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neu expression on survival of stage I-IIIA non-small cell lung cancer (NSCLC) patients radically resected
Biology
184
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Long-term effect of HER-2/neu expression on survival of stage I-IliA non-small cell lung cancer (NSCLC) patients radically resected
G. Selvaggi, S. Novello, G. Ferrari, P. Borasio, C. Mossetti, F. Ardissone, P. Lausi, G.V. Scagliotti. University of Torino, Orbassano;
S. Luigi Hospital, Orbassano; University of Torino, Department of Clinical and Biological Sciences, Orbassano, Italy In this study we investigated the long-term prognostic value of HER2/neu expression in radically resected NSCLC and its correlation with overall survival. Surgical specimens obtained from patients (n = 120) who underwent radical resection for NSCLC between January 1991 and February 1992 (squamous cell carcinoma 60 [46%], adenocarcinoma 48 [37%] and 22 large cell carcinoma [17%]) and surgically staged as stage I = 41 (31%), II = 37 (29%) and IliA = 52 (40%) were investigated for the presence and distribution of HER-2/neu using an avidin-biotin complex immunohistochemical technique. Obtained data were correlated to clinical and histopathologic variables. Normal bronchial tissue was used as a negative control. A semiquantitative four-stage grading was used (0%, 1%-5%, 6%-20%, >20% positive cells) and an average number of 1.500 cells/section was considered. Normal bronchial tissue was completely negative for HER-2/neu expression while 21/120 (16%) tumour specimens were positive (range 1%->20%). HER-2/neu positivity did not differ significantly among surgical stages (17% stage I, 24% stage II, 10% stage IliA) and histotypes (17% squamous cell carcinoma, 16% adenocercinoma, 18% large cell carcinoma). Using a cut-off value of 5% positive cells, 15 (12.5%) tumour specimens were above this value and median survival time (85 vs. 179 weeks) and 5-year survival rate (6% vs. 43%) (p < .01) were significantly lower in patients with >5% HER-2/neu positive turnouts. After a minimum follow up period of 2 years following radical surgery the survival probability was always significantly lower in the >5% HER-2/neu positive subgroup (3-yr survival rate: 14% vs. 53%, p < .009; 4-yr survival rate: 6% vs. 47%, p < .007). Our findings suggest that in NSCLC the negative impact of HER-2/neu expression on survival is maintained also in the long-term follow up of radically resected patients and therefore it could be a valuable prognostic factor as well as a potential target for biologic therapies
[•
Role of p53 positivity on the survival of patients with lung cancer as assessed by a systematic review of the literature M. Paesmans, E. Steels, T. Berghmans, F. Branle, A. Burniat, L. Ghisdal, J.J. Lafitte, F. Lemaitre, C. Mascaux, A.P. Meert, F. Vallot, J.P. Sculler. For the European Lung Cancer Working Party (ELCWP);
Institut Jules Bordet, Bruxelles, Belgium p53 status (protein expression or gene abnormalities) is one of the most studied "new" biological markers in lung cancer but its role is highly controversial and justifies the conduct of a systematic review of the literature. We selected, for our review, published studies with the objective to aggregate the available survival results after a methodological assessment of the trial design, methods (including laboratory) and analysis using a scale specifically designed by ELCWP. To be eligible, a trial had to deal with p53 assessment on lung cancer (primary site) only and to mention a survival comparison for p53 status. Among 74 studies, 30 identified P53 positivity as a poor prognostic factor and 56 provided sufficient data to allow survival results aggregation, both features being however associated (p = 0.005) but having no impact on the methodological score as on the laboratory technique. The studies were categorized according to histology, disease stage, therapeutic decision and laboratory technique with the following results for the relevant groups (all relative to non-small cell lung cancer); see the Table. Conclusion: We found consistently, in each a priori planned subgroup, that p53 positivity is associated with a poorer prognosis but our results are only univariate and subject to publication bias that we could not avoid.
