Lung cancer 2011 talk-for 17-10-2011-17-10-2011

• 1. From metagenes to biomarkers, a possibleapproach for prognosis and personalized medication through GWAS of lung cancer (NSCLC)Cheng Luo, Ph. D University of Tartu

2. 1, Metagene MAGEs obtained from GWAS (or geneexpression profiling of whole genome) of lungcancer are immunotherapy target.2, The prognosis influenced by EGFR and VEGFmutation and variation.3, COX-2/Keap1/Nrf2/ARE(s) antioxidant pathwaysin lung cancer (NSCLC).4, Acetaldehyde test in A549, an antioxidant assayfrom in vivo to in vitro. 3. 1, Metagenes obtained from GWASare possible to be genetic /diagnosis /prognosis biomarkers, andimmunotherapy target 4. Metagenes associated to survivalsGenes up-regulated in NSCLCGene ID adj. p-value Fold changeSPP1 2,76E-17 14,52MMP15,20E-1110,74KRT6A 3,85E-0510,39KRT17 1,86E-069,44CTHRC1 2,76E-17 7,62S100A22,21E-057,44MMP11 1,29E-11 7,14TMPRSS4 1,04E-08 6,85CDC201,59E-136,82KRT6B7,41E-046,15Genes down-regulated in NSCLCGene ID adj. p-value Fold changeSFTPC 7,13E-1024,43FCN3 6,77E-1716,71FABP4 3,28E-14 13,54C19ORF595,43E-20 12,51TMEM1001,65E-2112,11CYP4B1 1,35E-12 11,41CLIC5 9,37E-2110,99SFTPD 4,18E-0810,75SFTA2_HUMAN 4,02E-07 10,59CLEC3B 9,66E-22 10,23Urgard E. et al. 2011 5. MAGE family, PTGS2 (COX-2), Nre2l2 (Nrf2) and SOD1 (ARE genes) 6. Metagene: MAGEA9B,MAGEA10 (melanoma,ageing scenery) Expression anddemethylation ofMAGE genes occuronly in cancer cells(except testis)MAGE expression isknown to be activatedby promoterdemethylation. 7. Dendrogram Some of mouse andrepresentation of thehuman MAGE gene smultiple alignment are identicalbetween MAGEconserved domains ofhuman and mouse MAGEproteins.Kufer P. et al., 2002, HeterogeneousExpression of MAGE-AGenes in OccultDisseminated Tumor CellsA Novel Multimarker Reverse Transcription-Polymerase Chain Reaction for Diagnosis ofMicrometastatic DiseaseChromez P. et al 2001, An Overviewof the MAGE Gene Family with theIdentification of All HumanMembers of the Family 8. MAGE genes: transcriptional factorsBarker PA et al: The MAGE proteins: emerging roles in cell cycle progression, apoptosis, andneurogenetic disease. J Neurosci Res 67:705-712, 2002.Ohman FK et al: The melanoma antigen genes any clues to their functions in normaltissues? Exp Cell Res 265(2): 185-194, 2001.Chromez, et al 2001, An Overview of the MAGE Gene Family with the Identification of AllHuman Members of the Family , Cancer Research 9. Frequent MAGE Mutations in Human Melanoma:Distribution of mutations in the MAGE genesCaballero OL et al. Frequent MAGE Mutations in Human Melanoma, 2010 PLOS One 10. Swiss CohortCaballero OL et al. Frequent MAGE Mutations in Human Melanoma, 2010 PLOS One 11. Australiacohort 12. Globalcohorts byliteratures 13. Chicago cohortKufer P et al. 2002, Cancer Research 14. Genetic make-up of MAGE genes ofNSCLC of Estonia cohorts? 15. MAGE Expressions Mediated by Demethylation of MAGE Promoters Induce Progression of Non-small Cell Lung CancerYANAGAWA N. et al. 2011Correlation between MAGE expression and overall survival of 67 NSCLC patients using the Kaplan-Meier method.The patients with any expression had poorer prognosis than those with no expression. 16. MAGE (Melanomaassociated antigen)participate theapoptosis!!!Nasarre P. et al., 2010, Guidance molecules in lung cancer. Cell AdhMigr. 4(1):130-45. 