1. Molecular analysis of sporadic and neurofibromatosis 1 (NF1)-associated malignant peripheral nerve sheath tumors Mark A. Watson, Arie Perry, and David H. Gutmann Washington University School of Medicine *Supported by funding from the Department of Defense and Washington University/Siteman Cancer Center

2. MPNST Devastating tumor Poor response to therapy Poor patient survival Patients with neurofibromatosis 1 (NF1) harbor 10% lifetime risk of developing MPNST

3. NF1 loss seen in 76% MPNST by FISH Neurofibromin loss seen in 9/9 MPNST NF1 inactivation is critical event in MPNST pathogenesis

4. p16 deletion in MPNSTs J. Neuropath Exp. Neurol., 2002 Tumorp16 deletion p16 HD Schwannoma 0/5 0/5 Plexiform neurofibroma 0/13 0/13 MPNST 16/20 9/20 (75%) (45%) Synovial sarcoma 5/7 0/7 (71%) Fibrosarcoma 5/6 0/6 (83%) Hemangiopericytoma 8/12 0/12 (67%) CEP9 p16

5. EGFR amplification in MPNSTs J. Neuropath Exp. Neurol., 2002 Tumor EGF-R amplification Schwannoma 0/5 Plexiform neurofibroma 0/13 MPNST 5/19 (26%) Synovial sarcoma 0/7 Fibrosarcoma 0/6 Hemangiopericytoma 0/13 MB DG206 WU33 KL DP 19 157021575628621866968235 511 Benign Neurofibromas Malignant Peripheral Nerve Sheath Tumors EGF-R DG102 CEP7 EGFR

6. Gene expression profiling 25 NF1-associated 17 sporadic tumors Genetic signature to distinguish NF1-associated from sporadic? Genetic signature to predict clinical behavior?

7. SSSSSSSSSsnssSsnsnNnnnNnnnsNNnsNsnnNNnnnNNnNn 4247 4263 4260 4236 4252 4261 4240 4274 4262 4257 4241 4258 4238 4239 4276 4270 4235 4244 4266 4269 4251 4230 4264 4271 4232 4242 4245 4231 4255 4234 4253 4233 4254 4237 4256 4265 4243 4259 4246 4250 4248 4275 4249 4268 4267 SIAT9 MGLL GCM1 SGCE SEC14L1 DDX21 Few, if any, gene expression signatures that distinguish between NF1-associated and sporadic MPNSTs

8. Survival 24 mo. (n=11) LocusLink IDSymbolNameFCP 5068PAPPancreatitis-associated protein-2.00.0005 4209MEF2DMADS box transcription enhancer factor 2, polypeptide D-1.40.0002 8450CUL4BCullin 4B1.70.0001 5532PPP3CBProtein phosphatase 3, catalytic subunit, beta2.00.0001 4664NAB1NGFI-A binding protein 12.20.0009 9122SLC16A4solute carrier family 16, member 43.90.0002

9. Sporadic (n=17) NF1 (n=25) EGFR Expression EGFR(-) EGFR(+)

10. Group A NCAM, MBP, L1CAM, PLP1, BLBP, GAP43, APO-D, RELN IGF2, FGFR1, MDK, CCNB1, CCNB2, CCNF, Ki67 EGFR(-) “more differentiated phenotype”

11. Conclusions 1.NF1 loss is observed in both NF1-associated and sporadic MPNST 2.CDKN2A inactivation is frequently observed in MPNST 3.EGFR amplification is observed in one-third MPNST 4.Expression profiling did not identify a molecular fingerprint that distinguishes NF1-associated from sporadic MPNST 5.Expression profiling identified a molecular fingerprint for a subset of more aggressive MPNSTs 6.Expression profiling identified a molecular fingerprint for a subset of MPNST with a more differentiated phenotype 7.Future prospective studies will be required to determine whether these patterns of gene expression predict clinical behavior