Group
N trials
N patients HR
95% CI
Stages I-II Stages I-IliA Surgical stages Stages Ill-IV All stages Squamous cell Adenocarcinoma Immunohistochemistry Ab 1801 Immunohistochemistry Ab DO7 Molecular biology
19 6 21 9 11 9 9 8
2580 551 2020 332 1032 518 905 1035
1.52 1.54 1.48 1.68 1.44 1.37 2.24 1.57
1.32-1.70 1.16-2.06 1.29-1.69 1.36-2.18 1.20-1.72 1.02-1.85 1.70-2.95 1.26-1.91
17
2072
1.25
1.10-1.43
14
1061
1.64
1.36-1.99
[•
Correlation of outcome of resected non-small-cell lung cancer (NSCLC) with p53 mutations detected by yeast functional assay
M. Monz61, A. O'Brate 1, J. S~lnchez2, E. Sancho 1, I. Rosas 1, C. Aracil 1, M. Guillot 1, J.J. Guti~rrez 1, I. Escobar I , J. Astudillo I , R. Rosell 1. I Hospital Germans Trias i Pujol, Badalona, (Barcelona);
2Free University of Madrid, Madrid, Spain Several studies of p53 mutations have reported varying results regarding their negative effect on survival of resected non-small-cell lung cancer (NSCLC) patients. This conflict could be due to the fact that the COOH-terminal domain was not examined in many studies, though it has been reported that COOH-terminal domain mutants fail to transactivate p21, Bax and IGF-BP3 transcriptionally. To elucidate this issue, we have used a yeast-based assay measuring the transactivation function of cloned p53 to test for p53 mutations. RNA was extracted from 68 surgical lung tissue samples, using RT-PCR. Exons 2-11 of the p53 gene were amplified and cloned in vector pRDI22 and transfected into yeast ylG397 grown in an adenine-poor medium. More than 100 colonies were examined. In this type of assay, the yeast changes color according to the p53 status; wild-type p53 produces white colonies, while mutations produce red and pink colonies. Plasmids from the red and pink colonies were extracted from the yeast and sequenced to confirm mutations. Mutant colonies were found in 34/68 (50%) patients. No differences were observed according to gender, smoker status, performance status, weight loss, primary site (except a slightly greater incidence in the upper right lobe). Similar frequencies were also observed in adenocarcinomas (10/25 [40%]) and squamous cell carcinomas (20/35 [57.1%]), and no differences were found according to TNM: 12/20 (60%) T2N0 and 5/10 (50%) T2N2. Disease-free survival was 33 months (95%CI, 25.1-40.8) for patients with mutant colonies and 43.7 months (95%CI, 13.1-74.2) for patients without mutant colonies. We will present sequencing data and potential relevance of missense mutations and null mutations, especially those identified in the COOH-terminal domain.
~-~
Gene therapy with intratumor (IT) injection of an adenovirus expressing the IL2 gene (rAd-lL2) in lung cancer: Results of a phase I study
B. Escudier, T. Le Chevalier, E. Angevin, F. Griscelli. Institut Gustave
Roussy, Villejuif, France Once a single IT injection of rAd-bGAL was proven safe, we performed a phase I study using a rAd-lL2. Nine patients (pts) with inoperable NSCLC received a single IT injection of rAd-lL2 (107, 108 and 109 pfu, 3 pts at each dose) without concomitant- therapy. Virus shedding (293 cell culture and rlL2PCR) was verified in body fluids after injection. Control biopsies were performed at days 7, 14 and 28 to detect the presence of rAd-lL2, and to measure both native and recombinant IL-2 mRNA expression using a highly sensitive quantitative RT-PCR
184
•
Long-term effect of HER-2/neu expression on survival of stage I-IliA non-small cell lung cancer (NSCLC) patients radically resected
G. Selvaggi, S. Novello, G. Ferrari, P. Borasio, C. Mossetti, F. Ardissone, P. Lausi, G.V. Scagliotti. University of Torino, Orbassano;
S. Luigi Hospital, Orbassano; University of Torino, Department of Clinical and Biological Sciences, Orbassano, Italy In this study we investigated the long-term prognostic value of HER2/neu expression in radically resected NSCLC and its correlation with overall survival. Surgical specimens obtained from patients (n = 120) who underwent radical resection for NSCLC between January 1991 and February 1992 (squamous cell carcinoma 60 [46%], adenocarcinoma 48 [37%] and 22 large cell carcinoma [17%]) and surgically staged as stage I = 41 (31%), II = 37 (29%) and IliA = 52 (40%) were investigated for the presence and distribution of HER-2/neu using an avidin-biotin complex immunohistochemical technique. Obtained data were correlated to clinical and histopathologic variables. Normal bronchial tissue was used as a negative control. A semiquantitative four-stage grading was used (0%, 1%-5%, 6%-20%, >20% positive cells) and an average number of 1.500 cells/section was considered. Normal bronchial tissue was completely negative for HER-2/neu expression while 21/120 (16%) tumour specimens were positive (range 1%->20%). HER-2/neu positivity did not differ significantly among surgical stages (17% stage I, 24% stage II, 10% stage IliA) and histotypes (17% squamous cell carcinoma, 16% adenocercinoma, 18% large cell carcinoma). Using a cut-off value of 5% positive cells, 15 (12.5%) tumour specimens were above this value and median survival time (85 vs. 179 weeks) and 5-year survival rate (6% vs. 43%) (p < .01) were significantly lower in patients with >5% HER-2/neu positive turnouts. After a minimum follow up period of 2 years following radical surgery the survival probability was always significantly lower in the >5% HER-2/neu positive subgroup (3-yr survival rate: 14% vs. 53%, p < .009; 4-yr survival rate: 6% vs. 47%, p < .007). Our findings suggest that in NSCLC the negative impact of HER-2/neu expression on survival is maintained also in the long-term follow up of radically resected patients and therefore it could be a valuable prognostic factor as well as a potential target for biologic therapies
[•
Role of p53 positivity on the survival of patients with lung cancer as assessed by a systematic review of the literature M. Paesmans, E. Steels, T. Berghmans, F. Branle, A. Burniat, L. Ghisdal, J.J. Lafitte, F. Lemaitre, C. Mascaux, A.P. Meert, F. Vallot, J.P. Sculler. For the European Lung Cancer Working Party (ELCWP);
Institut Jules Bordet, Bruxelles, Belgium p53 status (protein expression or gene abnormalities) is one of the most studied "new" biological markers in lung cancer but its role is highly controversial and justifies the conduct of a systematic review of the literature. We selected, for our review, published studies with the objective to aggregate the available survival results after a methodological assessment of the trial design, methods (including laboratory) and analysis using a scale specifically designed by ELCWP. To be eligible, a trial had to deal with p53 assessment on lung cancer (primary site) only and to mention a survival comparison for p53 status. Among 74 studies, 30 identified P53 positivity as a poor prognostic factor and 56 provided sufficient data to allow survival results aggregation, both features being however associated (p = 0.005) but having no impact on the methodological score as on the laboratory technique. The studies were categorized according to histology, disease stage, therapeutic decision and laboratory technique with the following results for the relevant groups (all relative to non-small cell lung cancer); see the Table. Conclusion: We found consistently, in each a priori planned subgroup, that p53 positivity is associated with a poorer prognosis but our results are only univariate and subject to publication bias that we could not avoid.
