Cos è EGFR? Epidermal Growth Factor Receptor. EGFR e il cancro. EGFR e il cancro. EGFR e il cancro. EGFR e il cancro

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1 Formazione interna Cos è EGFR? Epidermal Growth Factor Receptor Valutazione di EGFR con FISH in varie malattie neoplastiche V. Martin Recettore transmembrana di tipo TK. Codificato dal gene EGFR, che mappa in 7p1. Attivato attraverso il binding con lo specifico ligando (EGF). Innesca pathway downstreams con numerosi effetti tra cui: - la proliferazione cellulare - la capacità del tumore di invadere - l angiogenesi - la capacità del tumore di dare metastasi. nucleo membrana citoplasmatica Nei tumori solidi EGFR è deregolato in diversi modi: - a livello del recettore - a livello del gene che lo codifica membrana citoplasmatica ) mutazione genica Cellula tumorale nucleo ) mutazione genica ) mutazione genica 1

2 pattern normale (disomia) normale aneusomia cromosoma 7 aneusomia cromosoma 7

3 amplificazione gene EGFR amplificazione gene EGFR aneusomia cromosoma 7 & amplificazione gene EGFR SCORING SYSTEM ICP numero segnali gene EGFR R = numero segnali centromero7

4 CRITERI DI CLASSIFICAZIONE ICP n FISH PATTERN FREQUENZA ANOMALIA CATEGORIA copia bilanciata gene copie bilanciate gene - copie bilanciate gene > copie bilanciate gene Rapporto (R) gene EGFR/centromero LOSS/PERDITA DISOMIA/NORMALE BASSA POLISOMIA ALTA POLISOMIA AMPLIFICAZIONE GENICA Importanza di EGFR MoAb in tumori solidi Valore prognostico: correla con progressione tumorale e scarsa sopravvivenza Valore predittivo: - anticorpi monoclonali (MoAb) - inibitori tirosi kinasici (TKI) TKI Cellula tumorale Authors CARCINOMI COLORETTALI (CRC) Famaco anti-egfr (cetuximab) FDA approved EGFR-FISH CARCINOMA COLORETTALE -ICP- Ooi et al. 00 CRC Amplification when a definite cluster or more than 10 orange signals were found. 11/ () Sauer et al. Moroni et al. Sartore- Bianchi et al 007 Cappuzzo et al. 007 Frattini et al RC 1) Balanced if R > 0.8 but <1. ) Copy loss if R <0.8 ) Copy gain if R 1. Score EGFR gene/ nucleus. Increased EGFR CNG was defined as the presence of three or more signals per nucleus. Score EGFR gene/nucleus and use the cut off value. FISH + if >.5 and/or > 0 chr 7 polysomy FISH - if <=.5 and/or <= 0 chr 7 polysomy Score EGFR /nucleus and use the cut off value. FISH + if >.9 FISH - if <=.9 1) Loss if 1 copy of chr 7 in >50 of cells ) Disomy if copies of chr 7 in >50 of cells ) Low polysomy If or copies of chr 7 in >50 of cells ) Marked polysomy if > copies of chr 7 in >50 of cells 5) Amplification if R> in at least 10 of cells 7/8 (77) /8 (8) 7/8 (15) 9/0 (5) 8-9/ /58 /85 (50) /85 (50) 0/7 (0) /7 (11) 0/7 (0) 16/7 (59) 8/7 (0) Pazienti con amplificazione o alta Of the 9 patients with CNG 8 respondered and 1non responded to cetuximab or polisomia di EGFR panitumumab,suggesting a genetic basis of response to antiegfr treatment possono beneficiare del farmaco mirato (cetuximab o panitumumab) Patients with disomic or low polisomy of chr7 have a reduced likelihoood to responde to panitumumab Patients with EGFR CNG have an increased likelihoood to responde to cetuximab therapy Pazienti disomici o Patients whit amplification or marked polysomy have a increased likelihood to responde con bassa to cetuximab polisomia therapy (depending from K-ras and PTEN status) while the sono resistenti disomic one in generally are resistant 7 Risposta clinica a cetuximab PR NR D 0 FISH EGFR HP 1 A 6 Our unpublished data 1) Loss if 1 copy of chr 7 in >50 of cells ) Disomy if copies of chr 7 in >50 of cells ) Low polysomy If or copies of chr 7 in >50 of cells ) Marked polysomy if > copies of chr 7 in >50 of cells 5) Amplification if R> in at least 10 of cells 1/ () 10/ () 7/ (16) 16/ (7) 9/ (1) British Journal of Cancer 007;97: CNG: copy number gain

