A phase II study of CPT-11, a new derivative of camptothecin, for previously untreated non-small-cell lung cancer.

PURPOSE: Camptothecin-11 (CPT-11) is a new semisynthetic derivative of CPT, and has been shown to inhibit DNA topoisomerase I and to have a strong antitumor activity with low toxicity in murine tumors. To evaluate the effectiveness of CPT-11 in patients with non-small-cell lung cancer (NSCLC), a phase II study was conducted between April 1989 and February 1990. PATIENTS AND METHODS: Seventy-three patients were entered onto the study. All patients had had no previous therapy and had measurable disease. Their median age was 67 years (range, 34 to 75 years). Fifty-four patients had a performance status (PS) of 0 or 1 on the Eastern Cooperative Oncology Group (ECOG) scale, and 19 had a PS of 2. CPT-11 was given at a dose of 100 mg/m2 by intravenous 90-minute infusion once a week. The dose of CPT-11 was modified based on the WBC count obtained on the day of drug administration. RESULTS: Of 72 assessable patients, 23 (31.9%) showed a partial response (95% confidence interval, 20.2% to 43.6%). Of 40 patients with a stage IV disease, 13 (32.5%) responded. Response rates for patients with PS 0 or 1 and those with PS 2 did not differ (34.0% and 26.3%, respectively). The median duration of response in patients showing a PR was 15 weeks. The median survival time for all patients was 42 weeks. The major toxicities were leukopenia and diarrhea. Grade 3 or 4 leukopenia and diarrhea occurred in 18 patients (25%) and 15 patients (21%), respectively. These toxicities were unpredictable. Other toxicities of greater than or equal to grade 3 included nausea/vomiting (22%), anemia (15%), alopecia (4%) and pneumonitis (3%). One patient died of pulmonary toxicity (interstitial pneumonitis). CONCLUSIONS: CPT-11 is a very active agent for NSCLC with acceptable toxicities. Further trials in combination with other agents for this disease are warranted.

Restriction fragment length polymorphism of the human CYP2E1 (cytochrome P450IIE1) gene and susceptibility to lung cancer: possible relevance to low smoking exposure.

Polymorphic metabolism of certain chemical carcinogens may result in differences in susceptibility to cancers. Human CYP2E1 (cytochrome P450IIE1) is an enzyme involved in the metabolic activation of precarcinogens such as nitrosamines. We detected a restriction fragment length polymorphism (RFLP) of the human CYP2E1 gene for the restriction endonuclease Dra I. The distribution of this polymorphism was examined among lung cancer patients (n = 91), patients with cancer of the digestive tract (n = 45) and controls (n = 76). A significant difference in the distribution was observed between lung cancer patients and controls (chi 2 = 11.4 with 2 df; p < 0.005). On the other hand, there was no significant difference between patients between cancer of the digestive tract and controls (chi 2 = 4.87 with 2 df; NS). This finding suggests that the Dra I polymorphism of the CYP2E1 gene is associated with susceptibility to lung cancer. In addition, an association was found between the amount of lifelong smoking exposure and the distribution of the genotypes of the RFLP among lung cancer patients. The distribution pattern seemed deviated from that of controls especially in the population of low smoking exposure. Our Northern blot analysis data using RNA from human liver autopsy samples suggest that the Dra I polymorphism might be associated with the gene expression of CYP2E1 at mRNA level.

Increased levels of mitochondrial DNA in an etoposide-resistant human monocytic leukaemia cell line (THP-1/E).

Electron microscopic observations of THP-1/E (an etoposide-resistant human monocytic leukaemia cell line) showed a remarkable change of mitochondrial structure. Mitochondria were swollen and cristae were relatively intact. There was no difference in the activity of cytochrome oxidase, an enzyme which contains three subunits coded by mitochondrial DNA (mtDNA) between THP-1/E and THP-1 (the parent cell of THP-1/E). No measurable quantitative change of mitochondrial RNA was observed, but the level of mtDNA in THP-1/E was increased by a factor of about 4 compared with that of mtDNA in THP-1. These results suggest that, on acquisition of resistance to etoposide, some factors affect mitochondria, change its morphology and amplify its DNA.

Lung clearance mechanisms in obstructive airways disease.

