Phase I study of a recombinant adenovirus-mediated gene transfer in lung cancer patients.

BACKGROUND: Despite vigorous efforts at curbing tobacco consumption and aggressive combined-modality treatment programs, both the incidence of and the mortality from lung cancer have remained virtually unchanged in the last 10 years. More effective innovative therapies are clearly needed. The direct transfer into tumor cells of tumor suppressor genes or toxic gene products that specifically promote tumor cell death and spare nonmalignant cells is a potentially novel anticancer treatment approach that should be investigated. PURPOSE: On the basis of compelling preclinical data, we initiated a phase I study involving six patients with inoperable lung cancer and an endobronchial lesion accessible by bronchoscopy. Our purpose was to evaluate the feasibility, tolerance, and clinical, biologic, and immunologic effects of the intratumoral administration of a recombinant, replication-deficient adenovirus (rAd.RSV beta-gal), using the Rous sarcoma virus promoter to drive transcription of the Escherichia coli lacZ marker gene that encodes for the bacterial enzyme beta-galactosidase (beta-gal). METHODS: From June 1994 through April 1995, six patients (five males and one female) were enrolled in the trial. A single dose of recombinant virus suspension containing 10(7) or 10(8) plaque-forming units (PFU) was injected intratumorally into two successive cohorts of three patients. Eligible patients received concomitant chemotherapy. Patients were kept under isolation conditions from 3 days before the injection was given until virus excretion was undetectable. Biopsy specimens of the tumor and surrounding mucosa were collected on the 8th day and at 1, 2, and 3 months after injection. They were analyzed by cell culture, polymerase chain reaction (PCR), and beta-gal expression for the presence of recombinant adenovirus. So that the risk of virus recombination or complementation could be minimized, wildtype adenovirus carriers among the hospital staff (identified by PCR) were excluded from contact with patients who were potentially excreting recombinant virus. RESULTS: beta-gal was expressed in tumor biopsy specimens of three patients (one who received the 10(7) PFU dose level and two who received 10(8)). Bronchoalveolar lavage specimens collected immediately after injection were positive for recombinant adenovirus when analyzed in culture and by PCR. All biologic fluids were negative for recombinant virus as judged by PCR after day 12, with the exception of bronchoalveolar lavage specimens (positive PCR up to 90 days in two of three patients treated with 10(8) PFU). The blood samples obtained from the three patients treated with 10(8) PFU showed positive PCR results immediately after virus injection. Patients were kept in isolation for a median of 17 days. The most common toxic effects were moderate bleeding (occurring in two patients) during bronchoscopy and fever (seen in four patients). Endoscopic and clinically objective antitumor responses were seen in four patients, including one patient who showed a complete response by pathologic evaluation. The median survival for the patients was 12.5 months (range, 3-16+ months). Throughout the study, hospital staff remained negative for recombinant adenovirus infection. CONCLUSIONS: This ongoing phase I study has demonstrated that a recombinant adenovirus-mediated marker gene, such as rAd.RSV beta-gal, can be safely introduced into humans and that the gene product is expressed by lung tumor cells of the host.

Aerosol-mediated delivery of recombinant adenovirus to the airways of nonhuman primates.

At present, it is conceivable that gene therapy of the cystic fibrosis airway epithelium is possible using the direct transfer of a functional human cystic fibrosis transmembrane conductance regulator (CFTR) gene to a wide variety of patients' tracheo-bronchial cells. Here we describe a novel approach (aerosolization) to deliver a replication-deficient adenovirus carrying the CFTR gene (Ad.CFTR) to the airways. Results obtained in vitro and in Rhesus monkeys suggest that the delivery of recombinant adenovirus as an aerosol is feasible and is not associated with severe toxicity after single or double administration depending on the Ad.CFTR dose. This study supports the concept of aerosolization as a delivery method for adenovirus-mediated lung gene therapy.

Aerosol administration of a recombinant adenovirus expressing CFTR to cystic fibrosis patients: a phase I clinical trial.

