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Anti-Her2 MAb plus calicheamicin immunoconjugate linked on liposomal vinorelbine tartrate induces ADCC and apoptosis in HER2 positive NSCLC
Combined Modality Therapy day 1 and CDDP 75 mg/m2 day 1; Arm B, VNB 30 mg/m2 day 1 and 8 and CDDP 80 mg/m2 day 1. Cycles were repeated every 21 days. At the completion of neoadjuvant chemotherapy pts assessed to be resectable underwent thoracotomy, in unresectable pts radiation therapy was applicated. The minimal follow up was 24 months. The characteristics of 46 pts were as follows: Arm A 23 pts: median age 58.5 years, male/female 20/3, stage IIINlllB 10118, histology 18 squamous cell, 3 adenoca and 2 large cell. Arm B: median age 60.5 years, male/female 20/1, stage IllA/IIIB 10/18, histology 21 squamous cell and 2 adenoca. A total of 63169 cycles have been delivered in Arm A/B. No complete response was observed. 12 pts (52%)/12 pts (52%) had partial response, 6 pts (26%)/5 pts (22%) had stable, 5 pts (22%)/6 pts (26%) had progressive disease in Arm A/B. 8 pts (35%)/6 pts (26%) had a complete resection and the median survival was 455/451 days in Arm A/B. No significant difference in survival was found in Arm A comparing with Arm B. However, the median survival in nonresected pts was 201/422 in Arm A/B. The difference in survival between these groups was near the statistical significance (p = 0.0569). 26-35% of pts with stage IIIA-IIIB NSCLC could reach a complete resectability after neoadjuvant VNB + CDDP. No difference in survival was found between the two dose intensity regimens. The median survival in nonresected pts was longer in Arm B.
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Induction chemotherapy for stage III non-small cell lung cancer: The AASLC experience
R. Malayeri, G. Pohl, G. Krajnik, F. Eckersberger, W. Klepetko, R.M. Mi]ller, FM. Smolle-J~ttner, A. Maier, G. Ambrosch, A.M. R~ger, M. Woltsche, C. Sch~fer-Prokop, H. Huber, R. Pirker. Austrian Association for the Study of Lung Cancer, Department of Internal Medicine I, University of Vienna, Vienna, Austria Induction chemotherapy might improve survival in stage III non-small cell lung cancer. Here we report the experience of the Austrian Association for the Study of Lung Cancer (AASLC). We have treated 35 patients with NSCLC (17 IliA, 18 IIIB, 23 male, 12 female) with either VIP (vinorelbine 25 mg/m_ per day on days 1 & 8, ifosfamide 1000 mg/m_per day on days 1-3 and cisplatin 30 rag/re_per day on days 1-3) plus prophylactic G-CSF, or PIP (paclitaxel 175 mg/m_day 1, ifosfamide 1000 mg/m per day on days 1-3 and cisplatin 60 mg/m_day 1) plus prophylactic G-CSF. Both treatment protocols were repeated every 3 weeks. After 3 cycles, patients were assessed for response. Thirty-two patients are evaluable for response and toxicity. The response rate was 59% for all patients, 73% for those with stage IliA and 53% for those with stage IIIB. The response rates were 60% for VIP and 57% for PIP. Toxic/ties of these protocols were tolerable and manageable. Fifteen (47%) patients (9 IliA, 6 IIIB) underwent surgery and a complete tumor resection was possible in 12 of these patients. Patients with incomplete tumor resection, patients with residual positive N2 lymph nodes and patients who did not undergo surgery received local radiation. The 5-year survival rate of all patients was 32%. In conclusion, induction chemotherapy is feasible and shows promising long-term survival in patients with stage III NSCLC and should be further evaluated in randomised trials.
