Selective Detection and Quantification of DNA Lesions

Johns Hopkins University, Baltimore MD

Investigators

Abstract

Oxidative DNA damage is a biologically important and chemically complex process. A large variety (>50) of chemical modifications (lesions) are produced when DNA is exposed to oxidative conditions. DNA lesions are associated with aging and the development of disease. Consequently, they are of interest as "biomarkers" to help predict the onset of disease and progression of the aging process. The goal of this project is to develop facile, sensitive chemical methods for detecting lesions in cellular DNA. In addition, the PI will create chemical methods for lesion detection that will be employed by other scientists using commonly available instrumentation and reagents that can be produced by other laboratories and/or commercial suppliers. This chemistry will also enable a determination of whether there is a correlation between specific DNA damage and mutational hotspots. It is particularly challenging to detect specific lesions in telomeres or at exact positions in cellular DNA, because unlike single nucleotide polymorphisms it is not possible to produce more of DNA containing a lesion using the polymerase chain reaction. Hence, a very sensitive detection method is required. Broader Impact. Cell biologists, toxicologists, pharmacologists, and medicinal chemists will use the reagents developed during the course of this research. In addition, the potential use of such tools for identifying DNA lesion biomarkers would benefit society directly by potentially detecting genetically based diseases, such as cancer at an early stage where it is most treatable. The undergraduate students, graduate students, and postdoctoral fellows participating in this research will receive broad training in biochemistry, synthetic organic, physical organic and analytical chemistry. In addition, they will be exposed to biochemically important problems through their reading of the background literature related to DNA damage and the significance of its detection. The PI is actively involved in broadening the participation of groups that have traditionally been underrepresented in chemistry. He has been involved in creating a new undergraduate major in Chemical Biology at Morgan State University, and participates in a variety of undergraduate research symposia (e.g., Annual Biomedical Research Conference for Minority Students) where members of these groups are well represented.