Ad CFTR, a replication-deficient adenovirus expressing the human cystic fibrosis transmembrane conductance regulator (CFTR), was administered by aerosolization in a single escalating dose to three pairs (cohorts) of cystic fibrosis (CF) patients. Buffer only was administered to the nose and lungs 9-14 days before nasal instillation of virus followed the day after by aerosolization of Ad CFTR to the lung. Nasal doses (defined in terms of viral plaque forming units, pfu) were 10(5), 10(7), and 4 x 10(8), whereas aerosolized doses were 10(7), 10(8), 5.4 x 10(8) for each cohort, respectively. No acute toxic effects were observed in the first 4 weeks after virus treatment. Shedding of infectious Ad CFTR was never detected, whereas detection of vector DNA sequences and CFTR expression demonstrated DNA transfer to the nose and airways of patients. No significant deviations in immunological and inflammatory parameters were observed in serum and in bronchoalveolar lavage (BAL). Importantly, for all patients, the serum anti-adenovirus antibody levels did not change significantly from baseline and no antibodies against adenovirus were found in BAL.

Object features, object locations, and object files: which does selective attention activate and when?

The representation within which attention operates was investigated in 3 experiments. The task was similar to that of R. Egly, J. Driver, and R. D. Rafal (1994). Participants had to detect the presence of a target at 1 of 4 ends of 2 shapes, differing in color and form. A precue appeared at 1 of the 4 possible corners. The 2 shapes occupied either the same or different locations in the cuing and target displays. The results showed that the cued object location was attended whether or not space was task relevant, whereas the cued object features (color and form) were attended only when these were task relevant. Moreover, when object file continuity was maintained through continuous movement, attention was found to follow the cued object file as it moved while also accruing to the cued location.

Perception of dyadic relationship and emotional states in patients with affective disorder.

BACKGROUND: Previous research has shown that interpersonal processes play a significant role in the development and maintenance of affective disorders. In this study, this claim was further investigated by comparing the perception of the dyadic relationship and judgment of other's emotions in affective disorder patients. METHOD: The sample included 39 couples (n=39 couples) with one of the partners suffering from an affective disorder and currently either in an acute or remitted depressive state. All participants completed four instruments, measuring the perceived quality of the dyadic relationship and the perception of other's emotions as reflected by judgments of facial expressions line drawings. RESULTS: While the level of marital satisfaction was found to be lower in the acute than in the remitted group both for ill partners and their spouses, spouses in both the acute and remitted group tended to be more critical of their ill partners. Patients who were depressed judged facial expressions significantly less positively than did remitted patients. Judgments of negative emotions were highly correlated between partners in the acute group, but uncorrelated in the remitted group. Acutely depressed patients were less sensitive to invitation than remitted patients, while their spouses displayed the opposite pattern. CONCLUSION: The present results shed further light on the interpersonal dynamics between depressed patients and their spouses by underscoring differences between couples with a remitted vs. acutely depressed partner in their perception of the dyadic unit and their judgments of facial emotions. LIMITATIONS: Longitudinal research is needed, in which the same patients are tested during periods of remission and acute episodes, as well as research investigating the role of patient gender in the perception of facial expressions of emotions.

A salient distractor does not disrupt conjunction search.

Leading theories of attention posit that bottom-up and top-down factors simultaneously affect attentional priority in visual search. Recent evidence, however, suggests that subjects may rely exclusively on top-down guidance when searching for a target defined by a specific known feature (Bacon & Egeth, 1994). In the present experiment, we addressed this issue in a conjunction search task. We investigated how searching for a green O among green Ts and red Os is affected by the presence of a distractor with a unique shape (green X), color (blue O), or both (blue X). We showed that the salient distractor does not disrupt performance on target-present trials, but produces a large interference on target-absent trials. We conclude that salience-based and top-down processes are alternative modes of guidance, rather than joint contributors in the allocation of attentional priority.

Object-based selection under focused attention: a failure to replicate.

In a recent study, Lavie and Driver (1996) reported that object-based effects found with distributed attention disappear when attention is focused on a narrow area of the display. This finding stands in contrast with previous reports of object-based effects under conditions of focused attention (e.g., Atchley & Kramer, 1998; Egly, Driver, & Rafal, 1994). The present study was an attempt to replicate Lavie and Driver's finding, using similar task and stimuli. While Lavie and Driver's object-based effect in the distributed attention condition was replicated, its absence in the focused attention condition was not. In the two experiments reported in this paper, object-based effects were found under conditions of both distributed and focused attention, with no difference in the magnitude of the object-based effects in the two conditions. It is concluded that, in contrast with Lavie and Driver's claim, the initial spatial setting of attention does not influence object-based constraints on the distribution of attention.

Attending to an object's color entails attending to its location: support for location-special views of visual attention.