[40•
Anti-Her2 MAb plus calicheamicin immunoconjugate linked on liposomal vinorelbine tartrate induces ADCC and apoptosis in HER2 positive NSCLC
J. Giannios, E. Michailakis, P. Ginopulos. Dept. of Clinical Oncology, Regional Hospital of Patras, St. Andreas & Oncology DepL of Genera/ Hospita/ of Athens, Greece Overexpression of the HER2 proto-oncogene which encodes a 185 kDa protein frequently coincide with aggressive and chemoresistant NSCLC due to inhibition of PCD after chemotherapy. Also, it is associated with aneuploidy, p53 abnormalities, enhanced DNA repair and synthesis, cell growth, mitotic rate and tumourigenicity. Lung adenocarcinoma tissue was excised from a patient and tumour cells were isolated by the collagenase method. IHC has exhibited HER2
121
overexpression. We prepare immunoconjugates of anti-HER2 MAbs and calicheamicin which is an apoptotic antibiotic with up to 1000 fold greater potency than the most clinically used ant/cancer drugs. This immunoconjugate was linked onto pegylated DRV liposomes which contained vinorelbine tartrete. After treatment, we observed HER2 downregulation by IHC. TEM exhibited disruption of the microtubular cytoskeleton due to vinorelbine and MAb (Fc) directed killing of tumour cells by immune effector cells such as macrophages, neutrophils and lymphoid cells such as (K) and (NK) cells indicating antibody-dept, cellular cytotoxicity (ADCC). Biochemical assays such as MTT exhibited reduced metabolic activity, while BrdU has shown great reduction of DNA synthesis. Anti-ssDNA MAbs and binding of Annexin-V with phosphatidyl-serine IHC confirmed that apoptosis was the mechanism of growth inhibition. SEM and TEM showed violent membrane blebbing of apoptotic cells (zeiosis) and fragmentation of DNA creating a vacuolar nucleus due to calicheamicin damaging action after binding to the third end of oligopurine tracts causing strand breaks, respectively. Subsequently, tumour cells break into apoptotic bodies that adjacent cells recognize and phagocytose due to PS external/sat/on indicating a bystander killing effect (BKE). Concluding, by delivering intracellularly cytotoxic agents such as celicheamicin through conjugating to NSCLC associated ant/HER2 MAbs linked on liposomal vinorelbine tartrete, we can circumvent chemoresistance, induce ADCC, inhibit mitosis at metaphase and destroy tumourel DNA leading to irreversible D2 apoptotic signs with subsequent bystander killing of NSCLC.
delivery of adenovirus-p53 with 2[4-0-• Systemic methoxyestradiol inhibits lung metastasis in nude mice M. Kataoka, G. Schumacher, R.J. Cristiano, N.E. Atokinson, T. Fujiwara, N. Tanaka, J.A. Roth, T. Mukhopadhyay. Okayama University, Okayama, Japan, Charite Campus Virchow Kiln/k, Berlin, Germany, University of Texas MD Anderson Cancer Center, Houston, USA 2-methoxywstradiol is the normal mammalian metabolite of estradiol, which is known to has a function to inhibit the cancer cell growth by antitublin, antiangiogenesis activity, or stabilization of p53 protein. Low levels of gene expression following systemic delivery have impaired the effectiveness of tumor suppressor gene replacement in treating metastases. We asked whether combined treatment with 2-methxyestradiol (2-Me), which increases levels of wild-type p53 protein in cancer cells, and the systemic administration if an adenoviral vector expressing wildtype p53 (Ad-p53) would inhibit the growth of human metastatic lung cancer cells in vitro and in vivo. The expression of endogenous p53 protein of A549 (/non-small cell lung cancer cell line) increase 5-fold after 48 h of 2-Me treatment, accompanied by the induction of p21. The A549 cells were refractory to low doses (MOI = 1) of Ad-p53 mediated growth inhibition. However, when the Ad-p53 tranceduced cells were treated with 2-Me at non-inhibitory doses, their growth was inhibited 80% associated with induction of apoptosis. Intravenous injection of A549 cells makes the multiple lung metastasis in lungs of nude mice. The simultaneous administration of p53 and 2-Me resulted in a greater than additive reduction with the lung colony count reduced to 33% of its control value using this lung metastasis model. These results suggest that the synergistic effect of 2-Me and Ad-p53 in combination treatment may have application in the systemic treatment of cancer.
