University of California, Los Angeles

Funding for Human Subjects Research:

DOE: Office of Scientific and Technical Information (OSTI)

$6,736,000.00 for: Fiscal Year 2007
Percent of funding associated with the use of human subjects: 1-20

Information on Use of Human Subjects:

This project involves the use of multiple protocols/subprojects.
Number of protocols/subprojects associated with this project: 3

Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: University of California, Los Angeles
Most recent approval: 05/29/07
IRB approval number: 94-12-447-31

Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects: Fiscal Year 2007

Type(s) of Human Subjects Involvement:

Collection of personally identifiable bodily materials (blood or blood products, urine, cells, tissue, teeth, organs, excreta, etc):

• Using cells cultured in a laboratory.
• Using bodily materials collected specifically for this project.

(a. Objectives, b. Methodology, c. Ionizing Radiation, Radioactive Substances, or Chemical Substances to which human subjects are exposed, d. Involvement of Human Subjects [d.1. procedures used, d.2. risks if any])

Study objectives: To study the role of the Abl oncogene in the growth regulation of hematopoietic stem cells and its relationship to the process of leukemogenesis in mice and humans.

We want to study how leukemia develops, the genes and pathways that lead to leukemia. We do know that the Bcr/Abl gene causes leukemia if we put the gene in cells, then into mice. But we do not know what factors or conditions cause this gene (or leukemaia) to appear or develop, since it is not a normally occurring gene. By comparing a patient with leukemia with one that does not have leukemia, we may be able to get a better understanding of the mechanisms, development or molecular events (pathways) that occur during the development of leukemia.

Study Procedures: Blood or bone marrow samples are obtained from either volunteers or patients. When patient samples are obtained, a normal control sample is also processed at the same time so that any differences between the patient and normal control samples can be observed under the same conditions. Normal samples may be obtained and used to test conditions and reagents before patient samples are obtained. All samples are coded and carry no identification information.

Briefly the samples are processed as follows: Cells are diluted with calcium and magnesium free phosphate balanced salt solution (PBS) and ficoll-hypaque is underlayered. Cells are spun for 15 minutes at 2000 rpm at room temperature. Cells are collected at the interface of the ficoll-hypaque and PBS layers and washed with media before further processing.

Cells may be further separated into different cell types using magnetic bead separation or by Fluorescence Activated Cell Sorting (FACS) or a combination of both methods depending on the experiment. Genes of interest are transduced with constructs containing the genes of interest, i.e., Bcr/Abl and variations of that gene. Cells are either put in culture (1, 2) to determine if their growth and differentiation is different from control; injected into mice to see if leukemia develops (ARC #2005-073-02) (3); or analyzed molecularly (4, 5). Use of these different techniques will enable us to define the role by which different genes and proteins can initiate cancer.

None of the subjects will be treated, no procedures other than to draw blood or bone marrow will be done. They will only provide blood or bone marrow samples. For patients, samples will only be drawn when samples are required for their care, and then only from patient healthy enough to tolerate the procedure. Normal safety procedures for drawing blood and bone marrow will be followed. All studies with these samples will be either at the bench, cultured, or injected into mice. Nothing will be done with or to the subjects.

Analysis: Data generated from experiments are always compared to control samples, in this case normal volunteers. Growth, morphology, and differentiation are compared in cultured cells. If a colony assay is done, the number and cell types are compared. Further analysis is done using immunohistochemistry and FACS analysis. For studies when cells are injected into mice, the cells will have no oncogenes or the cells will contain transduced genes. An oncogene is a modified gene, or a set of nucleotides that codes for a protein, that increases the malignancy of a tumor cell. In these studies, the length of time for leukemia to appear will be compared, and the cells analyzed by FACS and histochemistry. Statistical analysis will be done if the sample population is large enough. However, in most cases, the data will be obtained using descriptive analysis showing the cell type and cell numbers.

1. Sawyers C.L., Callahan W., and Witte O.N. 1992. MYC Blocks Transformation of ABL Oncogenes. Cell 70(6): 901-10.

2. Gishizky M.L. and Witte O.N. 1992. Initiation of Deregulated Growth of Multipotent Progenitor Cells by BCR/ABL In Vitro. Science, 256:836-839

3. Gishizky M.L., Johnson-White J., and Witte O.N. 1993. Efficient transplantation of BCR-ABL-induced chronic myelogenous Leukemia-like Syndrome in Mice. PNAS 90(8): 3755-9.

