1. Literature Report 李丽婷 2014.3.19 Article source : Gut 2013;62:1415--1424

2. Content Introduction Methods Results Conclusion Prospect

3. Part1: Introduction

4. 1.Over 1390 000 diagnoses per annum 2.By 2030,the 10th leading cause of global mortality. 3.In the western world, over fivefold in 30 years 4.5-year survival: 15% 1.Over 1390 000 diagnoses per annum 2.By 2030,the 10th leading cause of global mortality. 3.In the western world, over fivefold in 30 years 4.5-year survival: 15% Introduction The basic background New therapy!!!

5. Molecularly targeted therapy (Abnormality : key driver) Molecularly targeted therapy (Abnormality : key driver) Inter-patient heterogeneity Therapy resistance Inter-patient heterogeneity Therapy resistance Not been broadly adopted ??? ( target at initial presentation ) The aim: 1.Determine the most frequently active cell signaling pathways 2.RTK array measuring phosphorylation status could be used to select the most effective and broadly applicable therapy strategies. 3.Apply this approach to human tissue samples as a proof of principle. The aim: 1.Determine the most frequently active cell signaling pathways 2.RTK array measuring phosphorylation status could be used to select the most effective and broadly applicable therapy strategies. 3.Apply this approach to human tissue samples as a proof of principle. Introduction

6. Part2: Methods

7. Materials 1.samples:(1)cancer cohort for gene expression profiling n=75 (2)cases for P-Erk immunostaining n=434 (3)patient samples for RTK array n= 46 2.Cell line:14 sorts 3.Inhibitor :EGFR/ErbB2 inhibitor,Mek inhibitor,other TIKs(EGFR TKI,Met TKI,FGFR TKI,Ret TKI) Experiments 1.GSEA and HOPACH clustering 2.Immunohistochemistry for P-ERK 3.RTK phosphorylation profiling 4.Proliferation assays 5.Apoptosis arrays 6.Protein phosphorylation status MAPK? RTK? In patients? Methods

9. Part3: Results

10. 1.MPAK :enrichment pathway (1)Enrichment scores( figure A)by HOPACH GSEA KEGG (2)50.7%(38/75)of samples enriched for overexpression of signaling pathway (3)MPKA pathway is enriched in 42.7%(32/75)of samples. NE: Normal oesophagus BE:Barrett’s oesophagus DYS;Dysplastic Barrett’s AC:Adenocarcinoma Results GSEA :Gene set enrichment analysis REGG : molecular signatures database HOPACH :cluster simples using pathway enrichment scores

11. 2.Phospho-Erk status confirms MAPK as a key pathway 148/434(34.3%) of cases expressed high levels of phosphorylated (2+,3+) Results

12. 3.Selective inhibition of activated RTKs inhibits MAPK signalling RTK array : Drug selection to ensure the maximal therapeutic effect. Results MKNI RTK activation profile Full RTK array In MKNI,with no active RTKs,TKIs or Mek inhibition did not inhibit proliferation until the uM range Proliferation dose-response of MKN1 to TKIs

13. 3.Selective inhibition of activated RTKs inhibits MAPK signalling RTK activation profile Proliferation dose response to TKIs MEK Cell cycle profile in response to TKIs Western blotting for downstream MAPK components Atk and ERK NOTE:EGFR/ErbB2for OE19,Met,MKN45,FGFR for KatoIII One:An obvious dominant RTK Therapy with a singer TKIs One:An obvious dominant RTK Therapy with a singer TKIs Results

14. 3.Selective inhibition of activated RTKs inhibits MAPK signalling RIK activation profile Proliferation dose-response to TKIs (Mek inhibition) Cell cycle profile in response to TKIs Western blotting for downstream MAPK components Atk and ERK NOTE:FGFR2for HSC39,EGFR,ErbB2,ErbB3,FGFR3,Met and RET for OE33 Two :Complex RTKs activation Results

15. SUMMARY Cell line: OE19,MKN45, KatoIII An obvious RTK activation Therapy with a single TKI Cell line: HSC39,OE33 Complex PTK activation HSC39:a combination of three TKIs OE33:dual agent treatment(TKI combination and Mek inhibitor Patient cohort Results

16. RTK activation in a patient cohort Patient Samples:n=46 RTK array Results : RTK activation NoOneTw oMultiple Cases8/46(17,4%)8/46(17.4%)13/46(28.3%)17/46(37.0%) If RTK array could help in drug selection in patients? RTK activation ? If RTK array could help in drug selection in patients? RTK activation ? Results

17. Part4:Conclusion

18. 1.The MAPK pathway is commonly activated in oesophago –gastric cancer secondary to activation of number of RTK. 2.The RTK activity profile,using RTK arrays, is useful to guide which TKI would offer therapeutic efficacy.(cell lines) 3.If multiple RTKs are active, inhibition of downstream signaling might offer an alternative clinical approach. (Mek inhibitor) 4. Using an RTK array in patients could help in drug selection to ensue the maximal therapeutic effect. Conclusion

19. Part5:Prospect

20. 1.Which are the most clinically relevant therapeutic targets? 2.Which is more effective, using a single inhibitor or a combination? 3.How do we translate this information into a therapeutic strategy with maximise outcomes and minimize toxicity? Prospect

21. 李丽婷 邮箱： lilypretty12@163.com lilypretty12@163.com