- Email: [email protected]
O-012 High frequency of epidermal growth factor receptor mutation in lung adenocarcinoma in Thailand
$8
Oral Sessions/Basic science/Ceil and molecular biology
from four different ceunthos and compared the mutational status with clinics pathological features and the presence of KRAS (oxen 2) mutafions EGFR and ERBB2 mutations were present in 22% (149 of 671) and in 1 6% (11 of 671) of pnmary NSCLC respectively and were somafic in origin EGFR mutations were signific~,'lfly (P <0 001) more frequent in never smokers. adenocarcinema histology. Oriental ethnicity (Japan and Taiwan) and female gender, but were not related to patient age. clinical stage, brenchicloaiveolar histologic features or patient survival The mutations were of three common types (in frame deletions in exert 19. a single missenso mutation in e~on 21 and in frame duplications/Insertions in e~on 20). Rare missonse mutations were also present in e~ons 18.20 and 21. All ERBB2 mutations were in frame insertions in oxen 20 and targeted the idonfical corresponding genomic region as did EGFR mutations. They were limited to adenocaranema histology. In 394 adonocar~noma cases ERBB2 mutafions preferenfially targeted Onental ethniclty (3.9%) compared to ether ethnicitles (0.7%). female gender (3.6%) compared to male gender (1 9%) and never smokers (4 1%) compared to smokers (1 4%) Mutations of KRAS were present in 8.% (54 of 671) of primary NSCLC KRAS mutations were significantly (P <0 001) more frequent in ever smokers, adenocarcinema histology and non-Oriental ethnicity (USA and Australia) Mutafions in more than one of these three genes were never present simultaneously in individual tumors EGFR and ERBB2 mutafions are the first molecular change known to specifically target lung cancers ans~ng in never smokers. The remarkable similanfies of mutations in both genes with regard to type and location and targeted patient subpopulations are unprecedented. In contrast KRAS mutafions target ever smokers. EGFR. ERBB2 and KRAS mutations sppoar to be mutually exclusive. Our findings suggest dfferent pathways to lung cancer in smokers and never smokers. [~"
High frequency of epidermal growth factor receptor rnutatlon In
lung adenocardnoma In Thailand V. Srluranpor'~ ~. C. Chantranuwat 2. N. Huspai ~. 1". Chalermchai ~. K Leungtaweaboon 4. P Lertsaguansinchal 5. N Voravud I . A Mu'drangura3 1Medical Oncotogy, Faculty of Medicine, Chulalongkom University, Thailand,
2Department ct Pathology, Faculty of Medtctne, Chula/ongkom Untversrty, Thailand, 3Department of Anatomy, Faculty of Medicine, Chulalongkom Un/vesity, That/and, ~Department of Surge~ Faculty of Medicine, Chulalongkom University, Thailand, 5Department of Radology, Faculty of Medicine, Chulalongkom University, Thailand Background: Unique somatic mutation of epidermal growth factor receptor gone (egtr) in nor. small cell lung cancer (NSCLC) has been shown to be tightly correlated to the response to b'oatment with tyresino kinase ichib~tors. Recent reports reveal a racial cifference in frequency of mutation of efgr Less than 1% of Caucasian lung cancer patients cisplayed egfr mutation in the tymsine kinase domain whereas 30% of Jabanese pafients governed these mutafions We sought to characterize the frequency and pattern of egfr mutations in lung cancer patients in Thailand Methods: Genomic DNA was isolated from manually microdissected tumor calls procured from archival paraffin embedded pathological specimens Isolated DNA was then subjected to PCR amplificafion of exert 19 and exert 21 of egfr. PCR products were examined by gel eloctTophoresls and direct sequencing. A panel of 5 patients who received gofit]nib with known responses was used to confrm the association between mutations and the response to treat]'nont. Additional ,56 samples of lung adonecarclnoma were analyzed Ibr egfr mutafion. Results: Deletions of oxen 19 wore detected in 3 patients who showed radiolographic response to gefitinib tTeatment but no mutation of both e~on lg and e~on 21 detected in 2 patients without objec0ve response Overall mutafion rate was 36 out of 61 (58. 1%) in lung adenocarcinema We found 29/60 (48 3%) e~on 19 delefions. 6/54 (11 1%) ~ o n 21 point mutations, and 1 (1 6%) doable-mutation of both e0¢ons ,tvnong oxen 19 mutations, deletion of sgtr nucleotide 2481 249b leacing to deletion of amino acid E746-ATb0 in the catalytic domain were detected in 22/30 (73 3%) of oxen 19 deletions The presence of mutations significantly assoaated with non smoker but net with female. BAC histology, stage, nodal metastasis, or extrathoraclc metastasis. Conclusions: We found a stnl~ngly h~gh prevalence of egtr mutafions in The lung adonocarclnoma patients which may e0(pialn the excellent response to tyresino kinase ichibitors in Asian pppulafion. Further evaluafions in larger lung cancer populafion and study of clinical benefit from tyroslne klnase inhibitor treatment guided by molecular stratification are warrant.
