8.0 On Study Evaluation 
8.1 Daily CBC, platelet count, differential, electrolytes, glucose, BUN, creatinine while 
hospitalized and 2 times per week until day 60 post-transplant for patients not 
receiving taxol or vinblastine. Patients treated with post-transplant taxol or vinblastine 
will have twice weekly CBC, platelet count, and differential performed. 
8.2 Three times per week calcium phosphate, magnesium, albumin, total protein, SGOT, 
SGPT, alkaline phosphatase, LDH, total bilirubin, PT, PIT, fibrinogen, TT while 
hospitalized then weekly until day 60 post-transplant. For patients receiving taxol or 
vinblastine, electrolytes, calcium, phosphate, albumin, BUN, creatinine, glucose and 
liver function tests will be obtained every 3 weeks. 
8.3 Weekly chest x-ray, urinalysis until hospital discharge. 
8.4 One purple top and one green top tube will be obtained two times a week while 
patients are hospitalized to study hematopoietic cells for the presence of the MDR-1 
provirus by PCR, and Pgp expression and function by immunohistochemistry and the 
rhodamine efflux assay. 
8.5 Bone marrow aspiration and biopsy will be obtained on Days 14, 28, and 42 post- 
transplant for PCR, clonogenic assays and FACS analysis of hematopoietic cells in all 
patients. 
8.6 Patients will undergo a restaging evaluation of their known sites of metastatic breast 
cancer one month after their discharge from the hospital. Patients with residual or 
progressive disease will be eligible for treatment with vinblastine or taxol after this 
restaging evaluation. 
8.7 For patients receiving taxol or vinblastine, blood samples will be obtained on three 
successive days prior to beginning chemotherapy and at the end of each cycle of 
therapy to study the hematopoietic cells for the presence of the MDR-1 provirus by 
quantitative PCR and to study Pgp expression by immunohistochemistry. Bone 
marrow aspirates and biopsies will be obtained after marrow recovery from cycles 1 
and 3 of taxol or vinblastine to study the effects of chemotherapy on proviral copy 
number. We will also study expression of the vector derived MDR-1 mRNA in bone 
marrow cells by RT-PCR, immunohistochemistry, FACS, and in situ hybridization 
techniques. 
8 . 8 Evaluation after Day 60 post-transplantation: Patients not receiving taxol or vinblastine 
therapy will return to NCI monthly. Monthly evaluation will include CBC, platelets, 
differential, electrolytes, liver function tests, BUN, creatinine, calcium, albumin, 
magnesium. Peripheral blood samples for PCR analysis will also be obtained (1 green 
and 1 purple top tube). 
8.9 Bone marrow aspirates will be obtained every three months in patients who received 
transduced marrow and who are receiving treatment with taxol or vinblastine. These 
samples will be analyzed for the MDR-1 provirus by PCR. FACS analysis and 
clonogenic assays with and without taxol will also be performed. PCR will be 
performed on the hematopoietic clones that survive incubation in taxol. 
8.10 Patients will undergo restaging of their metastatic breast cancer every 3 months. This 
will include a CXR every 3 months and CT and bone scans of sites of previous 
disease. 
8.11 A creatinine clearance will be obtained prior to hospital discharge. 
8.12 An audiogram will be obtained prior to and one month after ABMT. 
8.13 Biopsies will be done of accessible tumor (skin, lymph nodes, pleural effusions, bone 
marrow, or liver or lung disease accessible by CT - guided FNA) at the time of relapse 
to assay for the presence of the MDR-1 gene by PCR. 
8.14 Apheresis following taxol or vinblastine/G-CSF (during the nadir recovery period) 
will be performed on patients with circulating cells or bone marrow cells that contain 
the transferred MDR-1 cDNA. For patients in a clinical CR or not otherwise receiving 
taxol or vinblastine/G-CSF, and whose marrow or peripheral cells contain the MDR-1 
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Recombinant DNA Research, Volume 18 