4. Capra M., Nuciforo P.G., Confalonieri S., Quarto M., Bianchi M., Nebuloni M., Boldorini R., Pallotti F., Viale G., Gishizky M.L., Draetta G.F., Di Fiore P.P. 2006. Frequent Alternations in the Expression of Serine/Threonine Kinases in Human Cancers. Cancer Res. 66(16): 8147-54.

5. Satterthwaite A.B., Li Z., and Witte O.N. 1998. Btk function in B cell development and response. Seminars in Immunology, 10:309-316.

Cord Blood will be obtained from the UCLA Cord Blood/Reproductive Sciences Core Facility only. The blood is separated into cell types and put into culture to see what kinds of cells grow.

Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: University of California, Los Angeles
Most recent approval: 12/19/06
IRB approval number: 03-09-042-04

Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 53
Reporting period for number of human subjects: Fiscal Year 2007

Type(s) of Human Subjects Involvement:

Internal administration of radioactive substances to human subjects:

• For clinical research

(a. Objectives, b. Methodology, c. Ionizing Radiation, Radioactive Substances, or Chemical Substances to which human subjects are exposed, d. Involvement of Human Subjects [d.1. procedures used, d.2. risks if any])

Purpose: Change in tumor size as classified by RECIST (Response Evaluation Criteria In Solid Tumors) poorly correlates with histopathologic response to neoadjuvant therapy in patients with soft tissue sarcomas (STS). The aim of this study was to prospectively evaluate whether Positron-Emission-Tomography with F18-fluorodeoxyglucose (FDG-PET) allows for a more accurate evaluation of histopathologic response.

Experimental Design: 53 patients with resectable biopsy proven high grade STS and 12 patients with Osteosarcoma underwent a FDG-PET/CT scan prior to and after neoadjuvant treatment. In STS, relative changes in tumor FDG uptake and size from the baseline to the follow-up scan were calculated, and their accuracy for assessment of histopathologic response was compared by receiver operating characteristic (ROC) curve analysis. Histopathologic response was defined as e95 percent.

Results: In histopathologic responders (n=10,19 percent), reduction in tumor FDG-uptake was significantly greater than in non-responders (p<0.001), whereas no significant differences were found for tumor size (p=0.35). The area under the ROC curve for metabolic changes was 0.93, but only 0.60 for size changes (p=0.004). Using a 60 percent decrease in tumor FDG uptake as a threshold resulted in a sensitivity of 100 percent and a specificity of 71 percent for assessment of histopathologic response, whereas RECIST demonstrated a sensitivity of 25 percent and a specificity of 100 percent.

Conclusion: Quantitative FDG-PET was significantly more accurate than size based criteria at assessing histopathologic response to neoadjuvant therapy. FDG-PET should be considered as a modality to monitor treatment response in patients with high grade STS.

Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: University of California, Los Angeles
Most recent approval: 01/25/07
IRB approval number: 04-10-068-03

Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 19
Reporting period for number of human subjects: Fiscal Year 2007

Type(s) of Human Subjects Involvement:

Internal administration of radioactive substances to human subjects:

• For clinical research

(a. Objectives, b. Methodology, c. Ionizing Radiation, Radioactive Substances, or Chemical Substances to which human subjects are exposed, d. Involvement of Human Subjects [d.1. procedures used, d.2. risks if any])

Purpose: Since kinase inhibitors have a cytostatic rather than cytotoxic effect computed tomography (CT) measurements of treatment responses are likely to be unreliable. In contrast, measurements of glycolytic activity with positron emission tomography (PET) have been shown to predict treatment responses to cytostatic drugs in some other cancers. This study aims at monitoring the effects of kinase inhibitor therapy on tumor growth and glucose metabolism.

Experimental design: Participants undergo a maximum of three PET/CT scans: The first FDG-PET/CT scan was performed within four weeks before the start of treatment, the second scan was performed within two weeks, and the third scan within 12 weeks after the start of cancer treatment. Changes in glucose metabolism and tumor size are quantified. Patient outcome (survival, time to recurrence) will serve as the gold standard. Untill this day, nineteen patients (10 males, 9 female) have been enrolled. Until now, eight patients completed all three scans, nine patients received a baseline and early follow-up scan, and two patients received a pre-therapeutic scan. No adverse events related to PET/CT imaging have been noticed. Patient enrollment continues to allow for a statistically meaningful interpretation of the data.

A typical PET/CT scan protocol includes the injection of 0.21 mCi/kg of 18F-fluoro-2-deoxyglucose (FDG) followed by a 45- to 60-minute uptake period, after which the patient is positioned in the scanner. A standard spiral CT scan is acquired with and without contrast.