Project Identifier: LLNL-88-104
Project Title:
Molecular Analysis of the Spectra of Mutation in the Human hprt Gene
Principle Investigator:
Dr. Irene Jones
Project started in: 1988
Project Funding Information:
Project received funding in Fiscal Year 1995.
Project did not use human subjects in Fiscal Year 1995.
Explanation:
No new human subjects in FY 95. Research focused on analysis of results obtained previously. Funding ended 9/29/95.
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 22, 1995
IRB Approval Number: 88-104
Number of Human Subjects in the Last Reporting Period for this Project: 0
(Reporting periods vary.)
Type of Human Subjects Involvement:
Objective: The objective of this research is to determine the frequency and molecular character of genetic alterations (mutations) that can be detected in the lymphocytes of humans. This information has the potential in the future to aid studies to identify types of mutations that are associated with exposures to toxic agents, and thereby aid surveillance of populations for genetic consequences of exposure to toxic agents. In this study the goal is to understand the mutations that occur in the DNA of people that are healthy and have no unusual exposures to radiation or chemicals. The gene studied is the hypoxanthine phosphoribosyltransferase (hprt) gene; mutations of this gene in lymphocytes do not affect the health of the person, but do reflect genetic changes that have occurred. By understanding the frequency and molecular nature of genetic changes that occur as a part of normal life, in this study, changes that occur in people exposed to chemicals or radiation may be identified, in future studies.
Methodology. Each subject recruited for this study was asked to read the Experimental Subject's Bill of Rights, sign a Consent Form, and complete a personal health questionnaire, either by phone interview or on a paper copy, prior to donating blood for this study. This information identified whether the person had exposures that disqualified them as donors (due to exposures to known toxic agents outside the scope of the study) and determined whether they fall into the study groups of interest (smokers and nonsmokers, but not former smokers).
White blood cells were isolated from peripheral blood samples and cultured in the laboratory under conditions that select for the growth of mutant cells. These mutant cells were propagated to large numbers and then frozen. The RNA and DNA of these cells were extracted and analyzed to detect the genetic alteration responsible for the mutant character of the cells. On occasion due to scheduling conflicts, cells from samples were frozen and cell culture performed at a later date.
Involvement of Human Subjects.
Procedures:
Up to 50ml of heparinized blood per donation were collected by medical professionals.
The identity of subjects recruited by collaborators at other institutions was never given to the Livermore scientists. The identity of subjects who were recruited at Livermore was coded on blood samples and all derived materials in the laboratory so that donor identity was not accessible from observation of any materials in the laboratory or publications. The code is maintained by Irene M. Jones and kept in a file drawer in her office, which is locked during nonbusiness hours.
There was no exposure of subjects to radiation, radioactive substances, or chemical substances.
Risks. The risks of participation in this study were limited to those of venipuncture, i.e. temporary pain, bruising and/or soreness of the affected tissue or surrounding tissue, formation of scar tissue, infection, and fainting, as noted in the consent form.