17. MAGE genes in clinic Jheon S. et al. Lung cancer detection by a RT-nested PCR using MAGE A16 common primers, Lung Cancer 2004 Conclusion MAGE A16 RT-PCR showed high sensitivity andspecificity in various respiratory specimens, andfor lung cancer detection, based on sputum samples, MAGE isprobably the best of the known tumor markers. Thesatisfactory conclusion of workaimed at determining the causesof false positivity,should ensure that MAGE A16 RT-PCRbecomes an effectively diagnostic tool for lung cancer. TANG Yan, XIA Da Wen, LI Ya Rong, et al. 2010, Measurement of Lung Cancer Cells in the Sputum and Peripheral Blood by the MAGE A1-6 mRNA as Specific Markers 18. Selvan SR et al. 2010, Establishment of stable cell lines forpersonalized melanoma cell vaccineTable 4 Various combinations of six different TAA distributions in melanoma cell linesPresence Combination ofNo. ofPresence of Combination ofNo. of TAAs expressedTAAs expressed byof no. of TAAs by melanoma cellcell linesno. of TAAs melanoma cell lines cell lines(absence oflines (absence ofspecific TAAs) specific TAAs)6S-100/MAGE-1/Mel-5/HMB-45/Melan-A/Tyrosinase 93 (S-100/MAGE-1/Tyrosinase) Mel-5/HMB-45/Melan-A55 (S-100) MAGE-1/Mel-5/HMB-45/Melan-A/3 (S-100/MAGE-1/Mel-5) HMB-45/Melan-A/Tyrosinase4Tyrosinase193 (Mel-5/HMB-45/Melan-A) S-100/MAGE-1/Tyrosinase25 (MAGE-1) S-100/Mel-5/HMB-45/Melan-A/3 (S-100/Mel-5/Melan-A) MAGE-1/HMB-45/Tyrosinase1Tyrosinase85 (Mel-5)S-100/HMB-45/Melan-A/MAGE-3 (S-100/Mel-5/Tyrosinase) MAGE-1/HMB-45/Melan-A11/Tyrosinase 33 (S-100/MAGE-1/HMB-45) Mel-5/Melan-A/Tyrosinase15 (HMB-45) S-100/MAGE-1/Mel-5/Melan-A/3 (S-100/HMB-45/Tyrosinase) MAGE-1/Mel-5/Melan-A1Tyrosinase 13 (MAGE-1/Mel-5/Tyrosinase) S-100/HMB-45/Melan-A14 (S-100/MAGE-1) Mel-5/HMB-45/Melan-A/2 (MAGE-1/Mel-5/HMB-45/ Melan-A)S-100/Tyrosinase3Tyrosinase 20 2 (S-100/Mel-5/Melan-A/Tyrosinase)MAGE-1/HMB-45 14 (S-100/Mel-5) MAGE-1/HMB-45/Melan-A/Tyrosinase 62 (S-100//MAGE-1/Mel-5/HMB-45)Melan-A/Tyrosinase14 (S-100/HMB-45) MAGE-1/Mel-5/Melan-A/Tyrosinase/ 2 2 (S-100/MAGE-1/HMB-45/Melan-A)Mel-5/Tyrosinase 14 (MAGE-1/Mel-5) S-100/HMB-45/Melan-A/Tyrosinase 11 (S-100/Mel-5/HMB-45/Melan-A/Tyrosinase)MAGE-1 34 (MAGE-1/HMB-45) S-100/Mel-5/Melan-A/Tyrosinase 11 (S-100/MAGE-1/Mel-5/HMB-45/Melan-A)Tyrosinase 24 (Mel-5/HMB-45) S-100/MAGE-1/Melan-A/Tyrosinase 11 (S-100/MAGE-1/HMB-45/Melan-A/Tyrosinase)Mel-5 1Total cell lines 1060 (S-100/MAGE-1/Mel-5/ HMB-45/Melan-A/Tyrosinase)None 7TAA, tumor-associated an Total cell lines 106, TAA, tumor-associated an 19. Increased expression of MAGEA9B, MAGEA10 for longger survival of NSCLC in Estonia Cohort Two situations for the expression of MAGE genes incancers: the expression of MAGE genes may blockapoptosis, but as cancer-specific antigen, the expressionmay stimulate immune response, particularly cancer specific IgG that could attack cancer cells self, or byvaccination. High expression of meta gene wasassociated with increase in the survival time (Urgard E.et al 2011). This fits with immune theory. 20. 