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Seamless lung cancer service provision: 2) A multidisciplinary approach to care
SM. Davidson, R. Jones, J. McPhelim, H. Towers, F. Bryden, D. Millan, I. Colquhoun, S. McKay, R. Milroy. On Behalf of the West of Scotland Lung Cancer Research Group; Stobhill Hospital, Glasgow, UK It is well recognised that optimal care for lung cancer patients includes multidisciplinary input and national guidelines now recommend this approach (ref. 1, ref. 2). We have developed a dedicated lung cancer
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Induction chemotherapy for stage III non-small cell lung cancer: The AASLC experience
R. Malayeri, G. Pohl, G. Krajnik, F. Eckersberger, W. Klepetko, R.M. Mi]ller, FM. Smolle-J~ttner, A. Maier, G. Ambrosch, A.M. R~ger, M. Woltsche, C. Sch~fer-Prokop, H. Huber, R. Pirker. Austrian Association for the Study of Lung Cancer, Department of Internal Medicine I, University of Vienna, Vienna, Austria Induction chemotherapy might improve survival in stage III non-small cell lung cancer. Here we report the experience of the Austrian Association for the Study of Lung Cancer (AASLC). We have treated 35 patients with NSCLC (17 IliA, 18 IIIB, 23 male, 12 female) with either VIP (vinorelbine 25 mg/m_ per day on days 1 & 8, ifosfamide 1000 mg/m_per day on days 1-3 and cisplatin 30 rag/re_per day on days 1-3) plus prophylactic G-CSF, or PIP (paclitaxel 175 mg/m_day 1, ifosfamide 1000 mg/m per day on days 1-3 and cisplatin 60 mg/m_day 1) plus prophylactic G-CSF. Both treatment protocols were repeated every 3 weeks. After 3 cycles, patients were assessed for response. Thirty-two patients are evaluable for response and toxicity. The response rate was 59% for all patients, 73% for those with stage IliA and 53% for those with stage IIIB. The response rates were 60% for VIP and 57% for PIP. Toxic/ties of these protocols were tolerable and manageable. Fifteen (47%) patients (9 IliA, 6 IIIB) underwent surgery and a complete tumor resection was possible in 12 of these patients. Patients with incomplete tumor resection, patients with residual positive N2 lymph nodes and patients who did not undergo surgery received local radiation. The 5-year survival rate of all patients was 32%. In conclusion, induction chemotherapy is feasible and shows promising long-term survival in patients with stage III NSCLC and should be further evaluated in randomised trials.