Molecular Biology II1: CGH/FISH/Expression profiling analysis Wednesday, 6 July 2005 14:30-16:30
[0•3]
Global e x p r e ~ l o n profiling of lung pdmary tumom: CorrelaUon with pattlologlcal and genetic ¢haractedstlcs
B. Angulo I . E. Condo ~. M. TangI . J. Carretere I . R Medina I . F. Lppoz R.Ios2. M Sanche,z-Cespedes ~ 1Spanish Nabone/Cancer Centre (CNtO), Madrid,
Spare: 2Pathology Program, Hospttal 12 Octubre, Spatn Background: As a censequonee of the scientific and economical investment in cancer research new therapies, spea fically targeting these regulatory pathways abnormal in cancer cells are being developed for most tumor types, including lung cancer. Abnormalities in several tumor suppressor genes and oncogonos that are involved in growth regulatory signaling pathways, cell cycle. DNA repair and apeptosis are among the most common alterations and consfitute molecular targets for the design of novel and specific therapies On the other hand. global analysis of gone e0¢pression using cligonucleotide or cDNA microarrays sheds light on a now classificafion of primary tumors on the basis of their molecular prefles Aim: In the present work we have used cDNA microarrays to determine the associafion between the global e0¢pression probes and the genetic and histological characteristics of non small cell lung tumors. Methods: Over 100 primary tumors from patients diagnosed with non small cell lung cancer were provided by the CNIO Tumor Bank Network. Good~luallty DNA and total RNA was obtain from 100 and 60 of the tumor speamons. respectively. A mutational analysis of the K;~,AS, p53, LKB'/ and EGFR genes was performed using prrevicusly desorlbed protocols For the cONA microarrays analysis we used the CNIO OnceChip. a cDNA microamay that has been specially designed for looking at genes involved in cancer and includes more than 11.000 different genes Four micrograms of total RNA was amplified and hybridized against Human Universal Reference total RNA Results: We have performed a mutational analysis of about 100 non-small call lung tumors and we have detected p53 point mutations in 45% of all of them. whereas KT~,AS, LKBf and EGFR mutafions were present in 27. 20 and 10% of the lung adenecarclnomas induded in the study. Our preliminary unsupervised data analysis on the global e~pression profiles of 15 tumors was able to dis~minate among the sqJamous cell ca~nomas and adenocaranemas histological types and identifies several genes with charactenstic patterns of espression in each tumor histology. These genes were implicated in secretory, development and cell growth regulation. Morec~,~or. we used supervised analysis to determine the gone express~on profiles that characterizes these tumors carrying p53 mutations Several genes implicated in cell differentiation were identified, including tGFBP3, a p53-regulated target gene. and VEGF. both down-regulated in p53-mutant tumors We are currently per~rming the analysis in the remaining tumors Conclusions: Our preliminary study in 1,5 lung tumors incicate that ~pression probes can ciscnminate lung cancer subgroups Definitive data including the analysis of 60 nen-small cell lung tumors will be presented