Van der Heijden, Kurvink, de Lange, de Leeuw, and van der Geest (1996) argued that the results supporting the location-special view obtained by Tsal and Lavie (1988) were due to uncontrollable shifts of fixation, rather than reflecting the properties of the attentional system. In the present study, we present an improved variation of the Tsal and Lavie (1988) paradigm and reassert our claim that location is a special dimension. Subjects were presented with circular arrays of six letters of different colors. Three of the letters were enclosed by (Experiment 1) or superimposed on (Experiments 2, 3, and 4) different colored shapes. The subjects were instructed to report the (target) shape with a given color (e.g., report whether the red shape was a square, a circle, or a triangle) and then either freely report letters from the array (Experiments 1, 2, and 4) or identify a prespecified target letter (Experiment 3). In all four experiments, performance was substantially better for the letter that appeared in the location of the to-be-reported shape (location letter) than for the letter that shared its color (color letter). We conclude that attending to the stimulus color entails directing attention to its location.

Secretion of the hepatitis B virus surface antigen from mouse cells using an extra-chromosomal eucaryotic vector.

Recombinant DNA molecules which contained a subgenomic fragment of the hepatitis B virus (HBV) genome, the pML2 vector and the bovine papillomavirus type 1 (BPV) genome were constructed. The HBV fragment includes the entire transcription unit for the hepatitis B surface antigen (HBsAg). After propagation in Escherichia coli, the recombinant plasmids were cleaved with endonucleases SalI and PvuI to eliminate most of the bacterial sequences before transfection of mouse C127 cells. Foci were observed 10--14 days after transfection. Cells from selected foci were cloned and the supernatants were assayed for the presence of HBsAg. Most of the clones tested were found to secrete HBsAg particles into the growth medium. These particles appear to be similar to the 22 nm particles present in the serum of HBV chronic carriers. SDS-polyacrylamide gel electrophoresis revealed that the particles contain two polypeptides, probably representing the glycosylated and unglycosylated forms of the HBsAg major polypeptide. An analysis of DNA from the transformed clones revealed that they contain multiple extra-chromosomal copies of the recombinant, which, however, had suffered rearrangement.

[Aerosol administration of a replication defective recombinant adenovirus expressing normal human cDNA-CFTR in the respiratory tractus in patients with cystic fibrosis]. / Administration par aérosol d'un adénovirus recombinant déficient pour la réplication contenant cADN-CFTR humain normal dans le tractus respiratoire de patients atteints de mucoviscidose.

At present it is conceivable to think that gene therapy represents a way to treat or even prevent the respiratory manifestations of cystic fibrosis. Consistent to such a concept, there is sufficient evidence that Ad-CFTR, a recombinant replication-deficient adenovirus expressing the human cystic fibrosis transmembrane conductance regulator cDNA, can vectorize the expression of a functional CFTR (cystic fibrosis transmembrane conductance regulator) to the nasal and airway epithelia. The clinical protocol was designed to assess the safety of single escalating doses of a replication defective adenovirus expressing the cystic fibrosis transmembrane conductance regulator gene (Ad-CFTR) when administered to the tracheobronchial portion of the airways and whether biological efficacy of CFTR delivery could be demonstrated. Six cystic fibrosis patients received nasal instillation and subsequent aerosol (Optineb, Air Liquide, Paris, France) administration of Ad-CFTR the following day. Doses (pfu) applied to the nose were 10(5) (patients SG and PB), 10(7) (patients FP and EP) and 4 x 10(8) (patients DS and FG), while aerosolised doses were 10(7) (patients SG and PB), 10(8) (patients FP and EP) and 5.4 x 10(8) (patients DS and FG), respectively. No acute toxic effects, no increase in the titer of anti-adenovirus antibodies and no spreading or shedding of Ad-CFTR were detected. In one patient Ad-CFTR DNA was found in the urine and blood two days after aerosolisation. Ad-CFTR DNA was detected in nasal and bronchial brush samples, in BAL, in saliva and tonsils 21, 8, 14 and 4 days post virus administration, respectively. Ad-CFTR mRNA (RT-PCR on bronchial cells) and CFTR protein (immunochemistry on nasal and bronchial cells) were detected up to 14 days following Ad-CFTR administration. These results show that the nebulisation of Ad-CFTR is a possible approach for treating the respiratory manifestation of cystic fibrosis.