PROTOCOL THS 94-002 
REVISED 5/19/94 
PAGE II 
V 
7.0 EVALUATION DURING STUDY I 
7.1 Patients will have a CBC, platelet count, PT, PTT, SMA-12, electrolytes, and a chest x-ray prior to 
each course of therapy. Serum will be collected pre- and post-treatment for analysis of antibodies to 'I 
adenovirus proteins. i 
7.2 History and physical with performance status and weight should be recorded before each course. 
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7.3 The endobronchial tumors will be photographed bronchoscopically at the beginning of each course. 
Tumor measurements are to be recorded before each course for ail measurable tumors. 
7.4 All relevant information regarding drug dosage, tumor response, laboratory examinations, and 
treatment-related toxicities must be recorded before each treatment is given. 
7.5 Parameters to be Measured In Vitro 
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7.5.1 Fine needle aspirates or core biopsies wili be obtained of accessible local tumor. Cells in the ^ 
cytospins of pleural effusions will be obtained pre- and post-treatment when possibie. Tumor 
specimens wili be coliected 72 hrs. after injection of the adenovirus during the first treatment 
cycle. For endobronchial tumors bronchoscopic tumor and normal bronchial epithelial | 
biopsies will be obtained prior to the beginning of each course. Tissue will be fixed j 
immediately in 4% paraformaldehyde and 0.5% gluteraldehyde at 4°C. This will permit i 
extraction of DNA and RNA and permit in situ hybridization. 
7.5.2 Biopsies will be analyzed for incorporation of the transduced gene into the host genomic [ 
DNA and expression of the transduced gene at the RNA level by standard hybridization K 
techniques following polymerase chain reaction and by in situ hybridization. 
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7.5.3 All patients will be evaluable for response and toxicity following one course of therapy. j 
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7.6 An autopsy will be requested on all patients enrolled in the protocol who die. DNA will be extracted 
from tumor and normal tissues to determine if the adenovirus genes are present and expressed. 
PCR amplification of specific sequences will be used to determine this. 
7.7 A blood sample will be collected three times at one-half hour intervals following Injection of the 
adenovirus. These samples will provide leukocytes to analyze for uptake of adenovirus DNA. Serum 
will be tested for antibodies to adenovirus proteins. This will be done by western blot analysis 
performed by Microbiological /Associates, inc. (Rockville, MD). Patients will be tested monthly during 
treatment, monthly for the first three months following completion of treatment, every three months , 
for the remainder of the year following completion of treatment, and then at least yearly thereafter. 
7.8 Normal tissue samples will be collected during the follow-up visits and bronchoscopies. These will 
include samples of normal bronchial mucosa, leukocytes, and germ cells, if possible. These tissues 
will be analyzed for Incorporation of the adenovirus. 
8.0 CRITERIA FOR RESPONSE AND TOXICITY 
8.1 The graded toxicity scale used in this study has been previously described^*. Three patients will be 
Initially entered at each dose level. If one patient In the cohort of 3 develops grade 3 toxicity for any 
system except hematologic (grade 4 required), an additional 3 patients will be entered at that dose 
level. If 2 of the 6 patients develop grade 3 (grade 4 hematologic) or greater toxicities, the next ' 
lower dose level not causing these toxicities is declared the Maximum Tolerated Dose (MTD). If MTD i 
toxicity were to occur, patients could continue treatment at the next lower dose level. The MTD will i 
be determined separat^y for each phase of the study. If the MTD Is reached with the adenovirus 
plus cisplatin phase but not with the adenovirus alone phase, the study may continue with the 
adenovirus alone. If MTD toxicity occurs in a cohort of 3 patients, then the next 3 patients may 
Recombinant DNA Research, Volume 19 
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