~0~
Finding blomarkem specific for eady stages of lung cancer using SAGE data
T. Chan ~. C. MaoAulay 2, W. Lain 2, S. Lain 2, K. Lonorgar~. R. Ng ~.
1University of British Columbia, Vancouver, BC, Canada, 2The British Cotumbia Cancer Agency, Vancouver, BC, Canada Background: The 5-year survival rate of an advanced lung cancer patient is only about 10% However. if the cancer is caught early at the CIS (carcinoma in situ) stage, the 5-year survival rate is 90% As far as we know. up to new. no studios have isolated biomarkors Ibr this elusive CIS stage. Hero. we present a study to isolate genes spoafic for the CIS stages of lung cancer that are not present in bronchial metaplasia tissue. Methods: To perform our analysis, we used Lung SAGE libranes prc~idod by BE; Cancer Agency which include 1 Motaplasia library. 15 smoked 10. CIS/Metaplasia >2 and CIS/Invasive >,5 In addition to CIS selection, we also perlbrmod similar analyses on the Invaslve libraries (replace CIS with Invaslvo). Results: Our preliminary results show that the intersection of the top 2000 tags of the Normal vs. CIS and CIS vs. Invaslve sots yielded 79 unique tags. After applying fold change and z score filters, we obtained 18 tags. Out of
Oral Sessions/Basic science/Ceil and molecular biology
from four different ceunthos and compared the mutational status with clinics pathological features and the presence of KRAS (oxen 2) mutafions EGFR and ERBB2 mutations were present in 22% (149 of 671) and in 1 6% (11 of 671) of pnmary NSCLC respectively and were somafic in origin EGFR mutations were signific~,'lfly (P <0 001) more frequent in never smokers. adenocarcinema histology. Oriental ethnicity (Japan and Taiwan) and female gender, but were not related to patient age. clinical stage, brenchicloaiveolar histologic features or patient survival The mutations were of three common types (in frame deletions in exert 19. a single missenso mutation in e~on 21 and in frame duplications/Insertions in e~on 20). Rare missonse mutations were also present in e~ons 18.20 and 21. All ERBB2 mutations were in frame insertions in oxen 20 and targeted the idonfical corresponding genomic region as did EGFR mutations. They were limited to adenocaranema histology. In 394 adonocar~noma cases ERBB2 mutafions preferenfially targeted Onental ethniclty (3.9%) compared to ether ethnicitles (0.7%). female gender (3.6%) compared to male gender (1 9%) and never smokers (4 1%) compared to smokers (1 4%) Mutations of KRAS were present in 8.% (54 of 671) of primary NSCLC KRAS mutations were significantly (P <0 001) more frequent in ever smokers, adenocarcinema histology and non-Oriental ethnicity (USA and Australia) Mutafions in more than one of these three genes were never present simultaneously in individual tumors EGFR and ERBB2 mutafions are the first molecular change known to specifically target lung cancers ans~ng in never smokers. The remarkable similanfies of mutations in both genes with regard to type and location and targeted patient subpopulations are unprecedented. In contrast KRAS mutafions target ever smokers. EGFR. ERBB2 and KRAS mutations sppoar to be mutually exclusive. Our findings suggest dfferent pathways to lung cancer in smokers and never smokers. [~"
High frequency of epidermal growth factor receptor rnutatlon In
lung adenocardnoma In Thailand V. Srluranpor'~ ~. C. Chantranuwat 2. N. Huspai ~. 1". Chalermchai ~. K Leungtaweaboon 4. P Lertsaguansinchal 5. N Voravud I . A Mu'drangura3 1Medical Oncotogy, Faculty of Medicine, Chulalongkom University, Thailand,
2Department ct Pathology, Faculty of Medtctne, Chula/ongkom Untversrty, Thailand, 3Department of Anatomy, Faculty of Medicine, Chulalongkom Un/vesity, That/and, ~Department of Surge~ Faculty of Medicine, Chulalongkom University, Thailand, 5Department of Radology, Faculty of Medicine, Chulalongkom University, Thailand Background: Unique somatic mutation of epidermal growth factor receptor gone (egtr) in nor. small cell lung cancer (NSCLC) has been shown to be tightly correlated to the response to b'oatment with tyresino kinase ichib~tors. Recent reports reveal a racial cifference in frequency of mutation of efgr Less than 1% of Caucasian lung cancer patients cisplayed egfr mutation in the tymsine kinase domain whereas 30% of Jabanese pafients governed these mutafions We sought to characterize the frequency and pattern of egfr mutations in lung cancer patients in Thailand Methods: Genomic DNA was isolated from manually microdissected tumor calls procured from archival paraffin embedded pathological specimens Isolated DNA was then subjected to PCR amplificafion of exert 19 and exert 21 of egfr. PCR products were examined by gel eloctTophoresls and direct sequencing. A panel of 5 patients who received gofit]nib with known responses was used to confrm the association between mutations and the response to treat]'nont. Additional ,56 samples of lung adonecarclnoma were analyzed Ibr egfr mutafion. Results: Deletions of oxen 19 wore detected in 3 patients who showed radiolographic response to gefitinib tTeatment but no mutation of both e~on lg and e~on 21 detected in 2 patients without objec0ve response Overall mutafion rate was 36 out of 61 (58. 1%) in lung adenocarcinema We found 29/60 (48 3%) e~on 19 delefions. 6/54 (11 1%) ~ o n 21 point mutations, and 1 (1 6%) doable-mutation of both e0¢ons ,tvnong oxen 19 mutations, deletion of sgtr nucleotide 2481 249b leacing to deletion of amino acid E746-ATb0 in the catalytic domain were detected in 22/30 (73 3%) of oxen 19 deletions The presence of mutations significantly assoaated with non smoker but net with female. BAC histology, stage, nodal metastasis, or extrathoraclc metastasis. Conclusions: We found a stnl~ngly h~gh prevalence of egtr mutafions in The lung adonocarclnoma patients which may e0(pialn the excellent response to tyresino kinase ichibitors in Asian pppulafion. Further evaluafions in larger lung cancer populafion and study of clinical benefit from tyroslne klnase inhibitor treatment guided by molecular stratification are warrant.
