Effect of SNPs in p53 and p53 Response Elements on the Inflammatory Response to DNA Damage
This research study will investigate the role of SNPs in p53 and p53 response elements on
the inflammatory response to DNA damage. A total of 210 healthy participants aged 18 years
and older carrying one of the seven SNPs of interest and wild-type controls will be
identified and recruited from the Environmental Polymorphism Registry (EPR). The EPR is a
long-term project to collect and store up to 15,000 DNA samples for use in research studies
from individuals in the greater North Carolina Triangle Region.
This observational gene association study will recruit participants on the basis of genotype
and then observe the phenotype of each participant. The SNPs of interest are p53, as well
as four of its downstream target genes including FLT1, MDM2, TLR8 and RRM1. A maximum of
150 mLs of blood will be obtained from each participant during one visit lasting
approximately one hour. Cells from the donated blood samples will be examined for their
response to exposed environmental stress ex vivo.
The primary objective is to determine the association between seven SNPs and p53 target gene
expression after exposure to Nutlin or doxorubicin (chemotherapeutic agents) with outcome
measured by RT-PCR. The seven SNPs are p53 rs1042522, p53 rs1800371, MDM2 rs2279744, MDM2
rs769412, FLT1 C-677T, TLR8 rs3761624 and RMM1 rs1465952. The secondary objectives are to:
(1) to determine the p53 promoter occupancy measured by ChIP analysis for the following
SNPs: FLT1 C-677T, TLR8 rs3761624 and RMM1 rs1465952; (2) to measure apoptosis by Annexin
V-PI assay for SNPs p53 rs1042522 and p53 rs1800371; (3) to examine the cell cycle profile
analysis (FACS) by cytofluorometry for SNPs p53 rs1042522 and p53 rs1800371; and (4) to
determine DNA repair using Pulse Field Electrophoresis Gel (TAFE gels) for the following
SNPs p53 rs1042522 and p53 rs1800371.
We hope the results of this study lead to discovery of important information regarding the
role of SNPs located in p53 and p53 response elements in human disease, potentially
identifying new targets for future studies.
Michael A Resnick, Ph.D.
National Institute of Environmental Health Sciences (NIEHS)
United States: Federal Government
|NIEHS Clinical Research Unit (CRU)||Research Triangle Park, North Carolina|