HIGH-THROUGHPUT MUTATION PROFILING OF OSTEOSARCOMA PRIMARY TUMORS AND CELL LINES IDENTIFIES NEW MUTATIONS IN PREVIOUSLY.


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HIGH-THROUGHPUT MUTATION PROFILING OF OSTEOSARCOMA PRIMARY TUMORS AND CELL LINES IDENTIFIES NEW MUTATIONS IN PREVIOUSLY UNASSOCIATED ONCOGENES AND TUMOR SUPRESSOR GENES . Francis Hornicek, Laura MacConail, Levi Garraway, David Harmon,, Zhenfeng Duan. Edwin Choy MD PhD. Disclosures. none.
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HIGH-THROUGHPUT MUTATION PROFILING OF OSTEOSARCOMA PRIMARY TUMORS AND CELL LINES IDENTIFIES NEW MUTATIONS IN PREVIOUSLY UNASSOCIATED ONCOGENES AND TUMOR SUPRESSOR GENES Francis Hornicek, Laura MacConail, Levi Garraway, David Harmon,, Zhenfeng Duan Edwin Choy MD PhD

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Disclosures none

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Arndt and Crist 341 (5): 342,  July 29, 1999

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McDermott and Settleman, JCO, 2009

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Gene Amplification Point transformation Translocation

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What we did We composed groundworks and genotyping examines utilizing MALDI-TOF (network helped laser desorption/ionization-time of flight mass spectrometer; Sequonom iPLEX Genotyping) Systematically portray 1057 changes in 108 qualities crosswise over 100 osteosarcoma tumor tests and cell lines.

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What we didn\'t do Whole quality sequencing Copy number investigation RNA expression microRNA sequencing Translocation examination SNP investigation Germ-line transformation testing

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Tarkkanen, Canc Res 1995

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Results 108 qualities were dissected utilizing iPLEX Genotyping. At first watched 19 changes in 11 qualities in no less than one osteosarcoma test To approve these outcomes, we retested every one of the 19 transformations utilizing hME Genotyping affirmed 15 changes in 8 qualities

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Genes Analyzed PTPN11 IGF1R NOTCH1 PTEN RB1 SRC EPHA1 NF1 CEBPA CTNNB1 C-MYC FLT3 GATA1 TP53 VHL ALK LRP1B EPHA3 TFDP1 PDPK1 STK11 MINK1 AKT ABL1 ADAMTSL3 AML1/RUNX1 APC CDKN2A ABL1 BRAF CDK4 EGFR (166 transformations) ERBB2 ERB4 FGFR HRAS NRAS KRAS JAK2 KIT PDGFRA PIK3CA RET

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Assay: OM_v2_1121 Samples: 50, 76 Gene: CDH1 Mutation: A617T

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Results We recognized changes in qualities beforehand known to be adjusted in osteosarcomas: p53 (R273H, Y163C, R273C, and Y163C) RB1 (E137).

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Results We didn\'t discover accepted changes in: EGFR (166 transformations tried) PDGF KIT BRAF ALK MET RAS

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Results We likewise distinguished 7 changes in 5 qualities already unimplicated in the pathogenesis of osteosarcomas: PIK3CA (H1047R, E545K, and H701P) KRas (G12S) CUBN (I3189V) CDH1/E-cadherin (A617T) CTNNB1/B-catenin (N287S)

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Future Directions Complete quality sequencing of each of the five novel qualities. Planned genotyping for above novel transformations to better decide recurrence Explore viability of wnt, PI-3 Kinase, and farnesyltransferase inhibitors in osteosarcoma cell lines. Encourage portray osteosarcoma tests for downstream flagging components, i.e. phosphorylated S6, TCF responsive components, MAPKinase initiation, and so on

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Acknowledgments Broad Institute of MIT and Harvard Eric Lander David Altshuler Todd Golub MGH Sarcoma Molecular Biology Lab Zhenfeng Duan Francis Hornicek Dana Farber Cancer Institute Levi Garraway Laura MacConail Jennifer Hunter Yates Foundation David Harmon

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Edwin Choy echoy@partners.org

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