Presentation on theme: "INTEGRATED ANALYSIS OF THE MOLECULAR DEREGULATIONS OF THE MDM2:P53 AXIS IN DDLPS KATE LYNN BILL, JEANNINE GARNETT, CHAD CREIGHTON, XIAOYAN MA, ISABELLE."— Presentation transcript:
INTEGRATED ANALYSIS OF THE MOLECULAR DEREGULATIONS OF THE MDM2:P53 AXIS IN DDLPS KATE LYNN BILL, JEANNINE GARNETT, CHAD CREIGHTON, XIAOYAN MA, ISABELLE MEAUX, LAURENT DEBUSSCHE, THERESA NGUYEN, DAVIS INGRAM, SVETLANA BOLSHAKOV, ROMAN BELOUSOV, DINA LEV, ALEXANDER J. LAZAR, RAPHAEL POLLOCK MD ANDERSON CANCER CENTER, Houston Tx
Anaya et al. Ann Surg 2009; 16(3)667-75 Lahat et al. Ann Surg Oncol 2008; 15(6):1585-1593 WDLPS DDLPS WDLPS DDLPS Local recurrenceDisease specific survival DDLPS: marked propensity for local recurrence and dismal outcome
Disclosures Provided drug (SAR405838) for studies under MTA Sanofi reviewed this presentation – no significant changes Referred to as “inhibitor” in this talk
MDM2/p53 axis Wang, S. et al., Pharm Taxicol, 2009.
MDM2: potential target for cancer therapy Small molecule inhibitors: Nutlin RG7112 SAR405838 Wang, S. et al., Pharm Taxicol, 2009.
MDM2 Inhibitor: SAR405838 Orally bioavailable small molecule Single-digit nM binding to hMDM2, high selectivity and specificity Blocks interaction of MDM2 & P53 Inactive in p53 mut cell lines Phase I initiated mid-2012 Safety and efficacy Biomarker development MW: 562.51 (Formerly: SAR299155 or MI-77301)
MDM2 Amplification and Overexpression in DDLPS PA A 224 246 863 141 815 SW872 PLS-1 DDLPSS MDM2 β -actin gDNARNA * * * *
Systematic characterization of DDLPS cell lines 4 DDLPS cell lines were treated with SAR405838 Proteomics mRNA miRNA DNA copy number variation* DNA Sequencing* *Previously reported data
Experimental Outline two independent biological replicates 224 815246863 cell lines were either treated with 0.003% DMSO, 1 µM or 3 µM SAR405838 (24hrs) Analysis for differential gene expression/miRNA array
>2 fold upregulated or downregulated genes in at least 3 cell lines Top Deregulated Pathways: Ingenuity Pathway Analysis P53 signaling / Cell cycle / DNA damage
Upregulated mRNA in DDLPS Downregulated by the MDM2:p53 Axis UPREGULATED TRANSCRIPTS IN DEDIFFERENTIATED LIPOSARCOMA CELLS 523 DOWNREGULATED TRANSCRIPTS WITH MDM2i 238 183 ~35% of transcripts that are upregulated in DDLPS when compared with normal controls are downregulated in response to MDM2 inhibition These data suggest that a 1/3 of all upregulated transcripts in DDLPS are regulated by the MDM2:p53 axis ~35%
Downregulated mRNA in DDLPS Upregulated by the MDM2:p53 Axis UPREGULATED TRANSCRIPTS WITH MDM2i 234 DOWNREGULATED TRANSCRIPTS IN DEDIFFERENTIATED LIPOSARCOMA CELLS 411 19 ~5% of transcripts that are downregulated in DDLPS when compared with normal controls are upregulated in response to MDM2 inhibition These data suggest that a small number of transcripts that are downregulated in DDLPS are regulated in a MDM2/p53 axis-dependent manner ~5%
p53 and Mitotic Pathway Connections SAR405838 SAR405838 uM 0 0.1 0.3 1 3 10 Aurora A PTTG Actin
Proposed signaling pathway for PTTG1 action at the G2 DNA damage checkpoint DNA Damage PTTG1 p53 Aurora A G2 M Cytoplasm Nucleus
Top Canonical Pathways Potentially Affected by miRNAs in DDLPS
Potential miRNA Regulation of mRNA with SAR405838: ABI SOliD platform Top 18 miRNAs differentially expressed in 3 cell lines vs controls MaxA > 50 P < 0.05 FC > 2 Top 472 genes differentially expressed genes in 4 DDLPS cell lines treated with SAR405838 vs DMSO controls FC > 2 P < 0.05 ~20%
Top Molecular Pathways Affected by miRNAs in DDLPS 1.Cell Cycle: G2/M DNA Damage Checkpoint 2.p53 Signaling 3.Mitotic Roles of Polo-Like Kinase 1
Summary Gene expression arrays identified multiple targets / pathways of potential interest Identify on and off effects, regulatory loops Studies focus on validating specific pathway/targets
Acknowledgements Dina Lev Raphael E. Pollock Jeannine Garnett Chad Creighton David Pollock Svetlana Bolshakov XiaoYan Ma Davis Ingram Roman Belousov Theresa Nguyen Gonzalo Lopez Danielle Braggio Sanofi-Aventis Isabelle Meaux Laurent Debussche Cedric Barriere Amschwand Sarcoma Cancer Foundation
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