Poster Discussion 2 - Biology Genome Transcriptome Proteome Metastasome

Genome 39267: HDAC inhibitor/multidrug resistant cell lines –PCI induces acetylation, inhibits growth, enhances chemosensitivity 39309: EXT/osteochondromas –Biallelic inactivation of either EXT1 or EXT2 is less than expected for classic tumor suppressor model 39356: DNA demethylation/synovial sarc –VPA and Zebularine (Zeb) are synergistic for suppression of growth of synovial sarcoma SYO-1 cells

Genome 39393: CNV (SNP)/osteosarcoma –Recurrent amplification: 1p36, 12q13-15, 17p11.2, 20p12 –Recurrent deletion: 1q31, 6q16, 8q24.13, 10q23, 17p13 –New targets to look for contribution to tumorigenesis 39448*: Sleeping Beauty/osteosarcoma - forward genetic screen for new genomic loci

Osteosarcoma Development and Metastasis Gene Discovery Conditional sleeping beauty osteosarcoma New loci that when interrupted by transposon (common insertion sites CIS) activates oncogene or inactivates tumor suppressor to contribute to osteosarcoma formation or metastasis. New gene p53R270H/+ B Moriartity and D Largaespada Osteoblast precursor (Osx)-Cre recombinase T2/onc concatomer Non-directed Screen for New Pathways

Transcriptome 39218: DAXX mRNA/CASP8/p38 MAPK/sarcoma –82 primary sarcomas, SW1353 chondrosarcoma line –FAS-mediated apoptosis via FADD/CASP8 or DAXX/ASK1 –EGF-related association between CASP8 and DAXX may involve p38 MAPK pathway 39338: GGAA/EWS-FLI regulation –NR0B1 (high in ES) promoter with ggaa repeats –Transcription by EWS/FLI requires dual binding of FLI 39473: miRNA profiles/imatinib-treated GIST –GIST cell lines treated with imatinib –10x upregulation of 23 microRNA –Check out a number of them from chromosome 14q *: miRNA profiles/liposarcoma

Identification of differentially expressed microRNAs in liposarcoma subtypes 12 liposarcomas 14 well-differenttiated 17 de-differentiated 9 myxoid 5 round cell 4 pleiomorphic Lipomas, fat - 7/10 microRNAs Hierarchal clustering Myxoid Round cell De-differentiated Differential Expression miRNAs Discriminatory miRNAs unsupervised supervised - 62 Which ones? - talk with Caroline Caroline Gits et al

Cell Biology 38426: PRP-1/chondrosarcoma –PRP-1 isolated from neurosecretory cells of the hypothalamus; associated with myc expression leading to antiproliferation –Specific therapeutic window 39388: Anti-CD99 antibody/Ewings –CD99 membrane glycocylated protein that when stimulated leads to reactivation of p53 pathway –Effective in sarcomas that have intact TP53

Metastasome 39270: expression profiles/osteosarcoma –63 primary osteosarcoma patients (+/- mets) –Patterns predictive of metastatic tumors –RhoA pathway may be best discriminator 39312: EMMPRIN/osteosarcoma

Emmprin expression is associated with metastatic phenotype in osteosarcoma - 16 osteosarcoma cell lines versus normal osteoblast - RT-PCR - IHC in 52 prechemotherapy patient biopsies - siRNA knockdown of Emmprin in HOS and SaOS-2 - Gelatin zymography, VEGF ELISA, Matrigel invasion assay (HOS) - Co-culture SaOS-2 with osteoblast (hFOB) and fibroblast (MRC-5) MMPsVEGF (Cancer cells and stroma) EMMPRIN - Extracellular matrix metalloproteinase inducer EMMPRIN H Han et al. Knock down EMMPRIN?

Emmprin expression is associated with metastatic phenotype in osteosarcoma - 16 osteosarcoma cell lines versus normal osteoblast - RT-PCR - IHC in 52 prechemotherapy patient biopsies - siRNA knockdown of Emmprin in HOS and SaOS-2 - Gelatin zymography, VEGF ELISA, Matrigel invasion assay (HOS) - Co-culture SaOS-2 with osteoblast (hFOB) and fibroblast (MRC-5) MMPsVEGF (Cancer cells and stroma) EMMPRIN - Extracellular matrix metalloproteinase inducer EMMPRIN H Han et al. Knock down EMMPRIN? Make things better!