Molecular Biology II1: CGH/FISH/Expression profiling analysis Wednesday, 6 July 2005 14:30-16:30
[0•3]
Global e x p r e ~ l o n profiling of lung pdmary tumom: CorrelaUon with pattlologlcal and genetic ¢haractedstlcs
B. Angulo I . E. Condo ~. M. TangI . J. Carretere I . R Medina I . F. Lppoz R.Ios2. M Sanche,z-Cespedes ~ 1Spanish Nabone/Cancer Centre (CNtO), Madrid,
Spare: 2Pathology Program, Hospttal 12 Octubre, Spatn Background: As a censequonee of the scientific and economical investment in cancer research new therapies, spea fically targeting these regulatory pathways abnormal in cancer cells are being developed for most tumor types, including lung cancer. Abnormalities in several tumor suppressor genes and oncogonos that are involved in growth regulatory signaling pathways, cell cycle. DNA repair and apeptosis are among the most common alterations and consfitute molecular targets for the design of novel and specific therapies On the other hand. global analysis of gone e0¢pression using cligonucleotide or cDNA microarrays sheds light on a now classificafion of primary tumors on the basis of their molecular prefles Aim: In the present work we have used cDNA microarrays to determine the associafion between the global e0¢pression probes and the genetic and histological characteristics of non small cell lung tumors. Methods: Over 100 primary tumors from patients diagnosed with non small cell lung cancer were provided by the CNIO Tumor Bank Network. Good~luallty DNA and total RNA was obtain from 100 and 60 of the tumor speamons. respectively. A mutational analysis of the K;~,AS, p53, LKB'/ and EGFR genes was performed using prrevicusly desorlbed protocols For the cONA microarrays analysis we used the CNIO OnceChip. a cDNA microamay that has been specially designed for looking at genes involved in cancer and includes more than 11.000 different genes Four micrograms of total RNA was amplified and hybridized against Human Universal Reference total RNA Results: We have performed a mutational analysis of about 100 non-small call lung tumors and we have detected p53 point mutations in 45% of all of them. whereas KT~,AS, LKBf and EGFR mutafions were present in 27. 20 and 10% of the lung adenecarclnomas induded in the study. Our preliminary unsupervised data analysis on the global e~pression profiles of 15 tumors was able to dis~minate among the sqJamous cell ca~nomas and adenocaranemas histological types and identifies several genes with charactenstic patterns of espression in each tumor histology. These genes were implicated in secretory, development and cell growth regulation. Morec~,~or. we used supervised analysis to determine the gone express~on profiles that characterizes these tumors carrying p53 mutations Several genes implicated in cell differentiation were identified, including tGFBP3, a p53-regulated target gene. and VEGF. both down-regulated in p53-mutant tumors We are currently per~rming the analysis in the remaining tumors Conclusions: Our preliminary study in 1,5 lung tumors incicate that ~pression probes can ciscnminate lung cancer subgroups Definitive data including the analysis of 60 nen-small cell lung tumors will be presented
~0~
Finding blomarkem specific for eady stages of lung cancer using SAGE data
T. Chan ~. C. MaoAulay 2, W. Lain 2, S. Lain 2, K. Lonorgar~. R. Ng ~.
1University of British Columbia, Vancouver, BC, Canada, 2The British Cotumbia Cancer Agency, Vancouver, BC, Canada Background: The 5-year survival rate of an advanced lung cancer patient is only about 10% However. if the cancer is caught early at the CIS (carcinoma in situ) stage, the 5-year survival rate is 90% As far as we know. up to new. no studios have isolated biomarkors Ibr this elusive CIS stage. Hero. we present a study to isolate genes spoafic for the CIS stages of lung cancer that are not present in bronchial metaplasia tissue. Methods: To perform our analysis, we used Lung SAGE libranes prc~idod by BE; Cancer Agency which include 1 Motaplasia library. 15 smoked