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Literature Report
李丽婷 2014.3.19
Article source : Gut 2013;62:1415--1424
Content
Introduction
Methods
Results
Conclusion
Prospect
Part1: Introduction
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!!!
Molecularly targeted therapy (Abnormality : key driver)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.
Introduction
Part2: Methods
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= 462.Cell line:14 sorts3.Inhibitor :EGFR/ErbB2 inhibitor,Mek inhibitor,other TIKs(EGFR TKI,Met TKI,FGFR TKI,Ret TKI)
Experiments
1.GSEA and HOPACH clustering2.Immunohistochemistry for P-ERK3.RTK phosphorylation profiling4.Proliferation assays5.Apoptosis arrays6.Protein phosphorylation status
MAPK?
RTK?
In patients?
Methods
Methods
Part3: Results
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 oesophagusBE:Barrett’s oesophagusDYS;Dysplastic Barrett’sAC:Adenocarcinoma
Results
GSEA :Gene set enrichment analysisREGG : molecular signatures databaseHOPACH :cluster simples using pathway enrichment scores
2.Phospho-Erk status confirms MAPK as a key pathway
148/434(34.3%) of cases expressed high levels of phosphorylated (2+,3+)
Results
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
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 ERKNOTE:EGFR/ErbB2for OE19,Met ,MKN45,FGFR for KatoIII
One:An obvious dominant RTKTherapy with a singer TKIs
Results
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 ERKNOTE:FGFR2for HSC39,EGFR,ErbB2,ErbB3,FGFR3,Met and RET for OE33
Two :Complex RTKs activation
Results
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
RTK activation in a patient cohort
Patient Samples:n=46 RTK array Results : RTK activation
No One Tw o Multiple
Cases 8/46(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 ?
Results
Part4:Conclusion
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
Part5:Prospect
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
