SCID mice were injected with 5x106 MM1S luc-GFP cells and followed by blood cell count, body weight, bioluminescence imaging (BLI) and survival.
Results
A primary screen of over 3,000 small-molecules identified 45 compounds with potent activity on H929 and NAMALWA. Further validation of these hits in 6 cell lines highlighted 3 compounds of the rocaglate class as by far the most effective. A follow-up SAR screen including 40 rocaglate derivatives identified a compound called CMLD10509 with an IC50 below 10nM against several MM cell lines, but no cytotoxicity on human PBMCs, suggesting a useful therapeutic window for this agent in MM. To define the mode of action for CMLD10509 against MM, we performed RNA-seq on drug- vs. DMSO-treated cell lines. By GSEA analysis, the most enriched pathways were regulators of transcriptional activation and translation inhibition. To validate these findings we queried our compound’s signature against the LINCS Cloud database - a large catalog of gene-expression profiles collected from human cells exposed to chemical and genetic perturbations - and identified a positive correlation between our drug signature and translation inhibitors as well as KD of ribosome subunits. To better define the consequences of CMLD10509-induced translation inhibition, we performed a quantitative proteomic MS experiment. We identified 7312 proteins of which 54 were significantly down-regulated by CMLD10509 exposure (p<0.05 and FC>2), among them key oncoproteins in MM such as MYC, MDM2, CCND1, MCL1 and MAF (Fig. 1). Strikingly, down-regulated proteins were strongly associated with KEGG cancer pathways, representing the oncogenic translation program in MM. Correlation between TMT and RNA-seq (confirmed by immunoblot and qRT-PCR) validated a purely translational mechanism as responsible for their depletion in MM cells. Importantly, increased expression of these same rocaglate-sensitive genes was significantly enriched in MM patients in several datasets. Finally, CMLD10509 was both active and well tolerated in a xenograft model of MM (n=10 per group), with significantly lower BLI counts (p<0.001) and prolonged survival (median OS 35 vs. 47 days, p<0.001).
Conclusion
From a large-scale small molecule screen, we identified CMLD10509 as a potent compound, which selectively inhibits the translation initiation and impairs an oncogenic translation program supporting MM cells that includes MYC, MDM2, CCND1 and MCL1. As proof of concept for the potential of CMLD10509 as a novel therapeutic strategy, we found it to be highly active and well tolerated in a mouse model of MM.
Session topic: New biological markers in MM
Keyword(s): MYC, Myeloma, Targeted therapy
Abstract: S449
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:00 - 12:15
Location: Auditorium 2
Background
Despite significant therapeutic advances, Multiple Myeloma (MM) remains an incurable hematological malignancy. Therefore, there is a need for the development of new therapeutic options with novel mechanisms of action that have not been previously explored in MM.
Aims
To identify new potential therapies in MM, we screened a library of small molecules against lymphoid cell lines and defined the mode of action for the most active class of hits that we found. These compounds act as potent, highly selective inhibitors of translation initiation and therefore inhibit the oncogenic translation program of MM.
Methods
A chemically diverse small-molecule drug screen of over 3,000 compounds from the Boston University Center for Molecular Discovery was performed using H929 and NAMALWA cell lines. Follow-up validation and structure-activity relationship (SAR) studies were performed in several cell lines including H929, MM1S, MM1R, U266, RPMI and OPM2. RNA sequencing was performed using the NEBNext kit and Illumina HiSeq 2500. Quantitative proteomic analysis was performed by Tandem Mass Tag (TMT) with mass spectrometry (MS). For animal studies, SCID mice were injected with 5x106 MM1S luc-GFP cells and followed by blood cell count, body weight, bioluminescence imaging (BLI) and survival.
Results
A primary screen of over 3,000 small-molecules identified 45 compounds with potent activity on H929 and NAMALWA. Further validation of these hits in 6 cell lines highlighted 3 compounds of the rocaglate class as by far the most effective. A follow-up SAR screen including 40 rocaglate derivatives identified a compound called CMLD10509 with an IC50 below 10nM against several MM cell lines, but no cytotoxicity on human PBMCs, suggesting a useful therapeutic window for this agent in MM. To define the mode of action for CMLD10509 against MM, we performed RNA-seq on drug- vs. DMSO-treated cell lines. By GSEA analysis, the most enriched pathways were regulators of transcriptional activation and translation inhibition. To validate these findings we queried our compound’s signature against the LINCS Cloud database - a large catalog of gene-expression profiles collected from human cells exposed to chemical and genetic perturbations - and identified a positive correlation between our drug signature and translation inhibitors as well as KD of ribosome subunits. To better define the consequences of CMLD10509-induced translation inhibition, we performed a quantitative proteomic MS experiment. We identified 7312 proteins of which 54 were significantly down-regulated by CMLD10509 exposure (p<0.05 and FC>2), among them key oncoproteins in MM such as MYC, MDM2, CCND1, MCL1 and MAF (Fig. 1). Strikingly, down-regulated proteins were strongly associated with KEGG cancer pathways, representing the oncogenic translation program in MM. Correlation between TMT and RNA-seq (confirmed by immunoblot and qRT-PCR) validated a purely translational mechanism as responsible for their depletion in MM cells. Importantly, increased expression of these same rocaglate-sensitive genes was significantly enriched in MM patients in several datasets. Finally, CMLD10509 was both active and well tolerated in a xenograft model of MM (n=10 per group), with significantly lower BLI counts (p<0.001) and prolonged survival (median OS 35 vs. 47 days, p<0.001).