Group
N trials
N patients HR
95% CI
Stages I-II Stages I-IliA Surgical stages Stages Ill-IV All stages Squamous cell Adenocarcinoma Immunohistochemistry Ab 1801 Immunohistochemistry Ab DO7 Molecular biology
19 6 21 9 11 9 9 8
2580 551 2020 332 1032 518 905 1035
1.52 1.54 1.48 1.68 1.44 1.37 2.24 1.57
1.32-1.70 1.16-2.06 1.29-1.69 1.36-2.18 1.20-1.72 1.02-1.85 1.70-2.95 1.26-1.91
17
2072
1.25
1.10-1.43
14
1061
1.64
1.36-1.99
[•
Correlation of outcome of resected non-small-cell lung cancer (NSCLC) with p53 mutations detected by yeast functional assay
M. Monz61, A. O'Brate 1, J. S~lnchez2, E. Sancho 1, I. Rosas 1, C. Aracil 1, M. Guillot 1, J.J. Guti~rrez 1, I. Escobar I , J. Astudillo I , R. Rosell 1. I Hospital Germans Trias i Pujol, Badalona, (Barcelona);
2Free University of Madrid, Madrid, Spain Several studies of p53 mutations have reported varying results regarding their negative effect on survival of resected non-small-cell lung cancer (NSCLC) patients. This conflict could be due to the fact that the COOH-terminal domain was not examined in many studies, though it has been reported that COOH-terminal domain mutants fail to transactivate p21, Bax and IGF-BP3 transcriptionally. To elucidate this issue, we have used a yeast-based assay measuring the transactivation function of cloned p53 to test for p53 mutations. RNA was extracted from 68 surgical lung tissue samples, using RT-PCR. Exons 2-11 of the p53 gene were amplified and cloned in vector pRDI22 and transfected into yeast ylG397 grown in an adenine-poor medium. More than 100 colonies were examined. In this type of assay, the yeast changes color according to the p53 status; wild-type p53 produces white colonies, while mutations produce red and pink colonies. Plasmids from the red and pink colonies were extracted from the yeast and sequenced to confirm mutations. Mutant colonies were found in 34/68 (50%) patients. No differences were observed according to gender, smoker status, performance status, weight loss, primary site (except a slightly greater incidence in the upper right lobe). Similar frequencies were also observed in adenocarcinomas (10/25 [40%]) and squamous cell carcinomas (20/35 [57.1%]), and no differences were found according to TNM: 12/20 (60%) T2N0 and 5/10 (50%) T2N2. Disease-free survival was 33 months (95%CI, 25.1-40.8) for patients with mutant colonies and 43.7 months (95%CI, 13.1-74.2) for patients without mutant colonies. We will present sequencing data and potential relevance of missense mutations and null mutations, especially those identified in the COOH-terminal domain.
~-~
Gene therapy with intratumor (IT) injection of an adenovirus expressing the IL2 gene (rAd-lL2) in lung cancer: Results of a phase I study
B. Escudier, T. Le Chevalier, E. Angevin, F. Griscelli. Institut Gustave
Roussy, Villejuif, France Once a single IT injection of rAd-bGAL was proven safe, we performed a phase I study using a rAd-lL2. Nine patients (pts) with inoperable NSCLC received a single IT injection of rAd-lL2 (107, 108 and 109 pfu, 3 pts at each dose) without concomitant- therapy. Virus shedding (293 cell culture and rlL2PCR) was verified in body fluids after injection. Control biopsies were performed at days 7, 14 and 28 to detect the presence of rAd-lL2, and to measure both native and recombinant IL-2 mRNA expression using a highly sensitive quantitative RT-PCR