[40•
Anti-Her2 MAb plus calicheamicin immunoconjugate linked on liposomal vinorelbine tartrate induces ADCC and apoptosis in HER2 positive NSCLC
J. Giannios, E. Michailakis, P. Ginopulos. Dept. of Clinical Oncology, Regional Hospital of Patras, St. Andreas & Oncology DepL of Genera/ Hospita/ of Athens, Greece Overexpression of the HER2 proto-oncogene which encodes a 185 kDa protein frequently coincide with aggressive and chemoresistant NSCLC due to inhibition of PCD after chemotherapy. Also, it is associated with aneuploidy, p53 abnormalities, enhanced DNA repair and synthesis, cell growth, mitotic rate and tumourigenicity. Lung adenocarcinoma tissue was excised from a patient and tumour cells were isolated by the collagenase method. IHC has exhibited HER2
121
overexpression. We prepare immunoconjugates of anti-HER2 MAbs and calicheamicin which is an apoptotic antibiotic with up to 1000 fold greater potency than the most clinically used ant/cancer drugs. This immunoconjugate was linked onto pegylated DRV liposomes which contained vinorelbine tartrete. After treatment, we observed HER2 downregulation by IHC. TEM exhibited disruption of the microtubular cytoskeleton due to vinorelbine and MAb (Fc) directed killing of tumour cells by immune effector cells such as macrophages, neutrophils and lymphoid cells such as (K) and (NK) cells indicating antibody-dept, cellular cytotoxicity (ADCC). Biochemical assays such as MTT exhibited reduced metabolic activity, while BrdU has shown great reduction of DNA synthesis. Anti-ssDNA MAbs and binding of Annexin-V with phosphatidyl-serine IHC confirmed that apoptosis was the mechanism of growth inhibition. SEM and TEM showed violent membrane blebbing of apoptotic cells (zeiosis) and fragmentation of DNA creating a vacuolar nucleus due to calicheamicin damaging action after binding to the third end of oligopurine tracts causing strand breaks, respectively. Subsequently, tumour cells break into apoptotic bodies that adjacent cells recognize and phagocytose due to PS external/sat/on indicating a bystander killing effect (BKE). Concluding, by delivering intracellularly cytotoxic agents such as celicheamicin through conjugating to NSCLC associated ant/HER2 MAbs linked on liposomal vinorelbine tartrete, we can circumvent chemoresistance, induce ADCC, inhibit mitosis at metaphase and destroy tumourel DNA leading to irreversible D2 apoptotic signs with subsequent bystander killing of NSCLC.
delivery of adenovirus-p53 with 2[4-0-• Systemic methoxyestradiol inhibits lung metastasis in nude mice M. Kataoka, G. Schumacher, R.J. Cristiano, N.E. Atokinson, T. Fujiwara, N. Tanaka, J.A. Roth, T. Mukhopadhyay. Okayama University, Okayama, Japan, Charite Campus Virchow Kiln/k, Berlin, Germany, University of Texas MD Anderson Cancer Center, Houston, USA 2-methoxywstradiol is the normal mammalian metabolite of estradiol, which is known to has a function to inhibit the cancer cell growth by antitublin, antiangiogenesis activity, or stabilization of p53 protein. Low levels of gene expression following systemic delivery have impaired the effectiveness of tumor suppressor gene replacement in treating metastases. We asked whether combined treatment with 2-methxyestradiol (2-Me), which increases levels of wild-type p53 protein in cancer cells, and the systemic administration if an adenoviral vector expressing wildtype p53 (Ad-p53) would inhibit the growth of human metastatic lung cancer cells in vitro and in vivo. The expression of endogenous p53 protein of A549 (/non-small cell lung cancer cell line) increase 5-fold after 48 h of 2-Me treatment, accompanied by the induction of p21. The A549 cells were refractory to low doses (MOI = 1) of Ad-p53 mediated growth inhibition. However, when the Ad-p53 tranceduced cells were treated with 2-Me at non-inhibitory doses, their growth was inhibited 80% associated with induction of apoptosis. Intravenous injection of A549 cells makes the multiple lung metastasis in lungs of nude mice. The simultaneous administration of p53 and 2-Me resulted in a greater than additive reduction with the lung colony count reduced to 33% of its control value using this lung metastasis model. These results suggest that the synergistic effect of 2-Me and Ad-p53 in combination treatment may have application in the systemic treatment of cancer.
[•-•
Seamless lung cancer service provision: 2) A multidisciplinary approach to care
SM. Davidson, R. Jones, J. McPhelim, H. Towers, F. Bryden, D. Millan, I. Colquhoun, S. McKay, R. Milroy. On Behalf of the West of Scotland Lung Cancer Research Group; Stobhill Hospital, Glasgow, UK It is well recognised that optimal care for lung cancer patients includes multidisciplinary input and national guidelines now recommend this approach (ref. 1, ref. 2). We have developed a dedicated lung cancer