Conclusion
From a large-scale small molecule screen, we identified CMLD10509 as a potent compound, which selectively inhibits the translation initiation and impairs an oncogenic translation program supporting MM cells that includes MYC, MDM2, CCND1 and MCL1. As proof of concept for the potential of CMLD10509 as a novel therapeutic strategy, we found it to be highly active and well tolerated in a mouse model of MM.
Session topic: New biological markers in MM
Keyword(s): MYC, Myeloma, Targeted therapy
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:00 - 12:15
Location: Auditorium 2
Background
Despite significant therapeutic advances, Multiple Myeloma (MM) remains an incurable hematological malignancy. Therefore, there is a need for the development of new therapeutic options with novel mechanisms of action that have not been previously explored in MM.
Aims
To identify new potential therapies in MM, we screened a library of small molecules against lymphoid cell lines and defined the mode of action for the most active class of hits that we found. These compounds act as potent, highly selective inhibitors of translation initiation and therefore inhibit the oncogenic translation program of MM.
Methods
A chemically diverse small-molecule drug screen of over 3,000 compounds from the Boston University Center for Molecular Discovery was performed using H929 and NAMALWA cell lines. Follow-up validation and structure-activity relationship (SAR) studies were performed in several cell lines including H929, MM1S, MM1R, U266, RPMI and OPM2. RNA sequencing was performed using the NEBNext kit and Illumina HiSeq 2500. Quantitative proteomic analysis was performed by Tandem Mass Tag (TMT) with mass spectrometry (MS). For animal studies, SCID mice were injected with 5x106 MM1S luc-GFP cells and followed by blood cell count, body weight, bioluminescence imaging (BLI) and survival.
Results
A primary screen of over 3,000 small-molecules identified 45 compounds with potent activity on H929 and NAMALWA. Further validation of these hits in 6 cell lines highlighted 3 compounds of the rocaglate class as by far the most effective. A follow-up SAR screen including 40 rocaglate derivatives identified a compound called CMLD10509 with an IC50 below 10nM against several MM cell lines, but no cytotoxicity on human PBMCs, suggesting a useful therapeutic window for this agent in MM. To define the mode of action for CMLD10509 against MM, we performed RNA-seq on drug- vs. DMSO-treated cell lines. By GSEA analysis, the most enriched pathways were regulators of transcriptional activation and translation inhibition. To validate these findings we queried our compound’s signature against the LINCS Cloud database - a large catalog of gene-expression profiles collected from human cells exposed to chemical and genetic perturbations - and identified a positive correlation between our drug signature and translation inhibitors as well as KD of ribosome subunits. To better define the consequences of CMLD10509-induced translation inhibition, we performed a quantitative proteomic MS experiment. We identified 7312 proteins of which 54 were significantly down-regulated by CMLD10509 exposure (p<0.05 and FC>2), among them key oncoproteins in MM such as MYC, MDM2, CCND1, MCL1 and MAF (Fig. 1). Strikingly, down-regulated proteins were strongly associated with KEGG cancer pathways, representing the oncogenic translation program in MM. Correlation between TMT and RNA-seq (confirmed by immunoblot and qRT-PCR) validated a purely translational mechanism as responsible for their depletion in MM cells. Importantly, increased expression of these same rocaglate-sensitive genes was significantly enriched in MM patients in several datasets. Finally, CMLD10509 was both active and well tolerated in a xenograft model of MM (n=10 per group), with significantly lower BLI counts (p<0.001) and prolonged survival (median OS 35 vs. 47 days, p<0.001).
Conclusion
From a large-scale small molecule screen, we identified CMLD10509 as a potent compound, which selectively inhibits the translation initiation and impairs an oncogenic translation program supporting MM cells that includes MYC, MDM2, CCND1 and MCL1. As proof of concept for the potential of CMLD10509 as a novel therapeutic strategy, we found it to be highly active and well tolerated in a mouse model of MM.
Session topic: New biological markers in MM
Keyword(s): MYC, Myeloma, Targeted therapy
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