Using radioaerosol inhalation lung cinescintigraphy, pulmonary clearance mechanisms were studied in 21 patients with obstructive airways disease. In none of them did we find homogeneous deposition of inhaled radioaerosol in the lungs, or a steady, constant, axial, and cephalad transport of radioactivity in the major airways. Of the 21 patients, 14 showed temporary but frequent stopping and starting of radioactivity in the airways in the course of lung clearance; in ten there was reversal of flow; in five migration of radioactivity from one bronchus into the opposite, bypassing the trachea and often followed by shuttling between right and left bronchi; and in four there was spiral or zigzag transport of radioactivity. The overall lung retention ratio in the first 2 hr was not abnormal, but the airway deposition ratio was significantly above normal, and airway clearance efficiency was below. The alveolar deposition ratio was also significantly smaller in these patients.

Mucociliary clearance and transport in bronchiectasis: global and regional assessment.

Global and regional mucociliary clearance and transport in the lungs was studied in 20 patients with bronchiectasis by radioaerosol inhalation lung cine-scintigraphy and the quantitative analysis following inhalation of ultrasonically-generated 99mTc-tagged human serum albumin aerosol (mass median diameter; 1.93 microns with geometric s.d. of 1.52). In bronchiectatic lung regions, deposition of inhaled aerosol was diminished or inhomogeneous. Transport of inhaled radioactivity from the bronchiectatic regions was deranged in 95% of the patients (19/20). The following abnormal mucous transport patterns were regionally observed; stasis in 12 of the 20 patients (12/20), regurgitation or reversed transport in 14/20, straying in 8/20, spiral or zigzag transport in 1/20, and/or various combinations of these four abnormal transport patterns. When coughs occurred, regurgitation and stray became more marked in the bronchiectatic regions. These regional abnormalities in mucociliary transport seem to be responsible for the development of infections and hemoptysis in the bronchiectatic regions.

Cloning of 67-kDa laminin receptor cDNA and gene expression in normal and malignant cell lines of the human lung.

Cell-adhesive protein laminin and its specific receptor play an important role in the processes of cancer proliferation, invasion and metastasis. In the present study, we cloned the cDNAs of the 67-kDa laminin receptor both from a human lung cell line (IMR90) and from a human lung cancer cell line (SBC3), and determined the nucleotide sequences. In comparison with both cDNA sequences of the protein-coding region, three nucleotide differences were found. These differences in the secondary structure of the protein, however, were caused by nucleotide substitutions. It was also demonstrated that the level of 67-kDa-laminin receptor mRNA was higher in SBC3 than in IMR90.

Variety of laminin expressions in murine neoplastic cell-lines - neuroblastoma na cells produce only laminin-b2 chain.

In the present study, we investigated the expression of laminin in three murine neoplastic cell lines; 3LL-SA (Lewis lung carcinoma), NA (neuroblastoma) and F9 (teratocarcinoma). Both Western and Northern blot analyses demonstrated that parietal endoderm-like F9 expressed three laminin chains A, B1 and B2. On the other hand, 3LL-SA cells synthesized two laminin chains B1 and B2, and NA cells only B2 chain. The analyses of the restriction fragment length polymorphism indicated that the genes for coding regions of all chains were present and grossly intact both in 3LL-SA and in NA just as in F9. These findings suggest that expression of laminin seems to be transcriptionally regulated in each neoplastic cell line specifically. Since these cell lines produce different forms of laminin, they can be used for investigation of the multifunctions of laminin molecule.

The corrected collagen content in paraquat lungs.

Biochemical analysis of collagen in a fibrosing lung often betrays its collagen-rich appearance in histopathology. This is due to the liability of lung tissue to extreme change in weight or volume in the presence of edema, hemorrhage, or other lesions, producing a great error in the biochemical data obtained from it. On morphometry of arterial density in lung section, we established a method for calculating the degree to which a given lung is transformed from a standard state of expansion, according to which we may correct for the content of collagen per lung volume. This method was applied to 14 autopsy paraquat lungs with various grades of fibrosis. Increase in collagen was shown to begin at about the 20th day of intoxication and advance thereafter, a finding in good accordance with lung morphology. The longer the survival, the less uniform the intrapulmonary distribution of collagen; even a severely fibrosing lung retained non-fibrotic areas.

A phase II study of vinorelbine, a new derivative of vinca alkaloid, for previously untreated advanced non-small cell lung cancer. Japan Vinorelbine Lung Cancer Study Group.

