USDOE Human Subjects Research Database, Fiscal Year 2002

Thomas Jefferson University

Public Information Contact:

Dr. Eric Wickstrom
Thomas Jefferson University
233 S. 10th Street
219 Bluemle Life Sciences Building
Philadelphia, PA 19107-5541

Phone: (215) 955-4578
Fax: (215)9554580
E-mail: eric@tesla.jci.tju.edu

Institutional Review Board (IRB):

Human Subject Projects:

Number of Human Subjects projects reported: 1

TJU-00-ER63055

"Oncogene mRNA Imaging with Tc-99m-PNA-Peptides"

Project Identifier: TJU-00-ER63055

"Oncogene mRNA Imaging with Tc-99m-PNA-Peptides"

Principal Investigator: Dr. Eric Wickstrom, Thomas Jefferson University

Project started in: 2000

Project Funding Information:

Funding for Human Subjects Research:

DOE: Office of Biological and Environmental Research (OBER)

$404,099.00 (Est.) for: Fiscal Year 2002
total costs for year 2

Information on Use of Human Subjects:

This project does not involve the use of multiple protocols/subprojects.

Identifier or number: ER63055

Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Thomas Jefferson University
Most recent approval: 09/19/02
IRB approval number: 00.1138

Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 10
Reporting period for number of human subjects: Fiscal Year 2002

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])

a. Objectives
We hypothesize that antisense Tc-99m-PNA-peptides will be taken up by human breast cancer cells, hybridize to complementary mRNA targets, and permit imaging of oncogene mRNAs in human breast cancer cell xenografts in a mouse model, providing a proof-of-principle for noninvasive detection of precancerous and invasive breast cancer. Oncogenes cyclin D1, ERBB2, c-MYC, and tumor suppressor p53 will be probed. If successful, this technique will be tested in freshly excised normal and cancerous human breast cells, standardized against each individual’s white blood cells, and may be useful for diagnostic imaging of other solid tumors as well.
b. Methodology
1. We will prepare Tc-99m-PNA-peptides with antisense sequences known to hybridize specifically to cyclin D1, ERBB2, c-MYC, and p53 mRNAs, and specific for cell surface receptors for insulin-like growth factor 1 or peptide factors containing the sequence asparagine-glycine-arginine.
2. We will test the specificity of uptake and mRNA hybridization of our Tc-99m-PNA-peptides in normal and transformed human breast cancer cell lines in culture.
3. We will administer Tc-99m-PNA-peptides to cohorts of nude mice bearing human breast cancer cell line xenografts and determine the specificity and sensitivity of scintigraphic imaging of cyclin D1, ERBB2, c-MYC, and p53 mRNAs in the tumors, compared with reverse transcriptase/polymerase chain reaction measurements of those mRNAs in xenograft cells removed from the animal subjects.
4. Finally, we will test the specificity of uptake and mRNA hybridization of our Tc-99m-PNA-peptides in freshly excised normal and cancerous human breast cells, standardized against each individual’s white blood cells.
c. Ionizing radiation
Only excised cells will be exposed to Tc-99m labeled probes.
d. Involvement of Human Subjects
1. Patients with resectable mammary tumors will be asked for permission to analyze the tissues removed during surgery, and a blood sample. As soon as possible after removal from the chest wall, the tumor will be aspirated on a back table in the operating room, in Dr. Sauter’s laboratory, or in the pathology suite for specimen collection. Tumor levels of cyclin D1, ERBB2, c-MYC, and p53 mRNAs, measured by 1) gamma imaging of Tc-99m-PNA-peptides specific for cyclin D1, ERBB2, c-MYC, and p53 mRNAs, and 2) reverse transcriptase/polymerase chain reaction. As a baseline control, white blood cell levels of those mRNAs will be measured by reverse transcriptase/polymerase chain reaction. To attain the goal of 10 matched pairs of organotypic cultures of breast cancer cells and normal cells probed with Tc-99m antisense PNA to quantitate oncogene mRNAs, up to 30 patient samples will be collected and processed.
2. Risk to the subject due to blood draws and laboratory analysis of excised tumor sample is negligible, which is less than minimal. Despite the potential benefits for future patients, no benefit may be claimed for the subjects of this experiment. No members of vulnerable populations will be enrolled.
3. Collection of patient samples did not begin until the protocol was approved by the Cancer Clinical Research Review Committee and the Institutional Review Board. Written informed consent was obtained from all patients before their surgical samples were collected for analysis. Records and data will be kept confidential to protect patient privacy.