5 Authors TUMORI DEL POLMONE NON A PICCOLE CELLULE () Farmaco anti-egfr (gefitinib) TUMORI SQUAMOCELLULARI TESTA COLLO () Famaco anti-egfr (cetuximab) FDA approved Hirsh et al 00 Cappuzzo et al Tsao et al Hirsh et al (BAC subtype) ) trisomy ) polysomy ) amplification* low level amplification if R.1>R>.0 high level amplification if R >= ) low trisomy ) high trisomy ) low polysomy 5) high polysomy 6) amplification Similar outcome of patients with high gene copy number suggested a combination in two classes: amplification and/or high polysomy disomy and/or low polysomy ) low trisomy ) high trisomy ) low polysomy 5) high polysomy 6) amplification* >= 0 of cells displaying >= copies of the EGFR signals or with gene amplifications** <than 0 of cells displaying >= copies of the EGFR gene and 7 (0) 70 (8) (1) 17 (9) () 69 (67) 56 (5) high polysomy and amplified 6 () 55 (68) Patients whit high gene copy number trend toward poor prognosis. Pazienti trattati con gefitinib o erlotinib che After gefitinib treatment, + patients had a significant improvement in response, presentano time to progression and survival with respect to - patients. amplificazione genica o alta polisomia di EGFR hanno maggiori Increased EGFR copy number were associated with responsiveness to erlotinib but possibilità not with increased di survival. risposta e migliore sopravvivenza rispetto a quelli con Increased EGFR gene copy number is associated with improved survival after gefitinib disomia therapy. o bassa polisomia di EGFR. 5 (69) 0 of cells displaying >= copies of the EGFR signals or with After gefitinib treatment, + Cappuzzo gene amplification** patients had a significant improvement in 6 et al 007 response, time to progression and survival <than 0 of cells displaying >= copies of the EGFR gene and with respect to - patients.. 11 (1) TMA=tissue microarray Amplification*= definied as clustered unbalanced gain of the EGFR gene Amplification**=defined by the presence of tight gene clusters, or R>=, or >=15 copies of the gene per cell in >= 10 analyzed cell Authors Freier et 609 Amplification when >10 of cells showing more than 8 signals or 79 (1) No correlation with survival. al. 00 tight cluster. 1) normal : samples that did not met the criteria for gain or Koyonova 1080 amplification (of et al. ) gain : R> 1.5 and < in at least 10 of cells (0.) lariynx) ) amplification: R> or tight clusters in 10 of cells 11 (10) (only abstract available) Marholova aneusomy et al. 7 amplified (only abstract available) Morrison amplification et al of larynx 6.10 deletion Chung et FISH + : amplification and/or high polysomy (50) 86 al. 006 disomy and/or low polysomy (50) /1 abnormal >= 0 of cells displaying >= copies of the EGFR signals or EGFR GCN : Temam et with gene amplification** con 1 al 007 increased and 10 with <than 0 of cells displaying >= copies of the EGFR gene and decreased 10 disomic Amplification*= definied as clustered unbalanced gain of the EGFR gene Amplification**=defined by the presence of tight gene clusters, or R>=, or >=15 copies of the gene per cell in >= 10 analyzed cell Patients whose tumors had either gene amplification Pazienti or deletion con had a poorer survival alterazione del numero di copie geniche di EGFR hanno prognosi Patients with FISH+ tumours had a worse survival. Patients whose tumors had aberrant copy number were more advanced and had a sfavorevole poorer clinical outcome. GLIOBLASTOMI (GBM) Amplificazione di EGFR è marcatore diagnostico di GBM primario TUMORI DELLA MAMMELLA (BC) Authors and year Smith et al. 001 Korshunov.et al 00 Korshunov et al AA 111 GBM GBM (age <50 years) High grade smallcell glioma s Amplification if R>=1. in > 10 of tumor cells or > EGFR signals. 1) Amplification when R> in >10 of tumor cells or innumerable clusters of EGFR signals. ) Polysomy when >0 of nuclei containing >= of chr7 1) Amplification when R> in >10 of tumor cells or innumerable clusters of EGFR signals. ) Polysomy when >10 of nuclei containing >= of chr7 AA: 11/6 (17) GBM: 6/111 (1) 60/189 (1) 171 /189 (90) 6/11 (0) 67 /11(59) EGFR amplification is an independent predictor of prolonged survival in patients with GBM who were elder than Pazienti 60 years of age. con amplificazione di EGFR amplification is a strong indicator of adverse outcome for young adults (pts <50 years). EGFR hanno prognosi sfavorevole EGFR amplification confers shorter survival, unfavourable prognostic marker. Authors and year Kersting et al. 00 Kersting et al 006 Shien et al. 006 Corzo et al. Schiller et al. poster 006 Our unpublished data IDC Phylloid tumours DCMD LABC BLBC +MBC BLBC Amplification if >than signals per cell are present -Criteria of Steodl et al 00 (urinary bladder tumor)- Amplification if >than signals per cell are present. -Criteria of Steodl et al 00 (urinary bladder tumor)- Amplification if R>.0 1) Loss if 1 copy of chr 7 in >50 of cells ) Disomy if copies of chr 7 in >50 of cells ) Low polysomy If or copies of chr 7 in >50 of cells ) Marked polysomy if > copies of chr 7 in >50 of cells 5) Amplification if R> in at least 10 of cells 8/170 (.7) 6/57 (11) 0/ (0) 0/8 (0) 0/9(0) 0/18 (0) 7/18 (9) 0/18 (0) 6/18 () 5/18 (8) EGFR gene amplification is a rare event in invasive breast cancer. AA: anaplastico astrocytoma polysomy of chromosome 7 concomitant to EGFR gene amplification = pac probe dig-labelled for EGFR gene high grade ductal carcinoma with myoepithelial differentiation (DCMD) locally advanced breast cancer (LABC) basal like breast cancer (BLBC) methaplastic breast cancer (MBC) CONCLUSIONI HER!!! è un analisi complessa EGFR 5

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