To evaluate the effectiveness of vinorelbine (NVB) in patients with non-small cell lung cancer (NSCLC), a late Phase II study was conducted. A total of 80 patients with Stage III or IV NSCLC who had no previous therapy were entered into the study. Seventy-nine patients were eligible for response and toxicity. NVB was administered weekly by intravenous injection at a dose of 25 mg/m2 in 20 ml of saline and was generally administered in four cycles or more, unless patients had disease progression. Of the 79 eligible patients, 23 (29.1%) showed a partial response (95% confidence interval, 19.1-40.4%). The median duration of partial responses was 14.7+ weeks. The median survival time for all patients was 40.1+ weeks. The major toxicity was leukopenia. Grade 3 and 4 leukopenia occurred in 48 patients (60.8%). Other toxicities of grade 3 or more included anemia (6.3%), local cutaneous reaction (3.8%), pneumonitis (1.3%), nausea and vomiting (1.3%), mucositis (1.3%) and constipation (1.3%). The absolute dose-intensity of NVB was 22.33 mg/m2/week. A weekly schedule of intravenous administration of 25 mg/m2/week of NVB was reasonable for maintenance of activity, and acceptable for toxicity in the chemotherapy of advanced NSCLC.

Pattern of fibronectin distribution in human lung cancer.

The pattern of fibronectin (FN) distribution in human lung cancer was studied by indirect immunofluorescent staining, and by the peroxidase-antiperoxidase method in a total of 60 surgical specimens. They comprised 8 small cell carcinomas, 4 large cell carcinomas, 19 squamous cell carcinomas, 28 adenocarcinomas, and 1 adenoid cystic carcinoma. Of the 60 specimens 13 were FN-positive. They included 4 large cell carcinomas, 4 small cell carcinomas, 3 poorly differentiated squamous cell carcinomas, and 2 poorly differentiated and 1 moderately differentiated adenocarcinomas. On the other hand, none of the well differentiated carcinomas was FN-positive around tumor cells. Our data suggest that undifferentiated, or poorly differentiated carcinomas of the lung tend to be FN-positive.

Application of an air-filled tube for measuring intraesophageal pressure.

We devised a new method for measuring esophageal pressure (Pes) with use of a flexible tube without a balloon at a constant rate of airflow through the tube into the esophagus (balloonless method). A study with 133Xe showed that the air that accumulated in the esophagus did not interfere with the measurement of Pes. We measured dynamic compliance (Cdyn) and pulmonary resistance (RL) with the balloonless method in 19 subjects and obtained a static deflation pressure-volume curve (P-V curve) in 10 other subjects. Cdyn was 0.243 +/- 0.099 l/cmH2O and RL was 1.52 +/- 0.42 cmH2O.l-1.s. In 6 of the 10 subjects, a P-V curve was also obtained with the balloon tube (balloon method). K, the index of compliance in the exponential function V = V0(1-e-KP) where V0 is volume at infinite pressure, was 0.136 +/- 0.040 cmH2O-1 with the balloonless method and 0.153 +/- 0.023 cmH2O-1 with the balloon method. No statistically significant difference was found between these two values. In conclusion, Cdyn, RL, and the P-V curve can be obtained precisely with the balloonless method.

The use of granulocyte colony-stimulating factor to shorten the interval between cycles of mitomycin C, vindesine, and cisplatin chemotherapy in non-small-cell lung cancer.

We investigated the possibility of shortening the interval between courses of the commonly prescribed 28-day MVP (mitomycin C, vindesine, and cisplatin) regimen in patients with non-small-cell lung cancer (NSCLC). We conducted a nonrandomized phase II study using recombinant human granulocyte colony-stimulating factor (G-CSF, Chugai) to explore the possibility of shortening the cycle length to 21 days and compared the results with those obtained in historical controls who had received the standard 28-day regimen. A total of 40 patients, 37 of whom were evaluable, were entered in the 21-day treatment group of the trial and were compared with 38 historical controls who had received standard 28-day cycles of MVP at our institution. Patients in the 21-day group received mitomycin C at 8 mg/m2 on day 1, vindesine at 3 mg/m2 on days 1 and 8, and cisplatin at 80 mg/m2 on day 1, with the schedule being repeated every 21 days. Controls had received the same regimen, albeit at 28-day intervals. G-CSF was given s.c. to the patients in the 21-day group at a daily dose of 2 micrograms/kg from day 2 to day 21 of every MVP cycle. The administration of G-CSF to these patients accelerated neutrophil recovery as compared with that observed in the historical controls. Significant differences were found between the two groups in terms of mean neutrophil nadirs (2666/microliters in the first cycle and 1369/microliters in the second for the G-CSF group vs 416/microliters in the first cycle and 685/microliters in the second cycle for the control group; P < 0.0001) and the mean duration of neutropenia (< or = 1000/microliters; 1.0 day in the first cycle and 1.7 days in the second for the G-CSF group vs 8.0 days in the first cycle and 6.9 days in the second for the control group; P < 0.0001). This enabled 32 (86%) of 37 patients in the G-CSF group to complete > or = 2 cycles on schedule. In 10 patients, the bone marrow aspirates taken after G-CSF administration showed increases in band neutrophil and myelocyte percentages. In conclusion, MVP treatment of patients with NSCLC at 21-day intervals is possible with the support of G-CSF.

