Ms. Brindha P. Muniappan
MIT E18-666
50 Ames Street
Cambridge, MA 02139-4307
Phone: 617-253-6226
Fax: 617-258-5424
Email: bpmuni7@mit.edu
Projects are approved by an IRB located at: Massachusetts Institute of Technology.
The approving IRB operates under a Multiple Project Assurance (MPA) recognized by DOE or by the Department of Health and Human Services (HHS).
MPA number of the IRB: M-1377
Number of Human Subjects Projects reported: 1
Project Identifier: MIT-86-ER60448
Project Title:
Comparative Mutagenesis of Human Cells In Vivo and In Vitro
Principal Investigator:
Dr. William G. Thilly
Project started in: 1986
This project ended in Fiscal Year 1996.
Project Funding Information:
Project received funding in Fiscal Year 1996.
Project used human subjects in Fiscal Year 1996.
Funding Sources:
This is the amount spent on this grant betweeen 10/1/95 and 9/30/96. All costs associated with this grant are also associated with the use of human samples.
Project involves use of multiple protocols/subprojects.
Number of protocols/subprojects associated with this project: 2
Protocol/Subproject # 1
Protocol/Subproject Identifier: 1389A
IRB Review:
Type of Review: Full Board
Most Recent Approval: June 21, 1996
IRB Approval Number: 1389A
Number of Human Subjects who participated in this project/protocol during
06/21/95 - 06/21/96: 0
Type of Human Subjects Involvement:
a. Objectives
Our goal is to measure the pattern of point mutations, both spontaneous and due to particular mutagens, in human DNA sequences. Our research is based on the hypothesis that this pattern or mutational spectrum in human tissues will permit discovery of the primary causes of mutations in humans.
b. Methodology
Our methodology involves the use of Constant Denaturant Capillary Electrophoresis (CDCE) in combination with high fidelity (hifi) polymerase chain reaction (PCR). With this technique, we are able to measure any point mutation in a 100 base pairs (bp) DNA sequence. This technique has a current sensitivity of 2E-6 with human multicopy mitochondrial DNA. We are not only able to identify mutant DNA sequences, but also recover them for sequencing.
c. Human Subject Exposure
Human subjects are NOT exposed to any ionizing radiation, radioactive substance, or chemical substance.
d. Involvement of Human Subjects
1. B and T-lymphocytes are obtained from blood of healthy volunteers. Samples are identified by code to maintain confidentiality, and the identity of the code is only available to the principal investigator. Lymphocytes are isolated from 10 ml of blood by Ficol-Hypaque centrifugation. The lymphocytes are then either grown in culture (in vitro spectrum) or directly analyzed (in vivo spectrum). Lymphocytes are grown in culture with defined media, serum, crude IL-2, and irradiated stimulator cells.
The DNA is isolated from lymphocytes by Protease K, RNase, and restriction digestion followed by spooling of the DNA with ethanol. The DNA is then exposed to a combination of CDCE and hifi PCR followed by sequencing of the mutant DNA.
2. NO risks are taken on the part of the volunteers, except for the mild discomfort from the 10ml blood donation.
IRB Review:
Type of Review: Full Board
Most Recent Approval: February 17, 1996
IRB Approval Number: 2152
Number of Human Subjects who participated in this project/protocol during
02/17/95 - 02/17/96: 4
Type of Human Subjects Involvement:
a. Objectives
Our goal is to measure the pattern of point mutations, both spontaneous and due to particular mutagens, in human DNA segments. Our research is based on the hypothesis that this pattern or mutational spectrum in human tissues will permit discovery of the primary causes of mutations on an organ-by-organ basis.
b. Methodology
Our methodology involves the use of Constant Denaturant Capillary Electrophoresis (CDCE) in combination with high fidelity (hifi) polymerase chain reaction (PCR). With this technique we are able to measure any point mutations in a 100 base pair (bp) DNA sequence. This technique has a current sensitivity of 2E-6 with human multicopy mitochondrial DNA. We are not only able to identify mutant DNA sequences, but also recover them for sequencing.
c. Human Subject Exposure
Human subjects are NOT exposed to any ionizing radiation, radioactive substance, or chemical substance.
d. Involvement of Human Subjects:
1. Tissue samples from specific organs are obtained from laboratories of individual collaborators. These samples are excess tissue that would otherwise be discarded. They are obtained from surgical procedures unrelated to our research. No extra procedures are performed in order to obtain these tissues. Tissue samples are identified by code to maintain confidentiality, and the identity of the code is only available to the principal investigator. Only the sex, age, race, tissue type, and date of sample are available to researchers.
The DNA from each tissue is isolated by Protease K, RNAse, and restriction digestion followed by spooling of the DNA with ethanol. The DNA is then exposed to a combination of CDCE and hifi PCR followed by sequencing of the mutant DNA.
2. NO risks are taken on the part of the tissue donors and no extra procedures are performed on subjects in order to obtain tissue samples for our research.