5.3. MAGE-A3 vaccineMAGE-A3 is a tumor-associated antigen that is notexpressed in most normal cells [31,61]. Approximately 3550% of lung cancers express MAGE-A3, and expression hasbeen associated with poor prognosis [31]. This providedthe rationale for the development of a TCV comprisingMAGE-A3 recombinant protein combined with the AS02Bimmunoadjuvant.In a randomized phase II study, 182 patients withcompletely resected, MAGE-A3-expressing, stage IB or IINSCLC were randomly assigned to receive postoperativeMAGE-A3 vaccine or placebo (2:1) [62]. The HRs fordisease-free interval, disease-free survival (DFS) and overallsurvival were 0.74 (95% CI 0.441.20), 0.73 (95% CI 0.451.16) and 0.66 (95% CI 0.361.20), respectively, in favor ofthe MAGE-A3-treated patient group. The ongoing phase IIIMAGRIT (MAGE-A3 as Adjuvant, Non-Small Cell LungCancer Immunotherapy) trial, initiated in 2007, plans toenroll approximately 2300 MAGE-A3-expressing patientswith resected stage IB, II, or IIIA NSCLC who will berandomized to vaccine or placebo, again in the adjuvantsetting. The administration of adjuvant platinumbasedchemotherapy for the treatment of the current NSCLC isallowed between surgery and randomization. The study ispowered for DFS as the primary endpoint in the totalpopulation and in the cohort without postoperativechemotherapy [63].Mellstedt H. et al 2011 21. Immunotherapy in NSCLC 22. Lung Cancer Vaccine Shows Promisehttp://www.gsk.com/ 23. Other Metagene: NLRP2Inflammation,proliferation,tumorigenesis. COX-2 24. Metagene:CYP3A5Detoxification,antioxidation 25. Metagene: AKR1B1 26. Summary and next: Likely, MEGA gene families, NLRP2, CYP3A5, AKR1B1, KEAP1 are decisive genes for survival of lung cancer for Estonia cohort. MAGE genes mutation frequency and expression distribution (PCR and Sequencing) for NSCLC in stock (Tarmo has total RNA) of Estonia cohort. MEGA(s) are cancer/testis specific antigen (recognized by HLA-A1 of CTL), high expression of MEGA genes make them ideal targets for immunotherapy for NSCLC patients: (polysaccahride, or other immunojuvants containing syrup) spray and vaccine!!! Next: Analyze MAGE genes expression (lung tissue, or sputum and peripheral blood) for further determination of MAGE gene expression, methylation/demethylations impacts on NSCLC for Estonia Cohorts, and for personalized medication, or vaccine preparation. 27. 2, The prognosis influenced by the EGFRand VEGF mutation and other geneticmake-up, or variationToday major personal medication or translational medicineare from EGFR and VEGFEGFR and VEGF were not among 599 genes which were down-regulatedand 402 genes which were up-regulated in Urgard E. et all 2011. However, typically the genemutation, or methylation, or other tissue-derived molecular information ofEGFR and VEGF has been combined with individuals personal medical history,family history, and data from imaging, and other laboratory tests for bettereffective treatments for a wider variety of conditions. 28. EGFR (Her-1), and VEGF, a clinicexample of translational medicine To determine whether a solid tumor, such as for the lung (non small cell), head and neck, colon, pancreas, or breast, the mutation positive or negative for EGFR, helps to guide treatment and determine prognosis When one have been diagnosed with certain invasive cancers, more and more doctors want to see if EGFR is being over-expressed in the tumor, in order to choose what kind of chemotherapies. 29. EGF (R), VEGF and others promoteangiogenesis 30. Bifunctional inhibitor of VEGF and EGFR Ryan AJ. et al. 2005 31. If VEGF mutationor overexpressionInvassion Alitalo K. et al 2002 32. Gene mutation may increase survivalTKI:tyrosinekinaseinhibition,RR:responserate, TTP:time toprogression 33. VEGF expression associated with tumor growth and survival(mutation, and silence by methylation)(A) Overall survival in patients with VEGF-negative and VEGF-positivetumours (P