Flow through a venous valve and its implication for thrombus formation.

To elucidate the possible connection between the flow patterns in the pockets of venous valves and thrombus formation, detailed studies of the behavior of model particles and red cells flowing through a venous valve have been carried out using isolated transparent dog saphenous veins containing two-leaflet valves, and cinemicrographic techniques. It was found that large paired vortices, located symmetrically on both sides of the bisector plane of the valve leaflets, were present in each valve pocket under physiological flow conditions. Particles continually entered the valve pockets from the mainstream, spending long periods of time describing a series of spiral orbits of decreasing diameter, while moving away from the bisector plane, and eventually left the vortex, rejoining the mainstream. With concentrated suspensions of red cells, it was found that another smaller counter-rotating secondary vortex, driven by the large primary vortex existed deep in each valve pocket. The concentration of red cells in this secondary vortex remained appreciably lower than that in the mainstream. In such regions, fluid circulated with extremely low velocities, thus creating a very low shear field which allowed red cells to form aggregates. The results suggest that in some pathological states, the valve-pocket vortices could act as automatic traps and generators of thrombi in a fashion similar to that previously demonstrated in an annular vortex formed downstream from a sudden tubular expansion.

Role of the posterior hypothalamus in the development of acute brain swelling.

Destructive stereotaxic lesions were made in the posterior hypothalamus, unilaterally or bilaterally, in 26 dogs. In 21 dogs the intracranial pressure (ICP) was maintained in normal range, and in five dogs the ICP was artificially elevated to 300 to 400 mm h2o, preceding the procedures to the hypothlamus. Arterial oxygen and carbon dioxide pressure ((PaO2 and PaCO2) were maintained in the normal range. Before and after each procedure, systemic arterial pressure (SAP) was elevated by intravenous injection of norepinephrine (5 X 10(-3) mg/kg) to determined whether the ICP increases coincidentally with elevation of the systemic arterial pressure. The intracranial pressure/mean arterial pressure ratio of elevation by injection of norepinephrine was not significant regardless of the level of the ICP, or of uni-or bilateral lesions of the hypothalamus. The authors conclude that dysfunction of posterior hypothalamus does not play a specific role in the development of vasomotor paralysis leading to acute brain swelling, under conditons of normal or moderately raised ICP with normal PaCO2 and Pato2 levels.

Reduced topoisomerase II-mediated DNA cleavage in VP-16 resistant human leukemic cell line.

Drug resistance in the VP-16 resistant human leukemic cell line (THP-1/E) was studied the possible relevance of topoisomerase II activity. Strand-passing activity in crude nuclear extract from sensitive and resistant cells was comparable and equally sensitive to inhibition by VP-16. However, it was demonstrated that VP-16-mediated pBR322 DNA cleavage in the presence of nuclear extract from resistant cells was reduced to one-tenth of that from sensitive cells. These results suggested that the resistance of THP-1/E cells to VP-16 was due to reduced DNA cleavage activity.

Total respiratory system compliance after thoracoplasty.

Recent attention has been directed towards the deleterious effects of the sequelae of pulmonary tuberculosis on respiratory function. Thus, a study was undertaken to find out to what extent the deformity of thorax after thoracoplasty increases the mechanical work of breathing. Compliance of the total respiratory system (Crs) was measured with a pressure-type body plethysmograph in nine post-thoracoplasty tuberculosis patients, 15 patients with pulmonary tuberculosis and three patients with pleural diseases. Mean Crs in nine post-thoracoplasty tuberculosis patients was 0.0861 cmH2O-1, and that in six patients with moderately advanced pulmonary tuberculosis was 0.0841 cmH2O-1. Mean Crs (0.1301 cmH2O-1) in nine patients with a minimum lesion of tuberculosis was not different from the reference Crs values reported to date by several authors. Crs was significantly correlated with vital capacity, but not with functional residual capacity. There was no difference in specific Crs between post-thoracoplasty tuberculosis patients and patients with moderately advanced pulmonary tuberculosis. Crs of patients with a pleural disease was smaller than Crs of patients with a minimal lesion of pulmonary tuberculosis. The deformity of thorax after thoracoplasty caused a decrease of Crs. However, ventilation is not as severely impaired as anticipated from the degree of thorax deformity on chest X-ray film, providing that movement of the diaphragm is unrestricted.

Flow patterns at the major T-junctions of the dog descending aorta.

Using a novel technique developed in our own laboratory, an isolated transparent arterial segment containing the whole descending aorta and its four major branches was prepared from a dog. The flow patterns at each aortic T-junction were studied in detail under the conditions of steady flow by means of flow visualization and cinemicrographic techniques. It was found that a standing recirculation zone consisting of a pair of thin-layered spiral secondary flows located symmetrically about the common median plane of the aorta and side branches was formed at each T-junction over a wide range of flow conditions including the time-averaged estimated mean values of physiological flow rates and flow rate ratios. The results support the recent in vivo findings by other investigators that flow reversal occurs at some junctions of the dog abdominal aorta during each cardiac cycle. The flow patterns at the aortic T-junctions were very much similar to those previously observed in various glass model T-junctions. However, due to the particular anatomical structure of the vessel wall at each branching site (the curvature of the wall was very sharp at the flow divider, but gently rounded at the bend opposite to it) no recirculation zone was formed in the side branches. At a given flow rate ratio, the measured critical Reynolds numbers for the formation of spiral secondary flows and fully developed disturbed flows were much higher in aortic T-junctions than those in glass model T-junctions having equivalent branching angles and diameter ratios. These results indicate that, in the circulation, conditions at arterial T-junctions appear to be optimal for minimizing the formation of disturbed flows.

Glycosaminoglycans and glycoproteins in bronchoalveolar lavage fluid from patients with pulmonary alveolar proteinosis.

Bronchoalveolar lavage fluid from two cases of pulmonary alveolar proteinosis were analyzed for glycosaminoglycans and glycoproteins. The clinical courses of the two cases were entirely different. In one patient, signs and symptoms recurred despite repeated therapeutic bronchoalveolar lavages. In the other patient, three successive bronchoalveolar lavages brought about complete recovery. It was found that the bronchoalveolar lavage fluid from the former case contained various subtypes of glycosaminoglycans [hyaluronic acid, chondroitin sulfate A(C), dermatan sulfate, and heparan sulfate] and glycoprotein. On the other hand, the bronchoalveolar lavage fluid from the latter case contained glycoprotein, but no detectable amounts of glycosaminoglycans. There was only a slight qualitative difference in glycoprotein of bronchoalveolar lavage fluid between the two cases. The presence or absence of glycosaminoglycans in bronchoalveolar lavage fluid may be related to the prognosis of pulmonary alveolar proteinosis.

Difference in inhaled aerosol deposition patterns in the lungs due to three different sized aerosols.

Deposition patterns of inhaled aerosol in the lungs were studied in five normal subjects and 20 patients with lung disease by inhaling radioaerosols with three different particle size distributions. These aerosols were generated from BARC, UltraVent, and Mistogen-EN-142. Particle size distributions generated by these three nebulizers were 0.84, 1.04 and 1.93 microns in activity median aerodynamic diameter (AMAD) with its geometric standard deviation (sigma g) of 1.73, 1.71 and 1.52, respectively. Deposition patterns of inhaled aerosols were compared qualitatively and quantitatively by studying six different parameters: alveolar deposition ratio (ALDR), Xmax, Xmean, standard deviation (S.D.), skewness and kurtosis of the radioactive distribution in the lungs following inhalation. It has been found that aerosol deposition patterns varied with particle size. The unevenness of aerosol deposition, Xmax, Xmean and the number of 'hot spots' became more prominent with the increase in particle size, whereas values of ALDR and S.D. decreased as particle size increased. Knowing these deposition characteristics would facilitate a judicious application of aerosol inhalation to medical use.

Technegas for inhalation lung imaging.

Technegas, an aerosol generator recently devised in Australia, produces aerosol particulates called 'technegas' which have characteristics of both an aerosol and a gas. The majority of the particulate is below 200 nm in size as measured by electron microscopy. Four normal subjects and 31 patients with various lung diseases were studied by imaging the lungs following inhalation of technegas. The penetration of inhaled technegas to the lung periphery was excellent; the average alveolar deposition ratio (ALDR) was 85%. Comparative studies with lung images obtained either with an ultrasonic nebulizer or jet nebulizers also confirmed better penetration of inhaled technegas to the lung periphery. There was no significant statistical difference in the ALDRs between normals and patients. Aerosol studies were comparable to perfusion counterparts, and evaluation of regional ventilatory status was greatly facilitated. Because of the large ALDR and the low airway deposition ratio (ADR), actual imaging could be done not only immediately after aerosol inhalation but also some time later without losing too much radioactivity from the lungs. One disadvantage was that technegas immediately after generation was anoxic.